WorldWideScience

Sample records for repeated freeze-thaw stress

  1. Effect of repeated freezing-thawing on the Achilles tendon of rabbits.

    Science.gov (United States)

    Chen, Lianxu; Wu, Yanping; Yu, Jiakuo; Jiao, Zhaode; Ao, Yingfang; Yu, Changlong; Wang, Jianquan; Cui, Guoqing

    2011-06-01

    The increased use of allograft tissue in the reconstruction of anterior cruciate ligament has brought more focus to the effect of storage and treatment on allograft. The purpose of this study was to observe the effect of histology and biomechanics on Achilles tendon in rabbits through repeated freezing-thawing before allograft tendon transplantation. Rabbit Achilles tendons were harvested and processed according to the manufacture's protocol of tissue bank, and freezing-thawing was repeated three times (group 1) and ten times (group 2). Those received only one cycle were used as controls. Then, tendons in each group were selected randomly to make for histological observations and biomechanics test. Histological observation showed that the following changes happened as the number of freezing-thawing increased: the arrangement of tendon bundles and collagen fibrils became disordered until ruptured, cells disrupted and apparent gaps appeared between tendon bundle because the formation of ice crystals. There were significant differences between the experimental and control groups in the values of maximum load, energy of maximum load and maximum stress, whereas no significant differences existed in other values such as stiffness, maximum strain, elastic modulus, and energy density. Therefore, repeated freezing-thawing had histological and biomechanical effect on Achilles tendon in rabbits before allograft tendon transplantation. This indicates that cautions should be taken in the repeated freezing-thawing preparation of allograft tendons in clinical application.

  2. Effect of Repeated Freeze-Thaw Cycles on Beef Quality and Safety

    Science.gov (United States)

    Rahman, Mohammad Hafizur; Hossain, Mohammad Mujaffar; Rahman, Syed Mohammad Ehsanur; Hashem, Mohammad Abul

    2014-01-01

    The objectives of this study were to know the effect of repeated freeze-thaw cycles of beef on the sensory, physicochemical quality and microbiological assessment. The effects of three successive freeze-thaw cycles on beef forelimb were investigated comparing with unfrozen fresh beef for 75 d by keeping at −20±1℃. The freeze-thaw cycles were subjected to three thawing methods and carried out to know the best one. As the number of freeze-thaw cycles increased color and odor declined significantly before cook within the cycles and tenderness, overall acceptability also declined among the cycles after cook by thawing methods. The thawing loss increased and dripping loss decreased significantly (pcycles and then decreased. Cooking loss increased in cycle 1 and 3, but decreased in cycle 2. pH decreased significantly (pcycles. Moreover, drip loss, cooking loss and WHC were affected (pcycles. 2-Thiobarbituric acid (TBARS) value increased (pcycles and among the cycles by thawing methods. Total viable bacteria, total coliform and total yeast-mould count decreased significantly (pcycles in comparison to the initial count in repeated freeze-thaw cycles. As a result, repeated freeze-thaw cycles affected the sensory, physicochemical and microbiological qua- lity of beef, causing the deterioration of beef quality, but improved the microbiological quality. Although repeated freeze-thaw cycles did not affect much on beef quality and safety but it may be concluded that repeated freeze and thaw should be minimized in terms of beef color for commercial value and WHC and tenderness/juiciness for eating quality. PMID:26761286

  3. Giant panda (Ailuropoda melanoleuca) sperm morphometry and function after repeated freezing and thawing.

    Science.gov (United States)

    Santiago-Moreno, J; Esteso, M C; Pradiee, J; Castaño, C; Toledano-Díaz, A; O'Brien, E; Lopez-Sebastián, A; Martínez-Nevado, E; Delclaux, M; Fernández-Morán, J; Zhihe, Z

    2016-05-01

    This work examines the effects of subsequent cycles of freezing-thawing on giant panda (Ailuropoda melanoleuca) sperm morphometry and function, and assesses whether density-gradient centrifugation (DGC) can increase the number of freezing-thawing cycles this sperm can withstand. A sperm sample was collected by electroejaculation from a mature giant panda and subjected to five freezing-thawing cycles. Although repeated freezing-thawing negatively affected (P 60% of the sperm cells in both treatments showed acrosome integrity even after the fifth freezing cycle. In fresh semen, the sperm head length was 4.7 μm, the head width 3.6 μm, area 14.3 μm(2) and perimeter length 14.1 μm. The present results suggest that giant panda sperm trends to be resistant to repeated freezing-thawing, even without DGC selection. © 2015 Blackwell Verlag GmbH.

  4. Resilience of Floating Treatment Wetlands to Repeated Freeze-Thaw Cycles

    Science.gov (United States)

    Ortega, K.; Marchetto, P.; Magner, J.

    2017-12-01

    Floating treatment wetlands (FTWs), made of a matrix of recycled polyethylene terephthalate (PET) fibers, are currently being used as a method to reduce nutrient loading in lakes. The matrix encourages growth of biofilms, which uptake nutrients from the water. However, the usefulness of FTWs has only been assessed in areas where the lakes do not freeze over in the winter. Repeated freeze-thaw cycles were run on sections of the FTWs in a laboratory setting in order to test the resilience of the PET fibers over the FTWs' advertised fifteen-year lifespan. Preliminary findings suggest that the stresses caused by freezing and thawing of the surrounding water contribute to deterioration of the PET fibers, leading to production of microplastics. Estimations indicate that approximately 0.063% of a FTW's mass could be lost as microplastics over the course of its lifespan. Production of microplastics contributes to plastic pollution in the treatment water, possibly offsetting any nutrient remediation the FTWs perform.

  5. The Impact of Repeated Freeze-Thaw Cycles on the Quality of Biomolecules in Four Different Tissues.

    Science.gov (United States)

    Ji, Xiaoli; Wang, Min; Li, Lingling; Chen, Fang; Zhang, Yanyang; Li, Qian; Zhou, Junmei

    2017-10-01

    High-quality biosamples are valuable resources for biomedical research. However, some tissues are stored without being sectioned into small aliquots and have to undergo repeated freeze-thaw cycles throughout prolonged experimentation. Little is known regarding the effects of repeated freeze-thaw cycles on the quality of biomolecules in tissues. The aim of this study was to evaluate the impact of repeated freeze-thaw (at room temperature or on ice) cycles on biomolecules and gene expression in four different types of tissues. Each fresh tissue was sectioned into seven aliquots and snap-frozen before undergoing repeated freeze-thaw cycles at room temperature or on ice. Biomolecules were extracted and analyzed. Both relative and absolute quantification were used to detect the changes in gene expression. The results indicated that the impact of repeated freeze-thaw cycles on RNA integrity varied by tissue type. Gene expression, including the housekeeping gene, was affected in RNA-degraded samples according to absolute quantification rather than relative quantification. Furthermore, our results suggest that thawing on ice could protect RNA integrity compared with thawing at room temperature. No obvious degradation of protein or DNA was observed with repeated freeze-thaw cycles either at room temperature or on ice. This research provides ample evidence for the necessity of sectioning fresh tissues into small aliquots before snap-freezing, thus avoiding degradation of RNA and alteration of gene expression resulting from repeated freeze-thaw cycles. For frozen tissue samples that were already in storage and had to be used repeatedly during their lifecycle, thawing on ice or sectioned at ultralow temperature is recommended.

  6. Effect of haemolysis and repeated freeze-thawing cycles on wild boar serum antibody testing by ELISA

    Directory of Open Access Journals (Sweden)

    Boadella Mariana

    2011-11-01

    Full Text Available Abstract Background Monitoring wildlife diseases is needed to identify changes in disease occurrence. Wildlife blood samples are valuable for this purpose but are often gathered haemolysed. To maximise information, sera often go through repeated analysis and freeze-thaw cycles. Herein, we used samples of clean and haemolysed Eurasian wild boar (Sus scrofa serum stored at -20°C and thawed up to five times to study the effects of both treatments on the outcome of a commercial ELISA test for the detection of antibodies against Suid Herpesvirus 1 (ADV. Results The estimated prevalence of antibodies against ADV was 50-53% for clean and haemolysed sera. Hence, haemolysis did not reduce the mean observed serum antibody prevalence. However, 10 samples changed their classification after repeated freeze-thawing. This included 3 (15% of the clean sera and 7 (41% of the haemolysed sera. Conclusions We recommend (1 establishing more restrictive cut-off values when testing wildlife sera, (2 recording serum quality prior to sample banking, (3 recording the number of freezing-thawing cycles and (4 store sera in various aliquots to reduce repeated usage. For instance, sera with more than 3 freeze-thaw cycles and a haemolysis of over 3 on a scale of 4 should better be discarded for serum antibody monitoring. Even clean (almost not haemolysed sera should not go through more than 5 freeze-thaw cycles.

  7. Repeated freeze-thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons.

    Science.gov (United States)

    Suto, Kaori; Urabe, Ken; Naruse, Kouji; Uchida, Kentaro; Matsuura, Terumasa; Mikuni-Takagaki, Yuko; Suto, Mitsutoshi; Nemoto, Noriko; Kamiya, Kentaro; Itoman, Moritoshi

    2012-03-01

    Frozen bone-patellar tendon bone allografts are useful in anterior cruciate ligament reconstruction as the freezing procedure kills tissue cells, thereby reducing immunogenicity of the grafts. However, a small portion of cells in human femoral heads treated by standard bone-bank freezing procedures survive, thus limiting the effectiveness of allografts. Here, we characterized the survival rates and mechanisms of cells isolated from rat bones and tendons that were subjected to freeze-thaw treatments, and evaluated the influence of these treatments on the mechanical properties of tendons. After a single freeze-thaw cycle, most cells isolated from frozen bone appeared morphologically as osteocytes and expressed both osteoblast- and osteocyte-related genes. Transmission electron microscopic observation of frozen cells using freeze-substitution revealed that a small number of osteocytes maintained large nuclei with intact double membranes, indicating that these osteocytes in bone matrix were resistant to ice crystal formation. We found that tendon cells were completely killed by a single freeze-thaw cycle, whereas bone cells exhibited a relatively high survival rate, although survival was significantly reduced after three freeze-thaw cycles. In patella tendons, the ultimate stress, Young's modulus, and strain at failure showed no significant differences between untreated tendons and those subjected to five freeze-thaw cycles. In conclusion, we identified that cells surviving after freeze-thaw treatment of rat bones were predominantly osteocytes. We propose that repeated freeze-thaw cycles could be applied for processing bone-tendon constructs prior to grafting as the treatment did not affect the mechanical property of tendons and drastically reduced surviving osteocytes, thereby potentially decreasing allograft immunogenecity.

  8. NMR Pore Structure and Dynamic Characteristics of Sandstone Caused by Ambient Freeze-Thaw Action

    Directory of Open Access Journals (Sweden)

    Bo Ke

    2017-01-01

    Full Text Available For a deeper understanding of the freeze-thaw weathering effects on the microstructure evolution and deterioration of dynamic mechanical properties of rock, the present paper conducted the nuclear magnetic resonance (NMR tests and impact loading experiments on sandstone under different freeze-thaw cycles. The results of NMR test show that, with the increase of freeze-thaw cycles, the pores expand and pores size tends to be uniform. The experimental results show that the stress-strain curves all go through four stages, namely, densification, elasticity, yielding, and failure. The densification curve is shorter, and the slope of elasticity curve decreases as the freeze-thaw cycles increase. With increasing freeze-thaw cycles, the dynamic peak stress decreases and energy absorption of sandstone increases. The dynamic failure form is an axial splitting failure, and the fragments increase and the size diminishes with increasing freeze-thaw cycles. The higher the porosity is, the more severe the degradation of dynamic characteristics is. An increase model for the relationships between the porosity or energy absorption and freeze-thaw cycles number was built to reveal the increasing trend with the freeze-thaw cycles increase; meanwhile, a decay model was built to predict the dynamic compressive strength degradation of rock after repeated freeze-thaw cycles.

  9. Application of one-sided stress wave velocity measurement technique to evaluate freeze-thaw damage in concrete

    International Nuclear Information System (INIS)

    Lee, Joon Hyun; Park, Won Su

    1998-01-01

    It is well recognized that damage resulting from freeze-thaw cycles is a serious problems causing deterioration and degradation of concrete. In general, freeze-thaw cycles change the microstructure of the concrete ultimately leading to internal stresses and cracking. In this study, a new method for one-sided stress wave velocity measurement has been applied to evaluate freeze-thaw damage in concrete by monitoring the velocity change of longitudinal and surface waves. The freeze-thaw damage was induced in a 400 x 150 x 100 mm concrete specimen in accordance with ASTM C666 using s commercial testing apparatus. A cycle consisted of a variation of the temperature from -14 to 4 degrees Celsius. A cycle takes 4-5 hours with approximately equal times devoted to freezing-thawing. Measurement of longitudinal and surface wave velocities based on one-sided stress wave velocity measurement technique was made every 5 freeze-thaw cycle. The variation of longitudinal and surface wave velocities due to increasing freeze-thaw damage is demonstrated and compared to determine which one is more effective to monitor freeze-thaw cyclic damage progress. The variation in longitudinal wave velocity measured by one-sided technique is also compared with that measured by the conventional through transmission technique.

  10. Behavior of Plain Concrete of a High Water-Cement Ratio after Freeze-Thaw Cycles

    OpenAIRE

    Shang, Huai-Shuai; Yi, Ting-Hua; Song, Yu-Pu

    2012-01-01

    An experimental study of plain concrete specimens of water-cement ratio 0.55, subjected to 0, 15, 25, 40, 50 and 75 cycles of freeze-thaw was completed. The dynamic modulus of elasticity (DME), weight loss, compressive strength, tensile strength, flexural strength, cleavage strength and stress-strain relationships of plain concrete specimens suffering from freeze-thaw cycles were measured. The experimental results showed that the strength decreased as the freeze-thaw cycles were repeated. A c...

  11. The freeze-thaw stress response of the yeast Saccharomyces cerevisiae is growth phase specific and is controlled by nutritional state via the RAS-cyclic AMP signal transduction pathway.

    Science.gov (United States)

    Park, J I; Grant, C M; Attfield, P V; Dawes, I W

    1997-10-01

    The ability of cells to survive freezing and thawing is expected to depend on the physiological conditions experienced prior to freezing. We examined factors affecting yeast cell survival during freeze-thaw stress, including those associated with growth phase, requirement for mitochondrial functions, and prior stress treatment(s), and the role played by relevant signal transduction pathways. The yeast Saccharomyces cerevisiae was frozen at -20 degrees C for 2 h (cooling rate, less than 4 degrees C min-1) and thawed on ice for 40 min. Supercooling occurred without reducing cell survival and was followed by freezing. Loss of viability was proportional to the freezing duration, indicating that freezing is the main determinant of freeze-thaw damage. Regardless of the carbon source used, the wild-type strain and an isogenic petite mutant ([rho 0]) showed the same pattern of freeze-thaw tolerance throughout growth, i.e., high resistance during lag phase and low resistance during log phase, indicating that the response to freeze-thaw stress is growth phase specific and not controlled by glucose repression. In addition, respiratory ability and functional mitochondria are necessary to confer full resistance to freeze-thaw stress. Both nitrogen and carbon source starvation led to freeze-thaw tolerance. The use of strains affected in the RAS-cyclic AMP (RAS-cAMP) pathway or supplementation of an rca1 mutant (defective in the cAMP phosphodiesterase gene) with cAMP showed that the freeze-thaw response of yeast is under the control of the RAS-cAMP pathway. Yeast did not adapt to freeze-thaw stress following repeated freeze-thaw treatment with or without a recovery period between freeze-thaw cycles, nor could it adapt following pretreatment by cold shock. However, freeze-thaw tolerance of yeast cells was induced during fermentative and respiratory growth by pretreatment with H2O2, cycloheximide, mild heat shock, or NaCl, indicating that cross protection between freeze-thaw stress

  12. Freeze-thaw durability of air-entrained concrete.

    Science.gov (United States)

    Shang, Huai-Shuai; Yi, Ting-Hua

    2013-01-01

    One of the most damaging actions affecting concrete is the abrupt temperature change (freeze-thaw cycles). The types of deterioration of concrete structures by cyclic freeze-thaw can be largely classified into surface scaling (characterized by the weight loss) and internal crack growth (characterized by the loss of dynamic modulus of elasticity). The present study explored the durability of concrete made with air-entraining agent subjected to 0, 100, 200, 300, and 400 cycles of freeze-thaw. The experimental study of C20, C25, C30, C40, and C50 air-entrained concrete specimens was completed according to "the test method of long-term and durability on ordinary concrete" GB/T 50082-2009. The dynamic modulus of elasticity and weight loss of specimens were measured after different cycles of freeze-thaw. The influence of freeze-thaw cycles on the relative dynamic modulus of elasticity and weight loss was analyzed. The findings showed that the dynamic modulus of elasticity and weight decreased as the freeze-thaw cycles were repeated. They revealed that the C30, C40, and C50 air-entrained concrete was still durable after 300 cycles of freeze-thaw according to the experimental results.

  13. Freeze-Thaw Durability of Air-Entrained Concrete

    Directory of Open Access Journals (Sweden)

    Huai-Shuai Shang

    2013-01-01

    Full Text Available One of the most damaging actions affecting concrete is the abrupt temperature change (freeze-thaw cycles. The types of deterioration of concrete structures by cyclic freeze-thaw can be largely classified into surface scaling (characterized by the weight loss and internal crack growth (characterized by the loss of dynamic modulus of elasticity. The present study explored the durability of concrete made with air-entraining agent subjected to 0, 100, 200, 300, and 400 cycles of freeze-thaw. The experimental study of C20, C25, C30, C40, and C50 air-entrained concrete specimens was completed according to “the test method of long-term and durability on ordinary concrete” GB/T 50082-2009. The dynamic modulus of elasticity and weight loss of specimens were measured after different cycles of freeze-thaw. The influence of freeze-thaw cycles on the relative dynamic modulus of elasticity and weight loss was analyzed. The findings showed that the dynamic modulus of elasticity and weight decreased as the freeze-thaw cycles were repeated. They revealed that the C30, C40, and C50 air-entrained concrete was still durable after 300 cycles of freeze-thaw according to the experimental results.

  14. SLAPex Freeze/Thaw 2015: The First Dedicated Soil Freeze/Thaw Airborne Campaign

    Science.gov (United States)

    Kim, Edward; Wu, Albert; DeMarco, Eugenia; Powers, Jarrett; Berg, Aaron; Rowlandson, Tracy; Freeman, Jacqueline; Gottfried, Kurt; Toose, Peter; Roy, Alexandre; hide

    2016-01-01

    Soil freezing and thawing is an important process in the terrestrial water, energy, and carbon cycles, marking the change between two very different hydraulic, thermal, and biological regimes. NASA's Soil Moisture Active/Passive (SMAP) mission includes a binary freeze/thaw data product. While there have been ground-based remote sensing field measurements observing soil freeze/thaw at the point scale, and airborne campaigns that observed some frozen soil areas (e.g., BOREAS), the recently-completed SLAPex Freeze/Thaw (F/T) campaign is the first airborne campaign dedicated solely to observing frozen/thawed soil with both passive and active microwave sensors and dedicated ground truth, in order to enable detailed process-level exploration of the remote sensing signatures and in situ soil conditions. SLAPex F/T utilized the Scanning L-band Active/Passive (SLAP) instrument, an airborne simulator of SMAP developed at NASA's Goddard Space Flight Center, and was conducted near Winnipeg, Manitoba, Canada, in October/November, 2015. Future soil moisture missions are also expected to include soil freeze/thaw products, and the loss of the radar on SMAP means that airborne radar-radiometer observations like those that SLAP provides are unique assets for freeze/thaw algorithm development. This paper will present an overview of SLAPex F/T, including descriptions of the site, airborne and ground-based remote sensing, ground truth, as well as preliminary results.

  15. In Vivo Perturbation of Membrane-Associated Calcium by Freeze-Thaw Stress in Onion Bulb Cells 1

    Science.gov (United States)

    Arora, Rajeev; Palta, Jiwan P.

    1988-01-01

    Incipient freeze-thaw stress in onion bulb scale tissue is known to cause enhanced efflux of K+, along with small but significant loss of cellular Ca2+. During the post-thaw period, irreversibly injured cells undergo a cytological aberration, namely, `protoplasmic swelling.' This cellular symptom is thought to be caused by replacement of Ca2+ from membrane by extracellular K+ and subsequent perturbation of K+ transport properties of plasma membrane. In the present study, onion (Allium cepa L. cv Sweet Sandwich) bulbs were slowly frozen to either −8.5°C or −11.5°C and thawed over ice. Inner epidermal peels from bulb scales were treated with fluorescein diacetate for assessing viability. In these cells, membrane-associated calcium was determined using chlorotetracycline fluorescence microscopy combined with image analysis. Increased freezing stress and tissue infiltration (visual water-soaking) were paralleled by increased ion leakage. Freezing injury (−11.5°C; irreversible) caused a specific and substantial loss of membrane-associated Ca2+ compared to control. Loss of membrane-associated Ca2+ caused by moderate stress (−8.5°C; reversible) was much less relative to −11.5°C treatment. Ion efflux and Ca2+-chlorotetracycline fluorescence showed a negative relationship. Extracellular KCl treatment simulated freeze-thaw stress by causing a similar loss of membrane-associated calcium. This loss was dramatically reduced by presence of extracellular CaCl2. Our results suggest that the loss of membrane-associated Ca2+, in part, plays a role in initiation and progression of freezing injury. Images Fig. 1 Fig. 2 PMID:16666196

  16. Effect of freezing and thawing on UMTRA covers

    International Nuclear Information System (INIS)

    Rager, R.; Smith, G.; Brody, R.

    1988-01-01

    The proposed US Environmental Protection Agency (EPA) groundwater standards (40 CFR 192) require that Uranium Mill Tailings Remedial Action (UMTRA) Project remedial action designs meet low numerical limits for contaminants contained in water or vapor exiting the embankments. To meet the standards, a cover of compacted fine-grained soil is placed over UMTRA Project embankments. One of the functions of this cover is to limit infiltration. The hydraulic conductivity of this infiltration barrier must be low in order to reduce seepage from the cell to the extent necessary to comply with the proposed EPA groundwater standards. Analyses of infiltration barriers covered with rock erosion protection show that the infiltration barriers may become saturated. Accordingly, it is necessary to assure that freezing and thawing of the infiltration barrier materials do not affect the performance of the embankment. A study has been conducted to determine if the hydraulic conductivity of fine-grained clayey soils used or proposed for use in radon/infiltration barriers is affected by repeated freezing and thawing cycles. In addition, a procedure for determining the depth of frost penetration has been developed. Laboratory freeze-thaw tests were undertaken in order to determine if the saturated hydraulic conductivity of clay soils used in UMTRA Project radon/infiltration barriers was affected. The results indicate that an increase of an order of magnitude or more in saturated hydraulic conductivity may occur during repeated freeze-thaw cycles

  17. The cytoplasmic Cu,Zn superoxide dismutase of saccharomyces cerevisiae is required for resistance to freeze-thaw stress. Generation of free radicals during freezing and thawing

    DEFF Research Database (Denmark)

    Park, J I; Grant, C M; Davies, Michael Jonathan

    1998-01-01

    The involvement of oxidative stress in freeze-thaw injury to yeast cells was analyzed using mutants defective in a range of antioxidant functions, including Cu,Zn superoxide dismutase (encoded by SOD1), Mn superoxide dismutase (SOD2), catalase A, catalase T, glutathione reductase, gamma...

  18. Freeze-thaw-induced embolism in Pinus contorta: centrifuge experiments validate the 'thaw-expansion hypothesis' but conflict with ultrasonic emission data.

    Science.gov (United States)

    Mayr, Stefan; Sperry, John S

    2010-03-01

    *The 'thaw-expansion hypothesis' postulates that xylem embolism is caused by the formation of gas bubbles on freezing and their expansion on thawing. We evaluated the hypothesis using centrifuge experiments and ultrasonic emission monitoring in Pinus contorta. *Stem samples were exposed to freeze-thaw cycles at varying xylem pressure (P) in a centrifuge before the percentage loss of hydraulic conductivity (PLC) was measured. Ultrasonic acoustic emissions were registered on samples exposed to freeze-thaw cycles in a temperature chamber. *Freeze-thaw exposure of samples spun at -3 MPa induced a PLC of 32% (one frost cycle) and 50% (two cycles). An increase in P to -0.5 MPa during freezing had no PLC effect, whereas increased P during thaw lowered PLC to 7%. Ultrasonic acoustic emissions were observed during freezing and thawing at -3 MPa, but not in air-dried or water-saturated samples. A decrease in minimum temperature caused additional ultrasonic acoustic emissions, but had no effect on PLC. *The centrifuge experiments indicate that the 'thaw-expansion hypothesis' correctly describes the embolization process. Possible explanations for the increase in PLC on repeated frost cycles and for the ultrasonic acoustic emissions observed during freezing and with decreasing ice temperature are discussed.

  19. Freeze-Thaw Cycles and Soil Biogeochemistry: Implications for Greenhouse Gas emission

    Science.gov (United States)

    Rezanezhad, F.; Milojevic, T.; Oh, D. H.; Parsons, C. T.; Smeaton, C. M.; Van Cappellen, P.

    2016-12-01

    Freeze-thaw cycles represent a major natural climate forcing acting on soils at middle and high latitudes. Repeated freezing and thawing of soils changes their physical properties, geochemistry, and microbial community structure, which together govern the biogeochemical cycling of carbon and nutrients. In this presentation, we focus on how freeze-thaw cycles regulate carbon and nitrogen cycling and how these transformations influence greenhouse gas (GHG) fluxes. We present a novel approach, which combines the acquisition of physical and chemical data in a newly developed experimental soil column system. This system simulates realistic soil temperature profiles during freeze-thaw cycles. A high-resolution, Multi-Fiber Optode (MuFO) microsensor technique was used to detect oxygen (O2) continuously in the column at multiple depths. Surface and subsurface changes to gas and aqueous phase chemistry were measured to delineate the pathways and quantify soil respiration rates during freeze-thaw cycles. The results indicate that the time-dependent release of GHG from the soil surface is influenced by a combination of two key factors. Firstly, fluctuations in temperature and O2 availability affect soil biogeochemical activity and GHG production. Secondly, the recurrent development of a physical ice barrier prevents exchange of gaseous compounds between the soil and atmosphere during freezing conditions; removal of this barrier during thaw conditions increases GHG fluxes. During freezing, O2 levels in the unsaturated zone decreased due to restricted gas exchange with the atmosphere. As the soil thawed, O2 penetrated deeper into the soil enhancing the aerobic mineralization of organic carbon and nitrogen. Additionally, with the onset of thawing a pulse of gas flux occurred, which is attributed to the build-up of respiratory gases in the pore space during freezing. The latter implies enhanced anaerobic respiration as O2 supply ceases when the upper soil layer freezes.

  20. Evaluating the freeze-thaw durability of portland cement-stabilized-solidified heavy metal waste using acoustic measurements

    International Nuclear Information System (INIS)

    El-Korchi, T.; Gress, D.; Baldwin, K.; Bishop, P.

    1989-01-01

    The use of stress wave propagation to assess freeze-thaw resistance of portland cement solidified/stabilized waste is presented. The stress wave technique is sensitive to the internal structure of the specimens and would detect structural deterioration independent of weight loss or visual observations. The freeze-thaw resistance of a cement-solidified cadmium waste and a control was evaluated. The control and cadmium wastes both showed poor freeze-thaw resistance. However, the addition of cadmium and seawater curing increased the resistance to more cycles of freezing and thawing. This is attributed to microstructural changes

  1. Investigation of Freeze and Thaw Cycles of a Gas-Charged Heat Pipe

    Science.gov (United States)

    Ku, Jentung; Ottenstein, Laura; Krimchansky, Alexander

    2012-01-01

    The traditional constant conductance heat pipes (CCHPs) currently used on most spacecraft run the risk of bursting the pipe when the working fluid is frozen and later thawed. One method to avoid pipe bursting is to use a gas-charged heat pipe (GCHP) that can sustain repeated freeze/thaw cycles. The construction of the GCHP is similar to that of the traditional CCHP except that a small amount of non-condensable gas (NCG) is introduced and a small length is added to the CCHP condenser to serve as the NCG reservoir. During the normal operation, the NCG is mostly confined to the reservoir, and the GCHP functions as a passive variable conductance heat pipe (VCHP). When the liquid begins to freeze in the condenser section, the NCG will expand to fill the central core of the heat pipe, and ice will be formed only in the grooves located on the inner surface of the heat pipe in a controlled fashion. The ice will not bridge the diameter of the heat pipe, thus avoiding the risk of pipe bursting during freeze/thaw cycles. A GCHP using ammonia as the working fluid was fabricated and then tested inside a thermal vacuum chamber. The GCHP demonstrated a heat transport capability of more than 200W at 298K as designed. Twenty-seven freeze/thaw cycles were conducted under various conditions where the evaporator temperature ranged from 163K to 253K and the condenser/reservoir temperatures ranged from 123K to 173K. In all tests, the GCHP restarted without any problem with heat loads between 10W and 100W. No performance degradation was noticed after 27 freeze/thaw cycles. The ability of the GCHP to sustain repeated freeze/thaw cycles was thus successfully demonstrated.

  2. Evaluation, including effects of storage and repeated freezing and thawing, of a method for measurement of urinary creatinine

    DEFF Research Database (Denmark)

    Garde, A H; Hansen, Åse Marie; Kristiansen, J

    2003-01-01

    The aims of this study were to elucidate to what extent storage and repeated freezing and thawing influenced the concentration of creatinine in urine samples and to evaluate the method for determination of creatinine in urine. The creatinine method was based on the well-known Jaffe's reaction...... and measured on a COBAS Mira autoanalyser from Roche. The main findings were that samples for analysis of creatinine should be kept at a temperature of -20 degrees C or lower and frozen and thawed only once. The limit of detection, determined as 3 x SD of 20 determinations of a sample at a low concentration (6...

  3. Effect of freeze-thaw cycles on greenhouse gas fluxes from peat soils

    Science.gov (United States)

    Oh, H. D.; Rezanezhad, F.; Markelov, I.; McCarter, C. P. R.; Van Cappellen, P.

    2017-12-01

    The ongoing displacement of climate zones by global warming is increasing the frequency and intensity of freeze-thaw cycles in middle and high latitude regions, many of which are dominated by organic soils such as peat. Repeated freezing and thawing of soils changes their physical properties, geochemistry, and microbial community structure, which together govern the biogeochemical cycling of carbon and nutrients. In this presentation, we focus on how freeze-thaw cycles influence greenhouse gas fluxes from peat using a newly developed experimental soil column system that simulates realistic soil temperature profiles during freeze-thaw cycles. We measured the surface and subsurface changes to gas and aqueous phase chemistry to delineate the diffusion pathways and quantify soil greenhouse gas fluxes during freeze-thaw cycles using sulfur hexafluoride (SF6) as a conservative tracer. Three peat columns were assembled inside a temperature controlled chamber with different soil structures. All three columns were packed with 40 cm of undisturbed, slightly decomposed peat, where the soil of two columns had an additional 10 cm layer on top (one with loose Sphagnum moss and one with an impermeable plug). The results indicate that the release of SF6 and CO2 gas from the soil surface was influenced by the recurrent development of a physical ice barrier, which prevented gas exchange between the soil and atmosphere during freezing conditions. With the onset of thawing a pulse of SF6 and CO2 occurred, resulting in a flux of 3.24 and 2095.52 µmol/m2h, respectively, due to the build-up of gases in the liquid-phase pore space during freezing. Additionally, we developed a model to determine the specific diffusion coefficients for each peat column. These data allow us to better predict how increased frequency and intensity of freeze-thaw cycles will affect greenhouse gas emissions in northern peat soils.

  4. Quality changes and freezing time prediction during freezing and thawing of ginger

    OpenAIRE

    Singha, Poonam; Muthukumarappan, Kasiviswanathan

    2015-01-01

    Abstract Effects of different freezing rates and four different thawing methods on chemical composition, microstructure, and color of ginger were investigated. Computer simulation for predicting the freezing time of cylindrical ginger for two different freezing methods (slow and fast) was done using ANSYS ? Multiphysics. Different freezing rates (slow and fast) and thawing methods significantly (P?

  5. Separation of Contaminants in The Freeze/Thaw Process

    Directory of Open Access Journals (Sweden)

    Szpaczyński Janusz A.

    2017-06-01

    Full Text Available These studies examined the concept of concentration and purification of several types of wastewater by freezing and thawing. The experiments demonstrated that freezing of contaminated liquid contributed to concentration of contaminants in solution as well as significant concentration and agglomeration of solid particles. A high degree of purification was achieved for many parameters. The results of comparative laboratory tests for single and multiple freezing are presented. It was found that there was a higher degree of concentration of pollutants in wastewater frozen as man-made snow than in bulk ice. Furthermore, the hypothesis that long storage time of liquid as snow and sufficient temperature gradient metamorphism allows for high efficiency of the concentration process was confirmed. It was reported that the first 30% of the melted liquid volume contained over 90% of all impurities. It gives great opportunities to use this method to concentrate pollutants. The results revealed that the application of this process in full scale is possible. Significant agglomeration of solid particles was also noted. Tests with clay slurry showed that repeated freezing and thawing processes significantly improve the characteristics of slurry for sedimentation and filtration.

  6. Effect of freeze-thaw repetitions upon the supercooling release ability of ice-nucleating bacteria

    International Nuclear Information System (INIS)

    Tsuchiya, Yooko; Hasegawa, Hiromi; Sasaki, Kazuhiro

    2004-01-01

    We have studied the durability of ice-nucleating bacteria with a potent supercooling release capacity through repeated freeze-thaw cycles. Through experiment, we confirmed that UV sterilized Erwinia ananas maintains a superior supercooling release capacity at around -1degC through 2000 freeze-thaw cycles. We also found that γ-ray sterilization, which is more suitable than UV for large-scale sterilization treatment, has a similar effect at appropriately selected doses. (author)

  7. Sperm DNA damage in relation to lipid peroxidation following freezing-thawing of boar semen

    OpenAIRE

    Fraser, L.; Strzeżek, J.; Wasilewska, K.; Pareek, C.S.

    2017-01-01

    This study investigated the relationships between lipid peroxidation (LPO) and sperm DNA damage following freezing-thawing of boar semen in different extenders. The comet assay was used to measure the extent of sperm DNA damage in a cryoprotectant-free extender or in cryoprotectant-based extenders after single and repeated freezing and thawing. As well as an analysis of sperm motion characteristics, mitochondrial function, membrane integrity, and lipid peroxidation (LPO) were assessed simulta...

  8. Measuring influenza RNA quantity after prolonged storage or multiple freeze/thaw cycles.

    Science.gov (United States)

    Granados, Andrea; Petrich, Astrid; McGeer, Allison; Gubbay, Jonathan B

    2017-09-01

    In this study, we aim to determine what effects prolonged storage and repeated freeze/thaw cycles have on the stability of influenza A(H1N1)pdm09 (influenza A/H1N1)RNA. Cloned influenza A/H1N1 RNA transcripts were serially diluted from 8.0-1.0 log 10 copies/μl. RT-qPCR was used to measure RNA loss in transcripts stored at -80°C, -20°C, 4°C and 25°C for up to 84days or transcripts undergoing a total of 10 freeze/thaw cycles. Viral load was measured in clinical specimens stored at-80°C for three years (n=89 influenza A RNA extracts; n=35 primary specimens) and in 10 clinical specimens from the 2015/2016 influenza season that underwent 7 freeze/thaw cycles. RNA stored at -80°C, -20°C, 4°C and 25°C is stable for up to 56, 56, 21, and 7days respectively or up to 9 freeze/thaw cycles when stored at -80°C. There is no difference in viral load in clinical specimens that have been stored for up to three years at -80°C if they are re-extracted. Similarly, clinical specimens undergoing up to 7 freeze/thaw cycles are stable if they are re-extracted between cycles. Influenza specimens can be stored for up to three years at -80°C or undergo up to 7 freeze/thaw cycles without loss of RNA quantity if re-extracted. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. The effect of repeated freeze-thaw cycles on human muscle tissue visualized by postmortem computed tomography (PMCT)

    NARCIS (Netherlands)

    Klop, Anthony C.; Vester, Marloes E. M.; Colman, Kerri L.; Ruijter, Jan M.; van Rijn, Rick R.; Oostra, Roelof-Jan

    2017-01-01

    The aim of this study was to determine whether effects of repetitive freeze-thaw cycles, with various thawing temperatures, on human muscle tissue can be quantified using postmortem computed tomography (PMCT) technology. An additional objective was to determine the preferred thawing temperature for

  10. Relative Sensitivity of Photosynthesis and Respiration to Freeze-Thaw Stress in Herbaceous Species 1

    Science.gov (United States)

    Steffen, Kenneth L.; Arora, Rajeev; Palta, Jiwan P.

    1989-01-01

    The relative effect of a freeze-thaw cycle on photosynthesis, respiration, and ion leakage of potato leaf tissue was examined in two potato species, Solanum acaule Bitt. and Solanum commersonii Dun. Photosynthesis was found to be much more sensitive to freezing stress than was respiration, and demonstrated more than a 60% inhibition before any impairment of respiratory function was observed. Photosynthesis showed a slight to moderate inhibition when only 5 to 10% of the total electrolytes had leaked from the tissue (reversible injury). This was in contrast to respiration which showed no impairment until temperatures at which about 50% ion leakage (irreversible injury) had occurred. The influence of freeze-thaw protocol was further examined in S. acaule and S. commersonii, in order to explore discrepancies in the literature as to the relative sensitivities of photosynthesis and respiration. As bath cooling rates increased from 1°C/hour to about 3 or 6°C/hour, there was a dramatic increase in the level of damage to all measured cellular functions. The initiation of ice formation in deeply supercooled tissue caused even greater damage. As the cooling rates used in stress treatments increased, the differential sensitivity between photosynthesis and respiration nearly disappeared. Examination of agriculturally relevant, climatological data from an 11 year period confirmed that air cooling rates in the freezing range do not exceed 2°C/hour. It was demonstrated, in the studies presented here, that simply increasing the actual cooling rate from 1.0 to 2.9°C/hour, in frozen tissue from paired leaflet halves, meant the difference between cell survival and cell death. Images Figure 4 Figure 5 PMID:16666712

  11. Aroma improvement by repeated freeze-thaw treatment during Tuber melanosporum fermentation

    Science.gov (United States)

    Xiao, Deng-Rong; Liu, Rui-Sang; He, Long; Li, Hong-Mei; Tang, Ya-Ling; Liang, Xin-Hua; Chen, Tao; Tang, Ya-Jie

    2015-01-01

    The aroma attributes of sulfurous, mushroom and earthy are the most important characteristics of the aroma of Tuber melanosporum. However, these three aroma attributes are absent in the T. melanosporum fermentation system. To improve the quality of the aroma, repeated freeze-thaw treatment (RFTT) was adopted to affect the interplay of volatile organic compounds (VOCs). Using RFTT, not only was the score on the hedonic scale of the aroma increased from the “liked slightly” to the “liked moderately” grade, but the aroma attributes of sulfurous, mushroom and earthy could also be smelled in the T. melanosporum fermentation system for the first time. A total of 29 VOCs were identified, and 9 compounds were identified as the key discriminative volatiles affected by RFTT. Amino acid analysis revealed that methionine, valine, serine, phenylalanine, isoleucine and threonine were the key substrates associated with the biosynthesis of the 9 key discriminative VOCs. This study noted that amino acid metabolism played an important role in the regulation of the aroma of the T. melanosporum fermentation system. PMID:26607288

  12. Micromechanical properties of canine femoral articular cartilage following multiple freeze-thaw cycles.

    Science.gov (United States)

    Peters, Abby E; Comerford, Eithne J; Macaulay, Sophie; Bates, Karl T; Akhtar, Riaz

    2017-07-01

    Tissue material properties are crucial to understanding their mechanical function, both in healthy and diseased states. However, in certain circumstances logistical limitations can prevent testing on fresh samples necessitating one or more freeze-thaw cycles. To date, the nature and extent to which the material properties of articular cartilage are altered by repetitive freezing have not been explored. Therefore, the aim of this study is to quantify how articular cartilage mechanical properties, measured by nanoindentation, are affected by multiple freeze-thaw cycles. Canine cartilage plugs (n = 11) from medial and lateral femoral condyles were submerged in phosphate buffered saline, stored at 3-5°C and tested using nanoindentation within 12h. Samples were then frozen at -20°C and later thawed at 3-5°C for 3h before material properties were re-tested and samples re-frozen under the same conditions. This process was repeated for all 11 samples over three freeze-thaw cycles. Overall mean and standard deviation of shear storage modulus decreased from 1.76 ± 0.78 to 1.21 ± 0.77MPa (p = 0.91), shear loss modulus from 0.42 ± 0.19 to 0.39 ± 0.17MPa (p=0.70) and elastic modulus from 5.13 ± 2.28 to 3.52 ± 2.24MPa (p = 0.20) between fresh and three freeze-thaw cycles respectively. The loss factor increased from 0.31 ± 0.38 to 0.71 ± 1.40 (p = 0.18) between fresh and three freeze-thaw cycles. Inter-sample variability spanned as much as 10.47MPa across freezing cycles and this high-level of biological variability across samples likely explains why overall mean "whole-joint" trends do not reach statistical significance across the storage conditions tested. As a result multiple freeze-thaw cycles cannot be explicitly or statistically linked to mechanical changes within the cartilage. However, the changes in material properties observed herein may be sufficient in magnitude to impact on a variety of clinical and scientific studies of cartilage, and should be considered

  13. Research on strength attenuation law of concrete in freezing - thawing environment

    Science.gov (United States)

    Xiao, qianhui; Cao, zhiyuan; Li, qiang

    2018-03-01

    By rapid freezing and thawing method, the experiments of concrete have been 300 freeze-thaw cycles specimens in the water. The cubic compression strength value under different freeze-thaw cycles was measured. By analyzing the test results, the water-binder ratio of the concrete under freeze-thaw environments, fly ash and air entraining agent is selected dosage recommendations. The exponential attenuation prediction model and life prediction model of compression strength of concrete under freezing-thawing cycles considering the factors of water-binder ratio, fly ash content and air-entraining agent dosage were established. The model provides the basis for predicting the durability life of concrete under freezing-thawing environment. It also provides experimental basis and references for further research on concrete structures with antifreeze requirements.

  14. Effect of freeze-thaw cycles on load transfer between the biomineral and collagen phases in bovine dentin

    Energy Technology Data Exchange (ETDEWEB)

    Deymier-Black, A.C., E-mail: AlixDeymier2010@u.northwestern.edu [Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208 (United States); Almer, J.D., E-mail: almer@aps.anl.gov [Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439 (United States); Haeffner, D.R., E-mail: haeffner@aps.anl.gov [Advanced Photon Source, Argonne National Laboratory, Argonne, IL 60439 (United States); Dunand, D.C., E-mail: dunand@northwestern.edu [Department of Materials Science and Engineering, Northwestern University, Evanston, IL 60208 (United States)

    2011-10-10

    Stabilization of biological materials by freezing is widespread in the fields of medicine and biomaterials research and yet, in the case of hard biomaterials such as dentin, there is not a good understanding of how such treatments might affect the mechanical properties. The freezing and thawing may have a number of different effects on dentin including formation of cracks in the microstructure and denaturation of the collagen. Using high-energy synchrotron X-ray diffraction, the apparent moduli of bovine dentin samples were measured before and after various numbers of freeze-thaw cycles. It was determined that repeated freezing and thawing has no measurable effect on the hydroxyapatite or fibrillar apparent moduli up to 10 cycles. This confirms that the use of low temperature storage for stabilization of dentin is reasonable in cases where stiffness is a property of importance. Highlights: {yields} Studied the effect of freezing on the load transfer of HAP and fibrils in dentin. {yields} X-ray scattering measured HAP and fibril apparent moduli vs. freezing cycles. {yields} Apparent moduli did not vary significantly between 0 and 10 freeze thaw cycles. {yields} Residual strains imply no widespread cracking due to volumetric expansion of water. {yields} Dentin can be freeze-thawed with no significant effects on elastic properties.

  15. Quality Evaluation of Pork with Various Freezing and Thawing Methods

    Science.gov (United States)

    2014-01-01

    In this study, the physicochemical and sensory quality characteristics due to the influence of various thawing methods on electro-magnetic and air blast frozen pork were examined. The packaged pork samples, which were frozen by air blast freezing at −45℃ or electro-magnetic freezing at −55℃, were thawed using 4 different methods: refrigeration (4±1℃), room temperature (RT, 25℃), cold water (15℃), and microwave (2450 MHz). Analyses were carried out to determine the drip and cooking loss, water holding capacity (WHC), moisture content and sensory evaluation. Frozen pork thawed in a microwave indicated relatively less thawing loss (0.63-1.24%) than the other thawing methods (0.68-1.38%). The cooking loss after electro-magnetic freezing indicated 37.4% by microwave thawing, compared with 32.9% by refrigeration, 36.5% by RT, and 37.2% by cold water in ham. The thawing of samples frozen by electro-magnetic freezing showed no significant differences between the methods used, while the moisture content was higher in belly thawed by microwave (62.0%) after electro-magnetic freezing than refrigeration (54.8%), RT (61.3%), and cold water (61.1%). The highest overall acceptability was shown for microwave thawing after electro-magnetic freezing but there were no significant differences compared to that of the other samples. PMID:26761493

  16. Recent developments in novel freezing and thawing technologies applied to foods.

    Science.gov (United States)

    Wu, Xiao-Fei; Zhang, Min; Adhikari, Benu; Sun, Jincai

    2017-11-22

    This article reviews the recent developments in novel freezing and thawing technologies applied to foods. These novel technologies improve the quality of frozen and thawed foods and are energy efficient. The novel technologies applied to freezing include pulsed electric field pre-treatment, ultra-low temperature, ultra-rapid freezing, ultra-high pressure and ultrasound. The novel technologies applied to thawing include ultra-high pressure, ultrasound, high voltage electrostatic field (HVEF), and radio frequency. Ultra-low temperature and ultra-rapid freezing promote the formation and uniform distribution of small ice crystals throughout frozen foods. Ultra-high pressure and ultrasound assisted freezing are non-thermal methods and shorten the freezing time and improve product quality. Ultra-high pressure and HVEF thawing generate high heat transfer rates and accelerate the thawing process. Ultrasound and radio frequency thawing can facilitate thawing process by volumetrically generating heat within frozen foods. It is anticipated that these novel technologies will be increasingly used in food industries in the future.

  17. Experimental research on durability of recycled aggregate concrete under freeze- thaw cycles

    Science.gov (United States)

    Cheng, Yanqiu; Shang, Xiaoyu; Zhang, Youjia

    2017-07-01

    The freeze-thaw durability of recycled aggregate concrete has significance for the concrete buildings in the cold region. In this paper, the rapid freezing and thawing cycles experience on recycle aggregate concrete was conducted to study on the effects of recycle aggregate amount, water-binder ratio and fly ash on freeze-thaw durability of recycle aggregate concrete. The results indicates that recycle aggregate amount makes the significant influence on the freeze-thaw durability. With the increase of recycled aggregates amount, the freeze-thaw resistance for recycled aggregate concrete decreases. Recycled aggregate concrete with lower water cement ratio demonstrates better performance of freeze-thaw durability. It is advised that the amount of fly ash is less than 30% for admixture of recycled aggregates in the cold region.

  18. Three-Dimensional Microstructure of Biological Tissues during Freezing and Thawing

    Science.gov (United States)

    Ishiguro, Hiroshi; Horimizu, Takashi; Kataori, Akinobu; Kajigaya, Hiroshi

    Three-dimensional behavior of ice crystals and cells during the freezing and thawing of biological tissues was investigated microscopically in real time by using a confocal laser scanning microscope(CLSM) and a fluorescent dye, acridine orange (AO). Fresh tender meat (2nd pectoral muscles) of chicken was stained with the AO in physiological saline to distinguish ice crystals and cells by their different colors, and then frozen and thawed under two different thermal protocols: a) slow-cooling and rapid-warming and b) rapid-cooling and rapid-warming. The CLSM noninvasively produced optical tomograms of the tissues to clarify the pattern of freezing, morphology of ice crystals in the tissues, and the interaction between ice crystals and cells. Also, the tissues were morphologically investigated by pathological means after the freezing and thawing. Typical freezing pattern during the slow-cooling was extracellular-freezing, and those during the rapid-cooling were extracellular-freezing and intracellular freezing with a lot of fine ice crystals in the cells. Cracks caused by the extracellular and intracellular ice crystals remained in the muscle tissues after the thawing. The results obtained by using the CLSM/dye method were consistent with pathologically morphological changes in the tissues through freezing and thawing.

  19. Freeze-thaw performance testing of whole concrete railroad ties.

    Science.gov (United States)

    2013-10-01

    Freezing and thawing durability tests of prestressed concrete ties are normally performed according to ASTM C666 specifications. Small specimens are cut from the shoulders of concrete ties and tested through 300 cycles of freezing and thawing. Saw-cu...

  20. Resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing

    Directory of Open Access Journals (Sweden)

    Sidney Verza Jr.

    2009-10-01

    Full Text Available Objective: To study the resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing by using the fast liquid nitrogen vapor method. Material and Methods: Semen specimens were obtained from sixteen normal and oligozoospermic individuals who required disposal at the sperm bank. Five of them had testicular cancer. Specimens were thawed and an aliquot was removed for analysis. The remaining specimens were refrozen without removing the cryomedia. Repeated freeze-thaw cycles were performed until no motile sperm were observed. Sperm motility, number of motile spermatozoa and viability were determined after thawing. Resistance to cryoinjury was compared between groups and also after each refreezing cycle within groups. Results: Motile spermatozoa were recovered after five and two refreeze-thawing cycles in normozoospermic and oligozoospermic specimens, respectively. There were no significant differences in the recovery of motile spermatozoa between thaws within each group of normal and oligozoospermic specimens, but percentage motility and total number of motile spermatozoa were significantly lower in the oligozoospermic one. Specimens from men with cancer were exposed to six refreeze-thawing cycles. Although recovery of motile spermatozoa was significantly impaired after each thawing, there were no significant differences in the recovery of motile sperm between thaws in cancer and non-cancer groups. Conclusions: Human spermatozoa resist repeated cryopreservation using the fast liquid nitrogen vapor method. Normozoospermic specimens withstand refreezing for an average two cycles longer than oligozoospermic ones. Specimens from cancer patients seem to resist repeated cryoinjury similarly to non-cancer counterparts. Resistance to repeated cryoinjury was related to the initial semen quality.

  1. Comparison Study of Three Common Technologies for Freezing-Thawing Measurement

    Directory of Open Access Journals (Sweden)

    Xinbao Yu

    2010-01-01

    Full Text Available This paper describes a comparison study on three different technologies (i.e., thermocouple, electrical resistivity probe and Time Domain Reflectometry (TDR that are commonly used for frost measurement. Specially, the paper developed an analyses procedure to estimate the freezing-thawing status based on the dielectric properties of freezing soil. Experiments were conducted where the data of temperature, electrical resistivity, and dielectric constant were simultaneously monitored during the freezing/thawing process. The comparison uncovered the advantages and limitations of these technologies for frost measurement. The experimental results indicated that TDR measured soil dielectric constant clearly indicates the different stages of the freezing/thawing process. Analyses method was developed to determine not only the onset of freezing or thawing, but also the extent of their development. This is a major advantage of TDR over other technologies.

  2. Effect of freeze/thaw cycles on several biomarkers in urine from patients with kidney disease.

    Science.gov (United States)

    Zhang, Yinan; Luo, Yi; Lu, Huijuan; Wang, Niansong; Shen, Yixie; Chen, Ruihua; Fang, Pingyan; Yu, Hong; Wang, Congrong; Jia, Weiping

    2015-04-01

    Urine samples were collected from eleven randomly selected patients with kidney disease, including diabetic nephropathy, chronic nephritis, and nephritic syndrome. Urine samples were treated with one of four protocols for freezing and thawing: freeze directly and thaw directly; freeze directly and thaw by temperature gradient; freeze by temperature gradient and thaw directly; and freeze by temperature gradient and thaw by temperature gradient. After one to six freeze/thaw cycles at -20°C or -80°C, different biomarkers showed differential stabilities. The concentrations of total protein, calcium, and potassium did not change significantly after five freeze/thaw cycles at either -20°C or -80°C. Albumin could only sustain three freeze/thaw cycles at -20°C before it started to degrade. We recommend that urine be stored at -80°C as albumin and the organic ions could sustain five and six freeze/thaw cycles, respectively, using the simple "direct freeze and direct thaw" protocol. Furthermore, in most cases, gradient freeze/thaw cycles are not necessary for urine sample storage.

  3. Response of Soil Biogeochemistry to Freeze-thaw Cycles: Impacts on Greenhouse Gas Emission and Nutrient Fluxes

    Science.gov (United States)

    Rezanezhad, F.; Parsons, C. T.; Smeaton, C. M.; Van Cappellen, P.

    2014-12-01

    Freeze-thaw is an abiotic stress applied to soils and is a natural process at medium to high latitudes. Freezing and thawing processes influence not only the physical properties of soil, but also the metabolic activity of soil microorganisms. Fungi and bacteria play a crucial role in soil organic matter degradation and the production of greenhouse gases (GHG) such as CO2, CH4 and N2O. Production and consumption of these atmospheric trace gases are the result of biological processes such as photosynthesis, aerobic respiration (CO2), methanogenesis, methanotrophy (CH4), nitrification and denitrification (N2O). To enhance our understanding of the effects of freeze-thaw cycles on soil biogeochemical transformations and fluxes, a highly instrumented soil column experiment was designed to realistically simulate freeze-thaw dynamics under controlled conditions. Pore waters collected periodically from different depths of the column and solid-phase analyses on core material obtained at the initial and end of the experiment highlighted striking geochemical cycling. CO2, CH4 and N2O production at different depths within the column were quantified from dissolved gas concentrations in pore water. Subsequent emissions from the soil surface were determined by direct measurement in the head space. Pulsed CO2 emission to the headspace was observed at the onset of thawing, however, the magnitude of the pulse decreased with each subsequent freeze-thaw cycle indicating depletion of a "freeze-thaw accessible" carbon pool. Pulsed CO2 emission was due to a combination of physical release of gases dissolved in porewater and entrapped below the frozen zone and changing microbial respiration in response to electron acceptor variability (O2, NO3-, SO42-). In this presentation, we focus on soil-specific physical, chemical, microbial factors (e.g. redox conditions, respiration, fermentation) and the mechanisms that drive GHG emission and nutrient cycling in soils under freeze-thaw cycles.

  4. A pore-scale study of fracture dynamics in rock using X-ray micro-CT under ambient freeze-thaw cycling.

    Science.gov (United States)

    De Kock, Tim; Boone, Marijn A; De Schryver, Thomas; Van Stappen, Jeroen; Derluyn, Hannelore; Masschaele, Bert; De Schutter, Geert; Cnudde, Veerle

    2015-03-03

    Freeze-thaw cycling stresses many environments which include porous media such as soil, rock and concrete. Climate change can expose new regions and subject others to a changing freeze-thaw frequency. Therefore, understanding and predicting the effect of freeze-thaw cycles is important in environmental science, the built environment and cultural heritage preservation. In this paper, we explore the possibilities of state-of-the-art micro-CT in studying the pore scale dynamics related to freezing and thawing. The experiments show the development of a fracture network in a porous limestone when cooling to -9.7 °C, at which an exothermal temperature peak is a proxy for ice crystallization. The dynamics of the fracture network are visualized with a time frame of 80 s. Theoretical assumptions predict that crystallization in these experiments occurs in pores of 6-20.1 nm under transient conditions. Here, the crystallization-induced stress exceeds rock strength when the local crystal fraction in the pores is 4.3%. The location of fractures is strongly related to preferential water uptake paths and rock texture, which are visually identified. Laboratory, continuous X-ray micro-CT scanning opens new perspectives for the pore-scale study of ice crystallization in porous media as well as for environmental processes related to freeze-thaw fracturing.

  5. Behaviour of polycyclic aromatic hydrocarbons (PAH) in soils under freeze-thaw cycles

    Science.gov (United States)

    Zschocke, Anne; Schönborn, Maike; Eschenbach, Annette

    2010-05-01

    The arctic region will be one of the most affected regions by climate change due to the predicted temperature rise. As a result of anthropogenic actions as mining, exploration and refining as well as atmospheric transport pollutions can be found in arctic soils. Therefore questions on the behaviour of organic contaminants in permafrost influenced soils are of high relevance. First investigations showed that permafrost can act as a semi-permeable layer for PAH (Curtosi et al., 2007). Therefore it can be assumed that global warming could result in a mobilization of PAH in these permafrost influenced soils. On the other hand a low but detectable mineralization of organic hydrocarbons by microorganisms under repeated freeze-thaw cycles was analysed (Börresen et al. 2007, Eschenbach et al. 2000). In this study the behaviour and distribution of PAH under freezing and periodically freezing and thawing were investigated in laboratory column experiments with spiked soil materials. Two soil materials which are typical for artic regions, a organic matter containing melt water sand and a well decomposed peat, were homogeneously spiked with a composite of a crude oil and the PAH anthracene and benzo(a)pyrene. After 14days preincubation time the soil material was filled in the laboratory columns (40cm high and 10 cm in diameter). Based on studies by Chuvilin et al. (2001) the impact of freezing of the upper third of the column from the surface downwards was examined. The impact of freezing was tested in two different approaches the first one with a single freezing step and the second one with a fourfold repeated cycle of freezing and thawing which takes about 6 or 7 days each. The experimental design and very first results will be shown and discussed. In some experiments with the peat a higher concentration of anthracene and benzo(a)pyrene could be detected below the freezing front in the unfrozen part of the column. Whereas the concentration of PAH had slightly decreased in

  6. Effec t of Freeze-Thaw Cycles on Lipid Oxidation and Myowater in Broiler Chickens

    Directory of Open Access Journals (Sweden)

    S Ali

    2016-03-01

    Full Text Available ABSTRACT The present study was carried out to investigate the influence of freezing-thawing cycles (0, 2, 4 and 6 on lipid oxidation and myowater contents and distribution. Nine replicates of chicken breast meat samples were used for each cycle. Lipid oxidation was determined by measuring peroxide value, and malondialdehyde (MDA concentrations, which reflect thiobarbituric acid reactive substance (TBARS. Color was determined with a digital colorimeter. Muscle moisture contents were determined by drip loss and thawing loss, water holding capacity, and nuclear magnetic resonance (NMR. The results showed that, as the number of freeze-thaw cycles increased, meat redness decreased and MDA and peroxide values increased. Drip loss and thawing loss tended to decreasing as the number of freeze-thaw cycles increased. Water holding capacity also decreased as a function of increasing freeze-thaw cycles. NMR relaxometry profile showed freeze-thaw cycles change the water distribution of meat subjected to multiple freeze-thaw cycles. In conclusion, multiple freezing and thawing rate (6 cycles increased lipid oxidation, decreased myowater, and impaired the color of chicken meat.

  7. Monitoring Freeze Thaw Transitions in Arctic Soils using Complex Resistivity Method

    Science.gov (United States)

    Wu, Y.; Hubbard, S. S.; Ulrich, C.; Dafflon, B.; Wullschleger, S. D.

    2012-12-01

    The Arctic region, which is a sensitive system that has emerged as a focal point for climate change studies, is characterized by a large amount of stored carbon and a rapidly changing landscape. Seasonal freeze-thaw transitions in the Arctic alter subsurface biogeochemical processes that control greenhouse gas fluxes from the subsurface. Our ability to monitor freeze thaw cycles and associated biogeochemical transformations is critical to the development of process rich ecosystem models, which are in turn important for gaining a predictive understanding of Arctic terrestrial system evolution and feedbacks with climate. In this study, we conducted both laboratory and field investigations to explore the use of the complex resistivity method to monitor freeze thaw transitions of arctic soil in Barrow, AK. In the lab studies, freeze thaw transitions were induced on soil samples having different average carbon content through exposing the arctic soil to temperature controlled environments at +4 oC and -20 oC. Complex resistivity and temperature measurements were collected using electrical and temperature sensors installed along the soil columns. During the laboratory experiments, resistivity gradually changed over two orders of magnitude as the temperature was increased or decreased between -20 oC and 0 oC. Electrical phase responses at 1 Hz showed a dramatic and immediate response to the onset of freeze and thaw. Unlike the resistivity response, the phase response was found to be exclusively related to unfrozen water in the soil matrix, suggesting that this geophysical attribute can be used as a proxy for the monitoring of the onset and progression of the freeze-thaw transitions. Spectral electrical responses contained additional information about the controls of soil grain size distribution on the freeze thaw dynamics. Based on the demonstrated sensitivity of complex resistivity signals to the freeze thaw transitions, field complex resistivity data were collected over

  8. Effects of aging and freezing/thawing sequence on quality attributes of bovine and

    Directory of Open Access Journals (Sweden)

    Hyun-Wook Kim

    2017-02-01

    Full Text Available Objective The effects of aging and freezing/thawing sequence on color, physicochemical, and enzymatic characteristics of two beef muscles (Mm. gluteus medius, GM and biceps femoris, BF were evaluated. Methods Beef muscles at 3 d postmortem were assigned to four different combinations of aging and freezing/thawing sequence as follows; aging at 2°C for 3 wk (A3, never-frozen control, freezing at −28°C for 2 wk then thawing (F2, frozen/thawed-only, aging at 2°C for 3 wk, freezing at −28°C for 2 wk then thawing (A3F2, and freezing at −28°C for 2 wk, thawing then further aging at 2°C for 3 wk (F2A3. Results No significant interactions between different aging/freezing/thawing treatments and muscle type on all measurements were found. Postmortem aging, regardless of aging/freezing/thawing sequence, had no impact on color stability of frozen/thawed beef muscles (p<0.05. F2A3 resulted in higher purge loss than F2 and A3F2 treatments (p<0.05. A3F2 and F2A3 treatments resulted in lower shear force of beef muscles compared to F2 (p<0.05. Although there was no significant difference in glutathione peroxidase (GSH-Px activity, F2A3 had the highest β-N-acetyl glucominidase (BNAG activity in purge, but the lowest BNAG activity in muscle (p<0.05. GM muscle exhibited higher total color changes and purge loss, and lower GSH-Px activity than BF muscle. Conclusion The results from this present study indicate that different combinations of aging/freezing/thawing sequence would result in considerable impacts on meat quality attributes, particularly thaw/purge loss and tenderness. Developing a novel freezing strategy combined with postmortem aging will be beneficial for the food/meat industry to maximize its positive impacts on tenderness, while minimizing thaw/purge loss of frozen/thawed meat.

  9. Protein and solute distribution in drug substance containers during frozen storage and post-thawing: a tool to understand and define freezing-thawing parameters in biotechnology process development.

    Science.gov (United States)

    Kolhe, Parag; Badkar, Advait

    2011-01-01

    Active pharmaceutical ingredient for biotechnology-based drugs, commonly known as drug substance (DS), is often stored frozen for longer shelf-life. Freezing DS enhances stability by slowing down reaction rates that lead to protein instability, minimizes the risk of microbial growth, and eliminates the risk of transport-related stress. High density polyethylene bottles are commonly used for storing monoclonal antibody DS due to good mechanical stress/strain resistant properties even at low temperatures. Despite the aforementioned advantages for frozen storage of DS, this is not devoid of risks. Proteins are known to undergo ice-water surface denaturation, cryoconcentration, and cold denaturation during freezing. A systematic investigation was performed to better understand the protein and solute distribution along with potential of aggregate formation during freeze and thaw process. A significant solute and protein concentration gradient was observed for both frozen and thawed DS bottles. In case of thawed DS, cryoconcentration was localized in the bottom layer and a linear increase in concentration as a function of liquid depth was observed. On the other hand, for frozen DS, a "bell shaped" cryoconcentration distribution was observed between the bottom layers and centre position. A cryoconcentration of almost three-fold was observed for frozen DS in the most concentrated part when freezing was conducted at -20 and -40 °C and 2.5-fold cryoconcentration was observed in the thawed DS before mixing. The information obtained in this study is critical to design freeze thaw experiments, storage condition determination, and process improvement in manufacturing environment. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

  10. The impact of freeze-thawing on the friability of formed elemental sulfur

    Energy Technology Data Exchange (ETDEWEB)

    Clark, P.D.; Almond, P.

    1995-10-01

    Winter weather fluctuation tests were performed in a laboratory to determine the freeze-thaw effect on the particle size distribution and friability of sulfur samples of Polish Air Prills, Procor GX Granules, Petrosul, Sandvik Rotoform, and slate. Formed elemental sulfur is often stored outdoors and is subjected to temperature fluctuations which could potentially result in structural stress, particularly if the sulfur contained any moisture from rainwater or melting snow. Concern was expressed that sulfur stored under these conditions might have increased friability during shipping and handling. Results of stress level tests indicated that winter weather fluctuations did not effect the particle size distribution or friability of the samples. When sulfur samples were in contact with water for one month, there were was no significant change in overall friability. It was concluded that all premium product forms of solid elemental sulfur could be stored outdoors, even if subjected to freeze-thaw cycles. 4 tabs., 5 figs.

  11. Effect of Novel Quick Freezing Techniques Combined with Different Thawing Processes on Beef Quality

    Science.gov (United States)

    Yoo, Seon-Mi; Han, Gui-Jung

    2014-01-01

    This study investigated the effect of various freezing and thawing techniques on the quality of beef. Meat samples were frozen using natural convection freezing (NF), individual quick freezing (IQF), or cryogenic freezing (CF) techniques, followed by natural convection thawing (NCT) or running water thawing (RT). The meat was frozen until the core temperature reached -12℃ and then stored at -24℃, followed by thawing until the temperature reached 5℃. Quality parameters, such as the pH, water binding properties, CIE color, shear force, and microstructure of the beef were elucidated. Although the freezing and thawing combinations did not cause remarkable changes in the quality parameters, rapid freezing, in the order of CF, IQF, and NF, was found to minimize the quality deterioration. In the case of thawing methods, NCT was better than RT and the meat quality was influence on the thawing temperature rather than the thawing rate. Although the microstructure of the frozen beef exhibited an excessive loss of integrity after the freezing and thawing, it did not cause any remarkable change in the beef quality. Taken together, these results demonstrate that CF and NCT form the best combination for beef processing; however, IQF and NCT may have practical applications in the frozen food industry. PMID:26761674

  12. Detection of freeze-thaw weathering effect using X-ray micro computed tomography

    Science.gov (United States)

    Park, J.; Hyun, C.; Park, H.

    2011-12-01

    Physical weathering caused by repeated freeze-thaw action of water inside rock pores or cracks was artificially simulated in laboratory. The tests were conducted on three rock types, i.e. diorite, basalt, and tuff, which are the major rock types around King Sejong Station of Korea located in Barton Peninsula, King George Island, Antarctica. The temperature of freeze-thaw cycle was also set with simulated the air temperature of the station, i.e. the maximum temperature was + 10 °C and the minimum temperature was - 20 °C. Three cylindrical specimens composed of one for each rock type with 24.6 mm diameter and 14.5 ~ 17.7 mm length were prepared, and 2 mm diameter and 7 mm shallow depth hole was drilled on the center of the specimens. To exaggerate the effect of the freeze-thaw weathering, all tests were conducted under completely saturated condition. 50 cycles of the freeze-thaw test was carried, and X-ray micro computed tomography (CT) images of each rock specimen were obtained after every 10 cycles. Using X-ray micro CT images, 3D structure was rendered and pore and crack structures were extracted. The changes of porosity, absorption rate and pore and crack structure were detected. Porosity of all specimens was decreased linearly and absorption rate of all specimens was increased linearly as weathering processes; the pore connection and crack propagation was detected in 3D rendering pore and crack structure. The change of tuff specimen is the most remarkable among three rock types used in the research, because of its relatively high initial absorption rate and low strength. This work was supported by the National Research Foundation of Korea(NRF) grant funded by the Korea government(MEST) (No. 2011-0027520).

  13. Phenome data - Freeze-thaw stress - DGBY | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available ndard Gene Name in SGD Growth rate BM/BW Rate of growth in non-strees condition (Mutant/Wild-type) Growth ra...te (AM/AW)/(BM/BW) AM/AW represents rate of growth after exposure to freeze-thaw strees

  14. Prediction model for carbonation depth of concrete subjected to freezing-thawing cycles

    Science.gov (United States)

    Xiao, Qian Hui; Li, Qiang; Guan, Xiao; Xian Zou, Ying

    2018-03-01

    Through the indoor simulation test of the concrete durability under the coupling effect of freezing-thawing and carbonation, the variation regularity of concrete neutralization depth under freezing-thawing and carbonation was obtained. Based on concrete carbonation mechanism, the relationship between the air diffusion coefficient and porosity in concrete was analyzed and the calculation method of porosity in Portland cement concrete and fly ash cement concrete was investigated, considering the influence of the freezing-thawing damage on the concrete diffusion coefficient. Finally, a prediction model of carbonation depth of concrete under freezing-thawing circumstance was established. The results obtained using this prediction model agreed well with the experimental test results, and provided a theoretical reference and basis for the concrete durability analysis under multi-factor environments.

  15. The effect of the freezing curve type on bull spermatozoa motility after thawing

    Directory of Open Access Journals (Sweden)

    Martina Doležalová

    2015-01-01

    Full Text Available The objective of this work was to determine the effect of selected freezing curves on spermatozoa survivability after thawing, defined by its motility. The ejaculates of nine selected sires of the same age, breed, and frequency of collecting, bred under the same breeding conditions including handling, stabling, feeding system and feeding ratio composition, were repeatedly collected and evaluated. Sperm samples of each sire were diluted using only one extender and divided into four parts. Selected four freezing curves – the standard, commercially recommended three-phase curve; a two-phase curve; a slow three-phase curve; and a fast three-phase curve, differing in the course of temperature vs time, were applied. The percentage rate of progressive motile spermatozoa above head was determined immediately after thawing, and after 30, 60, 90, and 120 min of the thermodynamic test (TDT. Moreover, average spermatozoa motility (AMOT and spermatozoa motility decrease (MODE throughout the entire TDT were evaluated. Insemination doses frozen using the simpler two-phase curve demonstrated the highest motility values (+2.97% to +10.37%; P < 0.05–0.01 immediately after thawing and during the entire TDT. Concurrently, the highest AMOT (+4.37% to +8.82%; P < 0.01 was determined. The highest spermatozoa motility values were detected after thawing doses frozen by the two-phase freezing curve in eight out of nine sires. Simultaneously, a significant effect of sire individuality was clearly confirmed. Inter-sire differences of spermatozoa motility during TDT as well as AMOT and MODE were significant (P < 0.01. The findings describing both factors of interaction indicate the necessity of individual cryopreservation of the ejaculate to increase its fertilization capability after thawing.

  16. FREEZING AND THAWING TIME PREDICTION METHODS OF FOODS II: NUMARICAL METHODS

    Directory of Open Access Journals (Sweden)

    Yahya TÜLEK

    1999-03-01

    Full Text Available Freezing is one of the excellent methods for the preservation of foods. If freezing and thawing processes and frozen storage method are carried out correctly, the original characteristics of the foods can remain almost unchanged over an extended periods of time. It is very important to determine the freezing and thawing time period of the foods, as they strongly influence the both quality of food material and process productivity and the economy. For developing a simple and effectively usable mathematical model, less amount of process parameters and physical properties should be enrolled in calculations. But it is a difficult to have all of these in one prediction method. For this reason, various freezing and thawing time prediction methods were proposed in literature and research studies have been going on.

  17. DSC and TMA studies on freezing and thawing gelation of galactomannan polysaccharide

    International Nuclear Information System (INIS)

    Iijima, Mika; Hatakeyama, Tatsuko; Hatakeyama, Hyoe

    2012-01-01

    Research highlights: ► Locust bean gum forms hydrogels by freezing and thawing. ► Syneresis was observed when freezing and thawing cycle (n) increased. ► Dynamic Young's modulus increased with increasing n. ► Non-freezing water content restrained by hydrogels decreased with increasing n. ► Strong gel with densely packed network structure formed with increasing n. - Abstract: Among various kinds of polysaccharides known to form hydrogels, locust bean gum (LBG) consisting of a mannose backbone and galactose side chains has unique characteristics, since LBG forms hydrogels by freezing and thawing. In this study, effect of thermal history on gelation was investigated by differential scanning calorimetry (DSC) and thermomechanical analysis (TMA). Gel/sol ratio calculated by weighing method was found to be affected by sol concentration, freezing rate and the number of freezing and thawing cycle (n). Once LBG hydrogels are formed, they are thermally stable, although syneresis was observed when n increased. Dynamic Young's modulus (E′) of hydrogels measured by TMA in water increased with increasing n and decreasing freezing rate. Non-freezing water calculated from DSC melting peak of ice in the gel decreased with increasing n and decreasing freezing rate. Morphological observation of freeze-dried gels was carried out by scanning electron microscopy (SEM). The above results indicate that weak hydrogel having large molecular network structure transformed into strong gel with densely packed network structure by increasing n and decreasing freezing rate.

  18. Products and stability of phosphate reactions with lead under freeze-thaw cycling in simple systems

    International Nuclear Information System (INIS)

    Hafsteinsdottir, Erla G.; White, Duanne A.; Gore, Damian B.; Stark, Scott C.

    2011-01-01

    Orthophosphate fixation of metal contaminated soils in environments that undergo freeze-thaw cycles is understudied. Freeze-thaw cycling potentially influences the reaction rate, mineral chemical stability and physical breakdown of particles during fixation. This study determines what products form when phosphate (triple superphosphate [Ca(H 2 PO 4 ) 2 ] or sodium phosphate [Na 3 PO 4 ]) reacts with lead (PbSO 4 or PbCl 2 ) in simple chemical systems in vitro, and assesses potential changes in formation during freeze-thaw cycles. Systems were subjected to multiple freeze-thaw cycles from +10 deg. C to -20 deg. C and then analysed by X-ray diffractometry. Pyromorphite formed in all systems and was stable over multiple freeze-thaw cycles. Low temperature lead orthophosphate reaction efficiency varied according to both phosphate and lead source; the most time-efficient pyromorphite formation was observed when PbSO 4 and Na 3 PO 4 were present together. These findings have implications for the manner in which metal contaminated materials in freezing ground can be treated with phosphate. - Highlights: → Formation of lead phosphate products in cold environments is identified. → Potential change in formation during freeze-thaw cycling is assessed. → Lead phosphate reaction efficiency varies according to phosphate and lead source. → Pyromorphite formation is stable during 240 freeze-thaw cycles. - Pyromorphite, formed from Pb phosphate fixation, is stable during multiple freeze-thaw cycles but the efficiency of the fixation depends on the phosphate source and the type of Pb mineral.

  19. Evaluation and Comparison of Freeze-Thaw Tests and Air Void Analysis of Pervious Concrete

    DEFF Research Database (Denmark)

    Lund, Mia Schou Møller; Hansen, Kurt Kielsgaard; Kevern, John T.

    2016-01-01

    Pearl-Chain Bridge technology is an innovative precast arch bridge solution which uses pervious concrete as fill material. To ensure longevity of the bridge superstructure it is necessary that the per-vious concrete fill is designed to be freeze-thaw durable; however, no standards exist on how...... to eval-uate the freeze-thaw resistance of fresh or hardened pervious concrete and correspondingly what constitutes acceptable freeze-thaw durability. A greater understanding of the correlation between the freeze-thaw performance and the air void structure of pervious concrete is needed. In the present...... study six pervious concrete mixes were exposed to freeze-thaw testing, and their air void structure was analyzed using an automated linear-traverse method. It was found that there is a miscorrelation between these two test methods in their assumption of whether or not the large interconnected voids...

  20. Evaluation of freeze-thaw durability of pervious concrete by use of operational modal analysis

    DEFF Research Database (Denmark)

    Lund, M.S.M.; Hansen, K. K.; Brincker, R.

    2018-01-01

    It is well-known that laboratory testing of pervious concrete's freeze-thaw performance is too harsh and does not agree well with field observations. The most commonly used laboratory freeze-thaw test method for pervious concrete is similar to that used for conventional concrete even though...... the void structure of the two materials is completely different. In the present study, a new freeze-thaw test method for pervious concrete is suggested and tested on one baseline mix, with three different contents of entrained air. The evaluation of freeze-thaw damage on pervious concrete beams...... was evaluated from the decrease in mass and from operational modal analysis which provides an accurate determination of the change in natural frequencies with freeze-thaw exposure. Operational modal analysis was also used to determine the Young's modulus, shear modulus, and Poisson's ratio of the pervious...

  1. Influence of Freeze-Thaw Damage on the Steel Corrosion and Bond-Slip Behavior in the Reinforced Concrete

    Directory of Open Access Journals (Sweden)

    Fangzhi Zhu

    2016-01-01

    Full Text Available This paper mainly studies the behavior of steel corrosion in various reinforced concrete under freeze-thaw environment. The influence of thickness of concrete cover is also discussed. Additionally, the bond-slip behavior of the reinforced concrete after suffering the freeze-thaw damage and steel corrosion has also be presented. The results show that the freeze-thaw damage aggravates the steel corrosion in concrete, and the results become more obvious in the concrete after suffering serious freeze-thaw damage. Compared with the ordinary concrete, both air entrained concrete and waterproofing concrete possess better resistance to steel corrosion under the same freeze-thaw environment. Moreover, increasing the thicknesses of concrete cover is also an effective method of improving the resistance to steel corrosion. The bond-slip behavior of reinforced concrete with corroded steel decreases with the increase of freeze-thaw damage, especially for the concrete that suffered high freeze-thaw cycles. Moreover, there exists a good correlation between the parameters of bond-slip and freeze-thaw cycles. The steel corrosion and bond-slip behavior of reinforced concrete should be considered serious under freeze-thaw cycles environment, which significantly impact the durability and safety of concrete structure.

  2. Effect of moisture and freeze-thaw on mechanical properties of CRM asphalt mixture

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Nak-Seok; Cho, Kee-Ju [Kyonggi University, Suwon(Korea)

    2000-06-30

    This paper presents the experimental test results on moisture and freeze-thaw resistance of hot mix crumb rubber modified (CRM) asphalt concrete mixture. To compare the differences in mechanical properties of conventional and CRM asphalt concretes, various tests were conducted under different moisture conditions and freeze-thaw cycles. Marshall mix design was also performed to determine the optimum asphalt contents for the both asphalt concrete mixtures. Test results revealed that the moisture and freeze-thaw resistance of CRM asphalt mixture was superior to the conventional asphalt concrete. As a result, it is considered that the utilization of waste tires in asphalt pavements has the potential of minimizing the damage due to the moisture and freeze-thaw. (author). 9 refs., 4 tabs., 8 figs.

  3. Products and stability of phosphate reactions with lead under freeze-thaw cycling in simple systems

    Energy Technology Data Exchange (ETDEWEB)

    Hafsteinsdottir, Erla G., E-mail: erla.hafsteinsdottir@gmail.com [Department of Environment and Geography, Macquarie University, NSW 2109 (Australia); White, Duanne A., E-mail: duanne.white@mq.edu.au [Department of Environment and Geography, Macquarie University, NSW 2109 (Australia); Gore, Damian B., E-mail: damian.gore@mq.edu.au [Department of Environment and Geography, Macquarie University, NSW 2109 (Australia); Stark, Scott C., E-mail: scott.stark@aad.gov.au [Environmental Protection and Change, Australian Antarctic Division, Department of Sustainability, Environment, Water, Population and Communities, Tasmania 7050 (Australia)

    2011-12-15

    Orthophosphate fixation of metal contaminated soils in environments that undergo freeze-thaw cycles is understudied. Freeze-thaw cycling potentially influences the reaction rate, mineral chemical stability and physical breakdown of particles during fixation. This study determines what products form when phosphate (triple superphosphate [Ca(H{sub 2}PO{sub 4}){sub 2}] or sodium phosphate [Na{sub 3}PO{sub 4}]) reacts with lead (PbSO{sub 4} or PbCl{sub 2}) in simple chemical systems in vitro, and assesses potential changes in formation during freeze-thaw cycles. Systems were subjected to multiple freeze-thaw cycles from +10 deg. C to -20 deg. C and then analysed by X-ray diffractometry. Pyromorphite formed in all systems and was stable over multiple freeze-thaw cycles. Low temperature lead orthophosphate reaction efficiency varied according to both phosphate and lead source; the most time-efficient pyromorphite formation was observed when PbSO{sub 4} and Na{sub 3}PO{sub 4} were present together. These findings have implications for the manner in which metal contaminated materials in freezing ground can be treated with phosphate. - Highlights: > Formation of lead phosphate products in cold environments is identified. > Potential change in formation during freeze-thaw cycling is assessed. > Lead phosphate reaction efficiency varies according to phosphate and lead source. > Pyromorphite formation is stable during 240 freeze-thaw cycles. - Pyromorphite, formed from Pb phosphate fixation, is stable during multiple freeze-thaw cycles but the efficiency of the fixation depends on the phosphate source and the type of Pb mineral.

  4. Effect of waxy rice flour and cassava starch on freeze-thaw stability of rice starch gels.

    Science.gov (United States)

    Charoenrein, Sanguansri; Preechathammawong, Nutsuda

    2012-10-01

    Repeatedly frozen and thawed rice starch gel affects quality. This study investigated how incorporating waxy rice flour (WF) and cassava starch (CS) in rice starch gel affects factors used to measure quality. When rice starch gels containing 0-2% WF and CS were subjected to 5 freeze-thaw cycles, both WF and CS reduced the syneresis in first few cycles. However CS was more effective in reducing syneresis than WF. The different composite arrangement of rice starch with WF or CS caused different mechanisms associated with the rice starch gel retardation of retrogradation, reduced the spongy structure and lowered syneresis. Both swollen granules of rice starch and CS caused an increase in the hardness of the unfrozen and freeze-thawed starch gel while highly swollen WF granules caused softer gels. These results suggested that WF and CS were effective in preserving quality in frozen rice starch based products. Copyright © 2012 Elsevier Ltd. All rights reserved.

  5. Development of Steel Fiber-Reinforced Expanded-Shale Lightweight Concrete with High Freeze-Thaw Resistance

    Directory of Open Access Journals (Sweden)

    Mingshuang Zhao

    2018-01-01

    Full Text Available For the popularized structural application, steel fiber-reinforced expanded-shale lightweight concrete (SFRELC with high freeze-thaw resistance was developed. The experimental study of this paper figured out the effects of air-entraining content, volume fraction of steel fibers, and fine aggregate type. Results showed that while the less change of mass loss rate was taken place for SFRELC after 300 freeze-thaw cycles, the relative dynamic modulus of elasticity and the relative flexural strength presented clear trends of freeze-thaw resistance of SFRELC. The compound effect of the air-entraining agent and the steel fibers was found to support the SFRELC with high freeze-thaw resistance, and the mechanisms were explored with the aid of the test results of water penetration of SFRELC. The beneficial effect was appeared from the replacement of lightweight sand with manufactured sand. Based on the test results, suggestions are given out for the optimal mix proportion of SFRELC to satisfy the durability requirement of freeze-thaw resistance.

  6. Polyvinyl alcohol/chitosan/montmorillonite nanocomposites preparation by freeze/thaw cycles and characterization

    Directory of Open Access Journals (Sweden)

    Părpăriţă Elena

    2014-12-01

    Full Text Available Polyvinyl alcohol (PVA and chitosan (CS based hydrogels are often chosen to obtain hydrogels as being considered non-toxic for human body. The present study aims the preparation and physical chemical characterisation of hydrogels based PVA and CS by using an environmental friendly method i.e. freeze/thaw. In this method the only parameters affecting the hydrogels’ properties is the PVA concentration in solution, time and number of cycles of freezing / thawing. Repeated freezing and thawing cycles resulted in production of a highly elastic polyvinyl alcohol hydrogel with higher degree of crystallization. Adding chitosan in polyvinyl alcohol hydrogel is giving to the newly formed material, biocompatibility and antibacterial properties due to the free amino groups of chitosan. Higher mechanical and thermal characteristics of PVA/CS based hydrogels were obtained by addition of a small amount of inorganic nanoparticles (montmorillonite clay, C30B into the matrix (i.e. 1%. Scanning electron microscopy (SEM, Fourier transform infrared spectroscopy (FT-IR, near-infrared chemical imaging spectroscopy (NI-CI, X-ray diffraction (XRD, thermogravimetric analysis (TG, swelling and rheological measurements were used to characterize the polyvinyl alcohol/chitosan/montmorillonite properties. The swelling degree increased with decreasing chitosan content in hydrogels and the variation is opposite in nanocomposites, decreasing after introducing the nanoclay. The swelling behaviour was influenced by the presence of the nanoparticles. The plasticizer effect of the nanoparticles was reflected by obtaining a more compact hydrogel network with higher mechanical and thermal properties. The proposed materials can be a promising alternative in biomedical applications

  7. Characterization of lipid oxidation process of beef during repeated freeze-thaw by electron spin resonance technology and Raman spectroscopy.

    Science.gov (United States)

    Chen, Qingmin; Xie, Yunfei; Xi, Jinzhong; Guo, Yahui; Qian, He; Cheng, Yuliang; Chen, Yi; Yao, Weirong

    2018-03-15

    In this study, electron spin resonance (ESR) and Raman spectroscopy were applied to characterize lipid oxidation of beef during repeated freeze-thaw (RFT). Besides the conventional indexes including peroxide values (PV), thiobarbituric acid-reactive substances (TBARS) and acid values (AV) were evaluated, the radical and molecular structure changes were also measured by ESR and Raman spectroscopy. The results showed that PV, TBARS and AV were increased (PRaman intensity of ν(CC) stretching region (1655cm -1 ) was decreased during RFT. Furthermore, lower Raman intensity ratio of I 1655 /I 1442 , I 1655 /I 1745 that determine total unsaturation was also observed. Significant correlations (pRaman spectroscopy. Our result has proved that ESR and Raman spectroscopy showed great potential in characterizing lipid oxidation process of beef during RFT. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Dispersion Stability of O/W Emulsions with Different Oil Contents Under Various Freezing and Thawing Conditions.

    Science.gov (United States)

    Katsuki, Kazutaka; Miyagawa, Yayoi; Nakagawa, Kyuya; Adachi, Shuji

    2017-07-01

    Freezing and thawing of oil-in-water (O/W) emulsion-type foods bring about oil-water separation and deterioration; hence, the effects of freezing and thawing conditions on the destabilization of O/W emulsions were examined. The freezing rate and thawing temperature hardly affected the stability of the O/W emulsion. O/W emulsions having different oil fractions were stored at temperatures ranging from -30 to -20 °C and then thawed. The stability after thawing depended on the storage temperature, irrespective of the oil fraction of the emulsion. A good correlation was found between the time at which the stability began to decrease and the time taken for the oil to crystalize. These results indicated that the dominant cause for the destabilization of the O/W emulsion during freezing and thawing is the crystallization of the oil phase and that the effects of the freezing and thawing rates on the stability are insignificant. © 2017 Institute of Food Technologists®.

  9. A Peltier-based freeze-thaw device for meteorite disaggregation

    Science.gov (United States)

    Ogliore, R. C.

    2018-02-01

    A Peltier-based freeze-thaw device for the disaggregation of meteorite or other rock samples is described. Meteorite samples are kept in six water-filled cavities inside a thin-walled Al block. This block is held between two Peltier coolers that are automatically cycled between cooling and warming. One cycle takes approximately 20 min. The device can run unattended for months, allowing for ˜10 000 freeze-thaw cycles that will disaggregate meteorites even with relatively low porosity. This device was used to disaggregate ordinary and carbonaceous chondrite regoltih breccia meteorites to search for micrometeoroid impact craters.

  10. Effect of freezing and thawing rates on the post-thaw viability of boar spermatozoa frozen in FlatPacks and Maxi-straws.

    Science.gov (United States)

    Eriksson, B M; Rodriguez-Martinez, H

    2000-11-01

    The effects of different freezing and thawing rates on the post-thaw motility and membrane integrity of boar spermatozoa, processed as split samples in Maxi-straws or flat PET-plastic packages (FlatPack) were studied. A programmable freezing device was used to obtain freezing rates of either 20, 50 or 80 degrees C/min. Thawing of the samples was performed in a bath of circulating water; for 40s at 50 degrees C or 27s at 70 degrees C for Maxi-straws and 23s at 35 degrees C, 13s at 50 degrees C or 8s at 70 degrees C for the FlatPacks. Sperm motility was assessed both visually and with a computer assisted semen analysis (CASA) apparatus, while plasma membrane integrity was assessed using the fluorescent probes Calcein AM and ethidium homodimer-1. Temperature changes during freezing and thawing were monitored in both forms of packaging. Values for motile spermatozoa, sperm velocity and lateral head displacement variables were significantly (pstraws, with superior results at higher thawing rates. Freezing at 50 degrees C/min yielded better motility than 20 or 80 degrees C/min, although the effect was rather small. Neither freezing rate nor thawing rate had any effect on membrane integrity (p>0.05). A significant boar effect was seen for several parameters. The most striking difference in temperature courses between containers was a 4-5-fold lowering of the thawing rate, between -20 and 0 degrees C, in the center of the Maxi-straw, compared with the FlatPack. This is apparently due to the insulating effect of the thawed water in the periphery of the Maxi-straw. The improvement in sperm motility seen when using the FlatPack appears to be related to the rapid thawing throughout the sample, which decreases the risk of cell damage due to recrystallization during thawing. Since sperm motility patterns have been reported to be correlated with fertility both in vitro and in vivo it is speculated that the use of the FlatPack might improve the results when using frozen-thawed

  11. The combined effect of freeze thaw events and heavy metal pollution leads to distinct lethal synergy in Enchytraeus albidus

    DEFF Research Database (Denmark)

    Boas, Sara Wincentz; Slotsbo, Stine; Holmstrup, Martin

    Many anthropogenic activities negatively affect the environment and stress the organisms living here in various ways. Due to global warming it is likely that freeze-thaw events will replace permanent freezing of soils in arctic regions. Metals are some of the most common contaminants in soil...

  12. Brown bear sperm double freezing: Effect of elapsed time and use of PureSperm(®) gradient between freeze-thaw cycles.

    Science.gov (United States)

    Alvarez-Rodríguez, Manuel; Alvarez, Mercedes; López-Urueña, Elena; Martínez-Rodriguez, Carmen; Borragan, Santiago; Anel-López, Luis; de Paz, Paulino; Anel, Luis

    2013-12-01

    The use of sexed spermatozoa has great potential to captive population management in endangered wildlife. The problem is that the sex-sorting facility is a long distance from the semen collection place and to overcome this difficulty two freeze-thaw cycles may be necessary. In this study, effects of refreezing on brown bear electroejaculated spermatozoa were analyzed. We carried out two experiments: (1) to assess the effects of the two freezing-thawing cycles on sperm quality and to analyze three different elapsed times between freezing-thawing cycles (30, 90 and 180 min), and (2) to analyze the use of PureSperm between freezing-thawing cycles to select a more motile and viable sperm subpopulation which better survived first freezing. The motility, viability and undamaged acrosomes were significantly reduced after the second thawing respect to first thawing into each elapsed time group, but the elapsed times did not significantly affect the viability and acrosome status although motility was damaged. Our results with the PureSperm gradient showed higher values of viability in freezability of select sample (pellet) respect to the rest of the groups and it also showed a significant decrease in the number of acrosome damaged. In summary, the double freezing of bear semen selected by gradient centrifugation is qualitatively efficient, and thus could be useful to carry out a sex-sorting of frozen-thawed bear spermatozoa before to send the cryopreserved sample to a biobank. Given the low recovery of spermatozoa after applying a selection gradient, further studies will be needed to increase the recovery rate without damaging of the cell quality. Copyright © 2013 Elsevier Inc. All rights reserved.

  13. Influence of freeze-thaw cycling on the resilient modulues of PFBC materials

    Energy Technology Data Exchange (ETDEWEB)

    Wolfe, W.E.; Butalia, T.S.; Meek, B.L. [Ohio State University, Columbus, OH (United States). Dept. of Civil and Environmental Engineering and Geodetic Science

    1999-01-01

    The dynamic stress-strain characteristics of a Pressurized Fluidized Bed Combustion (PFBC) material, before and after freeze and thaw cycling, were studied to evaluate its suitability as a substitute for conventional road construction materials in the design of flexible pavement systems. Samples compacted in the laboratory at two different moisture contents (optimum and 8% above optimum) were cyclically load tested after being allowed to cure for various duration. The results of the cyclic tests are presented in terms of the Resilient Modulus, which is a measure of the elastic property of the soil supporting the roadway. The modulus of the samples compacted near the optimum moisture content compared satisfactorily with data available for conventional materials. Samples compacted at moisture contents higher than the optimum exhibited a significant reduction in a resilient modulus values after freeze-thaw cycling. This comparing indicates that properly compacted PFBC holds good promise as a subgrade material in the construction of low traffic volume roads. 13 refs., 5 figs., 1 tab.

  14. Identification of Soil Freezing and Thawing States Using SAR Polarimetry at C-Band

    Directory of Open Access Journals (Sweden)

    Thomas Jagdhuber

    2014-03-01

    Full Text Available The monitoring of soil freezing and thawing states over large areas is very challenging on ground. In order to investigate the potential and the limitations of space-borne SAR polarimetry at C-band for soil state survey, analyses were conducted on an entire winter time series of fully polarimetric RADARSAT-2 data from 2011/2012 to identify freezing as well as thawing states within the soil. The polarimetric data were acquired over the Sodankylä test site in Finland together with in situ measurements of the soil and the snow cover. The analyses indicate clearly that the dynamics of the polarimetric entropy and mean scattering alpha angle are directly correlated to soil freezing and thawing states, even under distinct dry snow cover. First modeling attempts using the Extended Bragg soil scattering model justify the observed trends, which indicate surface-like scattering during frozen soil conditions and multiple/volume scattering for thawed soils. Hence, these first investigations at C-band foster motivation to work towards a robust polarimetric detection of soil freezing and thawing states as well as their transition phase.

  15. Study on dewatering of chemical sludge by freeze-thaw process

    International Nuclear Information System (INIS)

    Xu Shikun; Liu Pin

    1993-01-01

    The treatment of radioactive sludge that is produced from treating radioactive waste water contains radioactively is different from that of non-radioactive sludge. The methods of immersing freeze and simulated two-step freeze have been studied for the elementary properties of simulated low-level radioactive sledge, the effect of freezing temperature, freeze time, and settling time on volume-reduction factor. Some parameters for design of freeze-thaw device are provided

  16. Effects of Freezing and Thawing Cycle on Mechanical Properties and Stability of Soft Rock Slope

    Directory of Open Access Journals (Sweden)

    Yanlong Chen

    2017-01-01

    Full Text Available To explore the variation laws of mechanical parameters of soft rock and the formed slope stability, an experiment was carried out with collected soft rock material specimens and freezing and thawing cycle was designed. Meanwhile, a computational simulation analysis of the freezing-thawing slope stability was implemented. Key factors that influence the strength of frozen rock specimens were analyzed. Results showed that moisture content and the number of freezing-thawing cycles influenced mechanical parameters of soft rock significantly. With the increase of moisture content, cohesion of frozen soft rock specimens presents a quadratic function decrease and the internal friction angle shows a negative exponential decrease. The stability coefficient of soft rock material slope in seasonal freeze soil area declines continuously. With the increase of freezing and thawing cycle, both cohesion and internal friction angle of soft rock decrease exponentially. The higher the moisture content, the quicker the reduction. Such stability coefficient presents a negative exponential reduction. After three freezing and thawing cycles, the slope stability coefficient only changes slightly. Findings were finally verified by the filed database.

  17. Effects of freeze-thaw cycles on microarthropods and nutrient availability in a sub-arctic soil

    DEFF Research Database (Denmark)

    Sjursen, Heidi; Michelsen, Anders; Holmstrup, Martin

    2005-01-01

    It is predicted that Arctic regions may experience an increase in mean temperature in the future. This will affect the frequency of severe climatic events such as summer droughts and freeze-thaw cycles. In order to understand the impact of recurring freezing and thawing on soil organisms...... content were examined. There was no conclusive evidence that recurring freeze-thaw events had a negative effect on the investigated soil faunal groups, and the treatment even seemed to stimulate the abundance of Acaridida. Respiration of soil subjected to 16 freeze-thaw cycles was low when kept at -2 °C...... and high when kept at +2 °C, indicating rapid response of microbial activity even after long exposure to low and fluctuating temperatures. Oribatida and Gamasida displayed a higher abundance in controls kept at -2 °C for up to 80 days, compared to controls at +2 °C and the freeze-thaw treatment...

  18. Analysis of the influence of pasteurization, freezing/thawing, and offer processes on human milk's macronutrient concentrations.

    Science.gov (United States)

    Vieira, Alan Araujo; Soares, Fernanda Valente Mendes; Pimenta, Hellen Porto; Abranches, Andrea Dunshee; Moreira, Maria Elisabeth Lopes

    2011-08-01

    The macronutrient concentrations of human milk could be influenced by the various processes used in human milk bank. To determine the effect of various process (Holder pasteurization, freezing and thawing and feeding method) on the macronutrient concentration of human milk. The samples of donated fresh human milk were studied before and after each process (Holder pasteurization, freezing and thawing and feeding method) until their delivery to newborn infants. Fifty-seven raw human milk samples were analyzed in the first step (pasteurization) and 228 in the offer step. Repeated measurements of protein, fat and lactose amounts were made in samples of human milk using an Infrared analyzer. The influence of repeated processes on the mean concentration of macronutrients in donor human milk was analyzed by repeated measurements ANOVA, using R statistical package. The most variable macronutrient concentration in the analyzed samples was fat (reduction of 59%). There was a significant reduction of fat and protein mean concentrations following pasteurization (5.5 and 3.9%, respectively). The speed at which the milk was thawed didn't cause a significant variation in the macronutrients concentrations. However, the continuous infusion delivery significantly reduced the fat concentration. When the influence of repeated processes was analyzed, the fat and protein concentrations varied significantly (reduction of 56.6% and 10.1% respectively) (Pmilk is submitted before delivery to newborn infants cause a reduction in the fat and protein concentration. The magnitude of this decrease is higher on the fat concentration and it needs to be considered when this processed milk is used to feed preterm infants. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

  19. Effects of Freezing and Thawing Cycle on Mechanical Properties and Stability of Soft Rock Slope

    OpenAIRE

    Chen, Yanlong; Wu, Peng; Yu, Qing; Xu, Guang

    2017-01-01

    To explore the variation laws of mechanical parameters of soft rock and the formed slope stability, an experiment was carried out with collected soft rock material specimens and freezing and thawing cycle was designed. Meanwhile, a computational simulation analysis of the freezing-thawing slope stability was implemented. Key factors that influence the strength of frozen rock specimens were analyzed. Results showed that moisture content and the number of freezing-thawing cycles influenced mech...

  20. Effect of multiple cycles of freeze-thawing on the RNA quality of lung cancer tissues.

    Science.gov (United States)

    Yu, Keke; Xing, Jie; Zhang, Jie; Zhao, Ruiying; Zhang, Ye; Zhao, Lanxiang

    2017-09-01

    RNA degradation is a major problem in tissue banking. We explored the effect of thawing flash-frozen biospecimens on the quality and integrity of RNA for genetic testing as well as for other cancer research studies. The histological quality of the frozen tumor sections was evaluated by using hematoxylin and eosin staining. RNA extraction from 60 lung cancer tissue samples subjected to various freeze/thaw cycles was performed using the RNeasy Plus isolation kit. RNA integrity was assessed by using an Agilent bioanalyzer to obtain RNA integrity numbers (RIN). Furthermore, RNA from different groups was used for fluorescence Reverse transcription-polymerase chain reaction (RT-PCR) analysis of the echinoderm microtubule-associated protein-like 4 and anaplastic lymphoma kinase (EML4-ALK) fusion gene mutation to verify whether it can be used for research or clinical testing. Highly variable RIN values were observed among the samples, which showed no correlation with the number of freeze/thaw cycles conducted. However, after 3 freeze/thaw cycles (each thaw event lasted for 10 min), an increasing number of changes in peak intensity in RINs were observed. After 5 freeze/thaw cycles, RNA integrity decreased to approximately 35%. After 3 freeze/thaw cycles, the RNA could still be used for RT-PCR analysis of EML4-ALK fusion gene mutations; whereas those subjected to 5 freeze/thaw cycles could not. Limited (cycles did not adversely affect the quality of RNA extracted from tumor tissues and subsequent RT-PCR analysis. Our data could be utilized in the establishment of a standardized procedure for tissue biospecimen collection and storage.

  1. L-Band Microwave Emission of Soil Freeze-Thaw Process in the Third Pole Environment

    NARCIS (Netherlands)

    Zheng, Donghai; van der Velde, R.; Su, Z.; Zeng, Y.

    2017-01-01

    Soil freeze-thaw transition monitoring is essential for quantifying climate change and hydrologic dynamics over cold regions, for instance, the Third Pole. We investigate the L-band (1.4 GHz) microwave emission characteristics of soil freeze-thaw cycle via analysis of tower-based brightness

  2. A Loss in the Plasma Membrane ATPase Activity and Its Recovery Coincides with Incipient Freeze-Thaw Injury and Postthaw Recovery in Onion Bulb Scale Tissue 1

    Science.gov (United States)

    Arora, Rajeev; Palta, Jiwan P.

    1991-01-01

    Plasma membrane ATPase has been proposed to be functionally altered during early stages of injury caused by a freeze-thaw stress. Complete recovery from freezing injury in onion cells during the postthaw period provided evidence in support of this proposal. During recovery, a simultaneous decrease in ion leakage and disappearance of water soaking (symptoms of freeze-thaw injury) has been noted. Since reabsorption of ions during recovery must be an active process, recovery of plasma membrane ATPase (active transport system) functions has been implicated. In the present study, onion (Allium cepa L. cv Downing Yellow Globe) bulbs were subjected to a freeze-thaw stress which resulted in a reversible (recoverable) injury. Plasma membrane ATPase activity in the microsomes (isolated from the bulb scales) and ion leakage rate (efflux/hour) from the same scale tissue were measured immediately following thawing and after complete recovery. In injured tissue (30-40% water soaking), plasma membrane ATPase activity was reduced by about 30% and this was paralleled by about 25% higher ion leakage rate. As water soaking disappeared during recovery, the plasma membrane ATPase activity and the ion leakage rate returned to about the same level as the respective controls. Treatment of freeze-thaw injured tissue with vanadate, a specific inhibitor of plasma membrane ATPase, during postthaw prevented the recovery process. These results indicate that recovery of freeze-injured tissue depends on the functional activity of plasma membrane ATPase. PMID:16668063

  3. Effect of fast freeze-thaw cycles on mechanical properties of ordinary-air-entrained concrete.

    Science.gov (United States)

    Shang, Huai-shuai; Cao, Wei-qun; Wang, Bin

    2014-01-01

    Freezing-thawing resistance is a very significant characteristic for concrete in severe environment (such as cold region with the lowest temperature below 0°C). In this study, ordinary-air-entrained (O-A-E) concrete was produced in a laboratory environment; the compressive strength, cubic compressive strength of C50, C40, C30, C25, and C20 ordinary-air-entrained concrete, tensile strength, and cleavage strength of C30 ordinary-air-entrained concrete were measured after fast freeze-thaw cycles. The effects of fast freeze-thaw cycles on the mechanical properties (compressive strength and cleavage strength) of ordinary-air-entrained concrete materials are investigated on the basis of the experimental results. And the concise mathematical formula between mechanical behavior and number of fast freeze-thaw cycles was established. The experiment results can be used as a reference in design, maintenance, and life prediction of ordinary-air-entrained concrete structure (such as dam, offshore platform, etc.) in cold regions.

  4. Freeze-thaw regime effects on carbon and nitrogen dynamics in sub-arctic heath tundra mesocosms

    DEFF Research Database (Denmark)

    Grogan, P.; Michelsen, A.; Ambus, P.

    2004-01-01

    phase, and in a subsequent equilibration phase. A single deep freeze treatment phase enhanced dissolved total and labelled N pools in the soil solution at initial thaw, and resulted in reduced pool sizes at the end of the equilibration phase. By contrast, a multiple freeze-thaw cycling treatment......, these results indicate that moderate freeze-thaw fluctuations may have minimal influences on microbial biomass pools, but nevertheless can have strong contrasting effects on the amounts, forms, and timing of N and organic C supply into the soil solution. Ecosystem losses via N2O effluxes were of greatest...... directly enhanced the dissolved labelled N pool, but did not significantly affect dissolved total N. Furthermore, both dissolved labelled N and dissolved total N pools were significantly enhanced in the equilibration period following multiple freeze-thaw, the latter due to a marked increase in soil...

  5. The Relationship between Mechanical Properties and Gradual Deterioration of Microstructures of Rock Mass Subject to Freeze-thaw Cycles

    Directory of Open Access Journals (Sweden)

    Haibo Jiang

    2018-01-01

    Full Text Available Under freeze-thaw cycles, the relationship between rock microstructure deterioration and its macroscopic mechanical characteristics has drawn extensive attention from engineers. With the objective to incorporate freeze-thaw cycle experiment into headrace tunnel engineering, in the present study two groups of andesite rock samples in different states are tested under the conditions of the lowest freezing temperature of –40 ℃ and the thawing temperature of 20 ℃. Damage detection was performed by magnetic resonance imaging for the interior microstructure of rock samples subject to different freeze-thaw cycles, and the relationship between the sample mechanical properties and gradual deterioration of rock microstructures was discussed. The results demonstrate evident influence of freeze-thaw cycle on the damage and deterioration of internal pore structure in andesite, and the rock uniaxial compressive strength and elasticity modulus exhibit a decreasing trend with the increase of freeze-thaw cycles. After 40 cycles, the strength of naturally saturated rock samples decreases by 39.4% (equivalent to 69.4 MPa and the elasticity modulus drops by 47.46% (equivalent to 3.27 GPa. For rock samples saturated by vacuum, 40 freeze-thaw cycles lead to a decrease of 36.86% (equivalent to 58.2 MPa in rock strength and a drop of 44.85% (equivalent to 2.83 GPa in elasticity modulus. Therefore, the test results quantitatively elucidate the substantial influence of freeze-thaw cycle on the damage and deterioration of internal structure in andesite.

  6. Laboratory studies on the effect of freezing and thawing exposure on bentonite buffer performance: Closed-system tests

    Energy Technology Data Exchange (ETDEWEB)

    Schatz, T.; Martikainen, J. [B and Tech Oy, Helsinki (Finland)

    2010-12-15

    This report presents a set of results from laboratory studies on the effect of freezing and thawing on compacted bentonite buffer material. In order to evaluate the effect of freezing and thawing on compacted bentonite buffer performance a series of experiments were conducted using closed, constant-volume cells as follows: Pre- and post-freezing swelling pressure measurements were performed on fully saturated MX-80 and Deponit CA-N bentonite samples, at dry density values of approximately 1.6 g/cm{sup 3}, over five freeze/thaw cycles from room temperature to -18 deg C with rapid (instantaneous) temperature exposure. Pressure measurements were performed on fully saturated MX-80 bentonite samples, at dry density values of 1.470 and 1.501 g/cm{sup 3}, during a temperature run from room temperature to -10 deg C with step-change temperature exposure and back from -10 deg C to room temperature under continuous temperature change exposure at 0.1 deg C/h. Pressure measurements were performed on fully saturated MX-80 bentonite samples, encompassing a range of dry density values from 0.940 to 1.534 g/cm{sup 3}, during repeated temperature runs from room temperature to -10 deg C and back with continuous temperature change exposure at 0.1 deg C/h. Pressure measurements were performed on a fully saturated Deponit CA-N bentonite sample, at a dry density of 1.484 g/cm{sup 3}, during a temperature run from room temperature to -10 deg C and back with continuous temperature change exposure at 0.1 deg C/h. In some cases, hydraulic conductivity measurements were performed before and after freeze/thaw exposure. In general, exposure to freezing temperatures, down to an average temperature of -10 deg C, results in the development of significant internal pressures in compacted bentonite samples, which is attributed to the formation of ice. The specific test results are summarised as follows: Increases in pressure by factors of 1.5 to 2.2 were observed for MX-80 samples at dry densities

  7. Cryopreserving turkey semen in straws and nitrogen vapour using DMSO or DMA: effects of cryoprotectant concentration, freezing rate and thawing rate on post-thaw semen quality.

    Science.gov (United States)

    Iaffaldano, N; Di Iorio, M; Miranda, M; Zaniboni, L; Manchisi, A; Cerolini, S

    2016-04-01

    1. This study was designed to identify a suitable protocol for freezing turkey semen in straws exposed to nitrogen vapour by examining the effects of dimethylacetamide (DMA) or dimethylsulfoxide (DMSO) as cryoprotectant (CPA), CPA concentration, freezing rate and thawing rate on in vitro post-thaw semen quality. 2. Pooled semen samples were diluted 1:1 (v:v) with a freezing extender composed of Tselutin diluent containing DMA or DMSO to give final concentrations of 8% or 18% DMA and 4% or 10% DMSO. The semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen (LN2) surface (1, 5 and 10 cm) for 10 min. Semen samples were thawed at 4°C for 5 min or at 50°C for 10 s. After thawing, sperm motility, viability and osmotic tolerance were determined. 3. Cryosurvival of turkey sperm was affected by DMSO concentration. Freezing rate affected the motility of sperm cryopreserved using both CPAs, while thawing rates showed an effect on the motility of sperm cryopreserved using DMA and on the viability of sperm cryopreserved using DMSO. Significant interactions between freezing rate × thawing rate on sperm viability in the DMA protocol were found. 4. The most effective freezing protocol was the use of 18% DMA or 10% DMSO with freezing 10 cm above the LN2 surface and a thawing temperature of 50°C. An efficient protocol for turkey semen would improve prospects for sperm cryobanks and the commercial use of frozen turkey semen.

  8. Cryopreservation of Iberian pig spermatozoa. Comparison of different freezing extenders based on post-thaw sperm quality.

    Science.gov (United States)

    De Mercado, Eduardo; Rodríguez, Ana; Gómez, Emilio; Sanz, Elena

    2010-03-01

    The aim of this study was to evaluate the cryoprotective effect of different freezing extenders against cryopreservation injuries on Iberian boar sperm. The sperm-rich fraction was collected and pooled from six sexually mature Iberian boars, and was frozen in different extenders containing glucose, lactose or fructose as sugar source and including Orvus ES Paste only in the freezing extender-2 (Glucose; Lactose and Fructose) or in both freezing extenders (Glucose2; Lactose2 and Fructose2). During the cryopreservation process, the supernatant was removed after the centrifugation step, then was extended with freezing extender-1 for the equilibration period and with freezing extender-2 immediately before freezing. Post-thaw sperm characteristics, such as plasma membrane integrity (SYBR-14/PI), mitochondrial function (Rhodamine 123) and acrosome integrity (NAR), were monitored. Overall sperm motility and the individual kinematic parameters of motile spermatozoa (assessed by the computer-aided sperm analysis system Sperm Class Analyzer [SCA]) were recorded in the different experimental treatments. Measurements were taken at 30 and 150 min post-thaw. The state of the acrosome after thawing did not show significant differences between the freezing extenders studied. Freezing-thawing caused a significant decrease (Pextenders. Furthermore, spermatozoa frozen with Orvus ES Paste in both freezing extenders exhibited lower (Pextender. The spermatozoa frozen with the Lactose extender and with Orvus ES Paste only in the second freezing extender showed a better evolution of the motility and kinematic characteristics (Pextenders studied in the present experiment affected the quality of frozen-thawed semen in Iberian boar.

  9. Influence of waxy rice flour substitution for wheat flour on characteristics of batter and freeze-thawed cake.

    Science.gov (United States)

    Jongsutjarittam, Nisachon; Charoenrein, Sanguansri

    2013-09-12

    This study aimed to improve the freeze-thawed cake properties by10-20% waxy rice flour (WRF) substitution for wheat flour (WF). Viscosity of WRF-substituted batters was lower; consequently, trapped air was less uniformly distributed than WF batter. After five freeze-thaw cycles, firmness and enthalpy of melting retrograded amylopectin of WF- and WRF-substituted cakes increased and the matrix surrounding the air pores from SEM images was denser than in fresh-baked cakes. Sensory evaluation showed an increase in firmness and a decrease in firmness acceptability of freeze-thawed cakes. However, freeze-thawed cake with WRF substitution had significantly less firmness, less dense matrix and more acceptability than WF cake. This could have been due to a low amylose content of WRF and the spread of ruptured waxy rice starch granules around swollen wheat starch granules as observed by CLSM. Thus, WRF could be used for WF substitution to improve the firmness in freeze-thawed cake. Copyright © 2013 Elsevier Ltd. All rights reserved.

  10. Alpha-ketoglutarate enhances freeze-thaw tolerance and prevents carbohydrate-induced cell death of the yeast Saccharomyces cerevisiae.

    Science.gov (United States)

    Bayliak, Maria M; Hrynkiv, Olha V; Knyhynytska, Roksolana V; Lushchak, Volodymyr I

    2018-01-01

    Stress resistance and fermentative capability are important quality characteristics of baker's yeast. In the present study, we examined protective effects of exogenous alpha-ketoglutarate (AKG), an intermediate of the tricarboxylic acid cycle and amino acid metabolism, against freeze-thaw and carbohydrate-induced stresses in the yeast Saccharomyces cerevisiae. Growth on AKG-supplemented medium prevented a loss of viability and improved fermentative capacity of yeast cells after freeze-thaw treatment. The cells grown in the presence of AKG had higher levels of amino acids (e.g., proline), higher metabolic activity and total antioxidant capacity, and higher activities of catalase, NADP-dependent glutamate dehydrogenase and glutamine synthase compared to control ones. Both synthesis of amino acids and enhancement of antioxidant system capacity could be involved in AKG-improved freeze-thaw tolerance in S. cerevisiae. Cell viability dramatically decreased under incubation of stationary-phase yeast cells in 2% glucose or fructose solutions (in the absence of the other nutrients) as compared with incubation in distilled water or in 10 mM AKG solution. The decrease in cell viability was accompanied by acidification of the medium, and decrease in cellular respiration, aconitase activity, and levels of total protein and free amino acids. The supplementation with 10 mM AKG effectively prevented carbohydrate-induced yeast death. Protective mechanisms of AKG could be associated with the intensification of respiration and prevention of decreasing protein level as well as with direct antioxidant AKG action.

  11. Cold stress improves the ability of Lactobacillus plantarum L67 to survive freezing.

    Science.gov (United States)

    Song, Sooyeon; Bae, Dong-Won; Lim, Kwangsei; Griffiths, Mansel W; Oh, Sejong

    2014-11-17

    The stress resistance of bacteria is affected by the physiological status of the bacterial cell and environmental factors such as pH, salts and temperature. In this study, we report on the stress response of Lactobacillus plantarum L67 after four consecutive freeze-thaw cycles. The cold stress response of the cold-shock protein genes (cspC, cspL and cspP) and ATPase activities were then evaluated. The cold stress was adjusted to 5 °C when the bacteria were growing at the mid-exponential phase. A comparative proteomic analysis was performed with two-dimensional gel electrophoresis (2D SDS-PAGE) and a matrix assisted laser desorption/ionization-mass spectrometer. Only 56% of the L. plantarum L67 cells without prior exposure to cold stress survived after four consecutive freeze-thaw cycles. However, 78% of the L. plantarum L67 cells that were treated with cold stress at 5 °C for 6 h survived after freeze-thaw conditions. After applying cold stress to the culture for 6h, the cells were then stored for 60 days at 5 °C, 25 °C and 35 °C separately. The cold-stressed culture of L. plantarum L67 showed an 8% higher viability than the control culture. After applying cold stress for 6h, the transcript levels of two genes (cspP and cspL) were up-regulated 1.4 (cspP) and 1.2 (cspL) times compared to the control. However, cspC was not up-regulated. A proteomic analysis showed that the proteins increased after a reduction of the incubation temperature to 5 °C. The importance of the expression of 13 other relevant proteins was also determined through the study. The exposure of L. plantarum cells to low temperatures aids their ability to survive through subsequent freeze-thaw processes and lyophilization. Copyright © 2014. Published by Elsevier B.V.

  12. The anti-cell death FNK protein protects cells from death induced by freezing and thawing

    International Nuclear Information System (INIS)

    Sudo, Kentaro; Asoh, Sadamitsu; Ohsawa, Ikuroh; Ozaki, Daiya; Yamagata, Kumi; Ito, Hiromoto; Ohta, Shigeo

    2005-01-01

    The FNK protein, constructed from anti-apoptotic Bcl-x L with enhanced activity, was fused with the protein transduction domain (PTD) of the HIV/Tat protein to mediate the delivery of FNK into cells. The fusion protein PTD-FNK was introduced into chondrocytes in isolated articular cartilage-bone sections, cultured neurons, and isolated bone marrow mononuclear cells to evaluate its ability to prevent cell death induced by freezing and thawing. PTD-FNK protected the cells from freeze-thaw damage in a concentration-dependent manner. Addition of PTD-FNK with conventional cryoprotectants (dimethyl sulfoxide and hydroxyethyl starch) increased surviving cell numbers around 2-fold compared with controls treated only with the cryoprotectants. Notably, PTD-FNK allowed CD34 + cells among bone marrow mononuclear cells to survive more efficiently (12-fold more than the control cells) from two successive freeze-thaw cycles. Thus, PTD-FNK prevented cell death induced by freezing and thawing, suggesting that it provides for the successful cryopreservation of biological materials

  13. [Effects and mechanism of freeze-thawing cycles on key processes of nitrogen cycle in terrestrial ecosystem].

    Science.gov (United States)

    Wang, Li-qin; Qi, Yu-chun; Dong, Yun-she; Peng, Qin; Guo, Shu-fang; He, Yun-long; Yan, Zhong-qing

    2015-11-01

    As a widespread natural phenomenon in the soil of middle and high latitude as well as high altitude, freeze-thawing cycles have a great influence on the nitrogen cycle of terrestrial ecosystem in non-growing season. Freeze-thawing cycles can alter the physicochemical and biological properties of the soil, which thereby affect the migration and transformation of soil nitrogen. The impacts of freeze-thawing cycles on key processes of nitrogen cycle in terrestrial ecosystem found in available studies remain inconsistent, the mechanism is still not clear, and the research methods also need to be further explored and innovated. So it is necessary to sum up and analyze the existing achievements in order to better understand the processes of soil nitrogen cycle subjected to freeze-thawing cycles. This paper reviewed the research progress in China and abroad about the effects and mechanisms of freeze-thawing cycles on key processes of nitrogen cycle in terrestrial ecosystem, including mineralization, immobilization, nitrification and denitrification, N leakage and gaseous loss, and analyzed the deficiencies of extant research. The possible key research topics that should be urgently paid more attention to in the future were also discussed.

  14. Experimental Study Regarding the Freezing and Thawing Dynamics of Spruce Wood

    Directory of Open Access Journals (Sweden)

    Maria - Bernadett SZMUTKU

    2013-03-01

    Full Text Available The article presents the results regarding theevolution of the temperature field in spruce wood(Picea abies L. during freezing at two differentfreezing rates: -100C/h (rapid freezing and -10C/h(slow freezing and then during thawing at +50C,+30°C, +50°C temperature.This approach aimed at simulating thetemperature variations which occur inside timberduring open air storage in winter in two situations(sudden vs. gradual drop of temperature, and thenwhen the timber enters the drying kiln, depending onthe temperature applied in the initial heating phase.The results clearly show that the freezing ratesignificantly influences the thawing time and speed,which increase by 13-17% in the case of slowly frozenwood (at -10C/min compared to rapidly frozen wood(at -100C/min. It was also established that theoptimum temperature in the heating-up phase whendrying frozen spruce is 300C instead of the usual500C. This value leads to much better dryinguniformity without significantly prolonging the dryingtime.

  15. The effect of freeze-thaw cycles on the hydraulic conductivity of compacted clay

    International Nuclear Information System (INIS)

    Waite, D.; Anderson, L.; Caliendo, J.; McFarland, M.

    1994-01-01

    A study was conducted to investigate the detrimental effects of freeze-thaw on the hydraulic conductivity of compacted clay. The purpose of this study was to determine the effect that molding water content has on the hydraulic conductivity of a compacted clay soil that is subjected to freeze-thaw cycles, and to determine the relationship between the number of freeze-thaw cycles and the hydraulic conductivity of the compacted clay soil. Clay soils compacted and frozen wet of optimum experienced an increase in hydraulic conductivity of approximately 140 fold. The hydraulic conductivity of clay compacted dry of optimum increased ten fold. These results are consistent with recent research which suggests that clay compacted wet of optimum experiences large increases in hydraulic conductivity while the hydraulic conductivity of clay compacted dry of optimum increases to a lesser extent. 12 refs., 9 figs

  16. Dimensional and ice content changes of hardened concrete at different freezing and thawing temperatures

    DEFF Research Database (Denmark)

    Johannesson, Björn

    2010-01-01

    Samples of concrete at different water-to-cement ratios and air contents subjected to freeze/thaw cycles with the lowest temperature at about -80 degrees C are investigated. By adopting a novel technique, a scanning calorimeter is used to obtain data from which the ice contents at different freeze...... temperatures can be calculated. The length change caused by temperature and ice content changes during test is measured by a separate experiment using the same types of freeze-thaw cycles as in the calorimetric tests. In this way it was possible to compare the amount of formed ice at different temperatures...... and the corresponding measured length changes. The development of cracks in the material structure was indicated by an ultra-sonic technique by measuring on the samples before and after the freeze-thaw tests. Further the air void structure was investigated using a microscopic technique in which air'bubble' size...

  17. Efeito do resfriamento do sêmen eqüino sobre sua congelabilidade Effect of freezing and thawing protocols on post-thaw quality of equine semen

    Directory of Open Access Journals (Sweden)

    R. Fürst

    2005-10-01

    Full Text Available Utilizaram-se 25 ejaculados de cinco garanhões da raça Mangalarga Marchador, para avaliar dois protocolos de congelamento. No primeiro tratamento, resfriou-se o sêmen até 5ºC (curva de resfriamento - CR antes do congelamento, no segundo, congelou-se o sêmen sem resfriamento (SC. Compararam-se duas formas de descongelamento, a 37ºC e a 75ºC/sete segundos. Os protocolos foram avaliados pelo teste de termo resistência (TTR - motilidade total e vigor e pela funcionalidade da membrana plasmática (teste hiposmótico e eosina nigrosina. A motilidade total no tempo zero do TTR foi melhor (PTwo freezing protocols and two thawing methods were evaluated on 25 ejaculates from five stallions of the Mangalarga Marchador breed. In the first freezing method, semen was cooled to 5ºC before freezing (CR; and in the second, semen at room temperature was frozen directly (SC. The two thawing methods were thawing semen at 37ºC for 30 seconds versus thawing at 75ºC for 7 seconds. Thawed semen was evaluated by the thermal resistance test (TRT- total motility and vigor and by integrity of sperm membranes (the hypo-osmotic test and the eosine-nigrosine test. Semen that was cooled before freezing had higher (P<0.05 motility immediately post-thaw than semen that was more abruptly frozen (46.7% versus 21.0% for semen thawed at 37ºC and 44.1% versus 24.5% for semen thawed at 75ºC, respectively. At both thawing temperatures, the percentage of live spermatozoa was higher (P<0.05 in CR treatment than in the SC method (71% versus 54.6% for semen thawed at 37ºC and 77.3% versus 54.1% for semen thawed at 75ºC, respectively. The CR treatment also resulted in better hypo-osmotic test results and better semen vigor than did the SC treatment. Semen thawed at 75ºC showed better (P<0.05 vigor than semen thawed at 37ºC, independent of the semen freezing method. In conclusion, there were substantial benefits on subsequent semen quality from cooling of semen before it

  18. Effect of exogenous carbon addition and the freeze-thaw cycle on soil microbes and mineral nitrogen pools1

    Science.gov (United States)

    Hu, Xia; Yin, Peng; Nong, Xiang; Liao, Jinhua

    2018-01-01

    To elucidate the alpine soil process in winter, the response mechanism of soil mineral nitrogen and soil microbes to exogenous carbon (0 mg C, 1 mg C, 2 mg C, 4 mg C and 8 mg C·g-1 dry soil) and the freeze-thaw cycle (-2 °C, -2 ∼ 2 °C, -20 ∼2°C) were studied by laboratory simulation. The freeze-thaw treatment had no significant effect on microbial biomass nitrogen and the number of bacteria. The soil mineral N pool, the number of fungi, and enzyme activities were obviously affected by the freeze-thaw cycle. A mild freeze-thaw cycle (-2∼2°C) significantly increased the number of fungi and catalase activity, while severe freeze-thaw cycle (-20∼2°C) obviously decreased invertase activity. The results suggested that both the freeze-thaw rate and freeze-thaw temperature amplitudes have a strong effect on soil microbial dynamics in the alpine zone in winter. The results showed that exogenous carbon addition significantly decreased soil NO3-N and NH4 +-N contents, increased soil microbial biomass, the number of microbes, and soil enzyme activities. The results showed that microbial growth in the eastern Tibetan Plateau was somewhat limited by available C. It may represent a larger potential pulse of soil nutrient for alpine plants in the next spring, and may be instrumental for plant community shifts under future climate change predictions due to the possible increased litter addition.

  19. Influence of Rapid Freeze-Thaw Cycling on the Mechanical Properties of Sustainable Strain-Hardening Cement Composite (2SHCC

    Directory of Open Access Journals (Sweden)

    Seok-Joon Jang

    2014-02-01

    Full Text Available This paper provides experimental results to investigate the mechanical properties of sustainable strain-hardening cement composite (2SHCC for infrastructures after freeze-thaw actions. To improve the sustainability of SHCC materials in this study, high energy-consumptive components—silica sand, cement, and polyvinyl alcohol (PVA fibers—in the conventional SHCC materials are partially replaced with recycled materials such as recycled sand, fly ash, and polyethylene terephthalate (PET fibers, respectively. To investigate the mechanical properties of green SHCC that contains recycled materials, the cement, PVA fiber and silica sand were replaced with 10% fly ash, 25% PET fiber, and 10% recycled aggregate based on preliminary experimental results for the development of 2SHCC material, respectively. The dynamic modulus of elasticity and weight for 2SHCC material were measured at every 30 cycles of freeze-thaw. The effects of freeze-thaw cycles on the mechanical properties of sustainable SHCC are evaluated by conducting compressive tests, four-point flexural tests, direct tensile tests and prism splitting tests after 90, 180, and 300 cycles of rapid freeze-thaw. Freeze-thaw testing was conducted according to ASTM C 666 Procedure A. Test results show that after 300 cycles of freezing and thawing actions, the dynamic modulus of elasticity and mass loss of damaged 2SHCC were similar to those of virgin 2SHCC, while the freeze-thaw cycles influence mechanical properties of the 2SHCC material except for compressive behavior.

  20. Gelatin-Filtered Consomme: A Practical Demonstration of the Freezing and Thawing Processes

    Science.gov (United States)

    Lahne, Jacob B.; Schmidt, Shelly J.

    2010-01-01

    Freezing is a key food processing and preservation technique widely used in the food industry. Application of best freezing and storage practices extends the shelf-life of foods for several months, while retaining much of the original quality of the fresh food. During freezing, as well as its counterpart process, thawing, a number of critical…

  1. The choice of a suitable oligosaccharide to prevent aggregation of PEGylated nanoparticles during freeze thawing and freeze drying

    NARCIS (Netherlands)

    Hinrichs, Wouter; Manceñido, F A; Sanders, N N; Braeckmans, K; De Smedt, S C; Demeester, J; Frijlink, H W

    2006-01-01

    In a previous Study we have shown that the oligosaccharide inulin can prevent aggregation of poly(ethylene glycol) (PEG) coated plasmid DNA/cationic liposome complexes ('' PEGylated lipoplexes '') during freeze thawing and freeze drying [Hinrichs et al., 2005. J. Control. Release 103, 465]. By

  2. Effect of Long-Term Freezing and Freeze–Thaw Cycles on Indigenous and Inoculated Microorganisms in Dewatered Blackwater

    DEFF Research Database (Denmark)

    Gunnarsdottir, Ragnhildur; Müller, Karoline; Jensen, Pernille Erland

    2012-01-01

    Wastewater treatment in many Arctic regions is inadequate, even nonexisting. Natural freezing of wastewater in those areas may be beneficial for reduction of microorganisms. The aim of this study was to investigate the effect of long-term freezing, and repeated freezing and thawing, on indigenous...... coliforms, fecal streptococci, and antibiotic-resistant (AR) bacteria, and inoculated Salmonella Enteriditis and E. coli bacteriophage ΦX174 in dewatered blackwater. At the end of the long-term freezing experiment (10 months), an MPN recovery study was done, including the microbial groups that had shown...... the largest reduction, using tryptone soy broth at incubation temperatures of 10 and 20 °C overnight for the coliforms and AR bacteria, and buffered peptone water at incubation temperature of 37 °C for 18–20 h for Salmonella. Fecal streptococci were more resistant to long-term freezing than the coliform group...

  3. Post Thawing Sperm Quality and Ca+2 Intensity Characters of Local Goat Sperm After Freezing by Simple Method Using Deep Freezing

    Directory of Open Access Journals (Sweden)

    Gatot Ciptadi

    2018-02-01

    Full Text Available The objective of this research was to determine the effect of the simple modified freezing method, 1°C/minute freezing rate with different diluter ration on a post-thawing quality of local goat sperm namely Peranakan Etawah (PE. This work is aimed to study the quality of post-thawing sperm and to characterize the calcium intensity profile of both fresh and post thawing goat sperm. The method used is the experimental design of a laboratory. Freezing semen was performed in 2 main temperatures of -45°C then -196°C respectively using Mr. Frosty (® System. Early Sperm characters of Ca+2 intensity was performed by Confocal Laser Scanning Microscope (CLSM through Fluo-3 staining and Ca++ intensity was analysis descriptively. The result showed that post-thawing qualities are considered as good as standard qualities, at least, more than 40% based on Indonesian National Standard (SNI, 2014. The different level diluents commercial of Andromeda used were influenced highly significant (P<0.01. The best diluents ration is 1:4 (v/v for final sperms stocked at -196°C. However freezing sperm conserved in -196°C is better than in -45°C. Meanwhile, the sperm characters of two condition showed the important variation of Ca+2 intensity, with the length of region measurement of 39.06±4.595 and 32.696±9.011 µm each.  It was concluded that the calcium intensity pattern was varied more and higher in fresh sperm than in freezing sperms. This simple modified method of a freezing system was considered as a feasible alternative method for goat semen in a reason for both for sperm post-thawing quality and practical purposes.

  4. Freeze-thaw resistance of concrete with marginal air content

    Science.gov (United States)

    2007-05-01

    Freeze-thaw resistance is a key durability factor for concrete pavements. Recommendations for the air void system parameters are normally 6 1 percent total air and a spacing factor of : < 0.20 millimeter (mm) (0.008 inch). However, it was observed...

  5. Diurnal Freeze-Thaw Cycles Modify Winter Soil Respiration in a Desert Shrub-Land Ecosystem

    Directory of Open Access Journals (Sweden)

    Peng Liu

    2016-07-01

    Full Text Available Winter soil respiration (Rs is becoming a significant component of annual carbon budgets with more warming in winter than summer. However, little is known about the controlling mechanisms of winter Rs in dryland. We made continuous measurements of Rs in four microsites (non-crust (BS, lichen (LC, moss (MC, and a mixture of moss and lichen (ML in a desert shrub-land ecosystem northern China, to investigate the causes of Rs dynamics in winter. The mean winter Rs ranged from 0.10 to 0.17 µmol CO2 m−2·s−1 across microsites, with the highest value in BS. Winter Q10 (known as the increase in respiration rate per 10 °C increase in temperature values (2.8–19 were much higher than those from the growing season (1.5. Rs and Q10 were greatly enhanced in freeze-thaw cycles compared to frozen days. Diurnal patterns of Rs between freeze-thaw and frozen days differed. Although the freeze-thaw period was relatively short, its cumulative Rs contributed significantly to winter Rs. The presence of biocrust might induce lower temperature, thus having fewer freeze-thaw cycles relative to bare soil, leading to the lower Rs for microsites with biocrusts. In conclusion, winter Rs in drylands was sensitive to soil temperature (Ts and Ts-induced freeze-thaw cycles. The temperature impact on Rs varied among soil cover types. Winter Rs in drylands may become more important as the climate is continuously getting warmer.

  6. Durability of cracked fibre reinforced concrete exposed to freeze-thaw and deicing salt

    DEFF Research Database (Denmark)

    Hansen, Ernst Jan De Place

    1998-01-01

    Durability studies are carried out by subjecting FRC-beams to combined mechanical and environmental load. Mechanical load is obtained by subjecting beams to 4-point bending until a predefined crack width is reached. Specimens sawn from the beams after unloading are exposed to freeze-thaw and deic......Durability studies are carried out by subjecting FRC-beams to combined mechanical and environmental load. Mechanical load is obtained by subjecting beams to 4-point bending until a predefined crack width is reached. Specimens sawn from the beams after unloading are exposed to freeze-thaw...... and deicing salt. The concrete has a water-powder ratio of 0.38 including both fly ash and silica fume. Both steel fibres (ZP, 0.4 vol%) and polypropylene fibres (PP, 1 vol%) are used as well as main reinforcement. The freeze-thaw test emphasizes the need for a critical evaluation of the mix design and mixing...

  7. Globally important nitrous oxide emissions from croplands induced by freeze-thaw cycles

    NARCIS (Netherlands)

    Wagner-Riddle, Claudia; Congreves, Katelyn A.; Abalos Rodriguez, Diego; Berg, Aaron A.; Brown, Shannon E.; Ambadan, Jaison Thomas; Gao, Xiaopeng; Tenuta, Mario

    2017-01-01

    Seasonal freezing induces large thaw emissions of nitrous oxide, a trace gas that contributes to stratospheric ozone destruction and atmospheric warming. Cropland soils are by far the largest anthropogenic source of nitrous oxide. However, the global contribution of seasonal freezing to nitrous

  8. Physical degradation of membrane electrode assemblies undergoing freeze/thaw cycling: Micro-structure effects

    Energy Technology Data Exchange (ETDEWEB)

    Kim, S. [Fuel Cell Dynamics and Diagnostics Laboratory, Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, University Park, PA 16802 (United States); Research and Development Division, Hyundai Motor Company, Yongin 446-912 (Korea); Mench, M.M. [Fuel Cell Dynamics and Diagnostics Laboratory, Department of Mechanical and Nuclear Engineering, The Pennsylvania State University, University Park, PA 16802 (United States)

    2007-11-22

    The objective of this work is to investigate physical damage of polymer electrolyte fuel cell (PEFC) materials subjected to freeze/thaw cycling. Effects of membrane electrode assembly micro-structures (catalyst layer cracking, membrane thickness, and membrane reinforcement) and diffusion media with micro-porous layers were analyzed by comparing scanning electron microscopy images of freeze/thaw cycled samples (-40 C/70 C) with those of virgin material and thermal cycled samples without freezing (5 C/70 C). Ex situ testing performed in this study has revealed a strong direction for the material choices in the PEFC and confirmed the previous computational model in the literature [S. He, M.M. Mench, J. Electrochem. Soc., 153 (2006) A1724-A1731; S. He, S.H. Kim, M.M. Mench, J. Electrochem. Soc., in press]. Specifically, the membrane electrode assemblies were found to be a source of water that can damage the catalyst layers under freeze/thaw conditions. Damage was found to occur almost exclusively under the channel, and not under the land (the graphite that touches the diffusion media). Conceptually, the best material to mitigate freeze-damage is a crack free virgin catalyst layer on a reinforced membrane that is as thin as possible, protected by a stiff diffusion media. (author)

  9. Freeze/thaw phenomena in concrete at low temperatures

    DEFF Research Database (Denmark)

    Johannesson, Björn

    2007-01-01

    Freeze/thaw damage in concrete is by general practice concluded to be a problem that can be avoided by using air-entraining agents to develop an air bubble structure in the hardened concrete together with the use of a relatively low water to cement ratio in mix. This fact is true for inner damage...

  10. Investigation of the impact of nanotechnology on the freeze-thaw durability of concrete containing d-cracking aggregates.

    Science.gov (United States)

    2015-05-01

    Freezing and thawing damage is the most common cause of distress in Kansas pavements. Many : locally available aggregates in Kansas do not meet current standards for use in concrete pavements because : of poor freeze-thaw durability. The use of nanot...

  11. Nuclear magnetic resonance analysis of freeze-thaw damage in natural pumice concrete

    Directory of Open Access Journals (Sweden)

    Wang, Xiaoxiao

    2016-06-01

    Full Text Available This paper presents an analysis of the damage propagation features of the pore structure of natural pumice lightweight aggregate concrete (LWC under freeze-thaw cyclic action. After freeze-thaw cycling, we conducted nuclear magnetic resonance (NMR tests on the concrete and acquired the porosity, distribution of transverse relaxation time T2, and magnetic resonance imaging (MRI results. The results showed the following. The T2 distribution of the LWC prior to freeze-thaw cycling presented four peaks representative of a preponderance of small pores. After 50, 100, 150, and 200 freeze-thaw cycles, the total area of the T2 spectrum and the porosity increased significantly. The MRI presented the changing spatial distribution of pores within the LWC during freeze-thaw cycling. Ultrasonic testing technology was applied simultaneously to analyze the NMR results, which verified that the new NMR technology demonstrated high accuracy and practicability for research regarding freeze-thaw concrete damage.En este trabajo se analiza la propagación de los daños que se producen en la estructura porosa de hormigón aligerado a base de piedra pómez natural sometido a la acción cíclica de hielo-deshielo. Después de realizarse los ensayos de hielo-deshielo, el hormigón se analizó mediante resonancia magnética nuclear (RMN, determinándose la porosidad y la distribución del tiempo de relajación transversal, T2, y registrándose las imágenes captadas por resonancia magnética. De acuerdo con los resultados obtenidos, antes de los ciclos de hielo-deshielo la distribución de T2 del hormigón aligerado presentaba cuatro picos, indicativos de un predominio de poros pequeños. Después de que se sometiera a 50, 100, 150, y 200 ciclos, se observó un aumento importante tanto de la porosidad como de la superficie total del espectro de T2. Las imágenes captadas por resonancia magnética evidenciaron la modificación de la distribución espacial de los poros del

  12. FREEZE-THAW AND FIRE RESISTANCE OF GEOPOLYMER MORTAR BASED ON NATURAL AND WASTE POZZOLANS

    Directory of Open Access Journals (Sweden)

    F.Nurhayat Degirmenci

    2017-12-01

    Full Text Available The purpose of this research was to investigate the resistance of pozzolan-based geopolymer mortars subjected to high temperatures and freeze-thaw cycles. Low calcium fly ash and granulated blast furnace slag as waste pozzolans and natural zeolite as a natural pozzolan were used as base materials for producing geopolymer mortar. The other purpose the research was to study the effect of alkaline activator ratio (Na₂SiO₃/NaOH on the performance of pozzolan-based geopolymer mortar specimens subjected to extreme temperatures. The influence of high temperatures on the properties of mortars was investigated at 300°C, 600°C, and 900°C. Fire and freeze-thaw and resistance of mortars were investigated in terms of visual appearance, weight loss and residual compressive strength. The minimal values of the residual compressive strength were obtained at 900°C for all mixtures. The residual compressive strength of all specimens was lower than the values obtained for specimens not subjected to any freeze-thaw resistance test, except those containing GGBS. The Na₂SiO₃/NaOH ratios of the alkaline activator solution used to prepare the geopolymer mortars have an effect on the weight losses and residual compressive strengths of the specimens subjected to high temperatures and freeze-thaw cycles. As the Na2SiO3/NaOH ratios increased, the weight and strength losses decreased.

  13. Effects of freeze-thaw on characteristics of new KMP binder stabilized Zn- and Pb-contaminated soils.

    Science.gov (United States)

    Wei, Ming-Li; Du, Yan-Jun; Reddy, Krishna R; Wu, Hao-Liang

    2015-12-01

    For viable and sustainable reuse of solidified/stabilized heavy metal-contaminated soils as roadway subgrade materials, long-term durability of these soils should be ensured. A new binder, KMP, has been developed for solidifying/stabilizing soils contaminated with high concentrations of heavy metals. However, the effects of long-term extreme weather conditions including freeze and thaw on the leachability and strength of the KMP stabilized contaminated soils have not been investigated. This study presents a systematic investigation on the impacts of freeze-thaw cycle on leachability, strength, and microstructural characteristics of the KMP stabilized soils spiked with Zn and Pb individually and together. For comparison purpose, Portland cement is also tested as a conventional binder. Several series of tests are conducted including the toxicity characteristic leaching procedure (TCLP), modified European Community Bureau of Reference (BCR) sequential extraction procedure, unconfined compression test (UCT), and mercury intrusion porosimetry (MIP). The results demonstrate that the freeze-thaw cycles have much less impact on the leachability and strength of the KMP stabilized soils as compared to the PC stabilized soils. After the freeze-thaw cycle tests, the KMP stabilized soils display much lower leachability, mass loss, and strength loss. These results are assessed based on the chemical speciation of Zn and Pb, and pore size distribution of the soils. Overall, this study demonstrates that the KMP stabilized heavy metal-contaminated soils perform well under the freeze-thaw conditions.

  14. Oil recovery from refinery oily sludge via ultrasound and freeze/thaw.

    Science.gov (United States)

    Zhang, Ju; Li, Jianbing; Thring, Ronald W; Hu, Xuan; Song, Xinyuan

    2012-02-15

    The effective disposal of oily sludge generated from the petroleum industry has received increasing concerns, and oil recovery from such waste was considered as one feasible option. In this study, three different approaches for oil recovery were investigated, including ultrasonic treatment alone, freeze/thaw alone and combined ultrasonic and freeze/thaw treatment. The results revealed that the combined process could achieve satisfactory performance by considering the oil recovery rate and the total petroleum hydrocarbon (TPH) concentrations in the recovered oil and wastewater. The individual impacts of five different factors on the combined process were further examined, including ultrasonic power, ultrasonic treatment duration, sludge/water ratio in the slurry, as well as bio-surfactant (rhamnolipids) and salt (NaCl) concentrations. An oil recovery rate of up to 80.0% was observed with an ultrasonic power of 66 W and an ultrasonic treatment duration of 10 min when the sludge/water ratio was 1:2 without the addition of bio-surfactant and salt. The examination of individual factors revealed that the addition of low concentration of rhamnolipids (treatment process. The experimental results also indicated that ultrasound and freeze/thaw could promote the efficiency of each other, and the main mechanism of oil recovery enhancement using ultrasound was through enhanced desorption of petroleum hydrocarbons (PHCs) from solid particles. Copyright © 2011 Elsevier B.V. All rights reserved.

  15. Compressive strength, chloride permeability, and freeze-thaw resistance of MWNT concretes under different chemical treatments.

    Science.gov (United States)

    Wang, Xingang; Rhee, Inkyu; Wang, Yao; Xi, Yunping

    2014-01-01

    This study investigated compressive strength, chloride penetration, and freeze-thaw resistance of multiwalled carbon nanotube (MWNT) concrete. More than 100 cylindrical specimens were used to assess test variables during sensitivity observations, including water-cement ratios (0.75, 0.5, and 0.4) and exposure to chemical agents (including gum arabic, propanol, ethanol, sodium polyacrylate, methylcellulose, sodium dodecyl sulfate, and silane). To determine the adequate sonication time for MWNT dispersal in water, the compressive strengths of MWNT concrete cylinders were measured after sonication times ranging from 2 to 24 minutes. The results demonstrated that the addition of MWNT can increase the compressive strength of concrete by up to 108%. However, without chemical treatment, MWNT concretes tend to have poor freeze-thaw resistance. Among the different chemical treatments, MWNT concrete treated with sodium polyacrylate has the best compressive strength, chloride resistance, and freeze-thaw durability.

  16. Application of Spaceborne Scatterometer for Mapping Freeze-Thaw State in Northern Landscapes as a Measure of Ecological and Hydrological Processes

    Science.gov (United States)

    McDonald, Kyle; Kimball, John; Zimmermann, Reiner; Way, JoBea; Frolking, Steve; Running, Steve

    1999-01-01

    Landscape freeze/thaw transitions coincide with marked shifts in albedo, surface energy and mass exchange, and associated snow dynamics. Monitoring landscape freeze/thaw dynamics would improve our ability to quantify the interannual variability of boreal hydrology and river runoff/flood dynamics. The annual duration of frost-free period also bounds the period of photosynthetic activity in boreal and arctic regions thus affecting the annual carbon budget and the interannual variability of regional carbon fluxes. In this study, we use the NASA scatterometer (NSCAT) to monitor the temporal change in the radar backscatter signature across selected ecoregions of the boreal zone. We have measured vegetation tissue temperatures, soil temperature profiles, and micrometeorological parameters in situ at selected sites along a north-south transect extending across Alaska from Prudhoe Bay to the Kenai Peninsula and in Siberia near the Yenisey River. Data from these stations have been used to quantify the scatterometer's sensitivity to freeze/thaw state under a variety of terrain and landcover conditions. Analysis of the NSCAT temporal response over the 1997 spring thaw cycle shows a 3 to 5 dB change in measured backscatter that is well correlated with the landscape springtime thaw process. Having verified the instrument's capability to monitor freeze/thaw transitions, regional scale mosaicked data are applied to derive temporal series of freeze/thaw transition maps for selected circumpolar high latitude regions. These maps are applied to derive areal extent of frozen and thawed landscape and demonstrate the utility of spaceborne radar for operational monitoring of seasonal freeze-thaw dynamics and associated biophysical processes for the circumpolar high latitudes.

  17. The Critical Depth of Freeze-Thaw Soil under Different Types of Snow Cover

    Directory of Open Access Journals (Sweden)

    Qiang Fu

    2017-05-01

    Full Text Available Snow cover is the most common upper boundary condition influencing the soil freeze-thaw process in the black soil farming area of northern China. Snow is a porous dielectric cover, and its unique physical properties affect the soil moisture diffusion, heat conduction, freezing rate and other variables. To understand the spatial distribution of the soil water-heat and the variable characteristics of the critical depth of the soil water and heat, we used field data to analyze the freezing rate of soil and the extent of variation in soil water-heat in a unit soil layer under bare land (BL, natural snow (NS, compacted snow (CS and thick snow (TS treatments. The critical depth of the soil water and heat activity under different snow covers were determined based on the results of the analysis, and the variation fitting curve of the difference sequences on the soil temperature and water content between different soil layers and the surface 5-cm soil layer were used to verify the critical depth. The results were as follows: snow cover slowed the rate of soil freezing, and the soil freezing rate under the NS, CS and TS treatments decreased by 0.099 cm/day, 0.147 cm/day and 0.307 cm/day, respectively, compared with that under BL. In addition, the soil thawing time was delayed, and the effect was more significant with increased snow cover. During freeze-thaw cycles, the extent of variation in the water and heat time series in the shallow soil was relatively large, while there was less variation in the deep layer. There was a critical stratum in the vertical surface during hydrothermal migration, wherein the critical depth of soil water and heat change gradually increased with increasing snow cover. The variance in differences between the surface layer and both the soil water and heat in the different layers exhibited “steady-rising-steady” behavior, and the inflection point of the curve is the critical depth of soil freezing and thawing. This critical

  18. Freeze-thaw stress of Alhydrogel ® alone is sufficient to reduce the immunogenicity of a recombinant hepatitis B vaccine containing native antigen.

    Science.gov (United States)

    Clapp, Tanya; Munks, Michael W; Trivedi, Ruchit; Kompella, Uday B; Braun, LaToya Jones

    2014-06-24

    Preventing losses in vaccine potency due to accidental freezing has recently become a topic of interest for improving vaccines. All vaccines with aluminum-containing adjuvants are susceptible to such potency losses. Recent studies have described excipients that protect the antigen from freeze-induced inactivation, prevent adjuvant agglomeration and retain potency. Although these strategies have demonstrated success, they do not provide a mechanistic understanding of freeze-thaw (FT) induced potency losses. In the current study, we investigated how adjuvant frozen in the absence of antigen affects vaccine immunogenicity and whether preventing damage to the freeze-sensitive recombinant hepatitis B surface antigen (rHBsAg) was sufficient for maintaining vaccine potency. The final vaccine formulation or Alhydrogel(®) alone was subjected to three FT-cycles. The vaccines were characterized for antigen adsorption, rHBsAg tertiary structure, particle size and charge, adjuvant elemental content and in-vivo potency. Particle agglomeration of either vaccine particles or adjuvant was observed following FT-stress. In vivo studies demonstrated no statistical differences in IgG responses between vaccines with FT-stressed adjuvant and no adjuvant. Adsorption of rHBsAg was achieved; regardless of adjuvant treatment, suggesting that the similar responses were not due to soluble antigen in the frozen adjuvant-containing formulations. All vaccines with adjuvant, including the non-frozen controls, yielded similar, blue-shifted fluorescence emission spectra. Immune response differences could not be traced to differences in the tertiary structure of the antigen in the formulations. Zeta potential measurements and elemental content analyses suggest that FT-stress resulted in a significant chemical alteration of the adjuvant surface. This data provides evidence that protecting a freeze-labile antigen from subzero exposure is insufficient to maintain vaccine potency. Future studies should

  19. Compressive Strength, Chloride Permeability, and Freeze-Thaw Resistance of MWNT Concretes under Different Chemical Treatments

    Directory of Open Access Journals (Sweden)

    Xingang Wang

    2014-01-01

    Full Text Available This study investigated compressive strength, chloride penetration, and freeze-thaw resistance of multiwalled carbon nanotube (MWNT concrete. More than 100 cylindrical specimens were used to assess test variables during sensitivity observations, including water-cement ratios (0.75, 0.5, and 0.4 and exposure to chemical agents (including gum arabic, propanol, ethanol, sodium polyacrylate, methylcellulose, sodium dodecyl sulfate, and silane. To determine the adequate sonication time for MWNT dispersal in water, the compressive strengths of MWNT concrete cylinders were measured after sonication times ranging from 2 to 24 minutes. The results demonstrated that the addition of MWNT can increase the compressive strength of concrete by up to 108%. However, without chemical treatment, MWNT concretes tend to have poor freeze-thaw resistance. Among the different chemical treatments, MWNT concrete treated with sodium polyacrylate has the best compressive strength, chloride resistance, and freeze-thaw durability.

  20. A practical method to detect the freezing/thawing onsets of seasonal frozen ground in Alaska

    Science.gov (United States)

    Chen, Xiyu; Liu, Lin

    2017-04-01

    Microwave remote sensing can provide useful information about freeze/thaw state of soil at the Earth surface. An edge detection method is applied in this study to estimate the onsets of soil freeze/thaw state transition using L band space-borne radiometer data. The Soil Moisture Active Passive (SMAP) mission has a L band radiometer and can provide daily brightness temperature (TB) with horizontal/vertical polarizations. We use the normalized polarization ratios (NPR) calculated based on the Level-1C TB product of SMAP (spatial resolution: 36 km) as the indicator for soil freeze/thaw state, to estimate the freezing and thawing onsets in Alaska in the year of 2015 and 2016. NPR is calculated based on the difference between TB at vertical and horizontal polarizations. Therefore, it is strongly sensitive to liquid water content change in the soil and independent with the soil temperature. Onset estimation is based on the detection of abrupt changes of NPR in transition seasons using edge detection method, and the validation is to compare estimated onsets with the onsets derived from in situ measurement. According to the comparison, the estimated onsets were generally 15 days earlier than the measured onsets in 2015. However, in 2016 there were 4 days in average for the estimation earlier than the measured, which may be due to the less snow cover. Moreover, we extended our estimation to the entire state of Alaska. The estimated freeze/thaw onsets showed a reasonable latitude-dependent distribution although there are still some outliers caused by the noisy variation of NPR. At last, we also try to remove these outliers and improve the performance of the method by smoothing the NPR time series.

  1. Investigation of freeze/thaw durability in polymer electrolyte fuel cells

    Energy Technology Data Exchange (ETDEWEB)

    Lim, Soo-Jin; Park, Gu-Gon; Sohn, Young-Jun; Yim, Sung-Dae; Yang, Tae-Hyun; Kim, Chang-Soo [Fuel Cell Research Center, Korea Institute of Energy Research, 102, Gajeong-ro, Yuseong-gu, Daejeon 305-343 (Korea, Republic of); Park, Jin-Soo [Department of Environmental Engineering, College of Engineering, Sanmyung University, 300 Anseo-dong, Dongnam-gu, Cheonam, Chungnam Province 330-720 (Korea, Republic of); Hong, Bo Ki [Fuel Cell Vehicle Team 1, Ecotechnology Center, Hyundai-Kia Motors Company, 104, Mabuk-dong, Giheung-gu, Yongin-si, Gyeonggi-do 446-912 (Korea, Republic of)

    2010-12-15

    This study aims to investigate the effect of different gas diffusion layers (GDLs) on freeze/thaw condition durability in polymer electrolyte fuel cells (PEFCs). Three kinds of GDLs-cloth, felt and paper type - with similar basic properties except thickness and bending stiffness were used. The changes in the properties and cell performance were investigated from the -30 to 70 C range of freeze/thaw cycles. The I-V performance degradation was observed to be negligible for the felt GDL whereas the cloth and paper GDLs showed a marked I-V performance loss. No distinctive correlation between the changes in electrochemical properties, such as active metal surface area, hydrogen crossover rates and decreased I-V performance, was observed except an increase in ohmic resistance revealed by ac-impedance spectroscopy. The physical destruction of electrodes was also shown by scanning electron microscope (SEM) analysis. The present study found that sufficient mechanical supporting force between the interfaces of materials enhances PEFC durability in sub-zero temperature conditions. (author)

  2. Investigation of the impact of nanotechnology on the freeze-thaw durability of concrete containing d-cracking aggregates : [technical summary].

    Science.gov (United States)

    2015-05-01

    Freezing and thawing damage is the most common cause of distress in : Kansas pavements. Many locally available aggregates in Kansas do not : meet current standards for use in concrete pavements because of poor : freeze-thaw durability. The use of nan...

  3. Effects of freezing-thawing on sensory descriptive profiles of cooked poultry breast meat

    Directory of Open Access Journals (Sweden)

    ZHUANG Hong

    2014-12-01

    Full Text Available Freezing is a common method used by consumers to extend meat shelf life and by researchers to allow for subsequent meat quality assessments and processing.However,the effects of freezing on the sensory quality of cooked poultry breast meat are not well documented.The objective of this study was to compare sensory quality profiles of fresh and frozen/thawed chicken breast fillets (pectoralis major.Breast fillets were removed from carcasses within 24 h postmortem and either cooked from a fresh state or placed in a -20℃ freezer.Frozen samples were thawed by three different methods:thawing during cooking directly from a frozen state (0 h,thawing in 20℃ water for 2 h prior to cooking (2 h,or thawing at 4℃ for 24 h prior to cooking (24 h.A control treatment with fillets cooked directly from a fresh state was used.Fillets were cooked to an endpoint temperature of 78℃ and sensory quality was evaluated by trained descriptive panelists using 0~15 universal intensity scales.Results show that there were not treatment differences (P>0.05 in the average intensity scores for any of the descriptive flavor attributes or for 5 of the descriptive texture attributes (cohesiveness,hardness,juiciness,wad size,and wetness of wad.However,the intensity scores for cohesiveness of mass,rate of breakdown,and chewiness were significantly different among the treatments (P<0.05.Cohesiveness of mass intensity scores for 0h and 24 h fillets were significantly higher than 2 h samples.Fillets cooked directly from a frozen state (0 h had significantly higher intensity scores for rate of breakdown and chewiness than fresh controls and 2 h samples,respectively.These results indicate that freezing-thawing does not affect sensory flavor quality;however,it may change the texture attributes of cooked chicken breast meat products.The effects on meat texture depend on thawing methods prior to cooking.

  4. Freeze-thaw resistance of concrete with marginal air content : final report

    Science.gov (United States)

    2006-12-01

    Freeze-thaw resistance is a key durability factor for concrete pavements. Recommendations for the air : void system parameters are normally: 6 1 percent total air, and spacing factor less than 0.20 : millimeters. However, it was observed that some...

  5. Role of freeze-thaw cycles and chlorpyrifos insecticide use on diffuse Cd loss and sediment accumulation

    Science.gov (United States)

    Wang, Fangli; Ouyang, Wei; Hao, Fanghua; Jiao, Wei; Shan, Yushu; Lin, Chunye

    2016-06-01

    Freeze-thaw cycles are predicted to increase in cold temperate regions. The potential influence of the interactions of freeze-thaw cycles and agrochemicals on the release of Cd into river water is unknown. In this study, the interactions of freeze-thaw cycles and chlorpyrifos (FC) on Cd mobility in soils were analysed. The spatial variability of soil Cd under long-term intensive tillage in a freeze-thaw agro-system was also identified. The temporal variation of sediment Cd was detected based on analysis of the sediment geochemistry. The results showed that FC increased soil Cd mobility, with an increase of approximately 10% in CaCl2-extractable Cd. The increased mobile fractions of water-soluble and exchangeable Cd originated from the decreased fraction of Fe-Mn-oxide-associated Cd and organic matter-bound Cd. The total Cd content in the surface soil followed the zonally decreasing trend of dry land > paddy land > natural land. The Cd concentrations and sedimentation rates of the sediment core generally increased from 1943 to 2013 due to agricultural exploration and farmland irrigation system construction, indicating an increase of the Cd input flux into water. The results provide valuable information about the soil Cd transport response to the influence of climatic and anthropogenic factors in cold intensive agro-systems.

  6. Change in behavior of uniaxial compression due to degradation of salt water and freezing and thawing for rock

    International Nuclear Information System (INIS)

    Yamamoto, Kiyohito; Kobayashi, Akira; Aoyama, Shigeyasu

    2007-01-01

    To investigate the effect of the degradation on the mechanical behavior, the degraded rock samples were prepared to the uniaxial compression test. The degradation methods are divided into two types. One type is submerged in the 10% saline water (10%NaCl) for 90 days, and another one is freezing and thawing for 240 cycles. The degraded Smaland-granites were preserved in saline water. Kurihashi-granodiorite, Tage-tuff and Funyu-tuff were imposed on freezing and thawing test to make degraded state. The damage parameters were identified from the stress-strain relation obtained from the uniaxial compression tests. The damage parameters are K υ , n υ , K d , n d and B 0 . K υ and n υ are related to expansive strain. K d , n d and B 0 are subject to behavior of Young's modulus. By investigating the change in the damage parameters of the degraded rock, the effect of the degradation was tried to infer. As the results, it was inferred using the damage parameters that the Smaland-granite becomes more expansive material and the damage occurs earlier due to saline water degradation. Moreover, it was considered that the Kurihashi-granodiorite and Tage-tuff become more expansive and the axial strain at the failure decreases by freezing and thawing degradation, however the axial strain of the Funyu-tuff at the failure becomes large. It was found the proposed damage parameters can be good index for volumetric strain behavior after degradation. (author)

  7. Freeze-thaw cycles induce content exchange between cell-sized lipid vesicles

    Science.gov (United States)

    Litschel, Thomas; Ganzinger, Kristina A.; Movinkel, Torgeir; Heymann, Michael; Robinson, Tom; Mutschler, Hannes; Schwille, Petra

    2018-05-01

    Early protocells are commonly assumed to consist of an amphiphilic membrane enclosing an RNA-based self-replicating genetic system and a primitive metabolism without protein enzymes. Thus, protocell evolution must have relied on simple physicochemical self-organization processes within and across such vesicular structures. We investigate freeze-thaw (FT) cycling as a potential environmental driver for the necessary content exchange between vesicles. To this end, we developed a conceptually simple yet statistically powerful high-throughput procedure based on nucleic acid-containing giant unilamellar vesicles (GUVs) as model protocells. GUVs are formed by emulsion transfer in glass bottom microtiter plates and hence can be manipulated and monitored by fluorescence microscopy without additional pipetting and sample handling steps. This new protocol greatly minimizes artefacts, such as unintended GUV rupture or fusion by shear forces. Using DNA-encapsulating phospholipid GUVs fabricated by this method, we quantified the extent of content mixing between GUVs under different FT conditions. We found evidence of nucleic acid exchange in all detected vesicles if fast freezing of GUVs at ‑80 °C is followed by slow thawing at room temperature. In contrast, slow freezing and fast thawing both adversely affected content mixing. Surprisingly, and in contrast to previous reports for FT-induced content mixing, we found that the content is not exchanged through vesicle fusion and fission, but that vesicles largely maintain their membrane identity and even large molecules are exchanged via diffusion across the membranes. Our approach supports efficient screening of prebiotically plausible molecules and environmental conditions, to yield universal mechanistic insights into how cellular life may have emerged.

  8. Application of Freezing and Thawing in Apple (Malus domestica) Juice Extraction.

    Science.gov (United States)

    Nadulski, Rafał; Kobus, Zbigniew; Wilczyński, Kamil; Zawiślak, Kazimierz; Grochowicz, Józef; Guz, Tomasz

    2016-09-28

    The paper presents the results of the research on the impact of enzymatic liquefaction, freezing and thawing on the efficiency of juice pressure extraction from apple pulp and quality of the obtained juices. The research was conducted using three types of pretreatment prior the pressing: crushing and enzymatic liquefaction in temperature of 25 °C, crushing and enzymatic liquefaction in temperature of 45 °C and crushing followed by freezing and thawing of the pulp. The study included three varieties of apples. The juice was obtained using a laboratory basket press. It was determined that the pretreatment of the pulp as well as the varietal characteristics of the fruits have a significant impact on the efficiency of the pressure extraction process. The enzymatic treatment of the pulp, irrespective of the temperature at which it was conducted, significantly increased the efficiency of the process. No effect of the temperature (25 or 45 °C) of enzymatic treatment on the efficiency of the pressure extraction process was found. Pretreatment of the pulp based on freezing and thawing contributes to the increase of efficiency of pressing in the case of two apple cultivars, that is, Idared and Red Delicious. It was showed that total phenolic content, antioxidant activity, the soluble solids content and juice acidity (pH) depend on the pretreatment of the pulp and the varietal characteristics of apples. Following the application of pretreatment of the pulp, an increase was observed in the content of polyphenols and in the antioxidant activity of the juices obtained. © 2016 Institute of Food Technologists®

  9. Numerical Simulation of the Freeze-Thaw Behavior of Mortar Containing Deicing Salt Solution.

    Science.gov (United States)

    Esmaeeli, Hadi S; Farnam, Yaghoob; Bentz, Dale P; Zavattieri, Pablo D; Weiss, Jason

    2017-02-01

    This paper presents a one-dimensional finite difference model that is developed to describe the freeze-thaw behavior of an air-entrained mortar containing deicing salt solution. A phenomenological model is used to predict the temperature and the heat flow for mortar specimens during cooling and heating. Phase transformations associated with the freezing/melting of water/ice or transition of the eutectic solution from liquid to solid are included in this phenomenological model. The lever rule is used to calculate the quantity of solution that undergoes the phase transformation, thereby simulating the energy released/absorbed during phase transformation. Undercooling and pore size effects are considered in the numerical model. To investigate the effect of pore size distribution, this distribution is considered using the Gibbs-Thomson equation in a saturated mortar specimen. For an air-entrained mortar, the impact of considering pore size (and curvature) on freezing was relatively insignificant; however the impact of pore size is much more significant during melting. The fluid inside pores smaller than 5 nm (i.e., gel pores) has a relatively small contribution in the macroscopic freeze-thaw behavior of mortar specimens within the temperature range used in this study (i.e., +24 °C to -35 °C), and can therefore be neglected for the macroscopic freeze-thaw simulations. A heat sink term is utilized to simulate the heat dissipation during phase transformations. Data from experiments performed using a low-temperature longitudinal guarded comparative calorimeter (LGCC) on mortar specimens fully saturated with various concentration NaCl solutions or partially saturated with water is compared to the numerical results and a promising agreement is generally obtained.

  10. Research on the Strength Variation of Root-Clay Systems under Freeze-Thaw Action

    Directory of Open Access Journals (Sweden)

    Lin Yang

    2017-01-01

    Full Text Available The aim of this paper is to study the influence of an effective root system of rhizome plants on the reinforcement of slope soil under freeze-thaw conditions. This study focused on the mechanical properties between roots and clay in the root system of four plant species from different regions of China (northeast, northern, central, and southern areas: Setaria viridis, Eleusine indica, Zoysia japonica, and Carex leucochlora. Based on the interfacial friction effects between the plant roots and the soil, pull-out tests and unconfined compressive strength tests were conducted on the reinforced soil system for varying numbers of freeze-thaw cycles. Several stages of the pull-out process of the root system in clay are explicitly proposed based on the interfacial friction test results. The results showed that the friction effect between Zoysia japonica roots and the soil was the most significant and that these roots had the best reinforcement effect. In contrast, the friction and reinforcement effects between Setaria viridis roots and the soil were the worst, and the resulting unconfined compressive strength was the smallest. However, the freeze-thaw resistance ability of the Setaria viridis and soil system was stronger than that of the Zoysia japonica system.

  11. Effects of different cryoprotectants and freezing methods on post-thaw boar semen quality.

    Science.gov (United States)

    Yang, Chung-Hsun; Wu, Ting-Wen; Cheng, Feng-Pang; Wang, Jiann-Hsiung; Wu, Jui-Te

    2016-03-01

    The current study aimed to investigate the effects of different concentrations of glycerol (0%, 1%, 2%, 3%, and 5%) and dimethylacetamide (DMA: 0%, 1%, 3%, and 5%) on post-sperm quality characteristics following semen freezing in dry ice (D) or liquid nitrogen (N). Semen was collected from Duroc boars and was allocated to 32 treatment groups for cryopreservation. Analysis of post-thaw semen quality and fertility after artificial insemination (AI) was used to examine the combinatorial effects of different treatments. The best scores for post-thaw sperm motility, sperm viability, and sperm acrosomal integrity were observed in semen frozen in: (a) dry ice in the presence of 5% glycerol and no DMA (16D-treatment); (b) dry ice in the presence of 3% glycerol and no DMA (9D-treatment); and (c) liquid nitrogen in the presence of 3% glycerol and 1% DMA (10N-treatment), with no significant difference observed among these three treatments. The farrowing rates after AI with post-thawed semen after 9D- and 10N-treatments were 33% and 50%, respectively. To summarize, the results of the present study indicated that the freezing extender containing 3% glycerol in combination with the straw-freezing method using dry ice produced the best post-thaw quality parameters of boar semen. Combinations of glycerol and DMA did not enhance the cryosurvival of boar spermatozoa. Copyright © 2016 Society for Biology of Reproduction & the Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

  12. Cavitation and water fluxes driven by ice water potential in Juglans regia during freeze-thaw cycles.

    Science.gov (United States)

    Charra-Vaskou, Katline; Badel, Eric; Charrier, Guillaume; Ponomarenko, Alexandre; Bonhomme, Marc; Foucat, Loïc; Mayr, Stefan; Améglio, Thierry

    2016-02-01

    Freeze-thaw cycles induce major hydraulic changes due to liquid-to-ice transition within tree stems. The very low water potential at the ice-liquid interface is crucial as it may cause lysis of living cells as well as water fluxes and embolism in sap conduits, which impacts whole tree-water relations. We investigated water fluxes induced by ice formation during freeze-thaw cycles in Juglans regia L. stems using four non-invasive and complementary approaches: a microdendrometer, magnetic resonance imaging, X-ray microtomography, and ultrasonic acoustic emissions analysis. When the temperature dropped, ice nucleation occurred, probably in the cambium or pith areas, inducing high water potential gradients within the stem. The water was therefore redistributed within the stem toward the ice front. We could thus observe dehydration of the bark's living cells leading to drastic shrinkage of this tissue, as well as high tension within wood conduits reaching the cavitation threshold in sap vessels. Ultrasonic emissions, which were strictly emitted only during freezing, indicated cavitation events (i.e. bubble formation) following ice formation in the xylem sap. However, embolism formation (i.e. bubble expansion) in stems was observed only on thawing via X-ray microtomography for the first time on the same sample. Ultrasonic emissions were detected during freezing and were not directly related to embolism formation. These results provide new insights into the complex process and dynamics of water movements and ice formation during freeze-thaw cycles in tree stems. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  13. Freezing and thawing resistance of aerial lime mortar with metakaolin\

    Czech Academy of Sciences Publication Activity Database

    Nunes, Cristiana Lara; Slížková, Zuzana

    2016-01-01

    Roč. 114, July (2016), s. 896-905 ISSN 0950-0618 R&D Projects: GA ČR(CZ) GBP105/12/G059 Institutional support: RVO:68378297 Keywords : freeze-thaw * lime * metakaolin * linseed oil * mortar * water-repellency Subject RIV: JN - Civil Engineering Impact factor: 3.169, year: 2016 http://www.sciencedirect.com/science/article/pii/S0950061816305645

  14. Stem water transport and freeze-thaw xylem embolism in conifers and angiosperms in a Tasmanian treeline heath.

    Science.gov (United States)

    Feild, Taylor S; Brodribb, Tim

    2001-05-01

    The effect of freezing on stem xylem hydraulic conductivity and leaf chlorophyll a fluorescence was measured in 12 tree and shrub species from a treeline heath in Tasmania, Australia. Reduction in stem hydraulic conductivity after a single freeze-thaw cycle was minimal in conifers and the vessel-less angiosperm species Tasmannia lanceolata (Winteraceae), whereas mean loss of conductivity in vessel-forming angiosperms fell in the range 17-83%. A positive linear relationship was observed between percentage loss of hydraulic conductivity by freeze-thaw and the average conduit diameter across all 12 species. This supports the hypothesis that large-diameter vascular conduits have a greater likelihood of freeze-thaw cavitation because larger bubbles are produced, which are more likely to expand under tension. Leaf frost tolerances, as measured by a 50% loss of maximum PSII quantum yield, varied from -6 to -13°C, indicating that these species were more frost-sensitive than plants from northern hemisphere temperate forest and treeline communities. There was no evidence of a relationship between frost tolerance of leaves and the resilience of stem water transport to freezing, suggesting that low temperature survival and the resistance of stem water transport to freezing are independently evolving traits. The results of this study bear on the ecological importance of stem freezing in the southern hemisphere treeline zones.

  15. Deicer-freeze-thaw resistance characteristics of Portland cement concrete for Wisconsin pavements

    Science.gov (United States)

    1995-12-01

    The objectives of the study were to identify PCC mixes that offer increased deicer-freeze-thaw durability and to identify mix characteristics that contribute to premature durability distress. PCC mixes covering a range of air contents and water-cemen...

  16. Use of the TLX ultracentrifuge for the isolation of different density lipoproteins and effects of freeze/thawing of human plasma before ultracentrifugation.

    Science.gov (United States)

    Charlton-Menys, Valentine; Chobotova, Jelena; Durrington, Paul N

    2008-01-01

    Isolation of different density lipoproteins by ultracentrifugation can require lengthy centrifugation times and freeze/thawing of plasma may influence recovery. We isolated a range of lipoproteins using a preparative ultracentrifuge and the TLX micro-ultracentrifuge and determined the effect of freeze/thawing of plasma beforehand. In fresh plasma, there was no significant difference in results for small-dense low-density lipoprotein apolipoprotein B (LDL apoB) (density >1.044 g/mL) or cholesterol at density >1.006 g/mL. Freeze/thawing had no effect on closely correlated results for small-dense LDL apoB (r=0.85; pTLX micro-ultracentrifuge is a reliable alternative to the preparative ultracentrifuge and freeze/thawing has only a small effect on small-dense LDL apoB or high-density lipoprotein cholesterol.

  17. Petroleum hydrocarbon biodegradation under seasonal freeze-thaw soil temperature regimes in contaminated soils from a sub-Arctic site.

    Science.gov (United States)

    Chang, Wonjae; Klemm, Sara; Beaulieu, Chantale; Hawari, Jalal; Whyte, Lyle; Ghoshal, Subhasis

    2011-02-01

    Several studies have shown that biostimulation in ex situ systems such as landfarms and biopiles can facilitate remediation of petroleum hydrocarbon contaminated soils at sub-Arctic sites during summers when temperatures are above freezing. In this study, we examine the biodegradation of semivolatile (F2: C10-C16) and nonvolatile (F3: C16-C34) petroleum hydrocarbons and microbial respiration and population dynamics at post- and presummer temperatures ranging from -5 to 14 °C. The studies were conducted in pilot-scale tanks with soils obtained from a historically contaminated sub-Arctic site in Resolution Island (RI), Canada. In aerobic, nutrient-amended, unsaturated soils, the F2 hydrocarbons decreased by 32% during the seasonal freeze-thaw phase where soils were cooled from 2 to -5 °C at a freezing rate of -0.12 °C d(-1) and then thawed from -5 to 4 °C at a thawing rate of +0.16 °C d(-1). In the unamended (control) tank, the F2 fraction only decreased by 14% during the same period. Biodegradation of individual hydrocarbon compounds in the nutrient-amended soils was also confirmed by comparing their abundance over time to that of the conserved diesel biomarker, bicyclic sesquiterpanes (BS). During this period, microbial respiration was observed, even at subzero temperatures when unfrozen liquid water was detected during the freeze-thaw period. An increase in culturable heterotrophs and 16S rDNA copy numbers was noted during the freezing phase, and the (14)C-hexadecane mineralization in soil samples obtained from the nutrient-amended tank steadily increased. Hydrocarbon degrading bacterial populations identified as Corynebacterineae- and Alkanindiges-related strains emerged during the freezing and thawing phases, respectively, indicating there were temperature-based microbial community shifts.

  18. EPINEPHRINE CONCENTRATION IN WILD BOAR (SUS SCROFA L. SERUM AFTER REPEATED ELISA TESTED FREEZE-THAWING CYCLES

    Directory of Open Access Journals (Sweden)

    Neška Vukšić

    2013-06-01

    Full Text Available Blood samples for determining of epinephrine concentration and biochemical parameters in the blood serum of wild boars were taken from 42 healthy wild boars, both sexes, during the hunting season. All animals in good condition, body weight 20 to 95 kg, were divided into two groups up to 50 kg (group A and up to 95 kg (Group B. Epinephrine concentration was determined by ELISA twice: one week after taking samples and a month after repeated freezing at -80°C. It was higher in relation to the reference value of domestic pigs and human (109.45 pg/ml in A and 119.54 pg/ml in B group. Repeated freezing and re-analysis after a month were resulted in lower concentrations of epinephrine (12% in young and 11.17% in adult animal, but without statistical significance (P>0.05. Biochemical analysis results’ show increased glucose and triglycerides concentrations compared to the reference values, while other indicators were observed within or slightly increased referring to the normal range. The correlation between glucose and epinephrine was not determined.

  19. A Study on Accelerated Corrosion Test by Combined Deteriorating Action of Salt Damage and Freeze-Thaw

    Energy Technology Data Exchange (ETDEWEB)

    Park, Sang-Soon; So, Byung-Tak [Sangmyung University, Cheonan (Korea, Republic of)

    2016-01-15

    In this study, the accelerated corrosion test by combined deteriorating action of salt damage and freeze-thaw was investigated. freeze-thaw cycle is one method for corrosion testing; corrosion initiation time was measured in four types of concrete samples, i.e., two samples mixed with fly ash (FA) and blast furnace slag (BS), and the other two samples having two water/cement ratio (W/C = 0.6, 0.35) without admixture (OPC60 and OPC35). The corrosion of rebar embedded in concrete occurred most quickly at the 30th freeze-thaw cycle. Moreover, a corrosion monitoring method with a half-cell potential measurement and relative dynamic elastic modulus derived from resonant frequency measures was conducted simultaneously. The results indicated that the corrosion of rebar occurred when the relative dynamic elastic modulus was less than 60%. Therefore, dynamic elastic modulus can be used to detect corrosion of steel bar. The results of the accelerated corrosion test exhibited significant difference according to corrosion periods combined with each test condition. Consequently, the OPC60 showed the lowest corrosion resistance among the samples.

  20. The detection of Giardia muris and Giardia lamblia cysts by immunofluorescence in animal tissues and fecal samples subjected to cycles of freezing and thawing.

    Science.gov (United States)

    Erlandsen, S L; Sherlock, L A; Bemrick, W J

    1990-04-01

    The effects of freezing and thawing on the detection of selected Giardia spp. cysts were investigated using immunofluorescence, bright field microscopy, and low voltage scanning electron microscopy (SEM). Giardia muris cysts were obtained from either animal carcasses, fecal pellets, or isolated cyst preparations, whereas Giardia lamblia cysts were isolated from fecal samples. These samples were stained using an immunofluorescence technique after 1-3 freezing (-16 C) and thawing (20 C) cycles. Cysts were detected successfully by immunofluorescence in all samples. However, in those samples subjected to freeze-thawing, the cyst walls often became distorted and then were not detectable by bright field microscopy. Low voltage SEM demonstrated that the filaments in the distorted cyst wall underwent rearrangements of interfilament spacing. Quantitation of cyst recovery after freezing and thawing demonstrated that a substantial loss occurred after 1 cycle of alternating temperature when low concentrations of cysts were used, but not with high concentrations of cysts. Cyst recovery, after 3 freezing and thawing cycles, was dramatically lowered irrespective of the initial cyst concentration. These results demonstrated that immunofluorescence was an effective technique for the detection of Giardia spp. cysts in frozen samples and would suggest that freezing and thawing of fecal samples could prevent the detection of cysts when only bright field microscopy was employed.

  1. Effect of multiple freeze-thaw cycles on the quality of instant sea cucumber: Emphatically on water status of by LF-NMR and MRI.

    Science.gov (United States)

    Tan, Mingqian; Lin, Zhuyi; Zu, Yinxue; Zhu, Beiwei; Cheng, Shasha

    2018-07-01

    Instant sea cucumber has become one popular product due to its convenience to eat, favourable taste and minimal loss of nutrients and bioactive components. However, there was rare information about the water dynamic of instant sea cucumber subjected to multiple freeze-thaw cycles. In this study, low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance image (MRI) were employed to investigate the effect of freeze-thaw cycles on water status of instant sea cucumber. Four water populations corresponding to strongly bound water, weakly bound water, immobile water and free water were observed in instant sea cucumber. With the increase of freeze-thaw cycles, the transverse relaxation time of immobile and free water increased, while the peak area of free water decreased significantly. MRI enabled the visualization of water migration of instant sea cucumber during multiple freeze-thaw cycles. Multiple freeze-thaw cycles also led to significant changes of other quality properties including thawing loss, WHC, color parameters, texture and protein content, and enlarge the interspace between fiber network in microstructure. Good correlations between T 22 , A 22 , A 23 and thaw loss, WHC, L*, hardness and collagen content (0.873 ≤ r ≤ 0.958) revealed LF-NMR may be an effective real-time monitoring method of these physicochemical parameters as a non-destructive technique. Copyright © 2018 Elsevier Ltd. All rights reserved.

  2. Results of molten salt panel and component experiments for solar central receivers: Cold fill, freeze/thaw, thermal cycling and shock, and instrumentation tests

    Energy Technology Data Exchange (ETDEWEB)

    Pacheco, J.E.; Ralph, M.E.; Chavez, J.M.; Dunkin, S.R.; Rush, E.E.; Ghanbari, C.M.; Matthews, M.W.

    1995-01-01

    Experiments have been conducted with a molten salt loop at Sandia National Laboratories in Albuquerque, NM to resolve issues associated with the operation of the 10MW{sub e} Solar Two Central Receiver Power Plant located near Barstow, CA. The salt loop contained two receiver panels, components such as flanges and a check valve, vortex shedding and ultrasonic flow meters, and an impedance pressure transducer. Tests were conducted on procedures for filling and thawing a panel, and assessing components and instrumentation in a molten salt environment. Four categories of experiments were conducted: (1) cold filling procedures, (2) freeze/thaw procedures, (3) component tests, and (4) instrumentation tests. Cold-panel and -piping fill experiments are described, in which the panels and piping were preheated to temperatures below the salt freezing point prior to initiating flow, to determine the feasibility of cold filling the receiver and piping. The transient thermal response was measured, and heat transfer coefficients and transient stresses were calculated from the data. Freeze/thaw experiments were conducted with the panels, in which the salt was intentionally allowed to freeze in the receiver tubes, then thawed with heliostat beams. Slow thermal cycling tests were conducted to measure both how well various designs of flanges (e.g., tapered flanges or clamp type flanges) hold a seal under thermal conditions typical of nightly shut down, and the practicality of using these flanges on high maintenance components. In addition, the flanges were thermally shocked to simulate cold starting the system. Instrumentation such as vortex shedding and ultrasonic flow meters were tested alongside each other, and compared with flow measurements from calibration tanks in the flow loop.

  3. A combined effect of freeze--thaw cycles and polymer concentration on the structure and mechanical properties of transparent PVA gels.

    Science.gov (United States)

    Gupta, Siddhi; Goswami, Sudipta; Sinha, Arvind

    2012-02-01

    Transparent poly(vinyl alcohol) (PVA) hydrogel films, derived from aqueous solutions of varying concentration, were synthesized by the cyclic freeze-thaw method (0°-37 °C). This study demonstrates a variation in the transparency, degree of crystallinity, wettability, swelling and mechanical properties of the hydrogels as a function of the solution concentration and the number of freeze-thaw cycles for a given average molecular weight (95,000 Da). The study manifests a strong control of the number of freeze-thaw cycles on the structure-property correlations of the synthesized transparent PVA hydrogels, revealing the possibility of obtaining a window of structural and process parameters for the physically cross-linked hydrogels, making them suitable for cell-gel interactions.

  4. Failure of cement hydrates: freeze-thaw and fracture

    Science.gov (United States)

    Ioannidou, Katerina; Del Gado, Emanuela; Ulm, Franz-Josef; Pellenq, Roland

    Mechanical and viscoelastic behavior of concrete crucially depends on cement hydrates, the ``glue'' of cement. Even more than the atomistic structure, the mesoscale amorphous texture of cement hydrates over hundreds of nanometers plays a crucial role for material properties. We use simulations that combine information of the nano-scale building units of cement hydrates and on their effective interactions, obtained from atomistic simulations and experiments, into a statistical physics framework for aggregating nanoparticles.Our mesoscale model was able to reconcile different experimental results ranging from small-angle neutron scattering, SEM, adsorption/desorption of N2, and water to nanoindentation and gain the new fundamental insights into the microscopic origin of the properties measured. Our results suggest that heterogeneities developed during the early stages of hydration persist in the structure of C-S-H, impacting the rheological and mechanical performance of the hardened cement paste. In this talk I discuss recent investigation on failure mechanism at the mesoscale of hardened cement paste such as freeze-thaw and fracture. Using correlations between local volume fractions and local stress we provide a link between structural and mechanical heterogeneities during the failure mechanisms.

  5. Microwave freeze-thaw technique of injectable drugs. A review from 1980 to 2014.

    Science.gov (United States)

    Hecq, J-D; Godet, M; Jamart, J; Galanti, L

    2015-11-01

    Microwave freeze-thaw treatment (MFTT) of injectable drugs can support the development of centralized intravenous admixtures services (CIVAS). The aim of the review is to collect information and results about this method. A systematic review of the scientific literature about injectable drug stability studies was performed. The data are presented in a table and describe name of the drug, producer, final concentration, temperature and time of freezing storage, type of microwave oven, thawing power, method of dosage and results after treatment or final long-term storage at 5±3 °C. From 1980 to 2014, 59 drugs were studied by MFTT and the results were presented in 49 publications. Forty papers were presented by 8 teams (2 to 18 by team). The temperatures of freezing storage vary from -70 °C to -10 °C, the time storage from 4 hours to 12 months, the thaw from low to full power. Dosages are mainly made by high performance liquid chromatography. Most of the 59 drugs are stable during and after treatment. Only 3 teams have tested the long-term stability after MFTT, the first for ganciclovir after 7 days, the second for ceftizoxime after 30 days and the third for 19 drugs after 11 to 70 days. This review can help CIVAS to take in charge the productions of ready-to-use injectable drugs. Copyright © 2015 Académie Nationale de Pharmacie. Published by Elsevier Masson SAS. All rights reserved.

  6. Effect of Multiple Freezing/Thawing Cycles on the Structural and Functional Properties of Waxy Rice Starch

    Science.gov (United States)

    Tao, Han; Yan, Juan; Zhao, Jianwei; Tian, Yaoqi; Jin, Zhengyu; Xu, Xueming

    2015-01-01

    The structural and functional properties of non-gelatinized waxy rice starch were investigated after 1, 3, 7, and 10 freezing/thawing cycles. Freezing caused an increasing damaged starch from 1.36% in native waxy rice starch to 5.77% in 10 freezing/thawing-treated starch (FTS), as evidenced by the cracking surface on starch granules. More dry matter concentration was leached, which was characterized by high amylopectin concentration (4.34 mg/mL). The leaching was accompanied by a decrease in relative crystallinity from 35.19% in native starch to 31.34% in 10 FTS. Freezing treatment also led to significant deviations in the functional characteristics, for instance decreased gelatinization temperature range, enthalpy, and pasting viscosities. The resistant starch content of 10FTS significantly decreased from 58.9% to 19%, whereas the slowly digested starch content greatly increased from 23.8% in native starch to 50.3%. The increase in susceptibility to enzyme hydrolysis may be attributed to porous granular surface, amylopectin leaching, and the decrease in the relative crystallinity caused by freezing water. PMID:26018506

  7. Effect of multiple freezing/thawing cycles on the structural and functional properties of waxy rice starch.

    Directory of Open Access Journals (Sweden)

    Han Tao

    Full Text Available The structural and functional properties of non-gelatinized waxy rice starch were investigated after 1, 3, 7, and 10 freezing/thawing cycles. Freezing caused an increasing damaged starch from 1.36% in native waxy rice starch to 5.77% in 10 freezing/thawing-treated starch (FTS, as evidenced by the cracking surface on starch granules. More dry matter concentration was leached, which was characterized by high amylopectin concentration (4.34 mg/mL. The leaching was accompanied by a decrease in relative crystallinity from 35.19% in native starch to 31.34% in 10 FTS. Freezing treatment also led to significant deviations in the functional characteristics, for instance decreased gelatinization temperature range, enthalpy, and pasting viscosities. The resistant starch content of 10FTS significantly decreased from 58.9% to 19%, whereas the slowly digested starch content greatly increased from 23.8% in native starch to 50.3%. The increase in susceptibility to enzyme hydrolysis may be attributed to porous granular surface, amylopectin leaching, and the decrease in the relative crystallinity caused by freezing water.

  8. Cryopreservation of boar semen. III: Ultrastructure of boar spermatozoa frozen ultra-rapidly at various stages of conventional freezing and thawing.

    Science.gov (United States)

    Bwanga, C O; Ekwall, H; Rodriguez-Martinez, H

    1991-01-01

    Ejaculated boar spermatozoa subjected to a conventional freezing and thawing process, were ultra-rapidly fixed, freeze-substituted and examined by electron microscopy to monitor the presence of real or potential intracellular ice and the degree of cell protection attained with the different extenders used during the process. Numerous ice crystal marks representing the degree of hydration of the cells were located in the perinuclear space of those spermatozoa not in proper contact with the extender containing glycerol (i.e. prior to freezing). The spermatozoa which were in proper contact with the extenders presented a high degree of preservation of the acrosomes, plasma membranes as well as the nuclear envelopes. No ice marks were detected in acrosomes before thawing, indicating that the conventional assayed cryopreservation method provided a good protection against cryoinjury. The presence of acrosomal changes (internal vesiculization, hydration and swelling) in thawed samples however, raises serious questions about the thawing procedure employed.

  9. Properties and toughening mechanisms of PVA/PAM double-network hydrogels prepared by freeze-thawing and anneal-swelling.

    Science.gov (United States)

    Ou, Kangkang; Dong, Xia; Qin, Chengling; Ji, Xinan; He, Jinxin

    2017-08-01

    It is well known that preparation method of hydrogels has a significant effect on their properties. In this paper, freeze-thawing and anneal-swelling were applied to prepare poly(vinyl alcohol)/polyacrylamide (PVA/PAM) double-network hydrogels with covalently and physically cross-linked networks. The properties of these hydrogels were investigated and compared to control hydrogels. Results indicated that hydrogels fabricated by freeze-thawing show larger pores size and higher swelling capacity than those made by anneal-swelling and control hydrogels. Hydrogels prepared by anneal-swelling exhibit higher mechanical strength, energy dissipation, fracture energy, gel fraction and crystallinity than those made by freeze-thawing and control hydrogels. Physical cross-linking plays a key role in formation of physical-chemical double-network. The toughening mechanism of double-network hydrogel is related to their chain-fracture behavior and elasticity. The results also indicated that appropriate methods can endow hydrogels with specific microstructures and properties which would broaden current hydrogels research and applications in biomedical fields. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Freeze-all policy: fresh vs. frozen-thawed embryo transfer.

    Science.gov (United States)

    Roque, Matheus; Valle, Marcello; Guimarães, Fernando; Sampaio, Marcos; Geber, Selmo

    2015-05-01

    To compare in vitro fertilization (IVF) outcomes between fresh embryo transfer (ET) and frozen-thawed ET (the "freeze-all" policy), with fresh ET performed only in cases without progesterone (P) elevation. Prospective, observational, cohort study. Private IVF center. A total of 530 patients submitted to controlled ovarian stimulation (COS) with a gonadotropin-releasing hormone-antagonist protocol, and cleavage-stage, day-3 ET. None. Ongoing pregnancy rates. A total of 530 cycles were included in the analysis: 351 in the fresh ET group (when P levels were ≤1.5 ng/mL on the trigger day); and 179 cycles in the freeze-all group (ET performed after endometrial priming with estradiol valerate, at 6 mg/d, taken orally). For the fresh ET group vs. the freeze-all group, respectively, the implantation rate was 19.9% and 26.5%; clinical pregnancy rate was 35.9% and 46.4%; and ongoing pregnancy rate was 31.1% and 39.7%. The IVF outcomes were significantly better in the group using the freeze-all policy, compared with the group using fresh ET. These results suggest that even in a select group of patients that underwent fresh ET (P levels ≤1.5 ng/mL), endometrial receptivity may have been impaired by COS, and outcomes may be improved by using the freeze-all policy. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  11. Effects of the freezing and thawing process on biomechanical properties of the human skull.

    Science.gov (United States)

    Torimitsu, Suguru; Nishida, Yoshifumi; Takano, Tachio; Koizumi, Yoshinori; Hayakawa, Mutsumi; Yajima, Daisuke; Inokuchi, Go; Makino, Yohsuke; Motomura, Ayumi; Chiba, Fumiko; Iwase, Hirotaro

    2014-03-01

    The aim of this study was to determine if biomechanical investigations of skull samples are reliable after skulls have been subjected to a freezing and thawing process. The skulls were obtained from 105 Japanese cadavers (66 males, 39 females) of known age that were autopsied in our department between October 2012 and June 2013. We obtained bone specimens from eight sites (four bilaterally symmetrical pairs) of each skull and measured the mass of each specimen. They were then classified into three groups (A, B, C) based on the duration of freezing of the experimental samples. The left-side samples were subjected to frozen storage (experimental group). The corresponding right-side samples were their controls. Bending tests were performed on the controls immediately after they were obtained. The experimental samples were preserved by refrigeration at -20 °C for 1 day (group A), 1 month (group B), or 3 months (group C). Following refrigeration, these samples were placed at 37 °C to thaw for 1 h and then were subjected to bending tests using a three-point-bending apparatus attached to a Handy force gauge. The device recorded the fracture load automatically when the specimen fractured. Statistical analyses revealed that there were no significant differences in sample fracture loads between the frozen preserved/thawed samples and the unfrozen controls for each of the cryopreservation intervals. We eliminated any possible sample mass bias by using controls from the same skull in each case. The results suggest that the freezing/thawing process has little effect on the mechanical properties of human skulls. Thus, frozen storage for up to 3 months is a good method for preserving human skulls. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  12. Simulation of Changes in the Near-Surface Soil Freeze/Thaw Cycle Using CLM4.5 With Four Atmospheric Forcing Data Sets

    Science.gov (United States)

    Guo, Donglin; Wang, Aihui; Li, Duo; Hua, Wei

    2018-03-01

    Change in the near-surface soil freeze/thaw cycle is critical for assessments of hydrological activity, ecosystems, and climate change. Previous studies investigated the near-surface soil freeze/thaw cycle change mostly based on in situ observations and satellite monitoring. Here numerical simulation method is tested to estimate the long-term change in the near-surface soil freeze/thaw cycle in response to recent climate warming for its application to predictions. Four simulations are performed at 0.5° × 0.5° resolution from 1979 to 2009 using the Community Land Model version 4.5, each driven by one of the four atmospheric forcing data sets (i.e., one default Climate Research Unit-National Centers for Environmental Prediction [CRUNCEP] and three newly developed Modern Era Retrospective-Analysis for Research and Applications, Climate Forecast System Reanalysis, and European Centre for Medium-Range Weather Forecasts Reanalysis Interim). The observations from 299 weather stations in both Russia and China are employed to validate the simulated results. The results show that all simulations reasonably reproduce the observed variations in the ground temperature, the freeze start and end dates, and the freeze duration (the correlation coefficients range from 0.47 to 0.99, and the Nash-Sutcliffe efficiencies range from 0.19 to 0.98). Part of the simulations also exactly simulate the trends of the ground temperature, the freeze start and end dates, and the freeze duration. Of the four simulations, the results from the simulation using the CRUNCEP data set show the best overall agreement with the in situ observations, indicating that the CRUNCEP data set could be preferentially considered as the basic atmospheric forcing data set for future prediction. The simulated area-averaged annual freeze duration shortened by 8.03 days on average from 1979 to 2009, with an uncertainty (one standard deviation) of 0.67 days caused by the different atmospheric forcing data sets. These

  13. Nitrogen deposition may enhance soil carbon storage via change of soil respiration dynamic during a spring freeze-thaw cycle period.

    Science.gov (United States)

    Yan, Guoyong; Xing, Yajuan; Xu, Lijian; Wang, Jianyu; Meng, Wei; Wang, Qinggui; Yu, Jinghua; Zhang, Zhi; Wang, Zhidong; Jiang, Siling; Liu, Boqi; Han, Shijie

    2016-06-30

    As crucial terrestrial ecosystems, temperate forests play an important role in global soil carbon dioxide flux, and this process can be sensitive to atmospheric nitrogen deposition. It is often reported that the nitrogen addition induces a change in soil carbon dioxide emission in growing season. However, the important effects of interactions between nitrogen deposition and the freeze-thaw-cycle have never been investigated. Here we show nitrogen deposition delays spikes of soil respiration and weaken soil respiration. We found the nitrogen addition, time and nitrogen addition×time exerted the negative impact on the soil respiration of spring freeze-thaw periods due to delay of spikes and inhibition of soil respiration (p nitrogen), 39% (medium-nitrogen) and 36% (high-nitrogen) compared with the control. And the decrease values of soil respiration under medium- and high-nitrogen treatments during spring freeze-thaw-cycle period in temperate forest would be approximately equivalent to 1% of global annual C emissions. Therefore, we show interactions between nitrogen deposition and freeze-thaw-cycle in temperate forest ecosystems are important to predict global carbon emissions and sequestrations. We anticipate our finding to be a starting point for more sophisticated prediction of soil respirations in temperate forests ecosystems.

  14. Effects of storage in different semen extenders on the pre-freezing and post-thawing quality of boar spermatozoa.

    Science.gov (United States)

    Dziekońska, A; Zasiadczyk, Ł; Lecewicz, M; Strzeżek, R; Koziorowska-Gilun, M; Fraser, L; Mogielnicka-Brzozowska, M; Kordan, W

    2015-01-01

    The aim of this study was to investigate the effects of storage of semen in different commercial extenders on the pre-freezing and post-thawing quality of boar spermatozoa. Semen was diluted in BTS, Androhep (AH) and Gedil (GD), stored for 24 h at 17°C, and then frozen in accordance with the cryopreservation protocol. Analyses of the quality of spermatozoa included: motility, normal apical ridge (NAR) acrosome, plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), measurements of ATP content and activity of superoxidase dismutase (SOD) and glutathione peroxidase (GPx). Prior to the freezing process, no significant effect of the extender on the sperm quality parameters was noted. After thawing the spermatozoa it was demonstrated that the type of extender used influenced PMI, MMP, ATP content and activity of GPx. In the AH extender the percentage of spermatozoa with PMI and ATP content in spermatozoa was significantly higher (Pextenders. In addition, semen stored in the AH was characterised by a statistically higher (Pboar spermatozoa stored for 24 hours in liquid state can be used. However, the type of extender used prior to freezing may have a significant effect on the post-thawing quality of the spermatozoa. The AH extender better secured the quality of thawed boar spermatozoa as compared with the BTS or GD.

  15. Effects of salinity on physicochemical properties of Alaska pollock surimi after repeated freeze-thaw cycles.

    Science.gov (United States)

    Kang, E J; Hunt, A L; Park, J W

    2008-06-01

    The effects of residual salt in surimi on physicochemical properties as affected by various freeze and thaw (FT) cycles were examined. Fresh Alaska pollock surimi was mixed with 4.0% sugar and 5.0% sorbitol, along with 8 combinations of salt (0.4%, 0.6%, 0.8%, and 1.0% NaCl) and sodium polyphosphate (0.25% and 0.5%), vacuum-packed, and stored at -18 degrees C until used. FT cycles (0, 6, and 9) were used to mimic long-term frozen storage. At the time of gel preparation, each treatment was appropriately adjusted to maintain 2% salt and 78% moisture. The pH decreased as residual salt increased during frozen storage. Salt extractable protein (SEP) decreased (P residual salt and phosphate concentration during frozen storage, whiteness value (L*- 3b*) decreased (P salt/0.5% phosphate and 0.6% salt/0.25% phosphate. Water retention ability (WRA) and texture significantly (P salt content (0.8% and 1.0%) after 9 FT cycles, indicating higher residual salt concentration can shorten the shelf life of frozen surimi. Our study revealed lower residual salt concentration and higher phosphate concentration are likely to extend the shelf life of frozen surimi.

  16. Development and commisioning of a test procedure for the investigation of the impact of freeze-thaw cycles on the sealing material of geothermal probes; Entwicklung und Inbetriebnahme eines Pruefverfahrens zur Bestimmung des Frost-Tau-Wechseleinflusses auf das Verpressmaterial von Erdwaermesonden

    Energy Technology Data Exchange (ETDEWEB)

    Anbergen, Hauke [Knabe Enders Duehrkop Ingenieure GmbH, Hamburg (Germany); Technische Univ. Darmstadt (Germany); Frank, Jens [Knabe Enders Duehrkop Ingenieure GmbH, Hamburg (Germany); Sass, Ingo [Technische Univ. Darmstadt (Germany)

    2011-10-24

    In order to exploit the full potential of near-surface geothermal probes, an operation at brine temperatures below the freezing point of water is necessary. This can result in a cyclic freezing and thawing of the surrounding sealing materials. Thus, such a material must have permanently a water permeability below defined limits even after the freeze-thaw stress. For this, test conditions had to be defined, and a measurement method has to be developed. For this purpose, a measuring cell was modified according to DIN 18130 so that freezing processes can be simulated under in-situ conditions using an axially integrated cooling pipe, and the water permeability can be measured as a function of the number of freeze-thaw cycles. The authors of the contribution under consideration report on the test procedure as well as on the results of a complete series of tests.

  17. The effects of freezing, storage, and thawing on cell compartment integrity and ultrastructure

    DEFF Research Database (Denmark)

    Prentø, P

    1997-01-01

    The effects of slow freezing and thawing on enzyme compartmentalization and ultrastructure were studied in rat liver slices frozen in dry ice, isopentane/ethanol-dry ice, or liquid nitrogen, and stored at -80 degrees C for 1-14 days. Non-frozen slices served as controls. Frozen liver slices were...

  18. Does Platelet-Rich Plasma Freeze-Thawing Influence Growth Factor Release and Their Effects on Chondrocytes and Synoviocytes?

    Directory of Open Access Journals (Sweden)

    Alice Roffi

    2014-01-01

    Full Text Available PRP cryopreservation remains a controversial point. Our purpose was to investigate the effect of freezing/thawing on PRP molecule release, and its effects on the metabolism of chondrocytes and synoviocytes. PRP was prepared from 10 volunteers, and a half volume underwent one freezing/thawing cycle. IL-1β, HGF, PDGF AB/BB, TGF-β1, and VEGF were assayed 1 hour and 7 days after activation. Culture media of chondrocytes and synoviocytes were supplemented with fresh or frozen PRP, and, at 7 days, proliferation, gene expression, and secreted proteins levels were evaluated. Results showed that in the freeze-thawed PRP the immediate and delayed molecule releases were similar or slightly lower than those in fresh PRP. TGF-β1 and PDGF AB/BB concentrations were significantly reduced after freezing both at 1 hour and at 7 days, whereas HGF concentration was significantly lower in frozen PRP at 7 days. In fresh PRP IL-1β and HGF concentrations underwent a significant further increase after 7 days. Similar gene expression was found in chondrocytes cultured with both PRPs, whereas in synoviocytes HGF gene expression was higher in frozen PRP. PRP cryopreservation is a safe procedure, which sufficiently preserves PRP quality and its ability to induce proliferation and the production of ECM components in chondrocytes and synoviocytes.

  19. A Micro-Scale Investigation on the Behaviors of Asphalt Mixtures under Freeze-Thaw Cycles Using Entropy Theory and a Computerized Tomography Scanning Technique

    Directory of Open Access Journals (Sweden)

    Huining Xu

    2018-01-01

    Full Text Available The thermodynamic behavior of asphalt mixtures is critical to the engineers since it directly relates to the damage in asphalt mixtures. However, most of the current research of the freeze-thaw damage of asphalt mixtures is focused on the bulk body from the macroscale and lacks a fundamental understanding of the thermodynamic behaviors of asphalt mixtures from the microscale perspective. In this paper, to identify the important thermodynamic behaviors of asphalt mixtures under freeze-thaw loading cycle, the information entropy theory, an X-ray computerized tomography (CT scanner and digital image processing technology are employed. The voids, the average size of the voids, the connected porosity, and the void number are extracted according to the scanned images. Based on the experiments and the CT scanned images, the information entropy evolution of the asphalt mixtures under different freeze-thaw cycles is calculated and the relationship between the change of information entropy and the pore structure characteristics is established. Then, the influences of different freezing and thawing conditions on the thermodynamic behaviors of asphalt mixtures are compared. The combination of information entropy theory and CT scanning technique proposed in this paper provides an innovative approach to investigate the thermodynamics behaviors of asphalt mixtures and a new way to analyze the freeze-thaw damage in asphalt mixtures.

  20. Stabilization of heavy metals in municipal sewage sludge by freeze-thaw treatment with a blend of diatomite, FeSO4, and Ca(OH)2.

    Science.gov (United States)

    Wang, Jing; Fu, Rongbing; Xu, Zhen

    2017-08-01

    In this work, the effects of diatomite with 15% FeSO 4 •7H 2 O and 7.5% Ca(OH) 2 on sludge stabilization were investigated using batch leaching tests. The influence of cell rupture caused by freezing and thawing on stabilization was also evaluated. The results indicated that the optimal diatomite percentage was 2%. Cell rupture by freezing and thawing reduced heavy metal leachability, followed by cell death and decrease of organic groups. The concentration of heavy metals in sludge leachate increased after cell rupture, indicating that the heavy metal leachability was reduced after freezing and thawings. Moreover, the stabilization effects were generally improved after freezing and thawing. As compared with the stabilization of the original sludge, the unstable fractions decreased and the residual fractions of the heavy metals increased in the stabilized sludge after cell rupture. This study developed a method to stabilize heavy metals in municipal sewage sludge. Diatomite combined with FeSO 4 ·7H 2 O and Ca(OH) 2 improved the treatment of sewage sludge contaminated by heavy metals. Cell lysis by freeze-thaw treatment reduced the risk of leaching heavy metals caused by cell death and decreased major organic groups in the sludge.

  1. Investigation of Seasonal Landscape Freeze/Thaw Cycles in Relation to Cloud Structure in the High Northern Latitudes

    Science.gov (United States)

    Smith, Cosmo

    2011-01-01

    The seasonal freezing and thawing of Earth's cryosphere (the portion of Earth's surface permanently or seasonally frozen) has an immense impact on Earth's climate as well as on its water, carbon and energy cycles. During the spring, snowmelt and the transition between frozen and non-frozen states lowers Earth's surface albedo. This change in albedo causes more solar radiation to be absorbed by the land surface, raising surface soil and air temperatures as much as 5 C within a few days. The transition of ice into liquid water not only raises the surface humidity, but also greatly affects the energy exchange between the land surface and the atmosphere as the phase change creates a latent energy dominated system. There is strong evidence to suggest that the thawing of the cryosphere during spring and refreezing during autumn is correlated to local atmospheric conditions such as cloud structure and frequency. Understanding the influence of land surface freeze/thaw cycles on atmospheric structure can help improve our understanding of links between seasonal land surface state and weather and climate, providing insight into associated changes in Earth's water, carbon, and energy cycles that are driven by climate change.Information on both the freeze/thaw states of Earth's land surface and cloud characteristics is derived from data sets collected by NOAA's Special Sensor Microwave/Imager (SSM/I), the Advanced Microwave Scanning Radiometer on NASA's Earth Observing System(AMSR-E), NASA's CloudSat, and NASA's SeaWinds-on-QuickSCAT Earth remote sensing satellite instruments. These instruments take advantage of the microwave spectrum to collect an ensemble of atmospheric and land surface data. Our analysis uses data from radars (active instruments which transmit a microwave signal toward Earth and measure the resultant backscatter) and radiometers (passive devices which measure Earth's natural microwave emission) to accurately characterize salient details on Earth's surface

  2. Cryopreservation of collared peccary (Pecari tajacu) semen using different freezing curves, straw sizes, and thawing rates.

    Science.gov (United States)

    Silva, M A; Peixoto, G C X; Castelo, T S; Lima, G L; Silva, A M; Oliveira, M F; Silva, A R

    2013-08-01

    The objective of this study was to verify the effect of different freezing curves, straw sizes, and thawing rates on the cryopreservation of collared peccary semen. Twelve ejaculates were obtained from captive adult males by electroejaculation, and evaluated for sperm motility, kinetic rating, viability, morphology, and functional membrane integrity. The ejaculates were diluted in a coconut water extender (ACP-116c) with egg yolk and glycerol, packaged into 0.25 mL or 0.50 mL plastic straws and cryopreserved in liquid nitrogen following a slow (-10 °C/min) or a fast (-40 °C/min) freezing curve. After one week, samples were thawed at 37 °C/1 min or 70 °C/8s and evaluated as reported for fresh semen, and also for kinematic parameters (computerized analysis). A significant decrease in sperm motility and kinetic rating was observed after glycerol addition at 5 °C and also after thawing for all the treatments (Pstraw size and thawing rate were taken as reference (P>0.05). In general, values for sperm characteristics found after thawing at 37 °C were better preserved than at 70 °C (Pstraws, which were similar for semen packaging (P>0.05). The evaluation of the kinematic parameters of sperm motility confirmed these results at values varying from 20% to 30% motile sperm for the samples tha wed at 37 °C, and values fewer than 12% motile sperm for samples thawed at 70 °C (Pstraws, but the thawing should be conducted at 37 °C/1 min. Copyright © 2013 Elsevier Inc. All rights reserved.

  3. Effect of sucrose and pectin addition on physical, chemical, thermal and rheological properties of frozen/thawed pineapple pulps

    Science.gov (United States)

    Conceição, Márcia Cavalcante; Fernandes, Tatiana Nunes; Prado, Mônica Elisabeth Torres; de Resende, Jaime Vilela

    2012-09-01

    Pectin (0-1.0 g/100 mL) and sucrose (0-20 g/100 mL) were added to pineapple pulp to improve their rheological properties, thermal properties and stability after freezing and thawing processes. The properties of the mixes were characterized before and after freezing and thawing. Samples were frozen at -20°C, and the freeze concentration was evaluated every 60 min. The thawing rate was evaluated at 19°C and quantified by photographic editing and image analysis software. The thawing rates and values for the freeze concentration were leveled out at pectin concentrations above 0.5 g/100 mL pectin, which indicated that pectin functions to maintain structural homogeneity during freezing. In the thawed samples, the plastic viscosity values were leveled out from pectin concentrations (0.25-0.75 g/100 mL) as the sucrose concentration increased when compared to unfrozen samples. The differences between the rheological parameters of the unfrozen and frozen/thawed pulps, the higher yield stress values after thawing were attributed to the size of suspended particles in the pulp. Applications can specify formulations of frozen products containing pectin, where these properties can be handled after thawing the product.

  4. Freeze/Thaw-Induced Deformation Monitoring and Assessment of the Slope in Permafrost Based on Terrestrial Laser Scanner and GNSS

    Directory of Open Access Journals (Sweden)

    Lihui Luo

    2017-02-01

    Full Text Available Most previous studies of the Qinghai-Tibet engineering corridor (QTEC have focused on the impacts of climate change on thaw-induced slope failures, whereas few have considered freeze-induced slope failures. Terrestrial laser scanning was used in combination with global navigation satellite systems to monitor three-dimensional surface changes between 2014 and 2015 on the slope of permafrost in the QTEC, which experienced two thawing periods and a freezing period. Soil temperature and moisture sensors were also deployed at 11 depths to reveal the hydrological–thermal dynamics of the active layer. We analyzed scanned surface changes in the slope based on comparisons of multi-temporal point cloud data to determine how the hydrological–thermal process affected active layer deformation during freeze–thaw cycles, thereby comprehensively quantifying the surface deformation. During the two thawing periods, the major structure of the slope exhibited subsidence trends, whereas the major structure of the slope had an uplift trend in the freezing period. The seasonal subsidence trend was caused by thaw settlement and the seasonal uplift trend was probably due to frost heaving. This occurred mainly because the active layer and the upper permafrost underwent a phase transition due to heat transfer. The ground movements occurred approximately in the soil temperature conduction direction between the top of the soil and the permafrost table. The elevation deformation range was mainly −0.20 m to 0.20 m. Surface volume increases with heaving after freezing could have compensated for the loss of thawing twice and still led to the upward swelling of the slope. Thus, this type of slope in permafrost is dominated by frost heave. Deformation characteristics of the slope will support enhanced decision making regarding the implementation of remote sensing and hydrological–thermal measurement technologies to monitor changes in the slopes in permafrost adjacent to

  5. Postharvest Ultrasound-Assisted Freeze-Thaw Pretreatment Improves the Drying Efficiency, Physicochemical Properties, and Macamide Biosynthesis of Maca (Lepidium meyenii).

    Science.gov (United States)

    Chen, Jin-Jin; Gong, Peng-Fei; Liu, Yi-Lan; Liu, Bo-Yan; Eggert, Dawn; Guo, Yuan-Heng; Zhao, Ming-Xia; Zhao, Qing-Sheng; Zhao, Bing

    2018-04-01

    A novel technique of ultrasound-assisted freeze-thaw pretreatment (UFP) was developed to improve the drying efficiency of maca and bioactive amide synthesis in maca. The optimal UFP conditions are ultrasonic processing 90 min at 30 °C with 6 freeze-thaw cycles. Samples with freeze-thaw pretreatment (FP), ultrasound pretreatment (UP), and UFP were prepared for further comparative study. A no pretreatment (NP) sample was included as a control. The results showed that UFP improved the drying efficiency of maca slices, showing the highest effective moisture diffusivity (1.75 × 10 -9 m 2 /s). This result was further supported by low-field nuclear magnetic resonance (LF-NMR) analysis and scanning electron microscopy (SEM). The rehydration capacity and protein content of maca slices were improved by UFP. More importantly, contents of bioactive macamides and their biosynthetic precursors were increased in 2.5- and 10-fold, respectively. In conclusion, UFP is an efficient technique to improve drying efficiency, physicochemical properties, and bioactive macamides of maca, which can be applied in the industrial manufacture of maca products. © 2018 Institute of Food Technologists®.

  6. Freezing and post-thaw apoptotic behaviour of cells in the presence of palmitoyl nanogold particles

    International Nuclear Information System (INIS)

    Thirumala, Sreedhar; Forman, Julianne M; Monroe, W Todd; Devireddy, Ram V

    2007-01-01

    The aim of this study was to evaluate the freezing response of HeLa and Jurkat cells in the presence of commercially available nanoparticles, NPs (Palmitoyl Nanogold[reg], Nanoprobes). The cells were incubated with NPs for either 5 min or 3 h, and a calorimeter technique was then used to generate the volumetric shrinkage response during freezing at 20 deg. C min -1 . Concomitantly, we also examined the effect of a commonly used cryoprotectant, dimethylsulfoxide, DMSO (10% v/v ratio) on the freezing response of HeLa and Jurkat cells. By fitting a model of water transport to the experimentally determined volumetric shrinkage data, the reference hydraulic conductivity, L pg (μm/min-atm) and activation energy, E Lp (kcal mol -1 ) were obtained. For HeLa cells, the values of L pg ranged from 0.08 to 0.23 μm/min-atm, while E Lp ranged from 10.9 to 37.4 kcal mol -1 . For Jurkat cells these parameter values ranged from 0.05 to 0.16 μm/min-atm and 9.5 to 35.9 kcal mol -1 . A generic optimal cooling rate equation was then used to predict the optimal rates of freezing HeLa and Jurkat cells in the presence and absence of DMSO and NPs. The post-thaw viability and apoptotic response of HeLa and Jurkat cells was further investigated by cooling cells at three rates in the presence and absence of DMSO and NPs using a commercially available controlled rate freezer. Jurkat cells treated in this manner demonstrated an increase in their adhesive properties after 18 h incubation and adhered strongly to the bottom of the culture plate. This observation prevented further analysis of Jurkat apoptotic and necrotic post-thaw responses. There was no significant effect of NPs or DMSO alone on HeLa cell viability prior to freezing. The post-thaw results from HeLa cells show that the NPs increased the measured post-freeze apoptotic response when cooled at 1 deg. C min -1 , suggesting a possible therapeutic use of NPs in cryodestructive procedures

  7. The influence of the curing conditions of concrete on durability after freeze-thaw accelerated testing

    Directory of Open Access Journals (Sweden)

    Al-Assadi, G.

    2015-12-01

    Full Text Available This work relates the curing conditions of concrete with the damage caused by rapid freeze-thaw cycles (ASTM C 666. The “potential” durability of concrete after testing is also studied. In countries with a continental climate, the curing of concrete in summer is performed under high-temperature and low-humidity conditions, and during the winter the concrete undergoes freezing and thawing. This paper shows the experimental results of the behaviour of concrete specimens cured under climatic summer conditions and then subjected to freeze-thaw cycles. Curing of the specimens includes conditions of good and bad practice in relation to wetting and protection of the concrete. Mechanical properties, cement hydration, volume and pore sizes, oxygen permeability, chloride diffusion and water penetration under pressure tests of the concrete are assessed. These tests were performed before and after the application of the freeze-thaw cycles. Statistical analysis of the correlation among variables is also included.Este trabajo relaciona las condiciones de curado del hormigón con los daños causados por ciclos hielo-deshielo (ASTM C 666. Estudia la durabilidad “potencial” del hormigón dañado por este ensayo. En países con clima continental, el curado del hormigón en verano se realiza con alta temperatura y baja humedad, y durante el invierno sufre condiciones de hielo-deshielo. Se presentan los resultados experimentales de probetas de hormigón curadas bajo condiciones climáticas del verano y luego sometidas a ciclos hielo-deshielo. El curado de las probetas incluye condiciones de buenas y malas prácticas en relación con el curado del hormigón. Se miden las propiedades mecánicas, grado de hidratación, volumen de poros y distribución de tamaños, permeabilidad al oxígeno, penetración de agua bajo presión y coeficiente de difusión de cloruros antes y después de la aplicación de los ciclos de hielo-deshielo. Se incluye un estudio estad

  8. Snow cover, freeze-thaw, and the retention of nutrients in an oceanic mountain ecosystem

    NARCIS (Netherlands)

    Wipf, Sonja; Sommerkorn, Martin; Stutter, Marc I.; Wubs, E. R. Jasper; van der Wal, René

    2015-01-01

    As the climate warms, winters with less snow and therefore more soil freeze-thaw cycles are likely to become more frequent in oceanic mountain areas. It is a concern that this might impair the soil's ability to store carbon and nutrients, and lead to increased leaching losses of dissolved C and

  9. Freeze-Thaw Performance and Moisture-Induced Damage Resistance of Base Course Stabilized with Slow Setting Bitumen Emulsion-Portland Cement Additives

    Directory of Open Access Journals (Sweden)

    Mojtaba Shojaei Baghini

    2015-01-01

    Full Text Available Freeze-thaw (FT cycles and moisture susceptibility are important factors influencing the geotechnical characteristics of soil-aggregates. Given the lack of published information on the behavior of cement-bitumen emulsion-treated base (CBETB under environmental conditions, especially freezing and thawing, this study investigated the effects of these additives on the CBETB performance. The primary goal was to evaluate the resistance of CBETB to moisture damage by performing FT, Marshall conditioning, and AASHTO T-283 tests and to evaluate the long-term stripping susceptibility of CBETB while also predicting the liquid antistripping additives to assess the mixture’s durability and workability. Specimens were stabilized with Portland cement (0%–6%, bitumen emulsion (0%–5%, and Portland cement-bitumen emulsion mixtures and cured for 7 days, and their short- and long-term performances were studied. Evaluation results of both the Marshall stability ratio and the tensile strength ratio show that the additions of additives increase the resistance of the mixtures to moisture damage. Results of durability tests performed for determining the resistance of compacted specimens to repeated FT cycles indicate that the specimen with the 4% cement-3% bitumen emulsion mixture significantly improves water absorption, volume changes, and weight losses. This indicates the effectiveness of this additive as a road base stabilizer with excellent engineering properties for cold regions.

  10. Laboratory freeze-thaw durability of pervious concrete with respect to curing time and addition of sand, slag, silica fume, and saltguard.

    Science.gov (United States)

    2016-11-22

    Concerns persist regarding pervious concrete durability in cold climates related to freeze-thaw : and exposure to salt. This study was conducted as an extension to previous work regarding pervious : concrete in Vermont, to further investigate freeze-...

  11. Freeze-thaw treatment effects on the dynamic mechanical properties of articular cartilage

    Directory of Open Access Journals (Sweden)

    Muldrew Ken

    2010-10-01

    Full Text Available Abstract Background As a relatively non-regenerative tissue, articular cartilage has been targeted for cryopreservation as a method of mitigating a lack of donor tissue availability for transplant surgeries. In addition, subzero storage of articular cartilage has long been used in biomedical studies using various storage temperatures. The current investigation studies the potential for freeze-thaw to affect the mechanical properties of articular cartilage through direct comparison of various subzero storage temperatures. Methods Both subzero storage temperature as well as freezing rate were compared using control samples (4°C and samples stored at either -20°C or -80°C as well as samples first snap frozen in liquid nitrogen (-196°C prior to storage at -80°C. All samples were thawed at 37.5°C to testing temperature (22°C. Complex stiffness and hysteresis characterized load resistance and damping properties using a non-destructive, low force magnitude, dynamic indentation protocol spanning a broad loading rate range to identify the dynamic viscoelastic properties of cartilage. Results Stiffness levels remained unchanged with exposure to the various subzero temperatures. Hysteresis increased in samples snap frozen at -196°C and stored at -80°C, though remained unchanged with exposure to the other storage temperatures. Conclusions Mechanical changes shown are likely due to ice lens creation, where frost heave effects may have caused collagen damage. That storage to -20°C and -80°C did not alter the mechanical properties of articular cartilage shows that when combined with a rapid thawing protocol to 37.5°C, the tissue may successfully be stored at subzero temperatures.

  12. Trehalose in glycerol-free freezing extender enhances post-thaw survival of boar spermatozoa

    Science.gov (United States)

    ATHURUPANA, Rukmali; TAKAHASHI, Daisen; IOKI, Sumire; FUNAHASHI, Hiroaki

    2015-01-01

    Cryopreservation of boar semen is still considered suboptimal due to lower fertility as compared with fresh samples when glycerol, a permeating cryoprotectant, is used. Trehalose is a non-permeable cryoprotectant and nonreducing disaccharide known to stabilize proteins and biologic membranes. The aim of this study was to evaluate the cryosurvival and in vitro penetrability of boar spermatozoa when glycerol was replaced with trehalose in a freezing extender. Ejaculated Berkshire semen samples were diluted in egg yolk-based freezing extender containing glycerol (100 mM) or trehalose (0, 50, 100, 150, 200 and 250 mM) and cryopreserved using a straw freezing procedure. Thawed samples were analyzed for motility, viability, mitochondrial membrane potential (MMP), and acrosome integrity. In experiment 2, penetrability of spermatozoa cryopreserved with 100 mM glycerol or trehalose was examined. Replacement of cryoprotectant glycerol (100 mM) with trehalose had no effect on sperm viability, but replacing it with 100 mM trehalose improved motility, MMP and acrosome integrity significantly. Sperm motility and MMP were considerably higher in 100 mM trehalose, whereas the acrosome integrity was substantially higher in 100–250 mM trehalose. The in vitro penetration rate was also significantly higher in spermatozoa cryopreserved with trehalose (61.3%) than in those cryopreserved with glycerol (43.6%). In conclusion, 100 mM non-permeable trehalose can be used to replace glycerol, a permeating cryoprotectant, for maintenance of better post-thaw quality of boar spermatozoa. PMID:25754239

  13. Soil respiration and carbon loss relationship with temperature and land use conversion in freeze-thaw agricultural area.

    Science.gov (United States)

    Ouyang, Wei; Lai, Xuehui; Li, Xia; Liu, Heying; Lin, Chunye; Hao, Fanghua

    2015-11-15

    Soil respiration (Rs) was hypothesized to have a special response pattern to soil temperature and land use conversion in the freeze-thaw area. The Rs differences of eight types of land use conversions during agricultural development were observed and the impacts of Rs on soil organic carbon (SOC) loss were assessed. The land use conversions during last three decades were categorized into eight types, and the 141 SOC sampling sites were grouped by conversion type. The typical soil sampling sites were subsequently selected for monitoring of soil temperature and Rs of each land use conversion types. The Rs correlations with temperature at difference depths and different conversion types were identified with statistical analysis. The empirical mean error model and the biophysical theoretical model with Arrhenius equation about the Rs sensitivity to temperature were both analyzed and shared the similar patterns. The temperature dependence of soil respiration (Q10) analysis further demonstrated that the averaged value of eight types of land use in this freeze-thaw agricultural area ranged from 1.15 to 1.73, which was lower than the other cold areas. The temperature dependence analysis demonstrated that the Rs in the top layer of natural land covers was more sensitive to temperature and experienced a large vertical difference. The natural land covers exhibited smaller Rs and the farmlands had the bigger value due to tillage practices. The positive relationships between SOC loss and Rs were identified, which demonstrated that Rs was the key chain for SOC loss during land use conversion. The spatial-vertical distributions of SOC concentration with the 1.5-km grid sampling showed that the more SOC loss in the farmland, which was coincided with the higher Rs in farmlands. The analysis of Rs dynamics provided an innovative explanation for SOC loss in the freeze-thaw agricultural area. The analysis of Rs dynamics provided an innovative explanation for SOC loss in the freeze-thaw

  14. Effect of Salvia miltiorrhiza polysaccharides on boar spermatozoa during freezing-thawing.

    Science.gov (United States)

    Shen, Tao; Jiang, Zhong-Liang; Liu, Hong; Li, Qing-Wang

    2015-08-01

    Salvia miltiorrhiza polysaccharides (SMPs) were extracted from S. miltiorrhiza in this study. The aim of the present study was to evaluate the effect of SMP on the motility of boar sperm, including the antioxidant effect of SMP on boar sperm and the effect of SMP on the in vivo fertilizing ability of frozen-thawed boar sperm. Fifty ejaculates from 5 Swagger boars were collected and diluted with an extender, which contained 3% glycerol (v/v) with five concentrations of SMP (0.2, 0.4, 0.6, 0.8, and 1.0mg/mL). The semen was frozen in 0.25mL straws at 1.0×10(9) cells/mL. Sixty gilts were inseminated using fresh semen, frozen semen with 0.4mg/mL of SMP and frozen semen without SMP. The results indicate that the addition of SMP to the extender results in a higher percentage of motile sperm post-thaw (Pextender (Pboar sperm from peroxidative damage and increase sperm motility and litter size during the process of freezing-thawing. The optimal concentration of SMP for the frozen extenders in this study was determined to be 0.4mg/mL. Copyright © 2015. Published by Elsevier B.V.

  15. Temporal Monitoring of the Soil Freeze-Thaw Cycles over a Snow-Covered Surface by Using Air-Launched Ground-Penetrating Radar

    KAUST Repository

    Jadoon, Khan

    2015-09-18

    We tested an off-ground ground-penetrating radar (GPR) system at a fixed location over a bare agricultural field to monitor the soil freeze-thaw cycles over a snow-covered surface. The GPR system consisted of a monostatic horn antenna combined with a vector network analyzer, providing an ultra-wideband stepped-frequency continuous-wave radar. An antenna calibration experiment was performed to filter antenna and back scattered effects from the raw GPR data. Near the GPR setup, sensors were installed in the soil to monitor the dynamics of soil temperature and dielectric permittivity at different depths. The soil permittivity was retrieved via inversion of time domain GPR data focused on the surface reflection. Significant effects of soil dynamics were observed in the time-lapse GPR, temperature and dielectric permittivity measurements. In particular, five freeze and thaw events were clearly detectable, indicating that the GPR signals respond to the contrast between the dielectric permittivity of frozen and thawed soil. The GPR-derived permittivity was in good agreement with sensor observations. Overall, the off-ground nature of the GPR system permits non-invasive time-lapse observation of the soil freeze-thaw dynamics without disturbing the structure of the snow cover. The proposed method shows promise for the real-time mapping and monitoring of the shallow frozen layer at the field scale.

  16. Temporal Monitoring of the Soil Freeze-Thaw Cycles over a Snow-Covered Surface by Using Air-Launched Ground-Penetrating Radar

    KAUST Repository

    Jadoon, Khan; Weihermller, Lutz; McCabe, Matthew; Moghadas, Davood; Vereecken, Harry; Lambot, Sbastien

    2015-01-01

    We tested an off-ground ground-penetrating radar (GPR) system at a fixed location over a bare agricultural field to monitor the soil freeze-thaw cycles over a snow-covered surface. The GPR system consisted of a monostatic horn antenna combined with a vector network analyzer, providing an ultra-wideband stepped-frequency continuous-wave radar. An antenna calibration experiment was performed to filter antenna and back scattered effects from the raw GPR data. Near the GPR setup, sensors were installed in the soil to monitor the dynamics of soil temperature and dielectric permittivity at different depths. The soil permittivity was retrieved via inversion of time domain GPR data focused on the surface reflection. Significant effects of soil dynamics were observed in the time-lapse GPR, temperature and dielectric permittivity measurements. In particular, five freeze and thaw events were clearly detectable, indicating that the GPR signals respond to the contrast between the dielectric permittivity of frozen and thawed soil. The GPR-derived permittivity was in good agreement with sensor observations. Overall, the off-ground nature of the GPR system permits non-invasive time-lapse observation of the soil freeze-thaw dynamics without disturbing the structure of the snow cover. The proposed method shows promise for the real-time mapping and monitoring of the shallow frozen layer at the field scale.

  17. Temporal Monitoring of the Soil Freeze-Thaw Cycles over a Snow-Covered Surface by Using Air-Launched Ground-Penetrating Radar

    Directory of Open Access Journals (Sweden)

    Khan Zaib Jadoon

    2015-09-01

    Full Text Available We tested an off-ground ground-penetrating radar (GPR system at a fixed location over a bare agricultural field to monitor the soil freeze-thaw cycles over a snow-covered surface. The GPR system consisted of a monostatic horn antenna combined with a vector network analyzer, providing an ultra-wideband stepped-frequency continuous-wave radar. An antenna calibration experiment was performed to filter antenna and back scattered effects from the raw GPR data. Near the GPR setup, sensors were installed in the soil to monitor the dynamics of soil temperature and dielectric permittivity at different depths. The soil permittivity was retrieved via inversion of time domain GPR data focused on the surface reflection. Significant effects of soil dynamics were observed in the time-lapse GPR, temperature and dielectric permittivity measurements. In particular, five freeze and thaw events were clearly detectable, indicating that the GPR signals respond to the contrast between the dielectric permittivity of frozen and thawed soil. The GPR-derived permittivity was in good agreement with sensor observations. Overall, the off-ground nature of the GPR system permits non-invasive time-lapse observation of the soil freeze-thaw dynamics without disturbing the structure of the snow cover. The proposed method shows promise for the real-time mapping and monitoring of the shallow frozen layer at the field scale.

  18. The effect of firing temperature on the irreversible expansion, water absorption and pore structure of a brick body during freeze-thaw cycles

    Directory of Open Access Journals (Sweden)

    Mikuláš ŠVEDA

    2013-12-01

    Full Text Available The paper deals with the monitoring of brick body in the process of volumetric freezing and thawing. The samples were fired at temperatures of 900, 1000 and 1060 °C. Attention is focused on monitoring of the irreversible expansion, water absorption and pore structure of a brick body. We found that in all cases the endpoints take place continuously, where the amount firing temperature plays a crucial role. The greatest influence of freeze/thaw cycles on the change of the pore structure was also observed at the lowest temperature. The change of the pore system during the freeze-thaw cycles occurs in such a way, that the pore volume of small pores further decreases and conversely, the pore volume of large pores increases. The knowledge gained can be used not only in the production of new but also in predicting the remaining durability of older clay roofing tiles. DOI: http://dx.doi.org/10.5755/j01.ms.19.4.2741

  19. The combinatorial effect of different Equex STM paste concentrations, cryoprotectants and the straw-freezing methods on the post-thaw boar semen quality.

    Science.gov (United States)

    Wu, T-W; Cheng, F-P; Chen, I-H; Yang, C-H; Tsai, M-Y; Chang, M-H; Wang, J-H; Wu, J-T

    2013-02-01

    This study was to evaluate the combinatorial effect (14 treatments, A-N) of different Equex STM paste concentrations, cryoprotectants and the straw-freezing method on the post-thaw boar semen quality. Two ejaculates were collected from each of nine boars (three boars from each of three breeds). Semen was diluted in extenders with different concentrations of Equex STM paste and different cryoprotectants [glycerol or dimethylacetamide (DMA)] before cryopreserving via liquid nitrogen or dry ice. Motility, viability, percentage of spermatozoa with intense acrosomal staining and with normal morphology of post-thaw sperm were evaluated. The qualities of thawed semen were best preserved in treatment H (extender with 0.5% Equex STM paste and 5% glycerol and freezing by dry ice) and were worst in treatment B (extender with 0% Equex STM paste and 5% DMA and freezing by dry ice). Significant difference (p 0.05). Moreover, statistical analysis suggests that no significant difference was present in semen quality among breed or individual donors (p > 0.05). These findings suggest that Equex STM paste improved the cryosurvival efficiency of boar sperm, and the favourable straw-freezing method changes between glycerol and DMA. © 2012 Blackwell Verlag GmbH.

  20. Impacts of the thawing-freezing process on runoff generation in the Sources Area of the Yellow River on the northeastern Qinghai-Tibet Plateau

    Science.gov (United States)

    Wu, Xiaoling; Xiang, Xiaohua; Qiu, Chao; Li, Li

    2018-06-01

    In cold regions, precipitation, air temperature and snow cover significantly influence soil water, heat transfer, the freezing-thawing processes of the active soil layer, and runoff generation. Hydrological regimes of the world's major rivers in cold regions have changed remarkably since the 1960s, but the mechanisms underlying the changes have not yet been fully understood. Using the basic physical processes for water and heat balances and transfers in snow covered soil, a water-heat coupling model for snow cover and its underlying soil layers was established. We found that freezing-thawing processes can affect the thickness of the active layer, storage capacity for liquid water, and subsequent surface runoffs. Based on calculations of thawing-freezing processes, we investigated hydrological processes at Qumalai. The results show that the water-heat coupling model can be used in this region to provide an understanding of the local movement of hydrological regimes.

  1. Investigation on the Mechanical Properties of a Cement-Based Material Containing Carbon Nanotube under Drying and Freeze-Thaw Conditions

    Directory of Open Access Journals (Sweden)

    Wei-Wen Li

    2015-12-01

    Full Text Available This paper aimed to explore the mechanical properties of a cement-based material with carbon nanotube (CNT under drying and freeze-thaw environments. Mercury Intrusion Porosimetry and Scanning Electron Microscopy were used to analyze the pore structure and microstructure of CNT/cement composite, respectively. The experimental results showed that multi-walled CNT (MWCNT could improve to different degrees the mechanical properties (compressive and flexural strengths and physical performances (shrinkage and water loss of cement-based materials under drying and freeze-thaw conditions. This paper also demonstrated that MWCNT could interconnect hydration products to enhance the performance of anti-microcracks for cement-based materials, as well as the density of materials due to CNT’s filling action.

  2. Advancing Methods for Estimating Soil Nitrous Oxide Emissions by Incorporating Freeze-Thaw Cycles into a Tier 3 Model-Based Assessment

    Science.gov (United States)

    Ogle, S. M.; DelGrosso, S.; Parton, W. J.

    2017-12-01

    Soil nitrous oxide emissions from agricultural management are a key source of greenhouse gas emissions in many countries due to the widespread use of nitrogen fertilizers, manure amendments from livestock production, planting legumes and other practices that affect N dynamics in soils. In the United States, soil nitrous oxide emissions have ranged from 250 to 280 Tg CO2 equivalent from 1990 to 2015, with uncertainties around 20-30 percent. A Tier 3 method has been used to estimate the emissions with the DayCent ecosystem model. While the Tier 3 approach is considerably more accurate than IPCC Tier 1 methods, there is still the possibility of biases in emission estimates if there are processes and drivers that are not represented in the modeling framework. Furthermore, a key principle of IPCC guidance is that inventory compilers estimate emissions as accurately as possible. Freeze-thaw cycles and associated hot moments of nitrous oxide emissions are one of key drivers influencing emissions in colder climates, such as the cold temperate climates of the upper Midwest and New England regions of the United States. Freeze-thaw activity interacts with management practices that are increasing N availability in the plant-soil system, leading to greater nitrous oxide emissions during transition periods from winter to spring. Given the importance of this driver, the DayCent model has been revised to incorproate freeze-thaw cycles, and the results suggests that including this driver can significantly modify the emissions estimates in cold temperate climate regions. Consequently, future methodological development to improve estimation of nitrous oxide emissions from soils would benefit from incorporating freeze-thaw cycles into the modeling framework for national territories with a cold climate.

  3. Fertility disturbances of dimethylacetamide and glycerol in rooster sperm diluents: Discrimination among effects produced pre and post freezing-thawing process.

    Science.gov (United States)

    Abouelezz, F M K; Sayed, M A M; Santiago-Moreno, J

    2017-09-01

    With avian sperm cryopreservation protocols, the most widely used cryoprotectants (CPAs) are the glycerol (GLY; in gradual freezing: in-straw freezing method), and the dimethylacetamide (DMA; in pellets by plunging into liquid nitrogen: in-pellet rapid freezing method). Use of both methods results in a small portion of thawed live sperm with lesser fertilizing ability compared with the semen samples immediately after collection. This study was conducted to assess the pre-freezing damage occurring to the sperm due to the interaction with the cryoprotectants (CPAs) GLY (8%) and DMA (5%), as well as the post-freezing damage resulting from both freezing methods Data for each treatment, in fresh and frozen-thawed samples, were compared for sperm motility, fertilizing capacity and sperm-egg penetration holes/germinal disc (SP holes/GD). Hens (n=50) were artificially inseminated (10 hens/treatment) six times with 3day intervals between inseminations. The treatment of fresh sperm with DMA led to a reduction (P<0.05) in the count of SP holes/GD (21.4) and the fertility rate (66.7%). The addition and elimination of GLY in fresh samples resulted in a lesser (P<0.05) number of SP holes/GD (11.8) and the fertility rate (i.e., 50.0%). The number of SP-holes/GD was least in frozen-thawed samples using both DMA and GLY (14.2 and 9.2, respectively). The fertility rate when using semen frozen with DMA in- pellets was greater (P<0.05) than with use of semen that had been frozen using GLY in straws (46.4% compared with 31.3%). The reduction in fertility compared with the control when semen was cryopreserved using GLY was 64.1%; the GLY addition and elimination was responsible for two thirds of this reduction. The reduction in fertility when using semen cryopreserved with DMA was 46.7%; half of the reduction was attributed to the treatment with DMA. In conclusion, the mechanical damage attributed to the process for reducing GLY concentrations was more harmful to sperm fertilizing

  4. Improvement of stress tolerance and leavening ability under multiple baking-associated stress conditions by overexpression of the SNR84 gene in baker's yeast.

    Science.gov (United States)

    Lin, Xue; Zhang, Cui-Ying; Bai, Xiao-Wen; Feng, Bing; Xiao, Dong-Guang

    2015-03-16

    During the bread-making process, industrial baker's yeast cells are exposed to multiple baking-associated stresses, such as elevated high-temperature, high-sucrose and freeze-thaw stresses. There is a high demand for baker's yeast strains that could withstand these stresses with high leavening ability. The SNR84 gene encodes H/ACA snoRNA (small nucleolar RNA), which is known to be involved in pseudouridylation of the large subunit rRNA. However, the function of the SNR84 gene in baker's yeast coping with baking-associated stresses remains unclear. In this study, we explored the effect of SNR84 overexpression on baker's yeast which was exposed to high-temperature, high-sucrose and freeze-thaw stresses. These results suggest that overexpression of the SNR84 gene conferred tolerance of baker's yeast cells to high-temperature, high-sucrose and freeze-thaw stresses and enhanced their leavening ability in high-sucrose and freeze-thaw dough. These findings could provide a valuable insight for breeding of novel stress-resistant baker's yeast strains that are useful for baking. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Temporal monitoring of the soil freeze-thaw cycles over snow-cover land by using off-ground GPR

    KAUST Repository

    Jadoon, Khan; Lambot, Sé bastien; Dimitrov, Marin; Weihermü ller, Lutz

    2013-01-01

    We performed off-ground ground-penetrating radar (GPR) measurements over a bare agricultural field to monitor the freeze-thaw cycles over snow-cover. The GPR system consisted of a vector network analyzer combined with an off-ground monostatic horn

  6. The interplay between inner and outer frost damage and its implication for accelerated freeze-thaw testing

    DEFF Research Database (Denmark)

    Hasholt, Marianne Tange

    2014-01-01

    In the present project salt frost scaling was registered during an accelerated freeze-thaw test (CEN/TS 12390-9). After the test, inner damage was evaluated by observing the crack patterns on fluorescence impregnated plane sections. The results indicate that the developments of inner and outer...

  7. Effect of Addition of Concentrated Proteins and Seminal Plasma Low Molecular Weight Proteins in Freezing and Thawing of Equine Semen

    Directory of Open Access Journals (Sweden)

    Fagundes, B.

    2011-07-01

    Full Text Available Difficulties in obtaining equine frozen semen with potential fertility are recognized. This study was designed to investigate the effect of seminal plasma on frozen/thawing of eight stallion semen from different breed using the following treatments: Seminal plasma with ten-fold concentrated proteins with molecular weight above 10 kDa on frozen extender; Part of seminal plasma with proteins under 10 kDa on frozen extender; Conventional freezing, using whole seminal plasma on frozen extender. Using the parameter of 30% of seminal motility post-thawing as index of good freezability, it was verified an increased percentage of stallions that presented good freezability when semen was frozen with seminal plasma containing ten-fold concentrated proteins with molecular weight above 10 kDa on frozen extender. These results, suggested the use of seminal plasma concentrated proteins from own stallion to freezing/thawing semen.

  8. Stability Evaluation of Volcanic Slope Subjected to Rainfall and Freeze-Thaw Action Based on Field Monitoring

    Directory of Open Access Journals (Sweden)

    Shima Kawamura

    2011-01-01

    Full Text Available Rainfall-induced failures of natural and artificial slopes such as cut slopes, which are subjected to freezing and thawing, have been frequently reported in Hokkaido, Japan. In particular, many failures occur intensively from spring to summer seasons. Despite numerous field studies, explanation of their mechanical behavior based on in situ data has not yet been completely achieved due to the difficulty in grasping failure conditions. This study aims at clarifying the aspects of in-situ volcanic slopes subjected to rainfall and freeze-thaw action. The changes in soil moisture, pore pressure, deformations, and temperatures in the slope were investigated using soil moisture meters, tensiometers, thermocouple sensors, clinometers, settlement gauges, an anemovane, a snow gauge, and a rainfall gauge. The data generated from these measures indicated deformation in the slope examined mainly proceeded during the drainage process according to changes in soil moisture. Based on this data, a prediction method for failures is discussed in detail.

  9. Computational thermo-hydro-mechanics for freezing and thawing multiphase geological media in the finite deformation range

    Science.gov (United States)

    Sun, W.; Na, S.

    2017-12-01

    A stabilized thermo-hydro-mechanical (THM) finite element model is introduced to investigate the freeze-thaw action of frozen porous media in the finite deformation range. By applying the mixture theory, frozen soil is idealized as a composite consisting of three phases, i.e., solid grain, unfrozen water and ice crystal. A generalized hardening rule at finite strain is adopted to replicate how the elasto-plastic responses and critical state evolve under the influence of phase transitions and heat transfer. The enhanced particle interlocking and ice strengthening during the freezing processes and the thawing-induced consolidation at the geometrical nonlinear regimes are both replicated in numerical examples. The numerical issues due to lack of two-fold inf-sup condition and ill-conditioning of the system of equations are addressed. Numerical examples for engineering applications at cold region are analyzed via the proposed model to predict the impacts of changing climate on infrastructure at cold regions.

  10. Micro-fractures produced in the Cadalso de los Vidrios granite (Madrid) subjected to Freeze-Thaw Durability Testing

    Science.gov (United States)

    Freire-Lista, D. M.; Varas-Muriel, M. J.; Fort, R.

    2012-04-01

    A specific leucogranite (fine to medium grain sized) from Cadalso de los Vidrios, Madrid, Spain, from where it takes the name of the stone variety, which is a traditional heritage building material used in Central Spain, was subjected to freezing-thaw durability tests or accelerated artificial ageing tests (according to Spanish standard EN 12371:2001) to assess its durability by means of ultrasonic velocity measurements (a non-destructive technique), and optical and fluorescence petrography using a polarized optical microscope (destructive technique), both techniques used before, during and after laboratory ageing tests, or in other words, what is determined is the improvement or deterioration in some properties. The measurement of the ultrasonic velocity in the leucogranite cubic test specimens along the freezing-thaw cycles shown that the velocity diminishes with the number of cycles, in relation to the decay that the stones were experiencing. This deterioration can be observed by the loss of crystalline minerals in the surface of the analyzed samples and by the micro-fractures appearance up to one centimeter deep, which have been detected by the petrographic techniques previously mentioned. The images taken by means of the fluorescence microscope clearly show the micro-fractures generated during the durability test. These images have been processed and analyzed by the UTHSCSA Image Tool program with the purpose of being able to quantify the degree of decay that this type of crystalline materials undergone, when subjected to a number of freezing-thaw test cycles. It is therefore an effective, reliable and complementary technique to that of the petrography analysis, both optical and fluorescence ones. In the first cycles of the ageing test, the micro-fractures propagate along crystals edges and during the last cycles of the test, intracrystalline micro-fractures are generated, which are developed in different ways depending on the mineralogy of the crystals. Thus

  11. Saccharomyces cerevisiae glycerol/H+ symporter Stl1p is essential for cold/near-freeze and freeze stress adaptation. A simple recipe with high biotechnological potential is given

    Directory of Open Access Journals (Sweden)

    Ferreira Célia

    2010-11-01

    Full Text Available Abstract Background Freezing is an increasingly important means of preservation and storage of microbial strains used for many types of industrial applications including food processing. However, the yeast mechanisms of tolerance and sensitivity to freeze or near-freeze stress are still poorly understood. More knowledge on this regard would improve their biotechnological potential. Glycerol, in particular intracellular glycerol, has been assigned as a cryoprotectant, also important for cold/near-freeze stress adaptation. The S. cerevisiae glycerol active transporter Stl1p plays an important role on the fast accumulation of glycerol. This gene is expressed under gluconeogenic conditions, under osmotic shock and stress, as well as under high temperatures. Results We found that cells grown on STL1 induction medium (YPGE and subjected to cold/near-freeze stress, displayed an extremely high expression of this gene, also visible at glycerol/H+ symporter activity level. Under the same conditions, the strains harbouring this transporter accumulated more than 400 mM glycerol, whereas the glycerol/H+ symporter mutant presented less than 1 mM. Consistently, the strains able to accumulate glycerol survive 25-50% more than the stl1Δ mutant. Conclusions In this work, we report the contribution of the glycerol/H+ symporter Stl1p for the accumulation and maintenance of glycerol intracellular levels, and consequently cell survival at cold/near-freeze and freeze temperatures. These findings have a high biotechnological impact, as they show that any S. cerevisiae strain already in use can become more resistant to cold/freeze-thaw stress just by simply adding glycerol to the broth. The combination of low temperatures with extracellular glycerol will induce the transporter Stl1p. This solution avoids the use of transgenic strains, in particular in food industry.

  12. CHARACTERISTICS OF INTERACTIONS BETWEEN SOME TEXTURE PROPERTIES AND COMPOSITION OF CARRAGEENAN GELS AS A RESULT OF ITS DEFINED DIVERSIFIED FREEZING AND THAWING TREATMENT

    Directory of Open Access Journals (Sweden)

    Katarzyna Kozłowicz

    2013-06-01

    Full Text Available Model samples of carrageenan gels based on water, milk and juice were air-blast frozen and frozen by immersion in glycol and in liquid nitrogen. The gel freezing rate was determined on the basis of the kinetics of freezing. Carrageenan gel samples were characterized by evaluation of its thawing drip loss and hardness determined with compression and penetration tests. Freezing in liquid nitrogen ensured the highest freezing rates. Thawing drip loss of gels significantly depended on the carrageenan content, pH of the solution, freezing method and freezing rate. The resulting relationships are linear functions with high determination coefficients. The results of compression and penetration tests prove the significant effect of the carrageenan content and pH on gel hardness. The higher carrageenan content in a sample, the higher compression force and penetration of the gel. Gel freezing resulted in lower hardness. Freezing conditions had a significant effect on the properties tested. The correlation between compression forces and penetration depending on the carrageenan content and the freezing method was described using regression equations with high determination coefficients. Gels based on milk and juice with 2.2% carrageenan content are recommended for immersion freezing at rates above 5.0 cm·h-1.

  13. Modification of ASTM C666 for testing resistance of concrete to freezing and thawing in sodium chloride solution.

    Science.gov (United States)

    1978-01-01

    Since 1961 the Research Council has used equipment manufactured by Conrad, Inc. for exposing concrete specimens to rapid cycles of freezing and thawing. In addition, the Materials Division of the Virginia Department of Highways and Transportation sen...

  14. Freeze-Thaw Cycle Test on Basalt, Diorite and Tuff Specimens with the Simulated Ground Temperature of Antarctica

    Science.gov (United States)

    Park, J.; Hyun, C.; Cho, H.; Park, H.

    2010-12-01

    Physical weathering caused by freeze-thaw action in cold regions was simulated with artificial weathering simulator in laboratory. Physical weathering of rock in cold regions usually depends on the temperature, rock type and moisture content. Then these three variables were considered in this study. The laboratory freeze-thaw tests were conducted on the three types of rocks, e.g. diorite, basalt and tuff, which are the major rock types around Sejong Station, King George Island, Antarctica. Nine core samples composed of three samples from each rock type were prepared in NX core, and 50 cycles of freeze-thaw test was carried out under dried and saturated water conditions. In this study, the physical weathering of rocks was investigated after each 10 cycles by measuring P-wave velocity, bulk density, effective porosity, Schmidt hardness and uniaxial compression strength(UCS). The experimental result of the diorite and the tuff specimens showed that P-wave velocity, bulk density, effective porosity, Schmidt hardness and UCS were gradually decreased as weathering progresses, but the result of the basalt specimens did not show typical trends due to the characteristics of irregular pore distribution and various pore sizes. Scanning electron microscopy(SEM) photographs of diorite, basalt and tuff specimens weathered in dried and saturated conditions were also acquired to investigate the role of water during physical weathering processes. The number and size of microcracks were increased as weathering progresses. This work was supported by the National Research Foundation of Korea(NRF) Grant(NRF-2010-0027753).

  15. Temporal monitoring of the soil freeze-thaw cycles over snow-cover land by using off-ground GPR

    KAUST Repository

    Jadoon, Khan

    2013-07-01

    We performed off-ground ground-penetrating radar (GPR) measurements over a bare agricultural field to monitor the freeze-thaw cycles over snow-cover. The GPR system consisted of a vector network analyzer combined with an off-ground monostatic horn antenna, thereby setting up an ultra-wideband stepped-frequency continuous-wave radar. Measurements were performed during nine days and the surface of the bare soil was exposed to snow fall, evaporation and precipitation as the GPR antenna was mounted 110 cm above the ground. Soil surface dielectric permittivity was retrieved using an inversion of time-domain GPR data focused on the surface reflection. The GPR forward model used combines a full-waveform solution of Maxwell\\'s equations for three-dimensional wave propagation in planar layered media together with global reflection and transmission functions to account for the antenna and its interactions with the medium. Temperature and permittivity sensors were installed at six depths to monitor the soil dynamics in the top 8 cm depth. Significant effects of soil dynamics were observed in the time-lapse GPR, temperature and permittivity data and in particular freeze and thaw events were clearly visible. A good agreement of the trend was observed between the temperature, permittivity and GPR time-lapse data with respect to five freeze-thaw cycles. The GPR-derived permittivity was in good agreement with sensor observations. The proposed method appears to be promising for the real-time mapping and monitoring of the frozen layer at the field scale. © 2013 IEEE.

  16. Effect of Freezing and Thawing on Activity of Cu and Zn in Black Soil of Northeast China Under Simulated Fertilization Using Pig Manure

    Directory of Open Access Journals (Sweden)

    XU Cong-long

    2015-06-01

    Full Text Available With the development of large-scale farms and the addition of Cu and Zn in feed additives, high Cu and Zn contents in feces of livestock and poultry occurred, and may cause soil pollution of the heavy metal especially Cu and Zn by livestock and poultry manure fertilization. Both fertilization of livestock and poultry manure and freeze-thaw action can alter the activity of heavy metals in soil, and furthermore change the environmental effect of heavy metals in soil. This paper aimed at the influence of freezing and thawing on the activities of Cu and Zn (exchangeable and carbonate in black soil of Northeast China under pig manure fertilization according to the northeast climate characteristics through laboratory simulation. The results showed that the contents of exchangeable Cu and Zn significantly increased and that of carbonate Cu and Zn slightly decreased under pig manure fertilization initial stage comparing with the control without fertilizer. The contents of exchangeable Cu and Zn slightly increased and that of carbonate Cu and Zn decreased for the higher fertilization comparing with lower fertilization. The contents of exchangeable Cu and Zn decreased and that of carbonate Cu and Zn significantly increased with the extension of fertilization time (one month compared with the early fertilization (one week. Moreover, the contents of exchangeable Cu and Zn under higher fertilization were all higher than that under lower fertilization, on the contrary, the contents of carbonate Cu and Zn under higher fertilization all were lower than that under lower fertilization. The contents of exchangeable and carbonate Cu and Zn all increased obviously with the freezing and thawing temperature decreased, and the contents of Cu and Zn for higher fertilization were higher than that for lower fertilization. In conclusion, the activities of Cu and Zn in black soil were disturbed by both pig manure with different fertilization amounts and fertilization

  17. Freeze-thaw processes and intense winter rainfall: The one-two punch for high streambank legacy sediment and nutrient loads from Mid-Atlantic watersheds

    Science.gov (United States)

    Inamdar, S. P.; Johnson, E. R.; Rowland, R. D.; Walter, R. C.; Merritts, D.

    2017-12-01

    Historic and contemporary anthropogenic soil erosion combined with early-American milldams resulted in large deposits of legacy sediments in the valley bottoms of Piedmont watersheds of the eastern US. Breaching of milldams subsequently yielded highly incised streams with exposed vertical streambanks that are vulnerable to erosion. Streambank erosion is attributed to fluvial scouring, freeze-thaw processes and mass wasting. While streambanks represent a large reservoir of fine sediments and nutrients, there is considerable uncertainty about the contribution of these sources to watershed nonpoint source pollution. Using high-frequency hydrologic, sediment, and turbidity data we show that freeze-thaw events followed by intense winter rainstorms can export unusually high concentrations of suspended sediment and particulate nutrients from watersheds. Data from a 12 ha forested, Piedmont, stream following an intense rain event (54 mm) on February 2016 yielded suspended sediment and particulate nutrient (organic carbon and nitrogen) concentrations and exports that exceeded those from tropical storms Irene, Lee, and Sandy that had much greater rainfall and discharge amounts, but which occurred later in the year. A similar response was also observed with regards to turbidity data for USGS stream monitoring locations at Brandywine Creek (813 km2) and White Clay Creek (153 km2). We hypothesize that much of the sediment export associated with winter storms is likely due to erosion of streambank sediments and was driven by the coupled occurrence of freeze-thaw conditions and intense rainfall events. We propose that freeze-thaw erosion represents an important and underappreciated mechanism in streams that "recharges" the sediment supply, which then is available for flushing by moderate to large storms. Future climate projections indicate increased intensification of storm events and increased variability of winter temperatures. Freeze-thaw cycles coupled with winter rain events

  18. Chloride Transport in OPC Concrete Subjected to the Freeze and Thaw Damage

    Directory of Open Access Journals (Sweden)

    Ki Yong Ann

    2017-01-01

    Full Text Available To predict the durability of a concrete structure under the coupling degradation consisting of the frosting and chloride attack, microstructural analysis of the concrete pore structure should be accompanied. In this study, the correlation between the pore structure and chloride migration for OPC concrete was evaluated at the different cement content in the concrete mix accounting for 300, 350, and 400 kg/m3 at 0.45 of a free water cement ratio. The influence of frosting damage on the rate of chloride transport was assessed by testing with concrete specimens subjected to a rapid freezing and thawing cyclic environment. As a result, it was found that chloride transport was accelerated by frost damage, which was more influential at the lower cement content. The microscopic examination of the pore structure showed that the freezing environment increased the volume of the large capillary pore in the concrete matrix.

  19. Evaluation of Serial Thawing-Refreezing on Human Spermatozoa Resistance Using Cryovials and Straws

    Directory of Open Access Journals (Sweden)

    Fatemeh Ghasemian

    2012-01-01

    Full Text Available Background: We designed this study to detect the cryoinjury rate on human sperm after serialfreezing and thawing, taking into consideration the effects of using cryovials and straws.Materials and Methods: In this experimental study, semen specimens obtained from 15 subjectswere divided into normozoospermic and oligozoospermic groups. Each of the normozoospermicand oligozoo spermic semen specimens were additionally divided into two groups: i. washed andii. unwashed. Specimens were repeatedly freeze-thawed by using cryovials and straws with thefast liquid nitrogen vapor method, until no motile sperm remained. Sperm motility, recovery, andmorphology rate were then determined after thawing, and compared between the groups whiletaking into consideration the effects of using cryovials and straws.Results: Motile spermatozoa were observed in all normozoospermic samples up to thaw 6 with bothcryovials and straws while in oligozoospermic specimens up to thaw 4 (straw and thaw 3 (cryovialin the freeze-thawing cycle. Normozoospermic sample analysis showed no significant difference inmorphology rate. There was a significant increase in motility and recovery percentages for washedsamples, which was observed with straws in compared to the unwashed groups. Oligozoospermicsample analysis indicated a significant increase in motility, recovery (p<0.01, and morphology(p<0.001 rates in washed specimens compared to unwashed specimens using straws. Theimportance of washing sperm was obvious for oligozoospermic specimens.Conclusion: Normozoospermic sperm resisted freezing longer than oligozoospermic sperm. Use ofstraws and cryovials made significant differences in motility, recovery, and morphology of sperm ineach thaw. This difference was slightly higher for oligozoospermic specimens. Results indicated thatthe percentage of motility was higher for washed normozoospermic specimens in each thaw whenstraws were used, whereas the percentage of motility, recovery, and

  20. Changes of Soil Aggregate Stability as a Result of the Effect of Freeze-thaw Cycles

    Directory of Open Access Journals (Sweden)

    Aneta Žabenská

    2015-01-01

    Full Text Available The objective of the present research was to assess the changes in soil erodibility during the non-vegetation period as one of the factors affecting the snowmelt erosion. The temperature fluctuation was simulated with the use of a climatic chamber ex situ. The soil surface was for simplicity reasons considered without any plant or snow cover. The paper deals with the rate of soil erodibility determination – the soil erodibility should increase due to the decrease of soil aggregate stability depending on the number of freeze-thaw cycles and initial soil moisture. Soil samples (taken from three sites were subjected to freeze-thaw cycles under laboratory conditions. Changes in soil agreggate stability were monitored as one of the main soil characteristics which determine the soil erodibility. Two methods were used to determine the soil macroaggregate stability (soil aggregate fraction 1–2 mm: standard single-sieve method of wet sieving (Kemper and Rosenau, 1986, and dry aggregate analysis using a set of flat sieves with a diameter of 1 mm and 0.5 mm. The results of each method are controversial. Intended hypothesis has not been clearly confirmed.

  1. Dendritic cells induce specific cytotoxic T lymphocytes against prostate cancer TRAMP-C2 cells loaded with freeze- thaw antigen and PEP-3 peptide.

    Science.gov (United States)

    Liu, Xiao-Qi; Jiang, Rong; Li, Si-Qi; Wang, Jing; Yi, Fa-Ping

    2015-01-01

    Prostate cancer is the most common cancer in men. In this study, we investigated immune responses of cytotoxic T lymphocytes (CTLs) against TRAMP-C2 prostate cancer cells after activation by dendritic cells (DCs) loaded with TRAMP-C2 freeze-thaw antigen and/or PEP-3 peptide in vitro. Bone marrow-derived DC from the bone marrow of the C57BL/6 were induced to mature by using the cytokine of rhGM-CSF and rhIL-4, and loaded with either the freeze-thaw antigen or PEP-3 peptide or both of them. Maturation of DCs was detected by flow cytometry. The killing efficiency of the CTLs on TRAMP-C2 cells were detected by flow cytometry, CCK8, colony formation, transwell migration, and wound-healing assay. The levels of the IFN-γ, TNF-β and IL-12 were measured by enzyme-linked immunosorbent assay (ELISA). Compared with the unloaded DCs, the loaded DCs had significantly increased expression of several phenotypes related to DC maturation. CTLs activated by DCs loaded with freeze-thaw antigen and PEP-3 peptide had more evident cytotoxicity against TRAMP-C2 cells in vitro. The secretion levels of IFN-γ, TNF-β and IL-12, secreted by DCs loaded with antigen and PEP-3 and interaction with T cells, were higher than in the other groups. Our results suggest that the CTLs activated by DCs loaded with TRAMP-C2 freeze-thaw antigen and PEP-3 peptide exert a remarkable killing efficiency against TRAMP-C2 cells in vitro.

  2. Decrease in glutathione content in boar sperm after cryopreservation. Effect of the addition of reduced glutathione to the freezing and thawing extenders.

    Science.gov (United States)

    Gadea, Joaquín; Sellés, Elena; Marco, Marco Antonio; Coy, Pilar; Matás, Carmen; Romar, Raquel; Ruiz, Salvador

    2004-08-01

    Although glutathione content in boar spermatozoa has been previously reported, the effect of reduced glutathione (GSH) on semen parameters and the fertilizing ability of boar spermatozoa after cryopreservation has never been evaluated. In this study, GSH content was determined in ejaculated boar spermatozoa before and after cryopreservation. Semen samples were centrifuged and GSH content in the resulting pellet monitored spectrophotometrically. The fertilizing ability of frozen-thawed boar sperm was also tested in vitro by incubating sperm with in vitro matured oocytes obtained from gilts. GSH content in fresh semen was 3.84 +/- 0.21 nM GSH/10(8) sperm. Following semen cryopreservation, there was a 32% decrease in GSH content (P boars and after various preservation protocols (P = 0.0102 ). The effect of addition of GSH to the freezing and thawing extenders was also evaluated. Addition of 5 mM GSH to the freezing extender did not have a significant effect on standard semen parameters or sperm fertilizing ability after thawing. In contrast, when GSH was added to the thawing extender, a dose-dependent tendency to increase in sperm fertilizing ability was observed, although no differences were observed in standard semen parameters. In summary, (i) there was a loss in GSH content after cryopreservation of boar semen; (ii) addition of GSH to the freezing extender did not result in any improvement in either standard semen parameters or sperm fertilizing ability; and (iii) addition of GSH to the thawing extender resulted in a significant increase in sperm fertilizing ability. Nevertheless, future studies must conclude if this is the case for all boars. Furthermore, since addition of GSH to the thawing extender did not result in an improvement in standard semen parameters, this suggests that during the thawing process, GSH prevents damage of a sperm property that is critical in the fertilization process but that is not measured in the routine semen analysis.

  3. Performance degradation and microstructure changes in freeze-thaw cycling for PEMFC MEAs with various initial microstructures

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Sang-Yeop; Kim, Hyoung-Juhn; Cho, EunAe; Lee, Kug-Seung; Lim, Tae-Hoon; Jang, Jong Hyun [Fuel Cell Center, Korea Institute of Science and Technology (KIST), Seoul 136-791 (Korea, Republic of); Hwang, In Chul [Corporate Research and Development Division, Hyundai-Kia Motors, Gyeonggi-do 446-912 (Korea, Republic of)

    2010-12-15

    When the temperature of a fuel cell vehicle is repeatedly reduced to subzero temperatures, volume changes by water/ice transformations and frost heave mechanism can cause microstructural changes in membrane-electrode assemblies (MEA), and a resultant permanent decrease in the performance of fuel cell stacks. In this study, five MEAs manufactured by different methods, were tested under repeated freeze-thaw (F-T) cycles between -20 C and 10 C, and the variations in their electrochemical and microstructural characteristics were analyzed according to the initial microstructures. When the MEAs were prepared by spraying catalyst inks on polymer membranes, no significant microstructural changes were observed. In the case of two supplied MEAs, void formations at the electrolyte/electrode interface or vertical cracks within the catalyst layers were observed after 120 F-T cycles. Void formation seems to be responsible for performance degradation as a result of ohmic loss, but the effect of cracks in the catalyst layers was not confirmed. In 120 F-T cycles, activation overpotentials and concentration overpotentials did not increase significantly for any of the MEAs, even although gradual decreases in the electrochemically active surface area of the platinum catalysts and changes in the porous structure were observed. (author)

  4. Use of ground-based radiometers for L-Band Freeze/Thaw retrieval in a boreal forest site

    Science.gov (United States)

    Roy, A.; Sonnentag, O.; Derksen, C.; Toose, P.; Pappas, C.; Mavrovic, A.; El Amine, M.; Royer, A.; Berg, A. A.; Rowlandson, T. L.; Barr, A.; Black, T. A.

    2017-12-01

    The boreal forest is the second largest land biome in the world and thus plays a major role in the global and regional climate systems. The extent, timing and duration of the seasonal freeze/thaw (F/T) state influences vegetation developmental stages (phenology) and, consequently, constitutes an important control on how boreal forest ecosystems exchange carbon, water and energy with the atmosphere. Recently, new L-Band satellite-derived F/T information has become available. However, disentangling the seasonally differing contributions from forest overstory and understory vegetation, and the ground surface to the satellite signal remains challenging. Here we present results from an ongoing campaign with two L-Band surface-based radiometers (SBR) installed on a micrometeorological tower at the Southern Old Black Spruce site (53.99°N / 105.12°W) in central Saskatchewan. One radiometer unit is installed on top of the tower viewing the multi-layer vegetation canopy from above. A second radiometer unit is installed within the multi-layer canopy, viewing the understory and the ground surface only. The objectives of our study are to (i) disentangle the L-Band F/T signal contribution of boreal forest overstory from the combined understory and ground surface contribution, and (ii) link the L-Band F/T signal to related boreal forest structural and functional characteristics. Analysis of these radiometer measurements made from September to November 2016 shows that when the ground surface is thawed, the main contributor to both radiometer signals is soil moisture. The Pearson correlation coefficient between brightness temperature (TB) at vertical polarization (V-pol) and soil permittivity is 0.79 for the radiometer above the canopy and 0.74 for the radiometer below the canopy. Under cold conditions when the soil was thawed (snow insulation) and the trees were frozen (below 0°C), TB at V-pol is negatively correlated with tree permittivity. The freezing tree contribution to

  5. Effects of curing methods and supplementary cementitious material use on freeze thaw durability of concrete containing d-cracking aggregates.

    Science.gov (United States)

    2013-12-01

    For concrete pavements in Kansas, the most effective method of increasing their sustainability is to : increase the service life. One of the principle mechanisms of concrete pavement deterioration in Kansas is : freezing and thawing damage. Some Kans...

  6.   Ultrasonic monitoring of fish thawing process optimal time of thawing and effect of freezing/thawing.

    Science.gov (United States)

    El Kadi, Youssef Ait; Moudden, Ali; Faiz, Bouazza; Maze, Gerard; Decultot, Dominique

    2013-01-01

    Fish quality is traditionally controlled by chemical and microbiological analysis. The non-destructive control presents an enormous professional interest thanks to the technical contribution and precision of the analysis to which it leads. This paper presents the results obtained from a characterisation of fish thaw-ing process by the ultrasonic technique, with monitoring thermal processing from frozen to defrosted states. The study was carried out on fish type red drum and salmon cut into fillets of 15 mm thickness. After being frozen at -20°C, the sample is enclosed in a plexiglas vessel with parallel walls at the ambient temperature 30°C and excited in perpendicular incidence at 0.5 MHz by an ultrasonic pulser-receiver Sofranel 5052PR. the technique of measurement consists to study the signals reflected by fish during its thawing, the specific techniques of signal processing are implemented to deduce informations characterizing the state of fish and its thawing process by examining the evolution of the position echoes reflected by the sample and the viscoelastic parameters of fish during its thawing. The obtained results show a relationship between the thermal state of fish and its acoustic properties, which allowed to deduce the optimal time of the first thawing in order to restrict the growth of microbial flora. For salmon, the results show a decrease of 36% of the time of the second thawing and an increase of 10.88% of the phase velocity, with a decrease of 65.5% of the peak-to-peak voltage of the signal reflected, thus a decrease of the acoustic impedance. This study shows an optimal time and an evolution rate of thawing specific to each type offish and a correlation between the acoustic behavior of fish and its thermal state which approves that this technique of ultrasonic monitoring can substitute the control using the destructive chemical analysis in order to monitor the thawing process and to know whether a fish has suffered an accidental thawing.

  7. Influence of freezing and thawing on the hydration characteristics, quality, and consumer acceptance of whole muscle beef injected with solutions of salt and phosphate.

    Science.gov (United States)

    Pietrasik, Z; Janz, J A M

    2009-03-01

    Effects of salt/phosphate injection level (112% or 125% pump), salt level (0.5% or 1.5% salt), and freezing/thawing on hydration characteristics, quality, and consumer acceptance of beef semitendinosus were investigated. All enhancement treatments decreased shear force by 25-35%, but negatively affected colour. Increased salt concentration yielded lower purge and cooking losses, and higher water holding capacity. The higher injection level reduced water binding properties, however, the loss in functionality with higher water addition was overcome with increased salt content. Freezing and subsequent thawing was generally detrimental to colour and water binding properties and tended to increase shear force. Freezing and subsequent thawing did not affect fluid release in steaks held for 1 day before analysis, but resulted in decreased water retention in samples held for 7 days. Holding vacuum packaged steaks for 7 days generally increased package purge and negatively affected colour parameters, although water binding characteristics were improved. Consumer panel results demonstrated a negative effect on juiciness and tenderness where meat subject to low salt/high injection was frozen then thawed - the low salt level was insufficient to maintain any positive effect of injection treatment. In general, salt/phosphate injection improved product acceptability and increased willingness to purchase.

  8. First delivery of healthy offspring after freezing and thawing of oocytes in Switzerland.

    Science.gov (United States)

    De Geyter, Maria; Steimann, Sabine; Holzgreve, Wolfgang; De Geyter, Christian

    2007-08-11

    The interest in long-term storage of uninseminated oocytes through cryopreservation has seen a recent upsurge, because it provides the potential to assist young women to postpone childbirth after having overcome a malignant disease or delaying childbirth until after management of a professional career. The low fertilisation rate of frozen/thawed oocytes in earlier feasibility trials can now be improved by using intracytoplasmic sperm injection (ICSI) for assisting the penetration of the spermatozoon through the oocyte's hardened zona pellucida. Another reason for the reported low success rates of oocyte cryopreservation in earlier studies may have been the low developmental potential of spare oocytes, which were available for experimental cryopreservation. Oocytes retrieved from supernumerary follicles in women treated with gonadotropins for ovulation induction and intrauterine insemination can be used for the optimisation of cryostorage of uninseminated oocytes. We intended to investigate to what extent the well-established and successful cryopreservation protocols for pronucleate oocytes are also applicable for the cryopreservation of uninseminated oocytes. We herewith report the first successful pregnancy and delivery of frozen/thawed oocytes in Switzerland, which were inseminated with ICSI. In unbiased treatment groups the freezing and thawing of uninseminated oocytes and pronucleate oocytes give comparable results, if the additional manipulation during ICSI was taken into account.

  9. Estimation Model of Soil Freeze-Thaw Erosion in Silingco Watershed Wetland of Northern Tibet

    OpenAIRE

    Kong, Bo; Yu, Huan

    2013-01-01

    The freeze-thaw (FT) erosion is a type of soil erosion like water erosion and wind erosion. Limited by many factors, the grading evaluation of soil FT erosion quantities is not well studied. Based on the comprehensive analysis of the evaluation indices of soil FT erosion, we for the first time utilized the sensitivity of microwave remote sensing technology to soil moisture for identification of FT state. We established an estimation model suitable to evaluate the soil FT erosion quantity in S...

  10. Development of a freeze-stable formulation for vaccines containing aluminum salt adjuvants.

    Science.gov (United States)

    Braun, LaToya Jones; Tyagi, Anil; Perkins, Shalimar; Carpenter, John; Sylvester, David; Guy, Mark; Kristensen, Debra; Chen, Dexiang

    2009-01-01

    Vaccines containing aluminum salt adjuvants are prone to inactivation following exposure to freeze-thaw stress. Many are also prone to inactivation by heat. Thus, for maximum potency, these vaccines must be maintained at temperatures between 2 degrees C and 8 degrees C which requires the use of the cold chain. Nevertheless, the cold chain is not infallible. Vaccines are subject to freezing during both transport and storage, and frozen vaccines are discarded (under the best circumstances) or inadvertently administered despite potentially reduced potency. Here we describe an approach to minimize our reliance on the proper implementation of the cold chain to protect vaccines from freeze-thaw inactivation. By including PEG 300, propylene glycol, or glycerol in a hepatitis B vaccine, particle agglomeration, changes in the fluorescence emission spectrum--indicative of antigen tertiary structural changes--and losses of in vitro and in vivo indicators of potency were prevented following multiple exposures to -20 degrees C. The effect of propylene glycol was examined in more detail and revealed that even at concentrations too low to prevent freezing at -10 degrees C, -20 degrees C, and -80 degrees C, damage to the vaccine could be prevented. A pilot study using two commercially available diphtheria, tetanus toxoid, and acellular pertussis (DTaP) vaccines suggested that the same stabilizers might protect these vaccines from freeze-thaw agglomeration as well. It remains to be determined if preventing agglomeration of DTaP vaccines preserves their antigenic activity following freeze-thaw events.

  11. Is there a relationship between the chromatin status and DNA fragmentation of boar spermatozoa following freezing-thawing?

    Science.gov (United States)

    Fraser, L; Strzezek, J

    2007-07-15

    In this study a radioisotope method, which is based on the quantitative measurements of tritiated-labeled actinomycin D ((3)H-AMD) incorporation into the sperm nuclei ((3)H-AMD incorporation assay), was used to assess the chromatin status of frozen-thawed boar spermatozoa. This study also tested the hypothesis that frozen-thawed spermatozoa with altered chromatin were susceptible to DNA fragmentation measured with the neutral comet assay (NCA). Boar semen was diluted in lactose-hen egg yolk-glycerol extender (L-HEY) or lactose ostrich egg yolk lipoprotein fractions-glycerol extender (L-LPFo), packaged into aluminum tubes or plastic straws and frozen in a controlled programmable freezer. In Experiment 1, the chromatin status and DNA fragmentation were measured in fresh and frozen-thawed spermatozoa from the same ejaculates. There was a significant increase in sperm chromatin destabilization and DNA fragmentation in frozen-thawed semen as compared with fresh semen. The proportions of spermatozoa labeled with (3)H-AMD were concurrent with elevated levels of sperm DNA fragmentation in K-3 extender, without cryoprotective substances, compared with L-HEY or L-LPFo extender. Regression analysis revealed that the results of the (3)H-AMD incorporation assay and NCA for frozen-thawed spermatozoa were correlated. Boars differed significantly in terms of post-thaw sperm DNA damage. In Experiment 2, the susceptibility of sperm chromatin to decondensation was assessed using a low concentration of heparin. Treatment of frozen-thawed spermatozoa with heparin revealed enhanced (3)H-AMD binding, suggesting nuclear chromatin decondensation. The deterioration in post-thaw sperm viability, such as motility, mitochondrial function and plasma membrane integrity, was concurrent with increased chromatin instability and DNA fragmentation. This is the first report to show that freezing-thawing procedure facilitated destabilization in the chromatin structure of boar spermatozoa, resulting in

  12. Predicting meat quality traits of ovine m. semimembranosus, both fresh and following freezing and thawing, using a hand held Raman spectroscopic device.

    Science.gov (United States)

    Fowler, Stephanie M; Schmidt, Heinar; van de Ven, Remy; Wynn, Peter; Hopkins, David L

    2015-10-01

    Complementary studies were conducted to determine the potential for a Raman spectroscopic hand held device to predict meat quality traits of fresh lamb m. semimembranosus (topside) after ageing and freezing/thawing. Spectra were collected from 80 fresh muscles at 24h and 5d PM, another 80 muscles were measured at 24h, 5d and following freezing/thawing. Shear force, cooking loss, sarcomere length, colour, particle size, collagen content, pH24, pHu, purge and thaw loss were also measured. Results indicated a potential to predict pHu (R(2)cv=0.59), pH24 (R(2)cv=0.48) and purge (R(2)cv=0.42) using spectra collected 24h PM. L* could be predicted using spectra collected 24h (R(2)cv=0.33) or 5d PM (R(2)cv=0.33). This suggests that Raman spectroscopy is suited to identifying carcases which deviate from the normal metabolic processes and related meat quality traits. Copyright © 2015. Published by Elsevier Ltd.

  13. Effects of curing methods and supplementary cementitious material use on freeze thaw durability of concrete containing d-cracking aggregates : [technical summary].

    Science.gov (United States)

    2013-12-01

    For concrete pavements in Kansas, the most effective method of increasing their sustainability is to increase the service life. One of the principle mechanism of concrete pavement deterioration in Kansas is freezing and thawing damage. Some Kansas li...

  14. Mechanism of freeze-thaw injury and recovery: A cool retrospective and warming up to new ideas.

    Science.gov (United States)

    Arora, Rajeev

    2018-05-01

    Understanding cellular mechanism(s) of freeze-thaw injury (FTI) is key to the efforts for improving plant freeze-tolerance by cultural methods or molecular/genetic approaches. However, not much work has been done in the last 25+ years to advance our understanding of the nature and cellular loci of FTI. Currently, two FTI lesions are predominantly implicated: 1) structural and functional perturbations in plasma membrane; 2) ROS-induced oxidative damage. While both have stood the test of time, many questions remain unresolved and other potentially significant lesions need to be investigated. Additionally, molecular mechanism of post-thaw recovery (PTR), a critical component of frost-survival, has not been well investigated. Mechanistic understanding of repair after reversible injury could expand the options for strategies to improve frost-hardiness. In this review, without claiming to be exhaustive, I have attempted to synthesize major discoveries from last several decades on the mechanisms of FTI and the relatively little research conducted thus far on PTR mechanisms. It is followed by proposing of hypotheses for mechanism(s) for irreversible FTI or PTR involving cytosolic calcium and ROS signaling. Perspective is presented on some unresolved questions and research on new ideas to fill the knowledge gaps and advance the field. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Hydraulic conductivity of compacted clay frozen and thawed in situ

    International Nuclear Information System (INIS)

    Benson, C.H.; Othman, M.A.

    1993-01-01

    A large specimen of compacted clay (diameter = 298 mm; thickness = 914 mm) was subjected to freeze-thaw in the field for 60 days. Afterward, the hydraulic conductivity was measured. The hydraulic conductivity of the entire specimen remained essentially unchanged, but increases in hydraulic conductivity of 1.5-2 orders of magnitude were observed above the freezing plane. The increase in hydraulic conductivity was highest at the top of the specimen and decreased with depth. Changes in hydraulic conductivity also occurred at depths 150 mm below the freezing plane, where desiccation occurred because of water redistribution. Numerous horizontal and vertical cracks formed in the soil mass. Dissection of the sample after permeation revealed that the cracks were laden with water. Cracking was greatest at the surface and became less frequent with depth. For depths greater than 150 mm below the freezing plane, cracking was absent. The frequency of cracks is consistent with principles of mechanistic models of soil freezing. The results of laboratory tests were used to predict the hydraulic conductivity of the large specimen. Tests were conducted on specimens subjected to various freeze-thaw cycles, temperature gradients, and states of stress. It was found that the predicted hydraulic conductivities were lower than those measured on the large specimen, but they closely resembled the trend in hydraulic conductivity with depth

  16. Moisture migration, microstructure damage and protein structure changes in porcine longissimus muscle as influenced by multiple freeze-thaw cycles.

    Science.gov (United States)

    Zhang, Mingcheng; Li, Fangfei; Diao, Xinping; Kong, Baohua; Xia, Xiufang

    2017-11-01

    This study investigated the effects of multiple freeze-thaw (F-T) cycles on water mobility, microstructure damage and protein structure changes in porcine longissimus muscle. The transverse relaxation time T 2 increased significantly when muscles were subjected to multiple F-T cycles (Pcycles caused sarcomere shortening, Z line fractures, and I band weakening and also led to microstructural destruction of muscle tissue. The decreased free amino group content and increased dityrosine in myofibrillar protein (MP) revealed that multiple F-T cycles caused protein cross-linking and oxidation. In addition, the results of size exclusion chromatography, circular dichroism spectra, UV absorption spectra, and intrinsic fluorescence spectroscopy indirectly proved that multiple F-T cycles could cause protein aggregation and degradation, α-helix structure disruption, hydrophobic domain exposure, and conformational changes of MP. Overall, repeated F-T cycles changed the protein structure and water distribution within meat. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Satellite remote sensing of landscape freeze/thaw state dynamics for complex Topography and Fire Disturbance Areas Using multi-sensor radar and SRTM digital elevation models

    Science.gov (United States)

    Podest, Erika; McDonald, Kyle; Kimball, John; Randerson, James

    2003-01-01

    We characterize differences in radar-derived freeze/thaw state, examining transitions over complex terrain and landscape disturbance regimes. In areas of complex terrain, we explore freezekhaw dynamics related to elevation, slope aspect and varying landcover. In the burned regions, we explore the timing of seasonal freeze/thaw transition as related to the recovering landscape, relative to that of a nearby control site. We apply in situ biophysical measurements, including flux tower measurements to validate and interpret the remotely sensed parameters. A multi-scale analysis is performed relating high-resolution SAR backscatter and moderate resolution scatterometer measurements to assess trade-offs in spatial and temporal resolution in the remotely sensed fields.

  18. Theoretical and numerical studies of crack initiation and propagation in rock masses under freezing pressure and far-field stress

    Directory of Open Access Journals (Sweden)

    Yongshui Kang

    2014-10-01

    Full Text Available Water-bearing rocks exposed to freezing temperature can be subjected to freeze–thaw cycles leading to crack initiation and propagation, which are the main causes of frost damage to rocks. Based on the Griffith theory of brittle fracture mechanics, the crack initiation criterion, propagation direction, and crack length under freezing pressure and far-field stress are analyzed. Furthermore, a calculation method is proposed for the stress intensity factor (SIF of the crack tip under non-uniformly distributed freezing pressure. The formulae for the crack/fracture propagation direction and length of the wing crack under freezing pressure are obtained, and the mechanism for coalescence of adjacent cracks is investigated. In addition, the necessary conditions for different coalescence modes of cracks are studied. Using the topology theory, a new algorithm for frost crack propagation is proposed, which has the capability to define the crack growth path and identify and update the cracked elements. A model that incorporates multiple cracks is built by ANSYS and then imported into FLAC3D. The SIFs are then calculated using a FISH procedure, and the growth path of the freezing cracks after several calculation steps is demonstrated using the new algorithm. The proposed method can be applied to rocks containing fillings such as detritus and slurry.

  19. Consumer Attitudes Toward Storing and Thawing Chicken and Effects of the Common Thawing Practices on Some Quality Characteristics of Frozen Chicken

    Science.gov (United States)

    Benli, Hakan

    2016-01-01

    In this study, a survey was conducted to both evaluate the consumers’ general attitudes for purchasing and storing the raw chicken and determine the thawing practices used for defrosting frozen chicken at home. About 75% of the consumers indicated purchasing chicken meat at least once a week or more. Furthermore, the majority (82.16%) of those who stored at least a portion of the raw chicken stated freezing the raw chicken meat at home. Freezing the chicken meat was considered to have no effect on the quality by 43.49% of the consumers while 56.51% thought that freezing had either negative or positive effects on the quality. The survey study indicated that top five most commonly used thawing practices included thawing on the kitchen counter, thawing in the refrigerator, thawing in the warm water, thawing in the microwave, and thawing under tap water. In addition, an experimental study was conducted to determine the effects of these most commonly used thawing practices on some quality characteristics of the chicken meat including pH, drip loss, cooking loss, color analysis and textural profile analysis. Although, L* value for thawing on the kitchen counter was the lowest, after cooking, none of the thawing treatments have a significant effect on the color values. Thawing in the microwave produced the highest drip loss of 3.47% while the lowest drip loss of 0.62% was observed with thawing in the refrigerator. On the other hand, thawing in the microwave and refrigerator caused the lowest cooking loss values of 18.29% and 18.53%, respectively. Nevertheless, there were no significant differences among textural parameter values of the defrosted and then cooked samples using the home based thawing practices, indicating similar quality characteristics among the samples. PMID:26732333

  20. Consumer Attitudes Toward Storing and Thawing Chicken and Effects of the Common Thawing Practices on Some Quality Characteristics of Frozen Chicken

    Directory of Open Access Journals (Sweden)

    Hakan Benli

    2016-01-01

    Full Text Available In this study, a survey was conducted to both evaluate the consumers’ general attitudes for purchasing and storing the raw chicken and determine the thawing practices used for defrosting frozen chicken at home. About 75% of the consumers indicated purchasing chicken meat at least once a week or more. Furthermore, the majority (82.16% of those who stored at least a portion of the raw chicken stated freezing the raw chicken meat at home. Freezing the chicken meat was considered to have no effect on the quality by 43.49% of the consumers while 56.51% thought that freezing had either negative or positive effects on the quality. The survey study indicated that top five most commonly used thawing practices included thawing on the kitchen counter, thawing in the refrigerator, thawing in the warm water, thawing in the microwave, and thawing under tap water. In addition, an experimental study was conducted to determine the effects of these most commonly used thawing practices on some quality characteristics of the chicken meat including pH, drip loss, cooking loss, color analysis and textural profile analysis. Although, L* value for thawing on the kitchen counter was the lowest, after cooking, none of the thawing treatments have a significant effect on the color values. Thawing in the microwave produced the highest drip loss of 3.47% while the lowest drip loss of 0.62% was observed with thawing in the refrigerator. On the other hand, thawing in the microwave and refrigerator caused the lowest cooking loss values of 18.29% and 18.53%, respectively. Nevertheless, there were no significant differences among textural parameter values of the defrosted and then cooked samples using the home based thawing practices, indicating similar quality characteristics among the samples.

  1. Consumer Attitudes Toward Storing and Thawing Chicken and Effects of the Common Thawing Practices on Some Quality Characteristics of Frozen Chicken.

    Science.gov (United States)

    Benli, Hakan

    2016-01-01

    In this study, a survey was conducted to both evaluate the consumers' general attitudes for purchasing and storing the raw chicken and determine the thawing practices used for defrosting frozen chicken at home. About 75% of the consumers indicated purchasing chicken meat at least once a week or more. Furthermore, the majority (82.16%) of those who stored at least a portion of the raw chicken stated freezing the raw chicken meat at home. Freezing the chicken meat was considered to have no effect on the quality by 43.49% of the consumers while 56.51% thought that freezing had either negative or positive effects on the quality. The survey study indicated that top five most commonly used thawing practices included thawing on the kitchen counter, thawing in the refrigerator, thawing in the warm water, thawing in the microwave, and thawing under tap water. In addition, an experimental study was conducted to determine the effects of these most commonly used thawing practices on some quality characteristics of the chicken meat including pH, drip loss, cooking loss, color analysis and textural profile analysis. Although, L* value for thawing on the kitchen counter was the lowest, after cooking, none of the thawing treatments have a significant effect on the color values. Thawing in the microwave produced the highest drip loss of 3.47% while the lowest drip loss of 0.62% was observed with thawing in the refrigerator. On the other hand, thawing in the microwave and refrigerator caused the lowest cooking loss values of 18.29% and 18.53%, respectively. Nevertheless, there were no significant differences among textural parameter values of the defrosted and then cooked samples using the home based thawing practices, indicating similar quality characteristics among the samples.

  2. Optimizing a Test Method to Evaluate Resistance of Pervious Concrete to Cycles of Freezing and Thawing in the Presence of Different Deicing Salts.

    Science.gov (United States)

    Tsang, Chehong; Shehata, Medhat H; Lotfy, Abdurrahmaan

    2016-10-28

    The lack of a standard test method for evaluating the resistance of pervious concrete to cycles of freezing and thawing in the presence of deicing salts is the motive behind this study. Different sample size and geometry, cycle duration, and level of submersion in brine solutions were investigated to achieve an optimized test method. The optimized test method was able to produce different levels of damage when different types of deicing salts were used. The optimized duration of one cycle was found to be 24 h with twelve hours of freezing at -18 °C and twelve hours of thawing at +21 °C, with the bottom 10 mm of the sample submerged in the brine solution. Cylinder samples with a diameter of 100 mm and height of 150 mm were used and found to produce similar results to 150 mm-cubes. Based on the obtained results a mass loss of 3%-5% is proposed as a failure criterion of cylindrical samples. For the materials and within the cycles of freezing/thawing investigated here, the deicers that caused the most damage were NaCl, CaCl 2 and urea, followed by MgCl 2 , potassium acetate, sodium acetate and calcium-magnesium acetate. More testing is needed to validate the effects of different deicers under long term exposures and different temperature ranges.

  3. Optimizing a Test Method to Evaluate Resistance of Pervious Concrete to Cycles of Freezing and Thawing in the Presence of Different Deicing Salts

    Directory of Open Access Journals (Sweden)

    Chehong Tsang

    2016-10-01

    Full Text Available The lack of a standard test method for evaluating the resistance of pervious concrete to cycles of freezing and thawing in the presence of deicing salts is the motive behind this study. Different sample size and geometry, cycle duration, and level of submersion in brine solutions were investigated to achieve an optimized test method. The optimized test method was able to produce different levels of damage when different types of deicing salts were used. The optimized duration of one cycle was found to be 24 h with twelve hours of freezing at −18 °C and twelve hours of thawing at +21 °C, with the bottom 10 mm of the sample submerged in the brine solution. Cylinder samples with a diameter of 100 mm and height of 150 mm were used and found to produce similar results to 150 mm-cubes. Based on the obtained results a mass loss of 3%–5% is proposed as a failure criterion of cylindrical samples. For the materials and within the cycles of freezing/thawing investigated here, the deicers that caused the most damage were NaCl, CaCl 2 and urea, followed by MgCl 2 , potassium acetate, sodium acetate and calcium-magnesium acetate. More testing is needed to validate the effects of different deicers under long term exposures and different temperature ranges.

  4. Optimizing a Test Method to Evaluate Resistance of Pervious Concrete to Cycles of Freezing and Thawing in the Presence of Different Deicing Salts

    Science.gov (United States)

    Tsang, Chehong; Shehata, Medhat H.; Lotfy, Abdurrahmaan

    2016-01-01

    The lack of a standard test method for evaluating the resistance of pervious concrete to cycles of freezing and thawing in the presence of deicing salts is the motive behind this study. Different sample size and geometry, cycle duration, and level of submersion in brine solutions were investigated to achieve an optimized test method. The optimized test method was able to produce different levels of damage when different types of deicing salts were used. The optimized duration of one cycle was found to be 24 h with twelve hours of freezing at −18 °C and twelve hours of thawing at +21 °C, with the bottom 10 mm of the sample submerged in the brine solution. Cylinder samples with a diameter of 100 mm and height of 150 mm were used and found to produce similar results to 150 mm-cubes. Based on the obtained results a mass loss of 3%–5% is proposed as a failure criterion of cylindrical samples. For the materials and within the cycles of freezing/thawing investigated here, the deicers that caused the most damage were NaCl, CaCl2 and urea, followed by MgCl2, potassium acetate, sodium acetate and calcium-magnesium acetate. More testing is needed to validate the effects of different deicers under long term exposures and different temperature ranges. PMID:28773998

  5. Effect of Freeze-Thaw Cycles on Soil Nitrogen Reactive Transport in a Polygonal Arctic Tundra Ecosystem at Barrow AK Using 3-D Coupled ALM-PFLOTRAN

    Science.gov (United States)

    Yuan, F.; Wang, G.; Painter, S. L.; Tang, G.; Xu, X.; Kumar, J.; Bisht, G.; Hammond, G. E.; Mills, R. T.; Thornton, P. E.; Wullschleger, S. D.

    2017-12-01

    In Arctic tundra ecosystem soil freezing-thawing is one of dominant physical processes through which biogeochemical (e.g., carbon and nitrogen) cycles are tightly coupled. Besides hydraulic transport, freezing-thawing can cause pore water movement and aqueous species gradients, which are additional mechanisms for soil nitrogen (N) reactive-transport in Tundra ecosystem. In this study, we have fully coupled an in-development ESM(i.e., Advanced Climate Model for Energy, ACME)'s Land Model (ALM) aboveground processes with a state-of-the-art massively parallel 3-D subsurface thermal-hydrology and reactive transport code, PFLOTRAN. The resulting coupled ALM-PFLOTRAN model is a Land Surface Model (LSM) capable of resolving 3-D soil thermal-hydrological-biogeochemical cycles. This specific version of PFLOTRAN has incorporated CLM-CN Converging Trophic Cascade (CTC) model and a full and simple but robust soil N cycle. It includes absorption-desorption for soil NH4+ and gas dissolving-degasing process as well. It also implements thermal-hydrology mode codes with three newly-modified freezing-thawing algorithms which can greatly improve computing performance in regarding to numerical stiffness at freezing-point. Here we tested the model in fully 3-D coupled mode at the Next Generation Ecosystem Experiment-Arctic (NGEE-Arctic) field intensive study site at the Barrow Environmental Observatory (BEO), AK. The simulations show that: (1) synchronous coupling of soil thermal-hydrology and biogeochemistry in 3-D can greatly impact ecosystem dynamics across polygonal tundra landscape; and (2) freezing-thawing cycles can add more complexity to the system, resulting in greater mobility of soil N vertically and laterally, depending upon local micro-topography. As a preliminary experiment, the model is also implemented for Pan-Arctic region in 1-D column mode (i.e. no lateral connection), showing significant differences compared to stand-alone ALM. The developed ALM-PFLOTRAN coupling

  6. Effect of Gynostemma Pentaphyllum polysaccharide on boar spermatozoa quality following freezing-thawing.

    Science.gov (United States)

    Hu, Jian-Hong; Li, Qing-Wang; Zhang, Ting; Jiang, Zhong-Liang

    2009-12-01

    Gynostemma Pentaphyllum Polysaccharide (GPP) was added at concentrations of 0.25, 0.5, 1.0, 1.5 and 2.0 mg/ml to the extenders used to freeze boar semen and its effects on the quality of frozen-thawed sperm were assessed. The sperm motility was significantly higher in the extenders containing 0.25 and 0.5 mg/ml GPP, as compared to other groups (Pextender supplemented with 0.5 mg/ml GPP favored the highest intact membrane and intact acrosome percentages in comparison with other groups (Pextender supplemented with 0.25 and 0.5 mg/ml GPP significantly improved SOD levels, compared to other groups (P>0.05). However, the extenders supplemented with GPP did not cause significant differences in levels of CAT and GSH-Px, compared to the control (P>0.05). In summary, GPP exhibited a dose-related response and the lower concentration produced greater protective effect. According to the standard semen quality parameters and antioxidant activities measured in this study, the concentration of 0.5 mg/ml GPP caused a beneficial cryoprotective effects on the quality of frozen-thawed boar semen. It is proposed that an extender containing 0.5 mg/ml GPP could be used as cryoprotective medium of better efficiency.

  7. Meat Quality of Chicken Breast Subjected to Different Thawing Methods

    Directory of Open Access Journals (Sweden)

    MR Oliveira

    2015-06-01

    Full Text Available Freezing is one of the methods to preserve and guarantee the quality of meat until it reaches the consumer. Even though freezing is classified as a mild form of preservation, it causes meat changes resulting from the formation of ice crystals that subsequently affect the tenderness and functionality of meat. The aim of this study was to evaluate the physicochemical and structural characteristics of chicken half breast submitted to fast freezing (-36 °C for 2 hours and thawed by five different methods (under refrigeration, in a microwave, in a oven with air circulation, placed in cold water, or at room temperature. After thawing, the following parameters were evaluated: moisture content, drip loss (syneresis, water activity (aw, and shear force. Samples were also histologically evaluated by light microscopy. The results indicated that, despite being submitted to fast freezing, thawing affected (p <0.05 most of the physicochemical and structural properties of the meat, except for aw. Thawing in cold water (packed in low-density polyethylene bags and placed in cold water at approximately 10 °C for 2 hours and 15 minutes presented the best results due to lesser damage to the cell structure, as shown by the lower drip loss, higher moisture content, and greater tenderness of the samples compared to those thawed using the other methods. Histological examination showed that muscle fiber structural features and organization were maintained. Thawing at low temperatures seems to cause less damage to the meat structure and allows maintaining of its properties. It was concluded that the meat quality is not related only with the freezing method, but also with the method and conditions used in thawing.

  8. Cost Optimization of Mortars Containing Different Pigments and Their Freeze-Thaw Resistance Properties

    Directory of Open Access Journals (Sweden)

    Sadık Alper Yıldızel

    2016-01-01

    Full Text Available Nowadays, it is common to use colored concrete or mortar in prefabricated concrete and reinforced concrete construction elements. Within the scope of this study, colored mortars were obtained with the addition of brown, yellow, black, and red pigments into the white cement. Those mixtures are examined for their compressive strength, unit weight, water absorption, and freeze-thaw resistance. Subsequent to comparison of these properties, a cost optimization has been conducted in order to compare pigment costs. The outcomes showed that the pore structure in architectural mortar applications plays an important role in terms of durability. And cost optimization results show that light colored minerals can be used instead of white cements.

  9. Freezing and thawing effects on fat, protein, and lactose levels of human natural milk administered by gavage and continuous infusion

    Directory of Open Access Journals (Sweden)

    Andrea D. Abranches

    2014-07-01

    Full Text Available OBJECTIVES: to analyze the changes in human milk macronutrients: fat, protein, and lactose in natural human milk (raw, frozen and thawed, after administration simulation by gavage and continuous infusion. METHOD: an experimental study was performed with 34 human milk samples. The infrared spectrophotometry using the infrared analysis equipment MilkoScan Minor(r (Foss, Denmark equipment was used to analyze the macronutrients in human milk during the study phases. The analyses were performed in natural (raw samples and after freezing and fast thawing following two steps: gavage and continuous infusion. The non-parametric Wilcoxon test for paired samples was used for the statistical analysis. RESULTS: the fat content was significantly reduced after administration by continuous infusion (p < 0.001 during administration of both raw and thawed samples. No changes in protein and lactose content were observed between the two forms of infusion. However, the thawing process significantly increased the levels of lactose and milk protein. CONCLUSION: the route of administration by continuous infusion showed the greatest influence on fat loss among all the processes required for human milk administration.

  10. Freezing and thawing effects on fat, protein, and lactose levels of human natural milk administered by gavage and continuous infusion.

    Science.gov (United States)

    Abranches, Andrea D; Soares, Fernanda V M; Junior, Saint-Clair G; Moreira, Maria Elisabeth L

    2014-01-01

    to analyze the changes in human milk macronutrients: fat, protein, and lactose in natural human milk (raw), frozen and thawed, after administration simulation by gavage and continuous infusion. an experimental study was performed with 34 human milk samples. The infrared spectrophotometry using the infrared analysis equipment MilkoScan Minor® (Foss, Denmark) equipment was used to analyze the macronutrients in human milk during the study phases. The analyses were performed in natural (raw) samples and after freezing and fast thawing following two steps: gavage and continuous infusion. The non-parametric Wilcoxon test for paired samples was used for the statistical analysis. the fat content was significantly reduced after administration by continuous infusion (praw and thawed samples. No changes in protein and lactose content were observed between the two forms of infusion. However, the thawing process significantly increased the levels of lactose and milk protein. the route of administration by continuous infusion showed the greatest influence on fat loss among all the processes required for human milk administration. Copyright © 2014 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. All rights reserved.

  11. Comparison of the effect of two freeze-thawing curves for porcine semen. Preliminary results

    Directory of Open Access Journals (Sweden)

    Caldevilla M

    2016-12-01

    Full Text Available Results obtained in fertility and litter size using frozen-thawed porcine semen are far from those obtained with natural service or artificial insemination of cooled semen. The objective of this study was to evaluate freeze-thawing of porcine semen comparing the traditional slow method to a rapid curve of temperature descent, using two cryoprotectants. Six males of proven fertility (n=6, r=2 were used. Semen was obtained using the gloved-hand technique and was transported to the laboratory at 17 ºC diluted in Androstar® plus. Samples were centrifuged 15 minutes at 800 g and re-diluted in: a 5% dimethylformamide, 11% lactose, 20% egg yolk, 0.5% Equex or b 3% glycerol, 11% lactose, 20% egg yolk, 0.5% Equex. The semen was frozen in 0.5 ml straws up to a final concentration of 300 millions sperm /ml using either a modified slow traditional Westendorff curve or a rapid curve. In both cases thawing was carried out at 37 ºC during 1 minute. Kinetic motility parameters were evaluated using a CASA system (ISAS v1, Proiser®, Spain. Sperm viability and acrosome status were evaluated using the FITC-PNA/PI stain. The results were analyzed using a factorial design (analysis ofvariance with two factors, with two levels for each one and using the male as a blocking factor. No interaction was observed between the parameters. No significant differences (p> 0.05 were observed between curves or between cryoprotectants neither in any of the kinetic motility parameters evaluated nor in sperm viability and acrosome status. No significant differences (p> 0.05 were observed between curves or between cryoprotectants in sperm morphology in thawed porcine semen. Taking into account the results obtained, the rapid curve would be the practical choice as it is, faster and more manageable for fieldwork in any pig farm.

  12. Improved Performance of Unpaved Roads During Spring Thaw

    National Research Council Canada - National Science Library

    Henry, Karen S; Olson, James P; Farrington, Stephen P; Lens, John

    2005-01-01

    Unpaved roads in Vermont are subject to deterioration from seasonal freezing and thawing, and many towns have roads that suffer chronic serviceability problems during the so-called "spring thaw," or mud season...

  13. Effect of Curing Conditions and Freeze-Thaw Cycles on the Strength of an Expansive Soil Stabilized with a Combination of Lime, Jaggery, and Gallnut Powder

    Directory of Open Access Journals (Sweden)

    Jijo James

    2018-01-01

    Full Text Available This investigation involved the utilization of the combination of lime, jaggery, and gallnut powder, adopted in South India traditionally. This combination of materials, used for the manufacture of lime-based mortars, was adopted in stabilization of an expansive soil. Three combinations of lime, jaggery, and gallnut powder (LJG in the ratios of 8 : 2 : 2, 8 : 2 : 1, and 8 : 1 : 2 were put into use. The effect of subjecting the combinations to alternate cycles of freeze-thaw (up to 3 cycles and three different curing conditions of air, moisture, and heat was also investigated. In addition, a mineralogical investigation for studying the reaction products was also carried out. The investigation proceeded with the determination of the unconfined compression strength (UCS of stabilized specimens of dimensions 38 mm × 76 mm, cured for periods of 3, 7, 14, and 28 days. The results of the investigation revealed that the addition of LJG resulted in an increase in the strength of the stabilized soil. Freeze-thaw cycles resulted in a reduction in strength with LJG821 proving to be the most optimal combination developing the maximum strength and least strength loss due to freeze-thaw cycles. Thermal curing proved to be the most optimal curing condition out of all curing conditions evaluated.

  14. Effect of freeze-thawing on the long-term stability of calcium levofolinate in 5% dextrose stored on polyolefin infusion bags.

    Science.gov (United States)

    Lebitasy, M; Hecq, J-D; Athanassopoulos, A; Vanbeckbergen, D; Jamart, J; Galanti, L

    2009-08-01

    Calcium levofolinate infusions could be prepared in advance by a centralized intravenous additive service (CIVAS) to improve safety and time management. To investigate the effect of freezing, microwave thawing and long-term storage at 5 +/- 3 degrees C on the stability of calcium levofolinate in 5% dextrose solution. Solutions of 250 mL of 5% dextrose in polyolefin bags (n = 5) containing approximately 400 mg of calcium levofolinate were prepared under aseptic conditions and frozen for 95 days at -20 degrees C. The solutions were then thawed using microwaves and stored at 5 +/- 3 degrees C for 1 month. The calcium levofolinate concentrations were measured by high performance liquid chromatography (HPLC). Visual inspection was performed and pH was measured periodically during the storage at 5 +/- 3 degrees C. Stability of the solution was defined as a concentration remaining superior to 90% of the initial concentration by regression analysis as recommended by the Food and Drug Administration (FDA). No colour change or precipitation in the solutions was observed. Calcium levofolinate infusions were stable when stored at 5 +/- 3 degrees C during 1 month after freeze-thaw treatment. Throughout this period, the lower confidence limit of the estimated regression line of concentration-time profile remained above 90% of the initial concentration. Slight change in pH values from 6.52 +/- 0.01 to 6.50 +/- 0.01 during storage time did not affect retention time on HPLC and has no clinical consequence, the solutions remaining in the acceptable range for perfusion (4 thawed before use. Such treatment extends long-term stability and releases pharmacist's time for major activities such as checking medication order errors.

  15. Improved cryopreservability of stallion sperm using a sorbitol-based freezing extender.

    Science.gov (United States)

    Pojprasath, T; Lohachit, C; Techakumphu, M; Stout, T; Tharasanit, T

    2011-06-01

    Cryopreservation of stallion semen is often associated with poor post-thaw sperm quality. Sugars are among the important components of a freezing extender and act as non-permeating cryoprotectants. This study aimed to compare the quality of stallion sperm frozen with glucose, fructose or sorbitol-containing freezing extenders. Semen was collected from six stallions of proven fertility and cryopreserved using a freezing extender containing different types of monosaccharide sugars (glucose, fructose or sorbitol). After thawing, the semen was examined for sperm motility, viability, acrosome integrity, plasma membrane functionality and sperm longevity. The fertility of semen frozen in the presence of sorbitol was also tested by artificial insemination. Sperm quality was significantly decreased following freezing and thawing (P sorbitol and glucose (P sorbitol-based extender when examined at 2 and 4 h post-thaw, all of these parameters plus plasma membrane functionality were improved for sperm frozen in the sorbitol extender than in the glucose extender when examined 10 min post-thaw. Two of four mares (50%) inseminated with semen frozen with a sorbitol-containing freezing extender became pregnant. It is concluded that different sugars have different abilities to protect against cryoinjury during freezing and thawing of stallion sperm. This study demonstrated that an extender containing sorbitol as primary sugar can be used to successfully cryopreserve equine sperm; moreover, the quality of frozen-thawed sperm appeared to be better than when glucose or fructose was the principle sugar in the freezing extender. Copyright © 2011 Elsevier Inc. All rights reserved.

  16. Effects of various freezing containers for vitrification freezing on mouse oogenesis.

    Science.gov (United States)

    Kim, Ji Chul; Kim, Jae Myeoung; Seo, Byoung Boo

    2016-01-01

    In the present study, various freezing containers were tested for mouse embryos of respective developmental stages; embryos were vitrified and then their survival rate and developmental rate were monitored. Mouse two cell, 8 cell, and blastula stage embryos underwent vitrification freezing-thawing and then their recovery rate, survival rate, development rate, and hatching rate were investigated. EM-grid, OPS, and cryo-loop were utilized for vitrification freezing-thawing of mouse embryos. It was found that recovery rate and survival rate were higher in the group of cryo-loop compared to those of EM-grid (p containers on vitrified embryos of respective developmental stages; it was demonstrated that higher developmental rate was shown in more progressed (or developed) embryos with more blastomeres. There was however, no difference in embryonic development rate was shown amongst containers. Taken together, further additional studies are warranted with regards to 1) manipulation techniques of embryos for various vitrification freezing containers and 2) preventive measures against contamination via liquid nitrogen.

  17. Long-term stability of temocillin in dextrose 5% and in sodium chloride 0.9% polyolefin bags at 5 ± 3°C after freeze-thaw treatment.

    Science.gov (United States)

    Rolin, C; Hecq, J-D; Tulkens, P; Vanbeckbergen, D; Jamart, J; Galanti, L

    2011-11-01

    The aim of this study was to investigate the stability of a mixture of temocillin 20mg/ml in 5% dextrose and in 0.9% sodium chloride polyolefin bags after freezing, microwave thawing and long-term storage at 5±3°C. The stability of ten polyolefin bags containing 20mg/ml of temocillin, five bags in 5% dextrose and five bags in 0.9% sodium chloride, prepared under aseptic conditions was studied after freezing for 1 month at -20°C, thawing in a microwave oven with a validated cycle, and stored at 5±3°C. Over 30 days, temocillin concentrations were measured by high-pressure liquid chromatography. Visual inspections, microscope observation, spectrophotometric measurements and pH measurements were also performed. No precipitation occurred in the preparations but minor colour change was observed. No microaggregate was observed with optical microscopy or revealed by a change of absorbance. Based on a shelf life of 95% residual potency, temocillin infusions were stable at least 11 days in 5% dextrose and 14 days in 0.9% sodium chloride after freezing and microwave thawing (corresponding at the period where 95% lower confidence limit of the concentration-time profile remained superior to 95% of the initial concentration). During this period, the pH values of drug solutions have been observed to decrease without affecting chromatographic parameters. Within these limits, temocillin in 5% dextrose and in 0.9% sodium chloride infusions may be prepared and frozen in advance by a centralized intravenous admixture service then thawed before use in clinical units. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  18. Repeated exposure to conditioned fear stress increases anxiety and delays sleep recovery following exposure to an acute traumatic stressor

    Directory of Open Access Journals (Sweden)

    Benjamin N Greenwood

    2014-10-01

    Full Text Available Repeated stressor exposure can sensitize physiological responses to novel stressors and facilitate the development of stress-related psychiatric disorders including anxiety. Disruptions in diurnal rhythms of sleep-wake behavior accompany stress-related psychiatric disorders and could contribute to their development. Complex stressors that include fear-eliciting stimuli can be a component of repeated stress experienced by humans, but whether exposure to repeated fear can prime the development of anxiety and sleep disturbances is unknown. In the current study, adult male F344 rats were exposed to either control conditions or repeated contextual fear conditioning for 22 days followed by exposure to either no, mild (10, or severe (100 acute uncontrollable tail shock stress. Exposure to acute stress produced anxiety-like behavior as measured by a reduction in juvenile social exploration and exaggerated shock-elicited freezing in a novel context. Prior exposure to repeated fear enhanced anxiety-like behavior as measured by shock-elicited freezing, but did not alter social exploratory behavior. The potentiation of anxiety produced by prior repeated fear was temporary; exaggerated fear was present 1 day but not 4 days following acute stress. Interestingly, exposure to acute stress reduced REM and NREM sleep during the hours immediately following acute stress. This initial reduction in sleep was followed by robust REM rebound and diurnal rhythm flattening of sleep / wake behavior. Prior repeated fear extended the acute stress-induced REM and NREM sleep loss, impaired REM rebound, and prolonged the flattening of the diurnal rhythm of NREM sleep following acute stressor exposure. These data suggest that impaired recovery of sleep / wake behavior following acute stress could contribute to the mechanisms by which a history of prior repeated stress increases vulnerability to subsequent novel stressors and stress-related disorders.

  19. Research on Reasons for Repeated Falling of Tiles in Internal Walls of Construction

    Science.gov (United States)

    Xu, LiBin; Chen, Shangwei; He, Xinzhou; Zhu, Guoliang

    2018-03-01

    In view of the quality problem of repeated falling of facing tiles in some construction, the essay had a comparative trial in laboratory on cement mortar which is often used to paste tiles, special tile mortar and dry-hang glue, and measured durability of tile adhesive mortar through freezing and thawing tests. The test results indicated that ordinary cement mortar cannot meet standards due to reasons like big shrinkage and low adhesive. In addition, the ten times of freezing and thawing tests indicated that ordinary cement mortar would directly shell and do not have an adhesive force, and moreover, adhesive force of special tile mortar would reduce. Thus, for tiles of large size which are used for walls, dry-hang techniques are recommended to be used.

  20. Effect of freezing, long-term storage and microwave thawing on the stability of a mixture of diclofenac and sodium bicarbonate in glucose 5% polyolefin bags.

    Science.gov (United States)

    François, J-H; Hecq, J-D; Vanbeckbergen, D; Jamart, J; Galanti, L

    2009-11-01

    Preparation in advance of intravenous solution could be efficient to improve quality assurance, security, time management and cost saving of drug delivery. The aim of this study was to investigate the stability of a mixture of diclofenac 75 mg/100 ml and sodium bicarbonate 42 mg/100 ml in 5% glucose polyolefin bags after freezing, long-term storage, and microwave thawing. The stability of five polyolefin bags containing approximately 75 mg/100ml of diclofenac and 42 mg/100ml of sodium bicarbonate in 5% glucose prepared under aseptic conditions was studied after freezing for 2 months at -20 degrees C, thawing in a microwave oven with a validated cycle, and stored at 5 + or - 3 degrees C. Diclofenac concentrations were measured by high-pressure liquid chromatography using a reversed-phase column, a mobile phase consisting of 40% of acetonitrile (v/v) in KH(2)PO(4) buffer 0.02 M, pH 8.40 + or - 0.05, and UV detection at 276.0 nm. Visual inspection, microscope observation, spectrophotometric measurements and pH measurement were also performed. No colour change or precipitation occurred in the preparations. No microaggregate was observed with optical microscopy or revealed by a change of absorbance. Based on a shelf-life of 90% residual potency, diclofenac was stable for at least 30 days after freezing and microwave thawing, period where 95% lower confidence limit of the concentration-time profile remained superior to 90% of the initial concentration. During this period, the pH values of drug solutions have not been altered. Within these limits, diclofenac in 5% glucose infusion may be prepared and frozen in advance by a centralized intravenous admixture service, then thawed before use in clinical units.

  1. PVA/CM-chitosan/honey hydrogels prepared by using the combined technique of irradiation followed by freeze-thawing

    International Nuclear Information System (INIS)

    Afshari, M.J.; Sheikh, N.; Afarideh, H.

    2015-01-01

    Hydrogels with three components, poly(vinyl alcohol) (PVA), carboxymethylate chitosan (CM-chitosan) and honey have been prepared by using radiation method and radiation followed by freeze-thawing cycles technique (combinational method). The solid concentration of the polymer solution is 15 wt% and the ratios of PVA/CM-chitosan/honey are 10/1.5/3.5, 10/2/3, 10/3/2, and 10/3.5/1.5. The applied irradiation doses are 25, 30 and 40 kGy. Various tests have been done to evaluate the hydrogel properties to produce materials to be used as wound dressing. The results show that combinational method improves the mechanical strength of hydrogels while it has no significant effect on the water evaporation rate of gels. The combinational method decreases the swelling of hydrogels significantly, albeit this parameter is still acceptable for wound dressing. Microbiological analyses show that the hydrogel prepared by both methods can protect the wound from Escherichia coli bacterial infection. The wound healing test shows the good performance of the gels in mice. - Highlights: • Hydrogels prepared by the combination of irradiation and freeze-thawing methods. • Hydrogels with improved mechanical strength prepared by the combinational method. • The prepared hydrogels had acceptable transparency and degree of swelling. • The water evaporation rates of these hydrogels were pretty low. • Presences of honey in the formulation of gels led to a higher tissue regeneration

  2. Enhancement the conditioning of waste activated sludge through a sequence of freeze/thaw-electro-Fenton process

    Directory of Open Access Journals (Sweden)

    Shahheidar Narjes

    2018-03-01

    Full Text Available Sludge conditioning is an important stage in sludge management. In the present study, a sequence of freeze/thaw-electro-Fenton process was designed and specific resistance filtration (SRF was monitored during sludge conditioning as an important factor in sludge dewaterability. Furthermore, protein and polysaccharide concentrations were measured during the experiments. Results showed that the lowest SRF value contributed to −10°C in freezing process which showed a reducing trend by decreasing solution pH. In addition, results revealed that solution pH less than 3 caused a significant improvement in sludge dewatering; so the lowest SRF has been registered at pH = 2. By increasing current intensity from 0.5 to 1A, SRF values were reduced and then followed by an enhancement with increasing current intensity to 3.2 A. The lowest SRF value (6.1 × 104 m/kg was obtained at H2O2 = 30 mg/L which was the best conditions for sludge dewatering.

  3. Freeze-thaw decellularization of the trabecular meshwork in an ex vivo eye perfusion model

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    Yalong Dang

    2017-08-01

    Full Text Available Objective The trabecular meshwork (TM is the primary substrate of outflow resistance in glaucomatous eyes. Repopulating diseased TM with fresh, functional TM cells might be a viable therapeutic approach. Decellularized TM scaffolds have previously been produced by ablating cells with suicide gene therapy or saponin, which risks incomplete cell removal or dissolution of the extracellular matrix, respectively. We hypothesized that improved trabecular meshwork cell ablation would result from freeze-thaw cycles compared to chemical treatment. Materials and Methods We obtained 24 porcine eyes from a local abattoir, dissected and mounted them in an anterior segment perfusion within two hours of sacrifice. Intraocular pressure (IOP was recorded continuously by a pressure transducer system. After 72 h of IOP stabilization, eight eyes were assigned to freeze-thaw (F ablation (−80 °C × 2, to 0.02% saponin (S treatment, or the control group (C, respectively. The TM was transduced with an eGFP expressing feline immunodeficiency viral (FIV vector and tracked via fluorescent microscopy to confirm ablation. Following treatment, the eyes were perfused with standard tissue culture media for 180 h. TM histology was assessed by hematoxylin and eosin staining. TM viability was evaluated by a calcein AM/propidium iodide (PI assay. The TM extracellular matrix was stained with Picro Sirius Red. We measured IOP and modeled it with a linear mixed effects model using a B-spline function of time with five degrees of freedom. Results F and S experienced a similar IOP reduction of 30% from baseline (P = 0.64. IOP reduction of about 30% occurred in F within 24 h and in S within 48 h. Live visualization of eGFP demonstrated that F conferred a complete ablation of all TM cells and only a partial ablation in S. Histological analysis and Picro Sirius staining confirmed that no TM cells survived in F while the extracellular matrix remained. The viability assay showed

  4. Impacts of the Air Temperature Rising on the Soil Freezing-thawing Processes and the Surface Fluxes on the Tibetan Plateau

    Science.gov (United States)

    Zheng, G.; Yang, D.

    2017-12-01

    The Tibetan Plateau (TP) is the highest plateau all over the world and plays an essential role on the global water cycle and the atmospheric circulation, because many large rivers originating there and it acts as a "heat source" to pump the Asian summer monsoon. During the past 50 years, the TP is among the most sensitive regions to the climatic warming. Many previous researches have been delved into the impacts of the permafrost degradation there. But the variations and the impacts of the changes of the seasonally frozen ground, which consists 50 % of the plateau region, have been less discussed. Thus, this study uses the geomorphology-based eco-hydrological model to simulate the long-term land surface processes on 37 after picked China Meteorological Administration stations. And, these stations uniformly locate within the seasonally frozen regions of the TP. The modelled freezing-thawing cycles have successfully reproduced the observations with the correlation squares of 0.8 (significance level p rate of 0.13 m/decade and 4.6 days/decade. The changes of the near-surface freezing-thawing cycles exert large influences on the flux exchanges between the land surface and the atmosphere. The advance (delay) of the freezing ending (starting) time has caused 13 % (p influence the following summer monsoon and redistribute the precipitation over the southeastern Asia. Also, as the incoming radiation and the latent heat keeping stable, less sensible heat fluxes would lead to more ground heat storage which provides a better thermal condition for the vegetation growth.

  5. Defined Combinations of Cryomedia and Thawing Extenders Influence the Viable X-Y Boar Sperm Ratio in Vitro.

    Science.gov (United States)

    Korchunjit, W; Kaeoket, K; Kitiyanant, Y; Taylor, J; Wongtawan, T

    It is believed that plasma membrane X- and Y-chromosome bearing sperm are different; therefore the freezing and thawing process may affect X- and Y-sperm differently. The objective of this study was to investigate the effect of cryomedia and thawing extenders on the survival of X and Y-sperm. Three different cryomedia and thawing extenders were compared. Viable motile sperm were separated using a swim-up technique. Real-time PCR was used to identify the sperm type. Using CryoA for freezing and Beltsville-Thawing-Solution (BTS) as the thawing extender yielded significantly higher numbers of viable motile Y sperm (64 percent) than control (48 percent) (P semen freezing with CryoC and thawing with Androstar Plus gave a significantly lower number of viable motile Y sperm (32 percent) than control (51 percent). Our results revealed that defined combinations of cryomedia and thawing extenders significantly altered the survival ratio of frozen-thawed X-Y sperm in vitro, which has potential implications for artificial insemination.

  6. Effect of addition of coconut water (Cocos nucifera) to the freezing media on post-thaw viability of boar sperm.

    Science.gov (United States)

    Bottini-Luzardo, María; Centurión-Castro, Fernando; Alfaro-Gamboa, Militza; Aké-López, Ricardo; Herrera-Camacho, José

    2013-01-01

    The aims of this experiment were to evaluate the addition of coconut water in natura to the freezing media, compare the effect of deionized water vs filtered water of coconut over the post-thaw seminal characteristics, and evaluate the effect of the deionized water and in natura coconut water on the seminal characteristics of boar sperm at different post-thaw times. Thirty-four ejaculates were used divided in three aliquots which received one of the following treatments (T): T1, LEY (bidistilled water, lactose, and egg yolk) and LEYGO (LEY + glycerol and Orvus ET paste); T2, LEY(A) (coconut deionized water, lactose, and egg yolk)-LEYGO(A); and T3, LEY(B) (in natura coconut water, lactose, and egg yolk)-LEYGO(B). Samples of boar semen were frozen according to the Westendorf method, thawed at 38°C, and evaluated at three incubation times (0, 30, and 60 min). Seminal characteristics assessed were motility (Mot), acrosomal integrity (AInt), membrane integrity (MInt), and mitochondrial activity (MAct). T1 showed a higher percentage of viable sperm than T3 (Mot 36.5 vs 5.4 %, AInt 61.8 vs 41.2 %, MInt 50.4 vs 41.3 %, and MAct 56.9 vs 50.5 %). T2 kept a higher percentage of viable sperm at all incubation times. In natura coconut water showed a detrimental effect over the viability of the frozen-thawed boar semen. Deionized coconut water improved the boar semen viability post-thaw, outperforming results of in natura coconut water.

  7. Methods of human body odor sampling: the effect of freezing.

    Science.gov (United States)

    Lenochova, Pavlina; Roberts, S Craig; Havlicek, Jan

    2009-02-01

    Body odor sampling is an essential tool in human chemical ecology research. However, methodologies of individual studies vary widely in terms of sampling material, length of sampling, and sample processing. Although these differences might have a critical impact on results obtained, almost no studies test validity of current methods. Here, we focused on the effect of freezing samples between collection and use in experiments involving body odor perception. In 2 experiments, we tested whether axillary odors were perceived differently by raters when presented fresh or having been frozen and whether several freeze-thaw cycles affected sample quality. In the first experiment, samples were frozen for 2 weeks, 1 month, or 4 months. We found no differences in ratings of pleasantness, attractiveness, or masculinity between fresh and frozen samples. Similarly, almost no differences between repeatedly thawed and fresh samples were found. We found some variations in intensity; however, this was unrelated to length of storage. The second experiment tested differences between fresh samples and those frozen for 6 months. Again no differences in subjective ratings were observed. These results suggest that freezing has no significant effect on perceived odor hedonicity and that samples can be reliably used after storage for relatively long periods.

  8. A freeze-stable formulation for DTwP and DTaP vaccines.

    Science.gov (United States)

    Xue, Honggang; Yang, Bangling; Kristensen, Debra D; Chen, Dexiang

    2014-01-01

    Inadvertent vaccine freezing often occurs in the cold chain and may cause damage to freeze‑sensitive vaccines. Liquid vaccines that contain aluminum salt adjuvants are particularly vulnerable. Polyol cryoprotective excipients have been shown to prevent freeze damage to hepatitis B vaccine. In this study, we examined the freeze-protective effect of propylene glycol on diphtheria-tetanus-pertussis-whole-cell (DTwP) and acellular (DTaP) vaccines. Pilot lots of DTwP and DTaP formulated with 7.5% propylene glycol underwent 3 freeze-thaw treatments. The addition of propylene glycol had no impact on pH, particle size distribution, or potency of the vaccines prior to freeze-thaw treatment; the only change noted was an increase in osmolality. The potencies and the physical properties of the vaccines containing cryoprotectant were maintained after freeze-thawing and for 3 months in accelerated stability studies. The results from this study indicate that formulating vaccines with propylene glycol can protect diphtheria-tetanus-pertussis vaccines against freeze damages.

  9. Fracture energy evolution of two concretes resistant to the action of freeze-thaw cycles

    Directory of Open Access Journals (Sweden)

    Enfedaque, A.

    2014-03-01

    Full Text Available The current standards that regulate use of structural concrete have highlighted the durability of concrete. However, how the fracture energy of concrete evolves under the action of freeze-thaw cycles is not well known. The fracture energy of two types of concrete, one with an air-entraining additive and the other with silica fume addition, is studied after four, 14 and 28 freeze-thaw cycles. The results obtained show that the concrete with an air-entraining additive was undamaged and that fracture energy grew slightly. In addition to this, they also showed that the concrete with silica fume addition suffered severe surface scaling and its fracture energy changed due to the greater fracture areas generated.La actual normativa que rige el empleo de hormigón estructural ha puesto enfásis en la durabilidad del hormigón. Sin embargo, no se conoce cómo evoluciona la energía de fractura del hormigón sometido a ciclos hielo- deshielo, lo cual es de vital importancia para asegurar la durabilidad y el correcto comportamiento mecánico de las estructuras de hormigón en entornos con heladas durante su vida útil. Se ha estudiado la evolución de la energía de fractura de un hormigón con aireante y de un hormigón con humo de sílice después de 4, 14 y 28 ciclos hielo-deshielo realizando ensayos de fractura. Los resultados muestran cómo el hormigón con aireante no sufre daño por los ciclos hielo-deshielo y cómo la energía de fractura del mismo aumenta ligeramente. El hormigón con humo de sílice se daña por los ciclos hielo-deshielo y reduce su energía de fractura al aumentar el area fracturada.

  10. In situ synthesis of bilayered gradient poly(vinyl alcohol)/hydroxyapatite composite hydrogel by directional freezing-thawing and electrophoresis method.

    Science.gov (United States)

    Su, Cui; Su, Yunlan; Li, Zhiyong; Haq, Muhammad Abdul; Zhou, Yong; Wang, Dujin

    2017-08-01

    Bilayered poly(vinyl alcohol) (PVA)/hydroxyapatite (HA) composite hydrogels with anisotropic and gradient mechanical properties were prepared by the combination of directional freezing-thawing (DFT) and electrophoresis method. Firstly, PVA hydrogels with aligned channel structure were prepared by the DFT method. Then, HA nanoparticles were in situ synthesized within the PVA hydrogels via electrophoresis. By controlling the time of the electrophoresis process, a bilayered gradient hydrogel containing HA particles in only half of the gel region was obtained. The PVA/HA composite hydrogel exhibited gradient mechanical strength depending on the distance to the cathode. The gradient initial tensile modulus ranging from 0.18MPa to 0.27MPa and the gradient initial compressive modulus from 0.33MPa to 0.51MPa were achieved. The binding strength of the two regions was relatively high and no apparent internal stress or defect was observed at the boundary. The two regions of the bilayered hydrogel also showed different osteoblast cell adhesion properties. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Long-term Stability of Esomeprazole in 5% Dextrose Infusion Polyolefin Bags at 5 degrees C +/- 3 degrees C after Microwave Freeze-thaw Treatment.

    Science.gov (United States)

    Hecq, Jean-daniel; Rolin, Catherine; Godet, Marie; Gillet, Patricia; Jamart, Jacques; Galanti, Laurence M

    2015-01-01

    To improve quality assurance, security, time management, and cost saving of drug delivery, preparation in advance of intravenous solutions has been developed for several infusion solutions. The objective of this study was to investigate the stability of esomeprazole 0.4 mg/mL and 0.8 mg/mL in 5% dextrose polyolefin bags after freezing, long-term storage, and microwave thawing. The stability of five polyolefin bags containing approximately 0.4 mg/mL of esomeprazole and five other bags containing approximately 0.8 mg/mL in 5% dextrose prepared under aseptic conditions was studied after freezing for 1 month at -20 degrees C, thawing in a microwave oven with a validated cycle, and stored at 5 degrees C +/- 3 degrees C. Esomeprazole concentration was measured by high-pressure liquid chromatography using a reversed-phase column C8, a mobile phase consisting of 35% of acetonitrile and 65% of Na2HPO4 buffer at pH 7.59 with HPO4 (2 M) and NaOH (0.5 M), and detection with a diode array detector at 280 nm. Visual, microscopic, and spectrophotometric observation and pH measurements were also performed. No precipitation occurred in the preparations but little change of color was observed. No microaggregate was observed with optical microscopy or revealed by a change of absorbance at 350, 410, and 550 nm. Based on a shelf life of 90% residual potency, esomeprazole solutions (0.4 and 0.8 mg/mL) were stable for at least 20 or 29 days, respectively, after a freezing and microwave thawing period, where 95% one-side lower confidence limit of the concentration-time profile remained superior to 90% of the initial concentration. During this period, the pH values of drug solutions have been observed to decrease without affecting chromatographic parameters. Within these limits, esomeprazole (0.4 and 0.8 mg/mL) in 5% dextrose infusions may be prepared and frozen in advance by a centralized intravenous admixture service, thawed, and stored at least 20 days at 5 degrees C +/- 3 degrees C

  12. Influence of freezing down to 77.15 K on structure and antioxidant power of some proteins

    Science.gov (United States)

    Rozanova, S. L.; Narozhnyi, S. V.; Nardid, O. A.

    2017-06-01

    The purpose of the present work was to investigate influence of different freeze-thawing protocols on structure and antioxidant properties of isolated proteins. In our experiments we have studied human serum albumin, human hemoglobin and cytochrome C derived from equine heart frozen down to 77.15 K with 1-2 deg/min and 300 deg/min rate with following thawing on a water bath at 293.15 K. Native proteins were assumed as a control. Influence of freeze-thawing protocols on protein structure was investigated using spectrophotometric and fluorescent assays. Antioxidant activities of isolated proteins were estimated by their ability to reduce ABTS+ radical. It has been established that unfolding derived from freeze-thawing exposure leads to protein antioxidant activity increasing while decreasing of such an activity may be connected with macromolecule aggregation. Character of freeze-thawing influence on antioxidant activity of proteins depends on molecule structure peculiarities and freezing protocols.

  13. Spatiotemporal changes of freezing/thawing indices and their response to recent climate change on the Qinghai-Tibet Plateau from 1980 to 2013

    Science.gov (United States)

    Wu, Tonghua; Qin, Yanhui; Wu, Xiaodong; Li, Ren; Zou, Defu; Xie, Changwei

    2018-05-01

    The spatial and temporal changes of the ground surface freezing indices (GFIs), ground surface thawing indices (GTIs), air freezing indices (AFIs), and air thawing indices (ATIs) in permafrost and seasonally frozen ground regions of the Qinghai-Tibet Plateau (QTP) were analyzed based on the daily ground surface and air temperatures from 69 meteorological stations using the Mann-Kendall test and Sen's slope estimate. The spatial patterns of the freezing indices (FIs) and thawing indices (TIs) are nearly negatively correlated. On the annual scale, the GFI and GTI are greater than the AFI and ATI in both permafrost and seasonally frozen ground regions. The marked upward and downward trends have been observed for the time series of TI and FI, respectively, since 1998 on the QTP. Moreover, GFI and AFI decrease more significantly in permafrost regions than in seasonally frozen ground regions; the increasing rate of GTI and ATI in the seasonally frozen ground regions is greater than that in the permafrost regions. In permafrost regions, the downward trend of FI is greater than the upward trend of TI. However, the upward trend of TI shows a more drastic change than the FI in the seasonally frozen ground regions. The results indicate that the warming in the permafrost regions is more pronounced in winter than in the other seasons. The summer warming is more pronounced than the other seasons in the seasonally frozen ground regions. The decreasing rate of AFI and GFI increases as the altitude rises, while they decrease with increasing ATI. The average decreasing rate of GFI is greater than that of the AFI in different altitudinal zones. The greatest decrease of FI occurs in permafrost regions in the hinterland of the QTP, which indicates the dominant winter warming in this region. The downward trend of FI and upward trend of TI are responsible for the reported permafrost degradation on the QTP.

  14. Effects of PVA, agar contents, and irradiation doses on properties of PVA/ws-chitosan/glycerol hydrogels made by γ-irradiation followed by freeze-thawing

    International Nuclear Information System (INIS)

    Yang Xiaomin; Zhu Zhiyong; Liu Qi; Chen Xiliang; Ma Mingwang

    2008-01-01

    Poly(vinyl alcohol) (PVA)/water soluble chitosan (ws-chitosan)/glycerol hydrogels were prepared by γ-irradiation and γ-irradiation followed by freeze-thawing, respectively. The effects of irradiation dose and the contents of PVA and agar on the swelling, rheological, and thermal properties of these hydrogels were investigated. The swelling capacity decreases while the mechanical strength increases with increasing PVA or agar content. Increasing the irradiation dose leads to an increase in chemical crosslinking density but a decrease in physical crosslinking density. Hydrogels made by irradiation followed by freeze-thawing own smaller swelling capacity but larger mechanical strength than those made by pure irradiation. The storage modulus of the former hydrogels decreases above 50 deg. C and above 70 deg. C it comes to the same value as that prepared by irradiation. The ordered association of PVA is influenced by both chemical and physical crosslinkings and by the presence of ws-chitosan and glycerol. These hydrogels are high sensitive to pH and ionic strength, indicating that they may be useful in stimuli-responsive drug release system

  15. Structural Changes in Cattle Immature Oocytes Subjected to Slow Freezing and Vitrification

    Directory of Open Access Journals (Sweden)

    H. Wahid*, M. Thein1, E.A. El-Hafez2, M.O. Abas3, K. Mohd Azam4, O. Fauziah5, Y. Rosnina and H. Hajarian

    2012-05-01

    Full Text Available This study was conducted to evaluate the effect of different cryopreservation methods (slow-freezing and vitrification on structural changes of bovine immature oocytes. Bovine ovaries were collected from local abattoirs. Cumulus-oocyte-complexes (COCs were retrieved using aspiration method from 2-6 mm follicles. In Experiment 1, selected oocytes were randomly divided into 4 treatment groups namely freezing solution-exposed, frozen-thawed, vitrification solution-exposed and vitrified-thawed and then oocytes abnormalities were examined under a stereomicroscope. In Experiment 2, oocytes were randomly allocated to the same grouping as experiment 1 plus control group. Following freezing or vitrification, all oocytes were fixed in glutaraldehyde and processed for transmission electron microscopy. In experiment 1, there was a higher incidence of abnormalities in the frozen-thawed and vitrified-warmed oocytes compared to those in freezing solution and vitrification solution-exposed groups (P<0.05. In experiment 2, there were marked alterations in the perivitelline space, microvilli and vesicles of frozen-thawed and vitrified-warmed oocytes characterized by loss of elasticity and integrity of cytoplasmic processes and microvilli following cooling and warming. In conclusion, ethylene glycol-based freezing and vitrification solutions are suitable choices for cryopreservation of immature oocytes and most organelles are able to retain their normal morphology following cryopreservation and thawing processes.

  16. Tolerance of brown bear spermatozoa to conditions of pre-freezing cooling rate and equilibration time.

    Science.gov (United States)

    López-Urueña, E; Alvarez, M; Gomes-Alves, S; Martínez-Rodríguez, C; Borragan, S; Anel-López, L; de Paz, P; Anel, L

    2014-06-01

    Specific protocols for the cryopreservation of endangered Cantabrian brown bear spermatozoa are critical to create a genetic resource bank. The aim of this study was to assess the effect of cooling rates and equilibration time before freezing on post-thawed brown bear spermatozoa quality. Electroejaculates from 11 mature bears were extended to 100 × 10(6) spermatozoa/mL in a TES-Tris-Fructose-based extender, cryopreserved following performance of the respective cooling/equilibration protocol each sample was assigned to, and stored at -196 °C for further assessment. Before freezing, after thawing, and after 1 hour's incubation post-thawing at 37 °C (thermal stress test), the quality of the samples was assessed for motility by computer-assisted semen analysis, and for viability (SYBR-14/propidium iodide), acrosomal status (peanut agglutinin-fluorescein isothiocyanate /propidium iodide), and sperm chromatin stability (SCSA) by flow cytometry. In experiment 1, three cooling rates (0.25 °C/min, 1 °C/min, and 4 °C/min) to 5 °C were assessed. After thawing, total motility (%TM) was higher and percentage of damaged acrosomes (%dACR) was lower (P bear sperm. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Monitoring of Freeze-Thaw Cycles in Concrete Using Embedded Sensors and Ultrasonic Imaging

    Science.gov (United States)

    Ranz, Javier; Aparicio, Sofía; Romero, Héctor; Casati, María Jesús; Molero, Miguel; González, Margarita

    2014-01-01

    This paper deals with the study of damage produced during freeze-thaw (F-T) cycles using two non-destructive measurement approaches—the first approach devoted to continuous monitoring using embedded sensors during the cycles, and the second one, performing ultrasonic imaging before and after the cycles. Both methodologies have been tested in two different types of concrete specimens, with and without air-entraining agents. Using the first measurement approach, the size and distribution of pores were estimated using a thermoporometrical model and continuous measurements of temperature and ultrasonic velocity along cycles. These estimates have been compared with the results obtained using mercury porosimetry testing. In the second approach, the damage due to F-T cycles has been evaluated by automated ultrasonic transmission and pulse-echo inspections made before and after the cycles. With these inspections the variations in the dimensions, velocity and attenuation caused by the accelerated F-T cycles were determined. PMID:24481231

  18. Effects of different extenders on DNA integrity of boar spermatozoa following freezing-thawing.

    Science.gov (United States)

    Hu, Jian-hong; Li, Qing-wang; Jiang, Zhong-liang; Li, Wen-ye

    2008-12-01

    The sperm-rich fraction, collected from eight mature Yorkshire boars, was frozen in an extender containing 9% LDL (w/v), 100mM trehalose, or 20% yolk (v/v), respectively. Sperm DNA integrity was assessed using the single-cell gel electrophoresis (SCGE). Other sperm quality characteristics such as motility, acrosome and membrane integrity were also monitored. The results showed that freezing-thawing caused an increase in sperm DNA fragmentation, and extender containing 9% LDL could significantly protect sperm DNA integrity (Pextender containing 100mM trehalose and 20% yolk (v/v). No significant difference in damaged DNA was detected between frozen and unfrozen semen samples for extender of 9% LDL and 100mM trehalose, but cryopreservation could increase the degree of DNA damage (Pboar sperm DNA damage and protecting DNA integrity. It can be suggested that evaluation of sperm DNA integrity, coupled with correlative and basic characteristics such as motility, acrosome integrity and membrane integrity, may aid in determining the quality of frozen boar semen.

  19. Alcohol Brine Freezing of Japanese Horse Mackerel (Trachurus japonicus) for Raw Consumption

    Science.gov (United States)

    Maeda, Toshimichi; Yuki, Atsuhiko; Sakurai, Hiroshi; Watanabe, Koichiro; Itoh, Nobuo; Inui, Etsuro; Seike, Kazunori; Mizukami, Yoichi; Fukuda, Yutaka; Harada, Kazuki

    In order to test the possible application of alcohol brine freezing to Japanese horse mackerel (Trachurus japonicus) for raw consumption, the quality and taste of fish frozen by direct immersion in 60% ethanol brine at -20, -25 and -30°C was compared with those by air freezing and fresh fish without freezing. Cracks were not found during the freezing. Smell of ethanol did not remain. K value, an indicator of freshness, of fish frozen in alcohol brine was less than 8.3%, which was at the same level as those by air freezing and fresh fish. Oxidation of lipid was at the same level as air freezing does, and lower than that of fresh fish. The pH of fish frozen in alcohol brine at -25 and -30°C was 6.5 and 6.6, respectively, which were higher than that by air freezing and that of fresh fish. Fish frozen in alcohol brine was better than that by air and at the same level as fresh fish in total evaluation of sensory tests. These results show that the alcohol brine freezing is superior to air freezing, and fish frozen in alcohol brine can be a material for raw consumption. The methods of thawing in tap water, cold water, refrigerator, and at room temperature were compared. Thawing in tap water is considered to be convenient due to the short thaw time and the quality of thawed fish that was best among the methods.

  20. Enhanced-Resolution Satellite Microwave Brightness Temperature Records for Mapping Boreal-Arctic Landscape Freeze-Thaw Heterogeneity

    Science.gov (United States)

    Kim, Y.; Du, J.; Kimball, J. S.

    2017-12-01

    The landscape freeze-thaw (FT) status derived from satellite microwave remote sensing is closely linked to vegetation phenology and productivity, surface energy exchange, evapotranspiration, snow/ice melt dynamics, and trace gas fluxes over land areas affected by seasonally frozen temperatures. A long-term global satellite microwave Earth System Data Record of daily landscape freeze-thaw status (FT-ESDR) was developed using similar calibrated 37GHz, vertically-polarized (V-pol) brightness temperatures (Tb) from SMMR, SSM/I, and SSMIS sensors. The FT-ESDR shows mean annual spatial classification accuracies of 90.3 and 84.3 % for PM and AM overpass retrievals relative surface air temperature (SAT) measurement based FT estimates from global weather stations. However, the coarse FT-ESDR gridding (25-km) is insufficient to distinguish finer scale FT heterogeneity. In this study, we tested alternative finer scale FT estimates derived from two enhanced polar-grid (3.125-km and 6-km resolution), 36.5 GHz V-pol Tb records derived from calibrated AMSR-E and AMSR2 sensor observations. The daily FT estimates are derived using a modified seasonal threshold algorithm that classifies daily Tb variations in relation to grid cell-wise FT thresholds calibrated using ERA-Interim reanalysis based SAT, downscaled using a digital terrain map and estimated temperature lapse rates. The resulting polar-grid FT records for a selected study year (2004) show mean annual spatial classification accuracies of 90.1% (84.2%) and 93.1% (85.8%) for respective PM (AM) 3.125km and 6-km Tb retrievals relative to in situ SAT measurement based FT estimates from regional weather stations. Areas with enhanced FT accuracy include water-land boundaries and mountainous terrain. Differences in FT patterns and relative accuracy obtained from the enhanced grid Tb records were attributed to several factors, including different noise contributions from underlying Tb processing and spatial mismatches between Tb

  1. Tracheid diameter is the key trait determining the extent of freezing-induced embolism in conifers.

    Science.gov (United States)

    Pittermann, Jarmila; Sperry, John

    2003-09-01

    We tested the hypotheses that freezing-induced embolism is related to conduit diameter, and that conifers and angiosperms with conduits of equivalent diameter will exhibit similar losses of hydraulic conductivity in response to freezing. We surveyed the freeze-thaw response of conifers with a broad range of tracheid diameters by subjecting wood segments (root, stem and trunk wood) to a freeze-thaw cycle at -0.5 MPa in a centrifuge. Embolism increased as mean tracheid diameter exceeded 30 microm. Tracheids with a critical diameter greater than 43 microm were calculated to embolize in response to freezing and thawing at a xylem pressure of -0.5 MPa. To confirm that freezing-induced embolism is a function of conduit air content, we air-saturated stems of Abies lasiocarpa (Hook.) Nutt. (mean conduit diameter 13.7 +/- 0.7 microm) by pressurizing them 1 to 60 times above atmospheric pressure, prior to freezing and thawing. The air saturation method simulated the effect of increased tracheid size because the degree of super-saturation is proportional to a tracheid volume holding an equivalent amount of dissolved air at ambient pressure. Embolism increased when the dissolved air content was equivalent to a mean tracheid diameter of 30 microm at ambient air pressure. Our centrifuge and air-saturation data show that conifers are as vulnerable to freeze-thaw embolism as angiosperms with equal conduit diameter. We suggest that the hydraulic conductivity of conifer wood is maximized by increasing tracheid diameters in locations where freezing is rare. Conversely, the narrowing of tracheid diameters protects against freezing-induced embolism in cold climates.

  2. Rational design of an influenza subunit vaccine powder with sugar glass technology : preventing conformational changes of haemagglutinin during freezing and freeze-drying

    NARCIS (Netherlands)

    Amorij, J-P; Meulenaar, J; Hinrichs, W L J; Stegmann, T; Huckriede, A; Coenen, F; Frijlink, H W

    2007-01-01

    The development of a stable influenza subunit vaccine in the dry state was investigated. The influence of various carbohydrates, buffer types and freezing rates on the integrity of haemagglutinin after freeze-thawing or freeze-drying was investigated with a range of analytical and immunological

  3. Laboratory and numerical experiments on water and energy fluxes during freezing and thawing in the unsaturated zone

    Science.gov (United States)

    Holländer, Hartmut; Montasir Islam, Md.; Šimunek, Jirka

    2017-04-01

    Frozen soil has a major effect in many hydrologic processes, and its effects are difficult to predict. A prime example is flood forecasting during spring snowmelt within the Canadian Prairies. One key driver for the extent of flooding is the antecedent soil moisture and the possibility for water to infiltrate into frozen soils. Therefore, these situations are crucial for accurate flood prediction during every spring. The main objective of this study was to evaluate the water flow and heat transport within HYDRUS-1D version 4.16 and with Hansson's model, which is a detailed freezing/thawing module (Hansson et al., 2004), to predict the impact of frozen and partly frozen soil on infiltration. We developed a standardized data set of water flow and heat transport into (partial) frozen soil by laboratory experiments using fine sand. Temperature, soil moisture, and percolated water were observed at different freezing conditions as well as at thawing conditions. Significant variation in soil moisture was found between the top and the bottom of the soil column at the starting of the thawing period. However, with increasing temperature, the lower depth of the soil column showed higher moisture as the soil became enriched with moisture due to the release of heat by soil particles during the thawing cycle. We applied vadose zone modeling using the results from the laboratory experiments. The simulated water content by HYDRUS-1D 4.16 showed large errors compared to the observed data showing by negative Nash-Sutcliffe Efficiency. Hansson's model was not able to predict soil water fluxes due to its unstable behavior (Šimunek et al., 2016). The soil temperature profile simulated using HYDRUS-1D 4.16 was not able to predict the release of latent heat during the phase change of water that was visible in Hansson's model. Hansson's model includes the energy gain/loss due to the phase change in the amount of latent energy stored in the modified heat transport equation. However, in

  4. A single-repeat R3-MYB transcription factor MYBC1 negatively regulates freezing tolerance in Arabidopsis

    International Nuclear Information System (INIS)

    Zhai, Hong; Bai, Xi; Zhu, Yanming; Li, Yong; Cai, Hua; Ji, Wei; Ji, Zuojun; Liu, Xiaofei; Liu, Xin; Li, Jing

    2010-01-01

    We had previously identified the MYBC1 gene, which encodes a single-repeat R3-MYB protein, as a putative osmotic responding gene; however, no R3-MYB transcription factor has been reported to regulate osmotic stress tolerance. Thus, we sought to elucidate the function of MYBC1 in response to osmotic stresses. Real-time RT-PCR analysis indicated that MYBC1 expression responded to cold, dehydration, salinity and exogenous ABA at the transcript level. mybc1 mutants exhibited an increased tolerance to freezing stress, whereas 35S::MYBC1 transgenic plants exhibited decreased cold tolerance. Transcript levels of some cold-responsive genes, including CBF/DREB genes, KIN1, ADC1, ADC2 and ZAT12, though, were not altered in the mybc1 mutants or the 35S::MYBC1 transgenic plants in response to cold stress, as compared to the wild type. Microarray analysis results that are publically available were investigated and found transcript level of MYBC1 was not altered by overexpression of CBF1, CBF2, and CBF3, suggesting that MYBC1 is not down regulated by these CBF family members. Together, these results suggested that MYBC1is capable of negatively regulating the freezing tolerance of Arabidopsis in the CBF-independent pathway. In transgenic Arabidopsis carrying an MYBC1 promoter driven β-glucuronidase (GUS) construct, GUS activity was observed in all tissues and was relatively stronger in the vascular tissues. Fused MYBC1 and GFP protein revealed that MYBC1 was localized exclusively in the nuclear compartment.

  5. Season-induced changes in bovine sperm motility following a freeze-thaw procedure.

    Science.gov (United States)

    Orgal, Shlomo; Zeron, Yoel; Elior, Nili; Biran, David; Friedman, Eran; Druker, Shaked; Roth, Zvi

    2012-01-01

    Decreased conception rate of dairy cows in the summer is mainly associated with the deleterious effects of environmental thermal stress on the female reproductive tract. Here, we suggest that decreased reproductive performance might be partially due to inferior-quality semen. Semen from five representative bulls was collected in summer (August to September) and winter (December to January) and evaluated with a computerized sperm-quality analyzer for bulls (SQA-Vb). No seasonal effect was found in fresh ejaculate, but sperm examined post-thawing showed lower velocity, motility and progressive motility (Pconception rate of dairy cows in summer.

  6. Estimation model of soil freeze-thaw erosion in Silingco watershed wetland of Northern Tibet.

    Science.gov (United States)

    Kong, Bo; Yu, Huan

    2013-01-01

    The freeze-thaw (FT) erosion is a type of soil erosion like water erosion and wind erosion. Limited by many factors, the grading evaluation of soil FT erosion quantities is not well studied. Based on the comprehensive analysis of the evaluation indices of soil FT erosion, we for the first time utilized the sensitivity of microwave remote sensing technology to soil moisture for identification of FT state. We established an estimation model suitable to evaluate the soil FT erosion quantity in Silingco watershed wetland of Northern Tibet using weighted summation method of six impact factors including the annual FT cycle days, average diurnal FT phase-changed water content, average annual precipitation, slope, aspect, and vegetation coverage. Finally, with the support of GIS, we classified soil FT erosion quantity in Silingco watershed wetland. The results showed that soil FT erosion are distributed in broad areas of Silingco watershed wetland. Different soil FT erosions with different intensities have evidently different spatial and geographical distributions.

  7. Changes in antibiotic sensitivity and cell surface hydrophobicity in Escherichia coli injured by heating, freezing, drying or gamma radiation

    International Nuclear Information System (INIS)

    Mackey, B.M.

    1983-01-01

    Escherichia coli cells exposed to mild heating, freezing and thawing, drying or γ-radiation were sensitised to hydrophobic antibiotics and sodium deoxycholate but not to small hydrophilic antibiotics. These stress treatments also caused increases in cell surface hydrophobicity broadly reflecting the degree of sensitivity to hydrophobic antibiotics. (Auth.)

  8. Response of seasonal soil freeze depth to climate change across China

    Science.gov (United States)

    Peng, Xiaoqing; Zhang, Tingjun; Frauenfeld, Oliver W.; Wang, Kang; Cao, Bin; Zhong, Xinyue; Su, Hang; Mu, Cuicui

    2017-05-01

    The response of seasonal soil freeze depth to climate change has repercussions for the surface energy and water balance, ecosystems, the carbon cycle, and soil nutrient exchange. Despite its importance, the response of soil freeze depth to climate change is largely unknown. This study employs the Stefan solution and observations from 845 meteorological stations to investigate the response of variations in soil freeze depth to climate change across China. Observations include daily air temperatures, daily soil temperatures at various depths, mean monthly gridded air temperatures, and the normalized difference vegetation index. Results show that soil freeze depth decreased significantly at a rate of -0.18 ± 0.03 cm yr-1, resulting in a net decrease of 8.05 ± 1.5 cm over 1967-2012 across China. On the regional scale, soil freeze depth decreases varied between 0.0 and 0.4 cm yr-1 in most parts of China during 1950-2009. By investigating potential climatic and environmental driving factors of soil freeze depth variability, we find that mean annual air temperature and ground surface temperature, air thawing index, ground surface thawing index, and vegetation growth are all negatively associated with soil freeze depth. Changes in snow depth are not correlated with soil freeze depth. Air and ground surface freezing indices are positively correlated with soil freeze depth. Comparing these potential driving factors of soil freeze depth, we find that freezing index and vegetation growth are more strongly correlated with soil freeze depth, while snow depth is not significant. We conclude that air temperature increases are responsible for the decrease in seasonal freeze depth. These results are important for understanding the soil freeze-thaw dynamics and the impacts of soil freeze depth on ecosystem and hydrological process.

  9. To transfer fresh or thawed embryos?

    DEFF Research Database (Denmark)

    Pinborg, Anja

    2012-01-01

    Worldwide freezing and thawing of embryos has been increasingly used since the first infant was born as a result of this technique in 1984. The use of frozen embryo replacement (FER) currently even exceeds the number of fresh cycles performed in some countries. This article discusses the pros...... and multiple pregnancies, thereby increasing the safety for mother and child. Finally the article describes the accumulating literature on perinatal and long-term child outcome after transfer of frozen/thawed embryos, including a discussion on the concerns regarding cryo techniques and their possible roles...

  10. Oxidative stress adaptation with acute, chronic, and repeated stress.

    Science.gov (United States)

    Pickering, Andrew M; Vojtovich, Lesya; Tower, John; A Davies, Kelvin J

    2013-02-01

    Oxidative stress adaptation, or hormesis, is an important mechanism by which cells and organisms respond to, and cope with, environmental and physiological shifts in the level of oxidative stress. Most studies of oxidative stress adaption have been limited to adaptation induced by acute stress. In contrast, many if not most environmental and physiological stresses are either repeated or chronic. In this study we find that both cultured mammalian cells and the fruit fly Drosophila melanogaster are capable of adapting to chronic or repeated stress by upregulating protective systems, such as their proteasomal proteolytic capacity to remove oxidized proteins. Repeated stress adaptation resulted in significant extension of adaptive responses. Repeated stresses must occur at sufficiently long intervals, however (12-h or more for MEF cells and 7 days or more for flies), for adaptation to be successful, and the levels of both repeated and chronic stress must be lower than is optimal for adaptation to acute stress. Regrettably, regimens of adaptation to both repeated and chronic stress that were successful for short-term survival in Drosophila nevertheless also caused significant reductions in life span for the flies. Thus, although both repeated and chronic stress can be tolerated, they may result in a shorter life. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. Freezing-thawing hysteresis phenomena of biological systems by the new method of proton magnetic resonance

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, Eiichiro; Nagashima, Nobuya (Ajinomoto Co. Inc., Kawasaki, Kanagawa (Japan))

    1982-09-01

    an automatic recording system was developed for unfrozen water content and spin-spin relaxation time measurements as continuous functions of temperature, by using a broad-line pulsed NMR spectrometer and a minicomputer. The advantages of this system are that the exact quantitative measurements can be done by calibrating the nonlinearity of the NMR sensitivity, and that for high sensitivity temperature measurement the thermocouple with special device is directly immersed in a sample. Three types of freezing-thawing hysteresis phenomena, (1) recrystallization of solute (hydroxy-L-proline, D-mannitol) and refreezing of released hydrated water molecules in frozen aqueous solutions, and (2) hysteresis as the characteristic feature of gels (gelatin, alpha sub(sl)-casein), and (3) supercooling of capillary water in water-insoluble materials (zein, yeast RNA, cellulose) were analysed. The usefulness of this system as an analytical instrument of hydration properties of biological materials is emphasized.

  12. Freezing-thawing hysteresis phenomena of biological systems by the new method of proton magnetic resonance

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, E.; Nagashima, N.

    1982-01-01

    An automatic recording system was developed for unfrozen water content and spin-spin relaxation time measurements as continuous functions of temperature, by using a broad-line pulsed NMR spectrometer and a mini-computer. The advantages of this system are that the exact quantitative measurements can be done by calibrating the nonlinearity of the NMR sensitivity, and that for high sensitivity temperature measurement the thermocouple with special device is directly immersed in a sample. Three types of freezing-thawing hysteresis phenomena, (1) recrystallization of solute(hydroxy-L-proline, D-mannitol) and refreezing of released hydrated water molecules in frozen aqueous solutions, and (2) hysteresis as the characteristic feature of gels(gelatin, J/sub s1/-casein), and (3) supercooling of capillary water in water-insoluble materials(zein, yeast RNA, cellulose) were analysed. The usefulness of this system as an analytical instrument of hydration properties of biological materials is emphasized.

  13. Freezing-thawing hysteresis phenomena of biological systems by the new method of proton magnetic resonance

    International Nuclear Information System (INIS)

    Suzuki, Eiichiro; Nagashima, Nobuya

    1982-01-01

    an automatic recording system was developed for unfrozen water content and spin-spin relaxation time measurements as continuous functions of temperature, by using a broad-line pulsed NMR spectrometer and a minicomputer. The advantages of this system are that the exact quantitative measurements can be done by calibrating the nonlinearity of the NMR sensitivity, and that for high sensitivity temperature measurement the thermocouple with special device is directly immersed in a sample. Three types of freezing-thawing hysteresis phenomena, (1) recrystallization of solute (hydroxy-L-proline, D-mannitol) and refreezing of released hydrated water molecules in frozen aqueous solutions, and (2) hysteresis as the characteristic feature of gels(gelatin, alpha sub(sl)-casein), and (3) supercooling of capillary water in water-insoluble materials(zein, yeast RNA, cellulose) were analysed. The usefulness of this system as an analytical instrument of hydration properties of biological materials is emphasized. (author)

  14. Effect of magnetic field on food freezing

    OpenAIRE

    村田, 圭治; 奥村, 太一; 荒賀, 浩一; 小堀, 康功

    2010-01-01

    [Abstract] This paper presents an experimental investigation on effects of magnetic field on food freezing process. Although purpose of food freezing is to suppress the deterioration of food, freezing breaks food tissue down, and some nutrient and delicious element flow out after thawing. Recently, a few of refrigeration equipments with electric and magnetic fields have attracted attention from food production companies and mass media. Water and tuna were freezed in magnetic field (100kH, 1.3...

  15. Transcriptome Analysis of Spartina pectinata in Response to Freezing Stress.

    Directory of Open Access Journals (Sweden)

    Gyoungju Nah

    Full Text Available Prairie cordgrass (Spartina pectinata, a perennial C4 grass native to the North American prairie, has several distinctive characteristics that potentially make it a model crop for production in stressful environments. However, little is known about the transcriptome dynamics of prairie cordgrass despite its unique freezing stress tolerance. Therefore, the purpose of this work was to explore the transcriptome dynamics of prairie cordgrass in response to freezing stress at -5°C for 5 min and 30 min. We used a RNA-sequencing method to assemble the S. pectinata leaf transcriptome and performed gene-expression profiling of the transcripts under freezing treatment. Six differentially expressed gene (DEG groups were categorized from the profiling. In addition, two major consecutive orders of gene expression were observed in response to freezing; the first being the acute up-regulation of genes involved in plasma membrane modification, calcium-mediated signaling, proteasome-related proteins, and transcription regulators (e.g., MYB and WRKY. The follow-up and second response was of genes involved in encoding the putative anti-freezing protein and the previously known DNA and cell-damage-repair proteins. Moreover, we identified the genes involved in epigenetic regulation and circadian-clock expression. Our results indicate that freezing response in S. pectinata reflects dynamic changes in rapid-time duration, as well as in metabolic, transcriptional, post-translational, and epigenetic regulation.

  16. Predictive modeling of freezing and thawing of frost-susceptible soils.

    Science.gov (United States)

    2015-09-01

    Frost depth is an essential factor in design of various transportation infrastructures. In frost : susceptible soils, as soils freezes, water migrates through the soil voids below the freezing line : towards the freezing front and causes excessive he...

  17. Breed differences of bull frozen-thawed semen.

    Science.gov (United States)

    Ntemka, A; Tsousis, G; Brozos, C; Kiossis, E; Boscos, C M; Tsakmakidis, I A

    2016-12-01

    The objective of this study was to investigate the quality of frozen-thawed semen from different bull breeds. Commercial frozen-thawed bull semen samples (26 per breed, 130 totally) of five breeds (Holstein [Η], Brown Swiss [BS], Limousin [L], Belgian Blue [BB], Blonde d' Aquitaine [BA]) were used. After thawing, each semen sample was subjected to thermal resistance test (TR) for 0.5 and 1 hr at 38°C and hypo-osmotic swelling test (HOST) for 1 hr at 150 mOsm at 37°C. Additionally, all samples were evaluated at times 0 hr (thawing), 0.5 hr (TR), 1 hr (TR) for kinetics by CASA [progressive, immotile, rapid, medium, slow moving spermatozoa, curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (VSL), linearity (LIN), straightness (STR), beat cross-frequency (BCF), amplitude of lateral head displacement (ALH), wobble (WOB)]. Moreover, directly after thawing, all semen samples were evaluated for morphometry, morphology, viability and DNA fragmentation. Statistical analysis was conducted using a mixed model for repeated measures. The results showed (a) higher VCL after thawing in H, L breeds compared to BB and BA, (b) higher VAP after thawing in L compared to BB, BA, (c) higher values of progressive spermatozoa after TR in H, BS compared to BB, BA, (d) higher values of rapid spermatozoa after thawing and 0.5 hr of TR in H, BS, L compared to BB, BA, (e) lower viability in BA after thawing compared to H, BS, BB, (f) lower morphological abnormalities in H compared to L, BB, (g) higher head length in Η compared to BB. No significant differences were observed in the results from HOST and DNA fragmentation between breeds. In conclusion, quality characteristics of frozen-thawed bull semen are dependent on the breed. Frozen semen from BB and BA breeds should be handled more carefully after thawing, as it is more sensitive to stress. © 2016 Blackwell Verlag GmbH.

  18. Controlled cooling versus rapid freezing of teratozoospermic semen samples: Impact on sperm chromatin integrity

    Directory of Open Access Journals (Sweden)

    Shivananda N Kalludi

    2011-01-01

    Full Text Available Aim: The present study evaluates the impact of controlled slow cooling and rapid freezing techniques on the sperm chromatin integrity in teratozoospermic and normozoospermic samples. Setting: The study was done in a university infertility clinic, which is a tertiary healthcare center serving the general population. Design: It was a prospective study designed in vitro. Materials and Methods: Semen samples from normozoospermic (N=16 and teratozoospermic (N=13 infertile men were cryopreserved using controlled cooling and rapid freezing techniques. The sperm chromatin integrity was analyzed in fresh and frozen-thawed samples. Statistical Analysis Used: Data were reported as mean and standard error (mean ± SEM of mean. The difference between two techniques was determined by a paired t-test. Results: The freeze-thaw induced chromatin denaturation was significantly (P<0.01 elevated in the post-thaw samples of normozoospermic and teratozoospermic groups. Compared to rapid freezing, there was no difference in the number of red sperms (with DNA damage by the controlled slow cooling method in both normozoospermic and teratozoospermic groups. Freeze-thaw induced sperm chromatin denaturation in teratozoospermic samples did not vary between controlled slow cooling and rapid freezing techniques. Conclusions: Since the controlled slow cooling technique involves the use of expensive instrument and is a time consuming protocol, rapid freezing can be a good alternative technique for teratozoospermic and normozoospermic samples when sperm DNA damage is a concern.

  19. Stress preconditioning of rooster semen before cryopreservation improves fertility potential of thawed sperm.

    Science.gov (United States)

    Feyzi, S; Sharafi, M; Rahimi, S

    2018-03-22

    Avian semen cryopreservation is not as successful as that seen in mammals. This failure is mostly attributed to unique physiological characteristics of poultry semen that make it susceptible to cryo-damages. Utilization of sublethal oxidative stress for preconditioning of sperm, as an innovative approach, improves the cryo-survival of sperm in certain mammalian species. The purpose of this study was to investigate the effects of preconditioning of rooster semen with sublethal oxidative stress [very low concentrations of nitric oxide (NO)] before cryopreservation on the quality and fertility potential of thawed sperm. Semen samples were collected from 20 roosters, twice a wk, and different concentrations of NO [0 (NO-0), 0.01 (NO-0.01), 0.1 (NO-0.1), 1 (NO-1), 10 (NO-10), and 100 μM (NO-100)] were used to investigate the effects of controlled induction of sublethal stress before semen cryopreservation on the thawed sperm performance. A significantly higher (P 0.05) affected by different concentrations of NO. Sperm exposed to NO-1 produced the highest percentage of viable spermatozoa (Annexin-/PI-), which was significantly different from the other samples. Finally, rate of fertility after artificial insemination was significantly higher (P < 0.05) following treatment with NO-1 compared to NO-0 and NO-0.1. Application of 1 μM NO as a sublethal oxidative stress before cryopreservation of sperm efficiently increased numerous quality indices of thawed sperm as well as its fertility potential.

  20. Optimized production of vanillin from green vanilla pods by enzyme-assisted extraction combined with pre-freezing and thawing.

    Science.gov (United States)

    Zhang, Yanjun; Mo, Limei; Chen, Feng; Lu, Minquan; Dong, Wenjiang; Wang, Qinghuang; Xu, Fei; Gu, Fenglin

    2014-02-19

    Production of vanillin from natural green vanilla pods was carried out by enzyme-assisted extraction combined with pre-freezing and thawing. In the first step the green vanilla pods were pre-frozen and then thawed to destroy cellular compartmentation. In the second step pectinase from Aspergillus niger was used to hydrolyze the pectin between the glucovanillin substrate and β-glucosidase. Four main variables, including enzyme amount, reaction temperature, time and pH, which were of significance for the vanillin content were studied and a central composite design (CCD) based on the results of a single-factor tests was used. Response surface methodology based on CCD was employed to optimize the combination of enzyme amount, reaction temperature, time, and pH for maximum vanillin production. This resulted in the optimal condition in regards of the enzyme amount, reaction temperature, time, and pH at 84.2 mg, 49.5 °C, 7.1 h, and 4.2, respectively. Under the optimal condition, the experimental yield of vanillin was 4.63% ± 0.11% (dwb), which was in good agreement with the value predicted by the model. Compared to the traditional curing process (1.98%) and viscozyme extract (2.36%), the optimized method for the vanillin production significantly increased the yield by 133.85% and 96%, respectively.

  1. Decoupled responses of soil bacteria and their invertebrate consumer to warming, but not freeze-thaw cycles, in the Antarctic Dry Valleys.

    Science.gov (United States)

    Knox, Matthew A; Andriuzzi, Walter S; Buelow, Heather N; Takacs-Vesbach, Cristina; Adams, Byron J; Wall, Diana H

    2017-10-01

    Altered temperature profiles resulting in increased warming and freeze-thaw cycle (FTC) frequency pose great ecological challenges to organisms in alpine and polar ecosystems. We performed a laboratory microcosm experiment to investigate how temperature variability affects soil bacterial cell numbers, and abundance and traits of soil microfauna (the microbivorous nematode Scottnema lindsayae) from McMurdo Dry Valleys, Antarctica. FTCs and constant freezing shifted nematode body size distribution towards large individuals, driven by higher mortality among smaller individuals. FTCs reduced both bacterial and nematode abundance, but bacterial cell numbers also declined under warming, demonstrating decoupled consumer-prey responses. We predict that higher occurrence of FTCs in cold ecosystems will select for large body size within soil microinvertebrates and overall reduce their abundance. In contrast, warm temperatures without FTCs could lead to divergent responses in soil bacteria and their microinvertebrate consumers, potentially affecting energy and nutrient transfer rates in soil food webs of cold ecosystems. © 2017 John Wiley & Sons Ltd/CNRS.

  2. Effect of different procedures of ejaculate collection, extenders and packages on DNA integrity of boar spermatozoa following freezing-thawing.

    Science.gov (United States)

    Fraser, L; Strzezek, J

    2007-06-01

    Whole ejaculate or sperm-rich fraction, collected from four sexually mature boars, was frozen in an extender containing lactose-hen egg yolk with glycerol (lactose-HEY-G) or extender containing lactose, lyophilized lipoprotein fractions isolated from ostrich egg yolk and glycerol (lactose-LPFo-G), and Orvus Es Paste, respectively. The sperm samples were also frozen in a standard boar semen extender (Kortowo-3), without the addition of cryoprotective substances. Sperm DNA integrity was assessed using a modified neutral comet assay. Sperm characteristics such as motility, plasma membrane integrity (SYBR-14/PI), mitochondrial function (rhodamine 123) and acrosome integrity were monitored. Freezing-thawing caused a significant increase (Pextender type. Sperm DNA fragmentation was significantly lower (Pextender exhibited lower (Pboar semen.

  3. Damage development, phase changes, transport properties, and freeze-thaw performance of cementitious materials exposed to chloride based salts

    Science.gov (United States)

    Farnam, Yaghoob

    Recently, there has been a dramatic increase in premature deterioration in concrete pavements and flat works that are exposed to chloride based salts. Chloride based salts can cause damage and deterioration in concrete due to the combination of factors which include: increased saturation, ice formation, salt crystallization, osmotic pressure, corrosion in steel reinforcement, and/or deleterious chemical reactions. This thesis discusses how chloride based salts interact with cementitious materials to (1) develop damage in concrete, (2) create new chemical phases in concrete, (3) alter transport properties of concrete, and (4) change the concrete freeze-thaw performance. A longitudinal guarded comparative calorimeter (LGCC) was developed to simultaneously measure heat flow, damage development, and phase changes in mortar samples exposed to sodium chloride (NaCl), calcium chloride (CaCl 2), and magnesium chloride (MgCl2) under thermal cycling. Acoustic emission and electrical resistivity measurements were used in conjunction with the LGCC to assess damage development and electrical response of mortar samples during cooling and heating. A low-temperature differential scanning calorimetry (LT-DSC) was used to evaluate the chemical interaction that occurs between the constituents of cementitious materials (i.e., pore solution, calcium hydroxide, and hydrated cement paste) and salts. Salts were observed to alter the classical phase diagram for a salt-water system which has been conventionally used to interpret the freeze-thaw behavior in concrete. An additional chemical phase change was observed for a concrete-salt-water system resulting in severe damage in cementitious materials. In a cementitious system exposed to NaCl, the chemical phase change occurs at a temperature range between -6 °C and 8 °C due to the presence of calcium sulfoaluminate phases in concrete. As a result, concrete exposed to NaCl can experience additional freeze-thaw cycles due to the chemical

  4. Greenhouse gas balance over thaw-freeze cycles in discontinuous zone permafrost

    Science.gov (United States)

    Wilson, R. M.; Fitzhugh, L.; Whiting, G. J.; Frolking, S.; Harrison, M. D.; Dimova, N.; Burnett, W. C.; Chanton, J. P.

    2017-02-01

    Peat in the discontinuous permafrost zone contains a globally significant reservoir of carbon that has undergone multiple permafrost-thaw cycles since the end of the mid-Holocene ( 3700 years before present). Periods of thaw increase C decomposition rates which leads to the release of CO2 and CH4 to the atmosphere creating potential climate feedback. To determine the magnitude and direction of such feedback, we measured CO2 and CH4 emissions and modeled C accumulation rates and radiative fluxes from measurements of two radioactive tracers with differing lifetimes to describe the C balance of the peatland over multiple permafrost-thaw cycles since the initiation of permafrost at the site. At thaw features, the balance between increased primary production and higher CH4 emission stimulated by warmer temperatures and wetter conditions favors C sequestration and enhanced peat accumulation. Flux measurements suggest that frozen plateaus may intermittently (order of years to decades) act as CO2 sources depending on temperature and net ecosystem respiration rates, but modeling results suggest that—despite brief periods of net C loss to the atmosphere at the initiation of thaw—integrated over millennia, these sites have acted as net C sinks via peat accumulation. In greenhouse gas terms, the transition from frozen permafrost to thawed wetland is accompanied by increasing CO2 uptake that is partially offset by increasing CH4 emissions. In the short-term (decadal time scale) the net effect of this transition is likely enhanced warming via increased radiative C emissions, while in the long-term (centuries) net C deposition provides a negative feedback to climate warming.

  5. Effective freezing rate for semen cryopreservation in endangered Mediterranean brown trout (Salmo trutta macrostigma) inhabiting the Biferno river (South Italy).

    Science.gov (United States)

    Iaffaldano, Nicolaia; Di Iorio, Michele; Manchisi, Angelo; Esposito, Stefano; Gibertoni, Pier Paolo

    2016-10-01

    This study was designed to determine: (i) the in vitro effects of different freezing rates on post-thaw semen quality of Mediterranean brown trout (Salmo trutta macrostigma) from the Biferno river; and (ii) the in vivo fertilization and hatching percentage of freezing rate giving rise to the best post-thaw semen quality. Pooled semen samples were diluted 1:3 (v:v) in a freezing extender composed of 300 mM glucose, 10% egg yolk and 10% dimethyl sulfoxide (DMSO). The extended semen was packaged in 0.25 ml plastic straws and frozen at different heights above the liquid nitrogen surface (1, 5 or 10 cm) for 10 min to give three different freezing rates. Semen samples were thawed at 30°C for 10 s. The variables assessed after thawing were sperm motility, duration of motility and viability. Our results clearly indicate a significant effect of freezing rate on post-thaw semen quality. Semen frozen 5 cm above the liquid nitrogen surface showed the best quality after freezing/thawing. Based on these in vitro data, 2 groups of 200 eggs were fertilized with fresh semen or semen frozen 5 cm above the liquid nitrogen surface. Fertilization and hatching rates recorded for eggs fertilized with frozen semen were significantly lower (25.4% and 22.5%, respectively) than the ones obtained using fresh semen (87.8% and 75.5%, respectively). An effective freezing protocol will allow for the creation of a sperm cryobank to recover the original population of Mediterranean brown trout in the Biferno river.

  6. Modeling conductive heat transfer during high-pressure thawing processes: determination of latent heat as a function of pressure.

    Science.gov (United States)

    Denys, S; Van Loey, A M; Hendrickx, M E

    2000-01-01

    A numerical heat transfer model for predicting product temperature profiles during high-pressure thawing processes was recently proposed by the authors. In the present work, the predictive capacity of the model was considerably improved by taking into account the pressure dependence of the latent heat of the product that was used (Tylose). The effect of pressure on the latent heat of Tylose was experimentally determined by a series of freezing experiments conducted at different pressure levels. By combining a numerical heat transfer model for freezing processes with a least sum of squares optimization procedure, the corresponding latent heat at each pressure level was estimated, and the obtained pressure relation was incorporated in the original high-pressure thawing model. Excellent agreement with the experimental temperature profiles for both high-pressure freezing and thawing was observed.

  7. Mix design for improved strength and freeze-thaw durability of pervious concrete fill in Pearl-Chain Bridges

    DEFF Research Database (Denmark)

    Lund, Mia Schou Møller; Kevern, John T.; Schaefer, Vernon R.

    2017-01-01

    different mixture proportions using two different sizes of granite coarse aggregate and at two different water-to-cement ratios. The specimens had an average void content of 24-28 %. Specimens containing air entraining and high-range water reducing admixtures were most workable, as determined by fresh...... density, and thus the easiest to place. The addition of a high-range water reducing admixture and lightweight sand (expanded shale) for internal curing improved the 28-day compressive strength and splitting tensile strength. The coarse aggregate gradation had a large influence on permeability; however......, all tested permeabilities were high enough to drain the rain from a 100-year rain event in Denmark. The air entraining agent dosage used was not sufficiently high to create the necessary protective air content in the cement paste, and the freeze-thaw durability of the specimens were generally poor...

  8. Effect of freezing method and frozen storage duration on instrumental quality of lamb throughout display.

    Science.gov (United States)

    Muela, E; Sañudo, C; Campo, M M; Medel, I; Beltrán, J A

    2010-04-01

    This study evaluated the effect of freezing method (FM) (air blast freezer, freezing tunnel, or nitrogen chamber) and frozen storage duration (FSD) (1, 3, or 6 months) on the instrumental measurements of quality of thawed lamb, aged for a total of 72 h, throughout a 10-d display period, compared to the quality of fresh meat. pH, colour, lipid oxidation, thawing, and cooking losses in Longissimus thoracis and lumborum muscle, were determined following standard methods. FM affected yellowness, FSD redness and thawing losses, and both affected oxidation (increased as freezing rate decreased and/or as storage duration increased). When compared with fresh meat, the main differences appeared on oxidation (where a significant interaction between treatment (3FM x 3FSD + fresh meat) with display duration was detected), and on total losses (thaw + cook losses). Oxidation was lower in fresh meat, but values were not significantly different from those stored frozen for 1 month. Fresh meat had smaller total losses than did thawed meat, but losses were not significantly different from meat frozen in the freezing tunnel and stored frozen for 1 month. Display duration had a greater effect on instrumental quality parameters than did FM or FSD. pH, b*, and oxidation increased, and L* and a* decreased with an increase in the number of days on display. In conclusion, neither freezing method nor frozen storage up to 6 months influenced extensively the properties of lamb when instrumental measurements of quality were measured in meat that had been displayed for 1d after thawing. The small deterioration shown in this study should not give consumers concerns about frozen meat. 2009 Elsevier Ltd. All rights reserved.

  9. High-resolution Mapping of Permafrost and Soil Freeze/thaw Dynamics in the Tibetan Plateau Based on Multi-sensor Satellite Observations

    Science.gov (United States)

    Zhang, W.; Yi, Y.; Yang, K.; Kimball, J. S.

    2016-12-01

    The Tibetan Plateau (TP) is underlain by the world's largest extent of alpine permafrost ( 2.5×106 km2), dominated by sporadic and discontinuous permafrost with strong sensitivity to climate warming. Detailed permafrost distributions and patterns in most of the TP region are still unknown due to extremely sparse in-situ observations in this region characterized by heterogeneous land cover and large temporal dynamics in surface soil moisture conditions. Therefore, satellite-based temperature and moisture observations are essential for high-resolution mapping of permafrost distribution and soil active layer changes in the TP region. In this study, we quantify the TP regional permafrost distribution at 1-km resolution using a detailed satellite data-driven soil thermal process model (GIPL2). The soil thermal model is calibrated and validated using in-situ soil temperature/moisture observations from the CAMP/Tibet field campaign (9 sites: 0-300 cm soil depth sampling from 1997-2007), a multi-scale soil moisture and temperature monitoring network in the central TP (CTP-SMTMN, 57 sites: 5-40 cm, 2010-2014) and across the whole plateau (China Meteorology Administration, 98 sites: 0-320 cm, 2000-2015). Our preliminary results using the CAMP/Tibet and CTP-SMTMN network observations indicate strong controls of surface thermal and soil moisture conditions on soil freeze/thaw dynamics, which vary greatly with underlying topography, soil texture and vegetation cover. For regional mapping of soil freeze/thaw and permafrost dynamics, we use the most recent soil moisture retrievals from the NASA SMAP (Soil Moisture Active Passive) sensor to account for the effects of temporal soil moisture dynamics on soil thermal heat transfer, with surface thermal conditions defined by MODIS (Moderate Resolution Imaging Spectroradiometer) land surface temperature records. Our study provides the first 1-km map of spatial patterns and recent changes of permafrost conditions in the TP.

  10. Optimized Production of Vanillin from Green Vanilla Pods by Enzyme-Assisted Extraction Combined with Pre-Freezing and Thawing

    Directory of Open Access Journals (Sweden)

    Yanjun Zhang

    2014-02-01

    Full Text Available Production of vanillin from natural green vanilla pods was carried out by enzyme-assisted extraction combined with pre-freezing and thawing. In the first step the green vanilla pods were pre-frozen and then thawed to destroy cellular compartmentation. In the second step pectinase from Aspergillus niger was used to hydrolyze the pectin between the glucovanillin substrate and β-glucosidase. Four main variables, including enzyme amount, reaction temperature, time and pH, which were of significance for the vanillin content were studied and a central composite design (CCD based on the results of a single-factor tests was used. Response surface methodology based on CCD was employed to optimize the combination of enzyme amount, reaction temperature, time, and pH for maximum vanillin production. This resulted in the optimal condition in regards of the enzyme amount, reaction temperature, time, and pH at 84.2 mg, 49.5 °C, 7.1 h, and 4.2, respectively. Under the optimal condition, the experimental yield of vanillin was 4.63% ± 0.11% (dwb, which was in good agreement with the value predicted by the model. Compared to the traditional curing process (1.98% and viscozyme extract (2.36%, the optimized method for the vanillin production significantly increased the yield by 133.85% and 96%, respectively.

  11. Directional freezing for the cryopreservation of adherent mammalian cells on a substrate

    Science.gov (United States)

    Braslavsky, Ido

    2018-01-01

    Successfully cryopreserving cells adhered to a substrate would facilitate the growth of a vital confluent cell culture after thawing while dramatically shortening the post-thaw culturing time. Herein we propose a controlled slow cooling method combining initial directional freezing followed by gradual cooling down to -80°C for robust preservation of cell monolayers adherent to a substrate. Using computer controlled cryostages we examined the effect of cooling rates and dimethylsulfoxide (DMSO) concentration on cell survival and established an optimal cryopreservation protocol. Experimental results show the highest post-thawing viability for directional ice growth at a speed of 30 μm/sec (equivalent to freezing rate of 3.8°C/min), followed by gradual cooling of the sample with decreasing rate of 0.5°C/min. Efficient cryopreservation of three widely used epithelial cell lines: IEC-18, HeLa, and Caco-2, provides proof-of-concept support for this new freezing protocol applied to adherent cells. This method is highly reproducible, significantly increases the post-thaw cell viability and can be readily applied for cryopreservation of cellular cultures in microfluidic devices. PMID:29447224

  12. Effects of freezing conditions on quality changes in blueberries.

    Science.gov (United States)

    Cao, Xuehui; Zhang, Fangfang; Zhao, Dongyu; Zhu, Danshi; Li, Jianrong

    2018-03-12

    Freezing preservation is one of the most effective methods used to maintain the flavour and nutritional value of fruit. This research studied the effects of different freezing conditions, -20 °C, -40 °C, -80 °C, and immersion in liquid nitrogen, on quality changes of freeze-thawed blueberries. The water distribution estimates of blueberries were measured based on low-field nuclear magnetic resonance (LF-NMR) analysis. The pectin content, drip loss, and fruit texture were also detected to evaluate quality changes in samples. The freezing curves of blueberry showed super-cooling points at -20 °C and - 40 °C, whereas super-cooling points were not observed at -80 °C or in liquid nitrogen. After freeze-thaw treatment, the relaxation time of the cell wall water (T 21 ), cytoplasm water and extracellular space (T 22 ), and vacuole water (T 23 ) were significantly shortened compared to fresh samples, which suggested a lower liquidity. Although the freezing speed for samples immersed in liquid nitrogen was faster than other treatments, samples treated at -80 °C showed better quality regarding vacuole water holding, drip loss, and original pectin content retention. This study contributed to understanding how freezing temperature affects the qualities of blueberries. The super-fast freezing rate might injure fruit, and an appropriate freezing rate could better preserve blueberries. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  13. Differentiation of fresh and frozen-thawed fish samples using Raman spectroscopy coupled with chemometric analysis.

    Science.gov (United States)

    Velioğlu, Hasan Murat; Temiz, Havva Tümay; Boyaci, Ismail Hakki

    2015-04-01

    The potential of Raman spectroscopy was investigated in terms of its capability to discriminate the species of the fish samples and determine their freshness according to the number of freezing/thawing cycles they exposed. Species discrimination analysis was carried out on sixty-four fish samples from six different species, namely horse mackerel (Trachurus trachurus), European anchovy (Engraulis encrasicolus), red mullet (Mullus surmuletus), Bluefish (Pomatamus saltatrix), Atlantic salmon (Salmo salar) and flying gurnard (Trigla lucerna). Afterwards, fish samples were exposed to different numbers of freezing/thawing cycles and separated into three batches, namely (i) fresh, (ii) once frozen-thawed (OF) and (iii) twice frozen-thawed (TF) samples, in order to perform the freshness analysis. Raman data collected were used as inputs for chemometric analysis, which enabled us to develop two main PCA models to successfully terminate the studies for both species discrimination and freshness determination analysis. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. A longer climate memory carried by soil freeze–thaw processes in Siberia

    International Nuclear Information System (INIS)

    Matsumura, Shinji; Yamazaki, Koji

    2012-01-01

    The climate memory of a land surface generally persists for only a few months, but analysis of surface meteorological data revealed a longer-term climate memory carried by soil freeze–thaw processes in Siberia. Surface temperature variability during the snowmelt season corresponds reasonably well with that in the summer of the following year, when most stations show a secondary autocorrelation peak. The surface temperature memory is thought to be stored as variations in the amount of snowmelt water held in the soil, and through soil freezing, which emerges as latent heat variations in the near-surface atmosphere during soil thawing approximately one year later. The ground conditions are dry in the longer-term climate memory regions, such as eastern Siberia, where less snow cover (higher surface air temperature) in spring results in less snowmelt water or lower soil moisture in the summer. Consequently, through soil freezing, it will require less latent heat to thaw in the summer of the following year, resulting in higher surface air temperature. In addition to soil moisture and snow cover, soil freeze–thaw processes can also act as agents of climate memory in the near-surface atmosphere. (letter)

  15. Effect of freeze-thaw cycling on grain size of biochar.

    Science.gov (United States)

    Liu, Zuolin; Dugan, Brandon; Masiello, Caroline A; Wahab, Leila M; Gonnermann, Helge M; Nittrouer, Jeffrey A

    2018-01-01

    Biochar may improve soil hydrology by altering soil porosity, density, hydraulic conductivity, and water-holding capacity. These properties are associated with the grain size distributions of both soil and biochar, and therefore may change as biochar weathers. Here we report how freeze-thaw (F-T) cycling impacts the grain size of pine, mesquite, miscanthus, and sewage waste biochars under two drainage conditions: undrained (all biochars) and a gravity-drained experiment (mesquite biochar only). In the undrained experiment plant biochars showed a decrease in median grain size and a change in grain-size distribution consistent with the flaking off of thin layers from the biochar surface. Biochar grain size distribution changed from unimodal to bimodal, with lower peaks and wider distributions. For plant biochars the median grain size decreased by up to 45.8% and the grain aspect ratio increased by up to 22.4% after 20 F-T cycles. F-T cycling did not change the grain size or aspect ratio of sewage waste biochar. We also observed changes in the skeletal density of biochars (maximum increase of 1.3%), envelope density (maximum decrease of 12.2%), and intraporosity (porosity inside particles, maximum increase of 3.2%). In the drained experiment, mesquite biochar exhibited a decrease of median grain size (up to 4.2%) and no change of aspect ratio after 10 F-T cycles. We also document a positive relationship between grain size decrease and initial water content, suggesting that, biochar properties that increase water content, like high intraporosity and pore connectivity large intrapores, and hydrophilicity, combined with undrained conditions and frequent F-T cycles may increase biochar breakdown. The observed changes in biochar particle size and shape can be expected to alter hydrologic properties, and thus may impact both plant growth and the hydrologic cycle.

  16. Effect of freezing temperature on the color of frozen salmon.

    Science.gov (United States)

    Ottestad, Silje; Enersen, Grethe; Wold, Jens Petter

    2011-09-01

    New freezing methods developed with the purpose of improved product quality after thawing can sometimes be difficult to get accepted in the market. The reason for this is the formation of ice crystals that can give the product a temporary color loss and make it less appealing. We have here used microscopy to study ice crystal size as a function of freezing temperature by investigating the voids in the cell tissue left by the ice crystals. We have also investigated how freezing temperature affects the color and the visible absorption spectra of frozen salmon. Freezing temperatures previously determined to be the best for quality after thawing (-40 to -60 °C) were found to cause a substantial loss in perceived color intensity during frozen state. This illustrated the conflict between optimal freezing temperatures with respect to quality after thawing against visual appearance during frozen state. Low freezing temperatures gave many small ice crystals, increased light scattering and an increased absorption level for all wavelengths in the visible region. Increased astaxanthin concentration on the other hand would give higher absorption at 490 nm. The results showed a clear potential of using visible interactance spectroscopy to differentiate between poor product coloration due to lack of pigmentation and temporary color loss due to light scattering by ice crystal. This type of measurements could be a useful tool in the development of new freezing methods and to monitor ice crystal growth during frozen storage. It could also potentially be used by the industry to prove good product quality. In this article we have shown that freezing food products at intermediate to low temperatures (-40 to -80 °C) can result in paler color during frozen state, which could affect consumer acceptance. We have also presented a spectroscopic method that can separate between poor product color and temporary color loss due to freezing. © 2011 Institute of Food Technologists®

  17. Patterns of nitrogen export from a seasonal freezing agricultural watershed during the thawing period.

    Science.gov (United States)

    Zhao, Qiang; Chang, Dan; Wang, Kang; Huang, Jiesheng

    2017-12-01

    The objectives of this study were to investigate water, ammonium nitrogen (NH 4 + -N), and nitrate nitrogen (NO 3 - -N) export processes during the thawing period in a watershed with heavy agricultural activities and to evaluate contributions of N (i.e., NO 3 - -N and NH 4 + -N) from different source areas under different climate conditions. Experiments were conducted within the 75km 2 agricultural Heidingzi watershed in northeast China. The thawing period was divided into four stages: early-melt, late-melt, rain-on-melt, and post-melt. Drainage regions (DRs) were separated into three types. The processes of water and N discharge from soil into rivers were monitored in these DRs during the thawing periods of 2014, 2015, and 2016. Results show that the processes of water and N discharge were not synchronous during the thawing period. Variations in discharge concentrations of NH 4 + -N and NO 3 - -N during the thawing period were mainly affected by the flushing effect, which was controlled by the physical state of the surface water (snow or ice) and the melt rate of frozen soil. Contributions of N export from the DRs varied under different land uses and climate conditions during the thawing period. NO 3 - -N export was mainly from maize fields. Thawing stages with high NO 3 - -N export were always accompanied by higher discharge rates. NH 4 + -N export mainly occurred during the early-melt and late-melt stages and from riverside rural regions. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Effects of straw mulching on soil evaporation during the soil thawing ...

    Indian Academy of Sciences (India)

    26

    Keywords: straw mulching, soil water evaporation, soil thawing period, freezing depth, soil liquid water .... moisture and the soil water evaporation process. The Songnen Plain ...... soils on soil infiltration and evaporation: Water Sci. Technol.

  19. Identification and expression analysis of cold and freezing stress responsive genes of Brassica oleracea.

    Science.gov (United States)

    Ahmed, Nasar Uddin; Jung, Hee-Jeong; Park, Jong-In; Cho, Yong-Gu; Hur, Yoonkang; Nou, Ill-Sup

    2015-01-10

    Cold and freezing stress is a major environmental constraint to the production of Brassica crops. Enhancement of tolerance by exploiting cold and freezing tolerance related genes offers the most efficient approach to address this problem. Cold-induced transcriptional profiling is a promising approach to the identification of potential genes related to cold and freezing stress tolerance. In this study, 99 highly expressed genes were identified from a whole genome microarray dataset of Brassica rapa. Blast search analysis of the Brassica oleracea database revealed the corresponding homologous genes. To validate their expression, pre-selected cold tolerant and susceptible cabbage lines were analyzed. Out of 99 BoCRGs, 43 were differentially expressed in response to varying degrees of cold and freezing stress in the contrasting cabbage lines. Among the differentially expressed genes, 18 were highly up-regulated in the tolerant lines, which is consistent with their microarray expression. Additionally, 12 BoCRGs were expressed differentially after cold stress treatment in two contrasting cabbage lines, and BoCRG54, 56, 59, 62, 70, 72 and 99 were predicted to be involved in cold regulatory pathways. Taken together, the cold-responsive genes identified in this study provide additional direction for elucidating the regulatory network of low temperature stress tolerance and developing cold and freezing stress resistant Brassica crops. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Effects of reduced glutathione on acrosin activity in frozen-thawed boar spermatozoa.

    Science.gov (United States)

    Estrada, Efrén; Rodríguez-Gil, Joan E; Rivera Del Álamo, Maria M; Peña, Alejandro; Yeste, Marc

    2017-02-01

    In pigs, acrosin activity in extended semen is correlated with reproductive performance and has recently been identified as a freezability marker. Reduced glutathione (GSH) is known to decrease sperm cryodamage and increase the reproductive performance of frozen-thawed boar spermatozoa. However, the effects of GSH on the acrosin activity of good and poor freezability ejaculates (GFE and PFE, respectively) is yet to be examined. The present study investigated how supplementing cryopreservation media with GSH affected acrosin activity in GFE and PFE, as well as the relationship between acrosin activity and reproductive performance in frozen-thawed boar spermatozoa. In addition, we examined whether the increase in fertility rates and litter sizes observed after the addition of 2mM GSH to cryopreservation extenders was related to acrosin activity. Supplementing freezing media with 2mM GSH partially counteracted the cryopreservation-related decrease in acrosin activity in GFE but not PFE. Acrosin activity was found to be significantly correlated with in vivo reproductive performance of frozen-thawed boar semen. In conclusion, the effects of adding GSH to freezing extenders on the acrosin activity of frozen-thawed boar spermatozoa rely on the intrinsic freezability of the ejaculate. Furthermore, the maintenance of proper acrosin activity could contribute to the increase in reproductive performance mediated by GSH.

  1. Deep-freezing of boar semen in plastic film 'cochettes'.

    Science.gov (United States)

    Eriksson, B M; Rodriguez-Martinez, H

    2000-03-01

    The motility and membrane integrity of spermatozoa from nine boars frozen with a programmable freezing machine in plastic bags, 'cochettes', and in 'maxi-straws', in total doses of 5 x 10(9) spermatozoa/5 ml with glycerol (3%) used as cryoprotectant, were assessed after thawing. A computer-based cell motion analyser was used to evaluate sperm motility, while the integrity of the plasmalemma was assessed with fluorescent supravital dyes (C-FDA/PI). The fertilizing capacity of the semen frozen in the two containers was investigated by inseminating (AI) gilts. Pregnancy was monitored by Doppler-ultrasound, and the numbers of corpora lutea and viable embryos counted at slaughter, between days 30 and 38 after AI. The cochettes sustained the overall procedure of freezing/thawing (FT), with 30 min post-thaw (PT) sperm motility being significantly higher than for straws, 46.9 vs. 39.5%. The only significant difference in motility patterns detected when comparing the packages was a higher sperm velocity (VCL) in cochettes at 30 min PT. However, percentages of FT-spermatozoa with intact membranes, detected with the supravital probes, were higher in maxi-straws than in cochettes, 46.8 vs. 43.0% (P straws and those frozen in cochettes. The results indicate that although the deep-freezing of AI-doses of boar semen in large plastic bags is feasible, problems such as their inconvenient size for storage and inconsistent thawing must be solved before this type of container can be used for the commercial cryopreservation of boar semen.

  2. Evaluation of the freeze-thaw/evaporation process for the treatment of produced waters. Final report, August 1992--August 1996

    Energy Technology Data Exchange (ETDEWEB)

    Boysen, J.E.; Walker, K.L.; Mefford, J.L.; Kirsch, J.R. [Resource Technology Corp., Laramie, WY (United States); Harju, J.A. [North Dakota Univ., Grand Forks, ND (United States). Energy and Environmental Research Center

    1996-06-01

    The use of freeze-crystallization is becoming increasingly acknowledged as a low-cost, energy-efficient method for purifying contaminated water. The natural freezing process can be coupled with natural evaporative processes to treat oil and gas produced waters year round in regions where subfreezing temperatures seasonally occur. The climates typical of Colorado`s San Juan Basin and eastern slope, as well as the oil and gas producing regions of Wyoming, are well suited for application of these processes in combination. Specifically, the objectives of this research are related to the development of a commercially-economic FTE (freeze-thaw/evaporation) process for the treatment and purification of water produced in conjunction with oil and natural gas. The research required for development of this process consists of three tasks: (1) a literature survey and process modeling and economic analysis; (2) laboratory-scale process evaluation; and (3) field demonstration of the process. Results of research conducted for the completion of these three tasks indicate that produced water treatment and disposal costs for commercial application of the process, would be in the range of $0.20 to $0.30/bbl in the Rocky Mountain region. FTE field demonstration results from northwestern New Mexico during the winter of 1995--96 indicate significant and simultaneous removal of salts, metals, and organics from produced water. Despite the unusually warm winter, process yields demonstrate disposal volume reductions on the order of 80% and confirm the potential for economic production of water suitable for various beneficial uses. The total dissolved solids concentrations of the FTE demonstration streams were 11,600 mg/L (feed), 56,900 mg/L (brine), and 940 mg/L (ice melt).

  3. Biophysical characterization of the Lactobacillus delbrueckii subsp. bulgaricus membrane during cold and osmotic stress and its relevance for cryopreservation.

    Science.gov (United States)

    Meneghel, Julie; Passot, Stéphanie; Dupont, Sébastien; Fonseca, Fernanda

    2017-02-01

    Freezing lactic acid bacteria often leads to cell death and loss of technological properties. Our objective was to provide an in-depth characterization of the biophysical properties of the Lactobacillus delbrueckii subsp. bulgaricus membrane in relation to its freeze resistance. Freezing was represented as a combination of cold and osmotic stress. This work investigated the relative incidence of increasing sucrose concentrations coupled or not with subzero temperatures without ice nucleation on the biological and biophysical responses of two strains with different membrane fatty acid compositions and freeze resistances. Following exposure of bacterial cells to the highest sucrose concentration, the sensitive strain exhibited a survival rate of less than 10 % and 5 h of acidifying activity loss. Similar biological activity losses were observed upon freeze-thawing and after osmotic treatment for each strain thus highlighting osmotic stress as the main source of cryoinjury. The direct measurement of membrane fluidity by fluorescence anisotropy was linked to membrane lipid organization characterized by FTIR spectroscopy. Both approaches made it possible to investigate the specific contributions of the membrane core and the bilayer external surface to cell degradation caused by cold and osmotic stress. Cold-induced membrane rigidification had no significant implication on bacterial freeze-thaw resistance. Interactions between extracellular sucrose and membrane phospholipid headgroups under osmotic stress were also observed. Such interactions were more evident in the sensitive strain and when increasing sucrose concentration, thus suggesting membrane permeabilization. The relevance of biophysical properties for elucidating mechanisms of cryoinjury and cryoprotection is discussed.

  4. NaCl stress impact on the key enzymes in glycolysis from Lactobacillus bulgaricus during freeze-drying.

    Science.gov (United States)

    Li, Chun; Sun, Jinwei; Qi, Xiaoxi; Liu, Libo

    2015-01-01

    The viability of Lactobacillus bulgaricus in freeze-drying is of significant commercial interest to dairy industries. In the study, L.bulgaricus demonstrated a significantly improved (p enzymes in glycolysis during 2% NaCl stress were studied. NaCl stress significantly enhanced (p enzymes (phosphofructokinase, pyruvate kinase, and lactate dehydrogenase) decreased during freeze-drying, and NaCl stress were found to improve activities of these enzymes before and after freeze-drying. However, a transcriptional analysis of the corresponding genes suggested that the effect of NaCl stress on the expression of the pfk2 gene was not obvious. The increased survival of freeze-dried cells of L. bulgaricus under NaCl stress might be due to changes in only the activity or translation level of these enzymes in different environmental conditions but have no relation to their mRNA transcription level.

  5. Long-term stability of morphine hydrochloride in 0.9% NaCl infusion polyolefin bags after freeze-thaw treatment and in polypropylene syringes at 5 degrees C + 3 degrees C.

    Science.gov (United States)

    Hecq, J-D; Godet, M; Gillet, P; Jamart, J; Galanti, L

    2014-01-01

    The aim of this study was to investigate the long-term stability of morphine hydrochloride in 0.9% NaCI infusion polyolefin bags and polypropylene syringes after storage at 5 degrees C + 3 degrees C and to evaluate the influence of initial freezing and microwave thawing on this stability. Ten polyolefin bags and five polypropylene syringes containing 100 mL of 1 mg/mL of morphine hydrochloride solution in 0.9% NaCI were prepared under aseptic conditions. Five polyolefin bags were frozen at -20 degrees C for 90 days before storage. Immediately after the preparation and after thawing, 2 mL of each bag were withdrawn for the initial concentration measurements. All polyolefin bags and polypropylene syringes were then refrigerated at 5 degrees C + 3 degrees C for 58 days during which the morphine concentrations were measured periodically by high-performance liquid chromatography using a reversed-phase column, naloxone as internal standard, a mobile phase consisting of 5% acetonitrile and 95% of KH2PO4 buffer (pH 3.50), and detection with diode array detector at 254 nm. Visual and microscopic observations and spectrophotometric and pH measurements were also performed. Solutions were considered stable if the concentration remained superior to 90% of the initial concentration. The degradation products peaks were not quantitatively significant and were resolved from the native drug. Polyolefin bag and polypropylene syringe solutions were stable when stored at 5 degrees C + 3 degrees C during these 58 days. No color change or precipitation in the solutions was observed. The physical stability was confirmed by visual, microscopic, and spectrophotometric inspection. There was no significant change in pH during storage. Freezing and microwave thawing didn't influence the infusion stability. Morphine hydrochloride infusions may be prepared in advance by centralized intravenous additive service, frozen in polyolefin bags, and microwave thawed before storage under refrigeration

  6. The effect of two packaging systems on the post-thaw characteristics of canine sperm.

    Science.gov (United States)

    Strzeżek, R; Polakiewicz, P; Kordan, W

    2015-01-01

    The aim of this study was to compare the effect of different packaging systems on some parameters of cryopreserved canine spermatozoa. The experimental material consisted of the sperm-rich fractions of ejaculates collected from four Beagle dogs. Semen samples for cryopreservation were stored in 0.25 ml plastic straws and two aluminum tubes with a total volume of 5.0 ml. Semen was frozen in static nitrogen vapor for 10 minutes (0.25 ml straws) or 15 and 20 minutes (aluminum tubes). Post-thaw assessments involved the determination of sperm motility parameters using a computer assisted sperm analyzer (CASA), sperm plasma membrane integrity (SPMI), mitochondrial membrane potential (MMP) and acrosome integrity (normal apical ridge, NAR). Regardless of the packaging system applied, no significant differences in total sperm motility (TMOT) or selected kinematic parameters were observed after freezing-thawing. However, spermatozoa frozen in 0.25 mL straws were characterized by improved functionality, in particular mitochondrial function, after thawing. The results indicate that large quantities of canine semen can be frozen in aluminum tubes. Further studies are required, however, to evaluate different freezing and thawing rates of aluminum tubes.

  7. A Multi-Environment Thermal Control System With Freeze-Tolerant Radiator

    Science.gov (United States)

    Chen, Weibo; Fogg, David; Mancini, Nick; Steele, John; Quinn, Gregory; Bue, Grant; Littibridge, Sean

    2013-01-01

    Future space exploration missions require advanced thermal control systems (TCS) to dissipate heat from spacecraft, rovers, or habitats operating in environments that can vary from extremely hot to extremely cold. A lightweight, reliable TCS is being developed to effectively control cabin and equipment temperatures under widely varying heat loads and ambient temperatures. The system uses freeze-tolerant radiators, which eliminate the need for a secondary circulation loop or heat pipe systems. Each radiator has a self-regulating variable thermal conductance to its ambient environment. The TCS uses a nontoxic, water-based working fluid that is compatible with existing lightweight aluminum heat exchangers. The TCS is lightweight, compact, and requires very little pumping power. The critical characteristics of the core enabling technologies were demonstrated. Functional testing with condenser tubes demonstrated the key operating characteristics required for a reliable, freeze-tolerant TCS, namely (1) self-regulating thermal conductance with short transient responses to varying thermal loads, (2) repeatable performance through freeze-thaw cycles, and (3) fast start-up from a fully frozen state. Preliminary coolant tests demonstrated that the corrosion inhibitor in the water-based coolant can reduce the corrosion rate on aluminum by an order of magnitude. Performance comparison with state-of-the-art designs shows significant mass and power saving benefits of this technology.

  8. Effect of green tea (Camellia sinensis) extract and pre-freezing equilibration time on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender.

    Science.gov (United States)

    Mehdipour, Mahdieh; Daghigh Kia, Hossein; Najafi, Abouzar; Vaseghi Dodaran, Hossein; García-Álvarez, Olga

    2016-12-01

    The aim of this study was to determine the effect of Camellia sinensis extract as antioxidant supplement and pre-freezing equilibration times in a soybean lecithin extender for freezing ram semen. In this study, a total of 20 ejaculates were collected from four Ghezel rams and diluted with extenders (1.5% soybean lecithin, 7% glycerol) containing no supplements (control) and Camellia sinensis extract (5, 10, and 15 mg/L) and cryopreserved, immediately after thermal equilibrium was reached at 5 °C (0 h), or 4 h after equilibration. Sperm motility characteristics, membrane integrity, abnormal morphology, mitochondria activity, apoptotic status, MDA and antioxidant activities (GPx, SOD and total antioxidant capacity (TAC)) were evaluated following freeze-thawing. Camellia sinensis extract at level 10 mg/L led to the highest total and progressive motilities percentages, in comparison to other treatments (P extract at level of 5 and 10 mg/L led to higher plasma membrane integrity, mitochondria activity and Total antioxidant capacity (TAC) in comparison to the level of 15 mg/L and control group (P extract at 10 mg/L level produced the highest percentage of live spermatozoa and the lowest apoptotic spermatozoa in comparison to all treatments (P concentration, 10 mg/L, compared to all treatments (P  0.05) were observed between equilibration times (0 h vs. 4 h) for sperm samples incubated with or without different concentrations of Camellia sinensis extract. In conclusion, addition of Camellia sinensis extract at level of 10 mg/L can improve post-thawing quality of ram semen cryopreserved in a soybean lecithin extender. However, further research is needed to standardize the process of Camellia sinensis extraction and specially for identifying which compounds are responsible of its beneficial effect on ram sperm cryopreservation. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Simulation of Soil Frost and Thaw Fronts Dynamics with Community Land Model 4.5

    Science.gov (United States)

    Gao, J.; Xie, Z.

    2016-12-01

    Freeze-thaw processes in soils, including changes in frost and thaw fronts (FTFs) , are important physical processes. The movement of FTFs affects soil water and thermal characteristics, as well as energy and water exchanges between land surface and the atmosphere, and then the land surface hydrothermal process. In this study, a two-directional freeze and thaw algorithm for simulating FTFs is incorporated into the community land surface model CLM4.5, which is called CLM4.5-FTF. The simulated FTFs depth and soil temperature of CLM4.5-FTF compared well with the observed data both in D66 station (permafrost) and Hulugou station (seasonally frozen soil). Because the soil temperature profile within a soil layer can be estimated according to the position of FTFs, CLM4.5 performed better in soil temperature simulation. Permafrost and seasonally frozen ground conditions in China from 1980 to 2010 were simulated using the CLM4.5-FTF. Numerical experiments show that the spatial distribution of simulated maximum frost depth by CLM4.5-FTF has seasonal variation obviously. Significant positive active-layer depth trends for permafrost regions and negative maximum freezing depth trends for seasonal frozen soil regions are simulated in response to positive air temperature trends except west of Black Sea.

  10. Comparative study of deterioration procedure in chemical-leavened steamed bread dough under frozen storage and freeze/thaw condition.

    Science.gov (United States)

    Wang, Pei; Yang, Runqiang; Gu, Zhenxin; Xu, Xueming; Jin, Zhengyu

    2017-08-15

    Successive freeze/thaw (FT) cycle was a widely used empirical approach to shorten the experimental period since it could accelerate frozen dough deterioration compared with frozen storage (FS). In order to compare the effect of FS and FT cycle on deterioration procedure of chemical-leavened steamed bread dough, kinetic studies of bread quality indices were performed and the relationships between bread quality and dough components were further established. Results showed that degradation of steamed bread loaf volume and firmness followed first-order kinetics during FS and zero-order kinetics during FT, respectively. Glutenin macropolymers (GMP) depolymerization and dough weight loss occurred steadily throughout FS and FT. Significant enhancement of damaged starch and crystallinity were observed at the later FS period and FT cycle. Multiple regression study led to the conclusion that dough weight loss contributed the most to the reduced bread loaf volume under FS whereas GMP depolymerization dominated under FT condition. Copyright © 2017 Elsevier Ltd. All rights reserved.

  11. Effect of reduced glutathione supplementation in semen extender on tyrosine phosphorylation and apoptosis like changes in frozen thawed Hariana bull spermatozoa.

    Science.gov (United States)

    Shah, Nadeem; Singh, Vijay; Yadav, Hanuman Prasad; Verma, Meena; Chauhan, Dharmendra Singh; Saxena, Atul; Yadav, Sarvajeet; Swain, Dilip Kumar

    2017-07-01

    To provide new insights into the mechanisms through which reduced glutathione (GSH) is able to protect spermatozoa, we tested the hypothesis that cryocapacitation and apoptosis like changes can contribute to the negative effect of freezing and thawing on bull spermatozoa, and that GSH prevent this damage. Having known protective effects of GSH in terms of a potent antioxidant, we evaluated capacitation, tyrosine phosphorylation and apoptosis like changes in bull spermatozoa after freezing and thawing in egg yolk tris glycerol extender containing (0.5m M-GSH-T1 & 1mM GSH-T2) and without GSH serving as the control (C). Forty ejaculates were collected from four Hariana bulls and were pooled due to non significant variations among the bull ejaculates for the evaluation of sperm attributes. Capacitation like changes, tyrosine phosphorylation, localization of tyrosine phosphorylated proteins, apoptosis like changes in terms of mitochondrial transmembrane potential and DNA fragmentation after final dilution, 4h of equilibration at 4°C and 24h after freezing and thawing were evaluated. GSH supplementation at 0.5mM showed significant reduction in B- and AR- pattern spermatozoa during all stages of semen freezing and thawing. Immunoblot revealed six proteins which were tyrosine phosphorylated and protein of 30 and 75kDa (p30, p75) were the major tyrosine phosphorylted proteins. On further analysis, the p30 showed differential variation in intensity in all the three groups after freezing and thawing. Positive immune reactivity for tyrosine phosphorylated proteins was found in neck, middle piece and post-acrosomal regions of spermatozoa. Addition of 0.5mM GSH decreased percentage of spermatozoa showing fragmented DNA and increased the percentage of spermatozoa having high transmembrane mitochondrial potential (P<0.05). This study demonstrates that GSH favours survival of bull spermatozoa by interfering with apoptotic and cryocapacitation pathways, and thereby protects the

  12. Effects of repetitive freeze–thawing cycles on T2 and T2* of the Achilles tendon

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Eric Y., E-mail: ericchangmd@gmail.com [Department of Radiology, VA San Diego Healthcare System, 3350 La Jolla Village Drive, San Diego, CA 92161 (United States); Department of Radiology, University of California, 200 West Arbor St., San Diego, CA 92103 (United States); Bae, Won C., E-mail: wbae@ucsd.edu [Department of Radiology, University of California, 200 West Arbor St., San Diego, CA 92103 (United States); Statum, Sheronda, E-mail: sherondastatum@msn.com [Department of Radiology, University of California, 200 West Arbor St., San Diego, CA 92103 (United States); Du, Jiang, E-mail: jiangdu@ucsd.edu [Department of Radiology, University of California, 200 West Arbor St., San Diego, CA 92103 (United States); Chung, Christine B., E-mail: cbchung@ucsd.edu [Department of Radiology, University of California, 200 West Arbor St., San Diego, CA 92103 (United States); Department of Radiology, VA San Diego Healthcare System, 3350 La Jolla Village Drive, San Diego, CA 92161 (United States)

    2014-02-15

    Introduction: In this study we sought to evaluate the effects of multiple freezing and thawing cycles on two MR parameters to study Achilles tendon, T2 and T2*. Materials and methods: Four fresh Achilles tendons were imaged on a 3T clinical scanner and again after 1, 2, 4, and 5 freeze–thaw cycles with spin-echo (SE) and ultrashort echo time (UTE) sequences. Regions of interest were manually drawn over the entire Achilles tendon and mono-exponential curves were used to determine T2 and T2* relaxation times. Results: There was no statistically significant difference in mean T2 or T2* values between the fresh specimens and after subsequent cycles of freeze–thaw treatment (p > 0.1). Linear regression between SE T2 values at baseline and after successive freeze–thaw cycles demonstrated moderate agreement (r = 0.60) whereas UTE T2* values at baseline and after successive-freeze thaw cycles demonstrated strong agreement (r = 0.92). Conclusion: These findings suggest that changes between specimens seen in vitro are due to factors other than frozen storage. Furthermore, our results suggest that there is stronger agreement between baseline (fresh) and successive freeze–thaw T2* values of tendon obtained with the UTE technique in comparison to T2 values obtained with a conventional clinical CPMG technique.

  13. Effects of repetitive freeze–thawing cycles on T2 and T2* of the Achilles tendon

    International Nuclear Information System (INIS)

    Chang, Eric Y.; Bae, Won C.; Statum, Sheronda; Du, Jiang; Chung, Christine B.

    2014-01-01

    Introduction: In this study we sought to evaluate the effects of multiple freezing and thawing cycles on two MR parameters to study Achilles tendon, T2 and T2*. Materials and methods: Four fresh Achilles tendons were imaged on a 3T clinical scanner and again after 1, 2, 4, and 5 freeze–thaw cycles with spin-echo (SE) and ultrashort echo time (UTE) sequences. Regions of interest were manually drawn over the entire Achilles tendon and mono-exponential curves were used to determine T2 and T2* relaxation times. Results: There was no statistically significant difference in mean T2 or T2* values between the fresh specimens and after subsequent cycles of freeze–thaw treatment (p > 0.1). Linear regression between SE T2 values at baseline and after successive freeze–thaw cycles demonstrated moderate agreement (r = 0.60) whereas UTE T2* values at baseline and after successive-freeze thaw cycles demonstrated strong agreement (r = 0.92). Conclusion: These findings suggest that changes between specimens seen in vitro are due to factors other than frozen storage. Furthermore, our results suggest that there is stronger agreement between baseline (fresh) and successive freeze–thaw T2* values of tendon obtained with the UTE technique in comparison to T2 values obtained with a conventional clinical CPMG technique

  14. Freeze and Thaw of CD4+CD25+Foxp3+ Regulatory T Cells Results in Loss of CD62L Expression and a Reduced Capacity to Protect against Graft-versus-Host Disease.

    Directory of Open Access Journals (Sweden)

    Mareike Florek

    Full Text Available The adoptive transfer of CD4+CD25+Foxp3+ regulatory T cells (Tregs in murine models of allogeneic hematopoietic cell transplantation (HCT has been shown to protect recipient mice from lethal acute graft-versus-host disease (GVHD and this approach is being actively investigated in human clinical trials. Here, we examined the effects of cryopreservation on Tregs. We found that freeze and thaw of murine and human Tregs is associated with reduced expression of L-selectin (CD62L, which was previously established to be an important factor that contributes to the in vivo protective effects of Tregs. Frozen and thawed murine Tregs showed a reduced capacity to bind to the CD62L binding partner MADCAM1 in vitro as well as an impaired homing to secondary lymphoid organs in vivo. Upon adoptive transfer frozen and thawed Tregs failed to protect against lethal GVHD compared with fresh Tregs in a murine model of allogeneic HCT across major histocompatibility barriers. In summary, the direct administration of adoptively transferred frozen and thawed Tregs adversely affects their immunosuppressive potential which is an important factor to consider in the clinical implementation of Treg immunotherapies.

  15. Enhancement of the proline and nitric oxide synthetic pathway improves fermentation ability under multiple baking-associated stress conditions in industrial baker's yeast.

    Science.gov (United States)

    Sasano, Yu; Haitani, Yutaka; Hashida, Keisuke; Ohtsu, Iwao; Shima, Jun; Takagi, Hiroshi

    2012-04-01

    During the bread-making process, industrial baker's yeast, mostly Saccharomyces cerevisiae, is exposed to baking-associated stresses, such as air-drying and freeze-thaw stress. These baking-associated stresses exert severe injury to yeast cells, mainly due to the generation of reactive oxygen species (ROS), leading to cell death and reduced fermentation ability. Thus, there is a great need for a baker's yeast strain with higher tolerance to baking-associated stresses. Recently, we revealed a novel antioxidative mechanism in a laboratory yeast strain that is involved in stress-induced nitric oxide (NO) synthesis from proline via proline oxidase Put1 and N-acetyltransferase Mpr1. We also found that expression of the proline-feedback inhibition-less sensitive mutant γ-glutamyl kinase (Pro1-I150T) and the thermostable mutant Mpr1-F65L resulted in an enhanced fermentation ability of baker's yeast in bread dough after freeze-thaw stress and air-drying stress, respectively. However, baker's yeast strains with high fermentation ability under multiple baking-associated stresses have not yet been developed. We constructed a self-cloned diploid baker's yeast strain with enhanced proline and NO synthesis by expressing Pro1-I150T and Mpr1-F65L in the presence of functional Put1. The engineered strain increased the intracellular NO level in response to air-drying stress, and the strain was tolerant not only to oxidative stress but also to both air-drying and freeze-thaw stresses probably due to the reduced intracellular ROS level. We also showed that the resultant strain retained higher leavening activity in bread dough after air-drying and freeze-thaw stress than that of the wild-type strain. On the other hand, enhanced stress tolerance and fermentation ability did not occur in the put1-deficient strain. This result suggests that NO is synthesized in baker's yeast from proline in response to oxidative stresses that induce ROS generation and that increased NO plays an important

  16. Enhancement of the proline and nitric oxide synthetic pathway improves fermentation ability under multiple baking-associated stress conditions in industrial baker's yeast

    Directory of Open Access Journals (Sweden)

    Sasano Yu

    2012-04-01

    Full Text Available Abstract Background During the bread-making process, industrial baker's yeast, mostly Saccharomyces cerevisiae, is exposed to baking-associated stresses, such as air-drying and freeze-thaw stress. These baking-associated stresses exert severe injury to yeast cells, mainly due to the generation of reactive oxygen species (ROS, leading to cell death and reduced fermentation ability. Thus, there is a great need for a baker's yeast strain with higher tolerance to baking-associated stresses. Recently, we revealed a novel antioxidative mechanism in a laboratory yeast strain that is involved in stress-induced nitric oxide (NO synthesis from proline via proline oxidase Put1 and N-acetyltransferase Mpr1. We also found that expression of the proline-feedback inhibition-less sensitive mutant γ-glutamyl kinase (Pro1-I150T and the thermostable mutant Mpr1-F65L resulted in an enhanced fermentation ability of baker's yeast in bread dough after freeze-thaw stress and air-drying stress, respectively. However, baker's yeast strains with high fermentation ability under multiple baking-associated stresses have not yet been developed. Results We constructed a self-cloned diploid baker's yeast strain with enhanced proline and NO synthesis by expressing Pro1-I150T and Mpr1-F65L in the presence of functional Put1. The engineered strain increased the intracellular NO level in response to air-drying stress, and the strain was tolerant not only to oxidative stress but also to both air-drying and freeze-thaw stresses probably due to the reduced intracellular ROS level. We also showed that the resultant strain retained higher leavening activity in bread dough after air-drying and freeze-thaw stress than that of the wild-type strain. On the other hand, enhanced stress tolerance and fermentation ability did not occur in the put1-deficient strain. This result suggests that NO is synthesized in baker's yeast from proline in response to oxidative stresses that induce ROS

  17. Enhancement of the proline and nitric oxide synthetic pathway improves fermentation ability under multiple baking-associated stress conditions in industrial baker's yeast

    Science.gov (United States)

    2012-01-01

    Background During the bread-making process, industrial baker's yeast, mostly Saccharomyces cerevisiae, is exposed to baking-associated stresses, such as air-drying and freeze-thaw stress. These baking-associated stresses exert severe injury to yeast cells, mainly due to the generation of reactive oxygen species (ROS), leading to cell death and reduced fermentation ability. Thus, there is a great need for a baker's yeast strain with higher tolerance to baking-associated stresses. Recently, we revealed a novel antioxidative mechanism in a laboratory yeast strain that is involved in stress-induced nitric oxide (NO) synthesis from proline via proline oxidase Put1 and N-acetyltransferase Mpr1. We also found that expression of the proline-feedback inhibition-less sensitive mutant γ-glutamyl kinase (Pro1-I150T) and the thermostable mutant Mpr1-F65L resulted in an enhanced fermentation ability of baker's yeast in bread dough after freeze-thaw stress and air-drying stress, respectively. However, baker's yeast strains with high fermentation ability under multiple baking-associated stresses have not yet been developed. Results We constructed a self-cloned diploid baker's yeast strain with enhanced proline and NO synthesis by expressing Pro1-I150T and Mpr1-F65L in the presence of functional Put1. The engineered strain increased the intracellular NO level in response to air-drying stress, and the strain was tolerant not only to oxidative stress but also to both air-drying and freeze-thaw stresses probably due to the reduced intracellular ROS level. We also showed that the resultant strain retained higher leavening activity in bread dough after air-drying and freeze-thaw stress than that of the wild-type strain. On the other hand, enhanced stress tolerance and fermentation ability did not occur in the put1-deficient strain. This result suggests that NO is synthesized in baker's yeast from proline in response to oxidative stresses that induce ROS generation and that increased NO

  18. Chilling- and Freezing-Induced Alterations in Cytosine Methylation and Its Association with the Cold Tolerance of an Alpine Subnival Plant, Chorispora bungeana.

    Directory of Open Access Journals (Sweden)

    Yuan Song

    Full Text Available Chilling (0-18°C and freezing (<0°C are two distinct types of cold stresses. Epigenetic regulation can play an important role in plant adaptation to abiotic stresses. However, it is not yet clear whether and how epigenetic modification (i.e., DNA methylation mediates the adaptation to cold stresses in nature (e.g., in alpine regions. Especially, whether the adaptation to chilling and freezing is involved in differential epigenetic regulations in plants is largely unknown. Chorispora bungeana is an alpine subnival plant that is distributed in the freeze-thaw tundra in Asia, where chilling and freezing frequently fluctuate daily (24 h. To disentangle how C. bungeana copes with these intricate cold stresses through epigenetic modifications, plants of C. bungeana were treated at 4°C (chilling and -4°C (freezing over five periods of time (0-24 h. Methylation-sensitive amplified fragment-length polymorphism markers were used to investigate the variation in DNA methylation of C. bungeana in response to chilling and freezing. It was found that the alterations in DNA methylation of C. bungeana largely occurred over the period of chilling and freezing. Moreover, chilling and freezing appeared to gradually induce distinct DNA methylation variations, as the treatment went on (e.g., after 12 h. Forty-three cold-induced polymorphic fragments were randomly selected and further analyzed, and three of the cloned fragments were homologous to genes encoding alcohol dehydrogenase, UDP-glucosyltransferase and polygalacturonase-inhibiting protein. These candidate genes verified the existence of different expressive patterns between chilling and freezing. Our results showed that C. bungeana responded to cold stresses rapidly through the alterations of DNA methylation, and that chilling and freezing induced different DNA methylation changes. Therefore, we conclude that epigenetic modifications can potentially serve as a rapid and flexible mechanism for C. bungeana

  19. PVA/CM-chitosan/honey hydrogels prepared by using the combined technique of irradiation followed by freeze-thawing

    Science.gov (United States)

    Afshari, M. J.; Sheikh, N.; Afarideh, H.

    2015-08-01

    Hydrogels with three components, poly(vinyl alcohol) (PVA), carboxymethylate chitosan (CM-chitosan) and honey have been prepared by using radiation method and radiation followed by freeze-thawing cycles technique (combinational method). The solid concentration of the polymer solution is 15 wt% and the ratios of PVA/CM-chitosan/honey are 10/1.5/3.5, 10/2/3, 10/3/2, and 10/3.5/1.5. The applied irradiation doses are 25, 30 and 40 kGy. Various tests have been done to evaluate the hydrogel properties to produce materials to be used as wound dressing. The results show that combinational method improves the mechanical strength of hydrogels while it has no significant effect on the water evaporation rate of gels. The combinational method decreases the swelling of hydrogels significantly, albeit this parameter is still acceptable for wound dressing. Microbiological analyses show that the hydrogel prepared by both methods can protect the wound from Escherichia coli bacterial infection. The wound healing test shows the good performance of the gels in mice.

  20. The contribution of apoptosis and necrosis in freezing injury of sea urchin embryonic cells.

    Science.gov (United States)

    Boroda, Andrey V; Kipryushina, Yulia O; Yakovlev, Konstantin V; Odintsova, Nelly A

    2016-08-01

    Sea urchins have recently been reported to be a promising tool for investigations of oxidative stress, UV light perturbations and senescence. However, few available data describe the pathway of cell death that occurs in sea urchin embryonic cells after cryopreservation. Our study is focused on the morphological and functional alterations that occur in cells of these animals during the induction of different cell death pathways in response to cold injury. To estimate the effect of cryopreservation on sea urchin cell cultures and identify the involved cell death pathways, we analyzed cell viability (via trypan blue exclusion test, MTT assay and DAPI staining), caspase activity (via flow cytometry and spectrophotometry), the level of apoptosis (via annexin V-FITC staining), and cell ultrastructure alterations (via transmission electron microscopy). Using general caspase detection, we found that the level of caspase activity was low in unfrozen control cells, whereas the number of apoptotic cells with activated caspases rose after freezing-thawing depending on cryoprotectants used, also as the number of dead cells and cells in a late apoptosis. The data using annexin V-binding assay revealed a very high apoptosis level in all tested samples, even in unfrozen cells (about 66%). Thus, annexin V assay appears to be unsuitable for sea urchin embryonic cells. Typical necrotic cells with damaged mitochondria were not detected after freezing in sea urchin cell cultures. Our results assume that physical cell disruption but not freezing-induced apoptosis or necrosis is the predominant reason of cell death in sea urchin cultures after freezing-thawing with any cryoprotectant combination. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Changes in Ultrastructure and Sensory Characteristics on Electro-magnetic and Air Blast Freezing of Beef during Frozen Storage

    Science.gov (United States)

    2015-01-01

    The ultrastructure in the beef muscle of the electro-magnetic resonance and air blast freezing during the frozen storage, and the changes in the quality characteristics after thawing were evaluated. The size of ice crystal was small and evenly formed in the initial freezing period, and it showed that the size was increased as the storage period was elapsed (pfreezing showed the size of ice crystal with a lower rate of increase than the air blast freezing during the frozen storage. The thawing loss of beef stored by the electro-magnetic resonance freezing was significantly lower than the air blast freezing during frozen storage (pfreezing was higher than the air blast on 8 month (pfreezing did not show the difference until 4 months, and it showed higher acceptability in comparison with the beef stored by the air blast freezing. Thus, it is considered that the freezing method has an effect on the change in the ultrastructure and quality characteristics of the beef. PMID:26761797

  2. Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos

    NARCIS (Netherlands)

    Wagtendonk-de Leeuw, van A.M.; Daas, den J.H.; Kruip, T.A.; Rail, W.F.

    1995-01-01

    Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii)

  3. Influence of cryoprotectants glycerol and amides, combined with antioxidants on quality of frozen-thawed boar sperm.

    Science.gov (United States)

    Buranaamnuay, K; Grossfeld, R; Struckmann, C; Rath, D

    2011-08-01

    The present study was undertaken to examine whether the cooling and freezing extenders containing a mixture of antioxidants (AOs) catalase, Na-pyruvate and mercaptoethanol and one of three types of cryoprotectants (CPs) would be able to improve the quality of frozen-thawed boar sperm. The collected semen, only the sperm-rich fraction, was diluted 1:1 with Androhep plus™ extender, stored at 15°C for 2 h and centrifuged. The centrifuged sperm pellet was re-suspended in lactose-egg yolk extender and divided into four groups for mixing with freezing extenders containing different kinds of CPs at 5°C: (I) glycerol (GLY) as control; (II) GLY with AOs; (III) dimethyl formamide (DMF) with AOs and (IV) dimethyl acetamide (DMA) with AOs. Processed sperm were packaged in 0.25-mL straws and frozen using a controlled rate freezer. After thawing, the diluted thawed sperm were incubated at 38°C for 10 min and was assessed for motility by CASA, membrane/acrosome integrity by FITC-PNA/PI and DNA integrity (DFI) by SCSA. All sperm parameters evaluated, except DFI, were negatively affected (P0.05). There was no difference in DFI among the studied groups (P>0.05). In conclusion, based on the present results, addition of AOs to cooling and freezing extenders and/or replacement of GLY with DMF or DMA could not improve quality of frozen-thawed boar sperm. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Seasonal Variation in the Hepatoproteome of the Dehydration- and Freeze-Tolerant Wood Frog, Rana sylvatica

    Directory of Open Access Journals (Sweden)

    Jon P. Costanzo

    2011-11-01

    Full Text Available Winter’s advent invokes physiological adjustments that permit temperate ectotherms to cope with stresses such as food shortage, water deprivation, hypoxia, and hypothermia. We used liquid chromatography (LC in combination with tandem mass spectrometry (MS/MS quantitative isobaric (iTRAQ™ peptide mapping to assess variation in the abundance of hepatic proteins in summer- and winter-acclimatized wood frogs (Rana sylvatica, a northerly-distributed species that tolerates extreme dehydration and tissue freezing during hibernation. Thirty-three unique proteins exhibited strong seasonal lability. Livers of winter frogs had relatively high levels of proteins involved in cytoprotection, including heat-shock proteins and an antioxidant, and a reduced abundance of proteins involved in cell proliferation, protein synthesis, and mitochondrial function. They also exhibited altered levels of certain metabolic enzymes that participate in the biochemical reorganization associated with aphagia and reliance on energy reserves, as well as the freezing mobilization and post-thaw recovery of glucose, an important cryoprotective solute in freezing adaptation.

  5. Effect of Achillea millefolium-loaded nanophytosome in the post-thawing sperm quality and oxidative status of rooster semen.

    Science.gov (United States)

    Najafi, Davood; Taheri, Ramezan Ali; Najafi, Abouzar; Rouhollahi, Abbas Abbas; Alvarez-Rodriguez, Manuel

    2018-04-17

    The aim of this study was to compare the effectiveness of antioxidants including Achillea millefolium extract (AmE) (n0t1.5: 1.5, n0t3: 3 and n0t4.5: 4.5 mg/L) and AmE loaded in nano phytosome (n1t1.5: 1.5, n1t3: 3 and n1t4.5: 4.5 mg/L) in the freezing of Ross 308 rooster semen. Sperm motility (CASA), membrane integrity (HOS test), viability, total abnormality and enzymatic parameters (SOD, CAT and GPx) were assessed after thawing. AmE-loaded nano phytosome at a concentration of 3 mg/l resulted in significantly (P rooster sperm motility, viability and oxidative stress values during the freezing procedure. Copyright © 2018 Elsevier Inc. All rights reserved.

  6. Supplementation of different concentrations of Orvus Es Paste (OEP) to ostrich egg yolk lipoprotein extender improves post-thaw boar semen quality.

    Science.gov (United States)

    Fraser, L; Jasiewicz, E; Kordan, W

    2014-01-01

    This study aimed to compare post-thaw quality of boar semen following freezing in an ostrich egg yolk lipoprotein (LPFo) extender supplemented with 0%, 0.25% and 0.50% Orvus Es Paste (OEP). Sperm assessments included total motility (TMOT), mitochondrial function (MF), plasma membrane integrity (PMI) and acrosome integrity (normal apical ridge, NAR). Considerable variations among boars and OEP treatments had a significant effect (P semen samples frozen in the absence of OEP. By contrast, lactose-LPFo-glycerol extender supplemented with either 0.25% OEP or 0.50% OEP markedly enhanced post-thaw sperm characteristics. In all boars, there were no marked differences in post-thaw sperm TMOT between the freezing extenders supplemented with 0.25% and 0.50% OEP. However, a decline in the percentage of post-thaw motile spermatozoa was more pronounced in the extender supplemented with 0.50% OEP following a 120-min incubation period. Furthermore, the proportions of frozen-thawed spermatozoa with MF, PMI and NAR acrosomes varied significantly among the boars in the OEP-supplemented extenders. The findings of this study indicate that different OEP concentrations, in the presence of ostrich egg yolk lipoproteins, could have varying effects on post-thaw sperm survival.

  7. Medical and social egg freezing

    DEFF Research Database (Denmark)

    Lallemant, Camille; Vassard, Ditte; Andersen, Anders Nyboe

    2016-01-01

    INTRODUCTION: Until recently, limited options for preserving fertility in order to delay childbearing were available. Although egg freezing and successful thawing is now possible, it remains unclear to what extent women are aware of the availability of this technique, their attitudes towards its...... use, or the circumstances under which this technique may be considered. MATERIAL AND METHODS: An online cross-sectional survey was designed to investigate knowledge and attitudes of women in Denmark and the UK on egg freezing and their potential intentions regarding the procedure. RESULTS: Data...... was collected from September 2012 to September 2013 and the responses of 973 women were analyzed. In total, 83% of women reported having heard of egg freezing, and nearly all considered it acceptable for medical indications, whilst 89% considered it acceptable for social reasons. Overall, 19% expressed active...

  8. Dryland soil hydrological processes and their impacts on the nitrogen balance in a soil-maize system of a freeze-thawing agricultural area.

    Directory of Open Access Journals (Sweden)

    Wei Ouyang

    Full Text Available Understanding the fates of soil hydrological processes and nitrogen (N is essential for optimizing the water and N in a dryland crop system with the goal of obtaining a maximum yield. Few investigations have addressed the dynamics of dryland N and its association with the soil hydrological process in a freeze-thawing agricultural area. With the daily monitoring of soil water content and acquisition rates at 15, 30, 60 and 90 cm depths, the soil hydrological process with the influence of rainfall was identified. The temporal-vertical soil water storage analysis indicated the local albic soil texture provided a stable soil water condition for maize growth with the rainfall as the only water source. Soil storage water averages at 0-20, 20-40 and 40-60 cm were observed to be 490.2, 593.8, and 358 m3 ha-1, respectively, during the growing season. The evapo-transpiration (ET, rainfall, and water loss analysis demonstrated that these factors increased in same temporal pattern and provided necessary water conditions for maize growth in a short period. The dry weight and N concentration of maize organs (root, leaf, stem, tassel, and grain demonstrated the N accumulation increased to a peak in the maturity period and that grain had the most N. The maximum N accumulative rate reached about 500 mg m-2d-1 in leaves and grain. Over the entire growing season, the soil nitrate N decreased by amounts ranging from 48.9 kg N ha-1 to 65.3 kg N ha-1 over the 90 cm profile and the loss of ammonia-N ranged from 9.79 to 12.69 kg N ha-1. With soil water loss and N balance calculation, the N usage efficiency (NUE over the 0-90 cm soil profile was 43%. The soil hydrological process due to special soil texture and the temporal features of rainfall determined the maize growth in the freeze-thawing agricultural area.

  9. Folic acid content in thermostabilized and freeze-dried space shuttle foods

    Science.gov (United States)

    Lane, H. W.; Nillen, J. L.; Kloeris, V. L.

    1995-01-01

    This study was designed to determine whether freeze-dried and thermostabilized foods on a space shuttle contain adequate folate and to investigate any effects of freeze-drying on folacin. Frozen vegetables were analyzed after three states of processing: thawed; cooked; and rehydrated. Thermostabilized items were analyzed as supplied with no further processing. Measurable folate decreased in some freeze-dried vegetables and increased in others. Folacin content of thermostabilized food items was comparable with published values. We concluded that although the folacin content of some freeze-dried foods was low, adequate folate is available from the shuttle menu to meet RDA guidelines.

  10. Effect of freezing on electrical properties and quality of thawed chicken breast meat

    Directory of Open Access Journals (Sweden)

    Ran Wei

    2017-04-01

    Full Text Available Objective The objective of this research was to study the electrical properties and quality of frozen-thawed chicken breast meat and to investigate the relationship between these parameters at different times of frozen storage. Methods Thawed samples of chicken breast muscles were evaluated after being kept in frozen storage at −18°C for different periods of time (1, 2, 3, 4, 5, 6, 7, and 8 months. Results The results showed that water-holding capacity (WHC and protein solubility decreased while thiobarbituric acid-reactive substances content increased with increasing storage time. The impedance module of samples decreased during 8-month frozen storage. Pearson correlation coefficients showed that the impedance change ratio (Q value was significantly (p<0.05 related to pH, color, WHC, lipid oxidation and protein solubility, indicating a good relationship between the electrical properties and qualities of frozen-thawed chicken breast meat. Conclusion Impedance measurement has a potential to assess the quality of frozen chicken meat combining with quality indices.

  11. Environmental stress induces trinucleotide repeat mutagenesis in human cells.

    Science.gov (United States)

    Chatterjee, Nimrat; Lin, Yunfu; Santillan, Beatriz A; Yotnda, Patricia; Wilson, John H

    2015-03-24

    The dynamic mutability of microsatellite repeats is implicated in the modification of gene function and disease phenotype. Studies of the enhanced instability of long trinucleotide repeats (TNRs)-the cause of multiple human diseases-have revealed a remarkable complexity of mutagenic mechanisms. Here, we show that cold, heat, hypoxic, and oxidative stresses induce mutagenesis of a long CAG repeat tract in human cells. We show that stress-response factors mediate the stress-induced mutagenesis (SIM) of CAG repeats. We show further that SIM of CAG repeats does not involve mismatch repair, nucleotide excision repair, or transcription, processes that are known to promote TNR mutagenesis in other pathways of instability. Instead, we find that these stresses stimulate DNA rereplication, increasing the proportion of cells with >4 C-value (C) DNA content. Knockdown of the replication origin-licensing factor CDT1 eliminates both stress-induced rereplication and CAG repeat mutagenesis. In addition, direct induction of rereplication in the absence of stress also increases the proportion of cells with >4C DNA content and promotes repeat mutagenesis. Thus, environmental stress triggers a unique pathway for TNR mutagenesis that likely is mediated by DNA rereplication. This pathway may impact normal cells as they encounter stresses in their environment or during development or abnormal cells as they evolve metastatic potential.

  12. Breeding of Freeze-tolerant Yeast and the Mechanisms of Stress-tolerance

    Science.gov (United States)

    Hino, Akihiro

    Frozen dough method have been adopted in the baking industry to reduce labor and to produce fresh breads in stores. New freeze-tolerant yeasts for frozen dough preparations were isolated from banana peel and identified. To obtain strains that have fermentative ability even after several months of frozen storage in fermented dough, we attempted to breed new freeze-tolerantstrain. The hybrid between S.cerevisiae, which is a isolated freeze-tolerant strain, and a strain isolated from bakers' yeast with sexual conjugation gave a good quality bread made from frozen dough method. Freeze-tolerant strains showed higher surviving and trehalose accumulating abilities than freeze-sensitive strains. The freeze tolerance of the yeasts was associated with the basal amount of intracellular trehalose after rapid degradation at the onset of the prefermentation period. The complicated metabolic pathway and the regulation system of trehalose in yeast cells are introduced. The trehalose synthesis may act as a metabolic buffer system which contribute to maintain the intracellular inorganic phosphate and as a feedback regulation system in the glycolysis. However, it is not known enough how the trehalose protects yeast cells from stress.

  13. Cellular biophysics during freezing of rat and mouse sperm predicts post-thaw motility.

    Science.gov (United States)

    Hagiwara, Mie; Choi, Jeung Hwan; Devireddy, Ramachandra V; Roberts, Kenneth P; Wolkers, Willem F; Makhlouf, Antoine; Bischof, John C

    2009-10-01

    Though cryopreservation of mouse sperm yields good survival and motility after thawing, cryopreservation of rat sperm remains a challenge. This study was designed to evaluate the biophysics (membrane permeability) of rat in comparison to mouse to better understand the cooling rate response that contributes to cryopreservation success or failure in these two sperm types. In order to extract subzero membrane hydraulic permeability in the presence of ice, a differential scanning calorimeter (DSC) method was used. By analyzing rat and mouse sperm frozen at 5 degrees C/min and 20 degrees C/min, heat release signatures characteristic of each sperm type were obtained and correlated to cellular dehydration. The dehydration response was then fit to a model of cellular water transport (dehydration) by adjusting cell-specific biophysical (membrane hydraulic permeability) parameters L(pg) and E(Lp). A "combined fit" (to 5 degrees C/min and 20 degrees C/min data) for rat sperm in Biggers-Whitten-Whittingham media yielded L(pg) = 0.007 microm min(-1) atm(-1) and E(Lp) = 17.8 kcal/mol, and in egg yolk cryopreservation media yielded L(pg) = 0.005 microm min(-1) atm(-1) and E(Lp) = 14.3 kcal/mol. These parameters, especially the activation energy, were found to be lower than previously published parameters for mouse sperm. In addition, the biophysical responses in mouse and rat sperm were shown to depend on the constituents of the cryopreservation media, in particular egg yolk and glycerol. Using these parameters, optimal cooling rates for cryopreservation were predicted for each sperm based on a criteria of 5%-15% normalized cell water at -30 degrees C during freezing in cryopreservation media. These predicted rates range from 53 degrees C/min to 70 degrees C/min and from 28 degrees C/min to 36 degrees C/min in rat and mouse, respectively. These predictions were validated by comparison to experimentally determined cryopreservation outcomes, in this case based on motility. Maximum

  14. Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis.

    Science.gov (United States)

    Anand, Mukul; Yadav, Sarvajeet

    2016-02-01

    The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis. A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck). Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process. Significantly (plive percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (pvitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats.

  15. Theoretical study of heat transfer with moving phase-change interface in thawing of frozen food

    International Nuclear Information System (INIS)

    Leung, M; Ching, W H; Leung, D Y C; Lam, G C K

    2005-01-01

    A theoretical solution was obtained for a transient phase-change heat transfer problem in thawing of frozen food. In the physical model, a sphere originally at a uniform temperature below the phase-change temperature is suddenly immersed in a fluid at a temperature above the phase-change temperature. As the body temperature increases, the phase-change interface will be first formed on the surface. Subsequently, the interface will absorb the latent heat and move towards the centre until the whole body undergoes complete phase change. In the mathematical formulation, the nonhomogeneous problem arises from the moving phase-change interface. The solution in terms of the time-dependent temperature field was obtained by use of Green's function. A one-step Newton-Raphson method was specially designed to solve for the position of the moving interface to satisfy the interface condition. The theoretical results were compared with numerical results generated by a finite difference model and experimental measurements collected from a cold water thawing process. As a good agreement was found, the theoretical solution developed in this study was verified numerically and experimentally. Besides thawing of frozen food, there are many other practical applications of the theoretical solution, such as food freezing, soil freezing/thawing, metal casting and bath quenching heat treatment, among others

  16. Theoretical study of heat transfer with moving phase-change interface in thawing of frozen food

    Science.gov (United States)

    Leung, M.; Ching, W. H.; Leung, D. Y. C.; Lam, G. C. K.

    2005-02-01

    A theoretical solution was obtained for a transient phase-change heat transfer problem in thawing of frozen food. In the physical model, a sphere originally at a uniform temperature below the phase-change temperature is suddenly immersed in a fluid at a temperature above the phase-change temperature. As the body temperature increases, the phase-change interface will be first formed on the surface. Subsequently, the interface will absorb the latent heat and move towards the centre until the whole body undergoes complete phase change. In the mathematical formulation, the nonhomogeneous problem arises from the moving phase-change interface. The solution in terms of the time-dependent temperature field was obtained by use of Green's function. A one-step Newton-Raphson method was specially designed to solve for the position of the moving interface to satisfy the interface condition. The theoretical results were compared with numerical results generated by a finite difference model and experimental measurements collected from a cold water thawing process. As a good agreement was found, the theoretical solution developed in this study was verified numerically and experimentally. Besides thawing of frozen food, there are many other practical applications of the theoretical solution, such as food freezing, soil freezing/thawing, metal casting and bath quenching heat treatment, among others.

  17. Behaviour of Prestressed CFRP Anchorages during and after Freeze-Thaw Cycle Exposure

    Directory of Open Access Journals (Sweden)

    Yunus Emre Harmanci

    2018-05-01

    Full Text Available The long-term performance of externally-bonded reinforcements (EBR on reinforced concrete (RC structures highly depends on the behavior of constituent materials and their interfaces to various environmental loads, such as temperature and humidity exposure. Although significant efforts have been devoted to understanding the effect of such conditions on the anchorage resistance of unstressed EBR, with or without sustained loading, the effect of a released prestressing has not been thoroughly investigated. For this purpose, a series of experiments has been carried out herein, with concrete blocks strengthened with carbon fiber-reinforced polymer (CFRP strips, both unstressed, as well as prestressed using the gradient anchorage. The gradient anchorage is a non-mechanical technique to anchor prestressed CFRP by exploiting the accelerated curing property of epoxy under higher temperatures and segment-wise prestress-force releasing. Subsequently, strengthened blocks are transferred into a chamber for exposure in dry freeze-thaw cycles (FTC. Following FTC exposure, the blocks are tested in a conventional lap-shear test setup to determine their residual anchorage resistance and then compared with reference specimens. Blocks were monitored during FTC by conventional and Fabry–Pérot-based fiber optic strain (FOS sensors and a 3D-digital image correlation (3D-DIC system during gradient application and lap-shear testing. Results indicate a reduction of residual anchorage resistance, stiffness and deformation capacity of the system after FTC and a change in the failure mode from concrete substrate to epoxy-concrete interface failure. It was further observed that all of these properties experienced a more significant reduction for prestressed specimens. These findings are presented with a complementary finite element model to shed more light onto the durability of such systems.

  18. Digestion of frozen/thawed food waste in the hybrid anaerobic solid-liquid system

    International Nuclear Information System (INIS)

    Stabnikova, O.; Liu, X.Y.; Wang, J.Y.

    2008-01-01

    The hybrid anaerobic solid-liquid (HASL) system, which is a modified two-phase anaerobic digester, is to be used in an industrial scale operation to minimize disposal of food waste at incineration plants in Singapore. The aim of the present research was to evaluate freezing/thawing of food waste as a pre-treatment for its anaerobic digestion in the HASL system. The hydrolytic and fermentation processes in the acidogenic reactor were enhanced when food waste was frozen for 24 h at -20 deg. C and then thawed for 12 h at 25 deg. C (experiment) in comparison with fresh food waste (control). The highest dissolved COD concentrations in the leachate from the acidogenic reactors were 16.9 g/l on day 3 in the control and 18.9 g/l on day 1 in the experiment. The highest VFA concentrations in the leachate from the acidogenic reactors were 11.7 g/l on day 3 in the control and 17.0 g/l on day 1 in the experiment. The same volume of methane was produced during 12 days in the control and 7 days in the experiment. It gave the opportunity to diminish operational time of batch process by 42%. The effect of freezing/thawing of food waste as pre-treatment for its anaerobic digestion in the HASL system was comparable with that of thermal pre-treatment of food waste at 150 deg. C for 1 h. However, estimation of energy required either to heat the suspended food waste to 150 deg. C or to freeze the same quantity of food waste to -20 deg. C showed that freezing pre-treatment consumes about 3 times less energy than thermal pre-treatment

  19. Acclimation increases freezing stress response of Arabidopsis thaliana at proteome level

    KAUST Repository

    Fanucchi, Francesca

    2012-06-01

    This study used 2DE to investigate how Arabidopsis thaliana modulates protein levels in response to freezing stress after sub-lethal exposure at - 10 °C, both in cold-acclimated and in non-acclimated plants. A map was implemented in which 62 spots, corresponding to 44 proteins, were identified. Twenty-two spots were modulated upon treatments, and the corresponding proteins proved to be related to photosynthesis, energy metabolism, and stress response. Proteins demonstrated differences between control and acclimation conditions. Most of the acclimation-responsive proteins were either not further modulated or they were down-modulated by freezing treatment, indicating that the levels reached during acclimation were sufficient to deal with freezing. Anabolic metabolism appeared to be down-regulated in favor of catabolic metabolism. Acclimated plants and plants submitted to freezing after acclimation showed greater reciprocal similarity in protein profiles than either showed when compared both to control plants and to plants frozen without acclimation. The response of non-acclimated plants was aimed at re-modulating photosynthetic apparatus activity, and at increasing the levels of proteins with antioxidant-, molecular chaperone-, or post-transcriptional regulative functions. These changes, even less effective than the acclimation strategy, might allow the injured plastids to minimize the production of non-useful metabolites and might counteract photosynthetic apparatus injuries. © 2012 Elsevier B.V. All rights reserved.

  20. Acclimation increases freezing stress response of Arabidopsis thaliana at proteome level

    KAUST Repository

    Fanucchi, Francesca; Alpi, Emanuele; Olivieri, Stefano; Cannistraci, Carlo; Bachi, Angela; Alpi, Amedeo; Alessio, Massimo

    2012-01-01

    This study used 2DE to investigate how Arabidopsis thaliana modulates protein levels in response to freezing stress after sub-lethal exposure at - 10 °C, both in cold-acclimated and in non-acclimated plants. A map was implemented in which 62 spots, corresponding to 44 proteins, were identified. Twenty-two spots were modulated upon treatments, and the corresponding proteins proved to be related to photosynthesis, energy metabolism, and stress response. Proteins demonstrated differences between control and acclimation conditions. Most of the acclimation-responsive proteins were either not further modulated or they were down-modulated by freezing treatment, indicating that the levels reached during acclimation were sufficient to deal with freezing. Anabolic metabolism appeared to be down-regulated in favor of catabolic metabolism. Acclimated plants and plants submitted to freezing after acclimation showed greater reciprocal similarity in protein profiles than either showed when compared both to control plants and to plants frozen without acclimation. The response of non-acclimated plants was aimed at re-modulating photosynthetic apparatus activity, and at increasing the levels of proteins with antioxidant-, molecular chaperone-, or post-transcriptional regulative functions. These changes, even less effective than the acclimation strategy, might allow the injured plastids to minimize the production of non-useful metabolites and might counteract photosynthetic apparatus injuries. © 2012 Elsevier B.V. All rights reserved.

  1. Effects of Diatomite and SBS on Freeze-Thaw Resistance of Crumb Rubber Modified Asphalt Mixture

    Directory of Open Access Journals (Sweden)

    Haibin Wei

    2017-01-01

    Full Text Available Asphalt mixture is susceptible to moisture damage under the effect of freeze-thaw (F-T cycles. In this paper, crumb rubber (CR was used to modify stone mastic asphalt (SMA and the effects of diatomite and styrene butadiene styrene (SBS on antifreezing performances of crumb rubber modified SMA (CRSMA were investigated. Regression analysis and modified grey model (MGM were used to construct the prediction models for properties of modified mixtures. CRSMA, CR and diatomite modified SMA (CRDSMA, and CR and SBS modified SMA (CRSSMA were prepared in laboratory, respectively. Process of F-T cycles was designed. Air void, indirect tensile strength (ITS, and indirect tensile stiffness modulus (ITSM were measured to evaluate the antifreezing performances of CRSMA, CRDSMA, and CRSSMA. Results indicate that air voids increase with the increasing of F-T cycles. ITS and ITSM all decrease with the increasing of F-T cycles. The addition of diatomite and SBS can reduce the air void and improve the ITS and ITSM of CRSMA. CRSSMA presents the lowest air void, highest tensile strength, and largest stiffness modulus, which reveals that CRSSMA has the best F-T resistance among three different kinds of mixtures. Moreover, MGM (1, 2 models present more favorable accuracy in prediction of air void and ITS compared with regression ones.

  2. Research Article: Effects of long-term simulated Martian conditions on a freeze-dried and homogenized bacterial permafrost community

    DEFF Research Database (Denmark)

    Hansen, Aviaja Anna; Jensen, Lars Liengård; Kristoffersen, Tommy

    2009-01-01

    Indigenous bacteria and biomolecules (DNA and proteins) in a freeze-dried and homogenized Arctic permafrost were exposed to simulated martian conditions that correspond to about 80 days on the surface of Mars with respect to the accumulated UV dose. The simulation conditions included UV radiation......, freeze-thaw cycles, the atmospheric gas composition, and pressure. The homogenized permafrost cores were subjected to repeated cycles of UV radiation for 3 h followed by 27 h without irradiation. The effects of the simulation conditions on the concentrations of biomolecules; numbers of viable, dead......, and cultured bacteria; as well as the community structure were determined. Simulated martian conditions resulted in a significant reduction of the concentrations of DNA and amino acids in the uppermost 1.5 mm of the soil core. The total number of bacterial cells was reduced in the upper 9 mm of the soil core...

  3. Ageing of rice husk biochar along a freeze-thaw cycles

    OpenAIRE

    Miao Wei; Cheng Xiao Ying; Meng Jun; Tang Liang

    2016-01-01

    In order to elucidate the aging possess of biochar, the experiment with treatment biochar with soil were performed. For accelerating aging process, freeing-thawing cycle were conducted to simulate the changing process of the physical and chemical properties of biochar and explore the roles of biochar in the changes of the soil nutrition. Aging treatment has a significant impact on the physical and chemical properties of biochar. The pH values, element composition, oxidation of the surface, ab...

  4. THAWING PROCEDURES FOR HOSPITAL-MADE ENTERAL FEEDINGS: SURVIVAL OF COLIFORM AND MESOPHILIC AEROBIC BACTERIA

    Directory of Open Access Journals (Sweden)

    KATHIA ROSSI ROLIM LOPES

    2009-07-01

    Full Text Available

    ABSTRACT: This study had the purpose to observe the effect of thawing procedures on survival of coliform and mesophilic aerobic bacteria in hospital-made enteral feedings. The samples are represented by three different lots. The tests were realized in three moments: immediately after the sample preparation and after freezing during 1 or 2 months. The thawing procedures were denominated convencional and alternative. The first, used by hospital, utilizes water bath at 50ºC, considering the time spent from the total thawing to its distribution in the infirmaries. The second was the fast thawing made by microwaving. The results showed that the reduction of the mesophiles and coliform was related to the time the samples were frozen. The results obtained indicate an advantage of the alternative method, which presented lower total and fecal coliform counts than the conventional one. KEYWORDS: Enteral feedings; thawing; food microbiology.

  5. Storage of human platelets by freezing

    Energy Technology Data Exchange (ETDEWEB)

    Kim, B K; Tanoue, K; Baldini, M G

    1976-01-01

    Prolonged, probably indefinite storage of viable and functional human platelets is now possible by freezing with dimethylsulfoxide (DMSO). The platelets have a nearly normal survival upon reinfusion and are capable of sustained hemostatic effectiveness in thrombocytopenic patients. Adaptation of the freezing technique for large-scale usage has more recently been achieved. The method is mainly based on the following principles: (1) use of plasma for suspension of the platelet concentrate; (2) gradual addition (0.5% every 2 min) of DMSO to a final concentration of 5% and its gradual removal; (3) a slow cooling rate of about 1/sup 0/C per min and rapid thawing (in 1 min); (4) use of a polyolefin plastic bag for freezing; (5) a washing medium of 20% plasma in Hanks' balanced salt solution; (6) final resuspension of the platelets in 50% plasma in Hanks' solution.

  6. Spermatic feasibility after 20 years of freezing and refreezing: case study

    Directory of Open Access Journals (Sweden)

    Paulo Franco Taitson

    2010-08-01

    Full Text Available The aim of the present work was to study the spermatic feasibility after 20 years of freezing, thawing and refreezing. A young man affected by testicular cancer sought the sperm freezing service. After 20 years, the samples were thawed and frozen again. After two days, thawing and analysis of the material took place. Comparison of the findings showed activity reduction from the first thawing to the fresh semen 59.09%. For the second thawing it was 77.27%. Considering the morphology, first thawing showed a reduction of 44.64% in relation to fresh semen. In the second thawing, the reduction was of 75%. Spermatic motility decreased by 64.28% in relation to fresh semen and 92.85% at the level of second thawing. The patient had gone through in vitro fertilization in 2007, obtaining success in the first one attempt.Este trabalho mostra a viabilidade espermática após 20 anos de congelamento seminal, descongelamento e novo congelamento. Jovem do sexo masculino afetado por câncer testicular procurou o serviço para congelamento de espermatozóides. Após vinte anos, as amostras foram descongeladas e congeladas novamente. Após dois dias, realizou-se um novo descongelamento e análise do material. As comparações dos resultados mostraram uma redução da atividade do primeiro descongelamento ao semen fresco em 59,09%. A partir do segundo descongelamento era de 77,27%. Considerando a morfologia, o primeiro descongelamento mostrou uma redução de 44,64% em relação ao sêmen fresco. No segundo descongelamento, a redução foi de 75%. A motilidade espermática diminuiu 64,28% em relação ao semen fresco e 92,85% a nível do segundo descongelamento. O paciente passou por fertilização in vitro em 2007, obtendo sucesso na primeira tentativa.

  7. Effect of different cryo-protectants on the viability of frozen/thawed semen from boars of the Piau breed.

    Science.gov (United States)

    Pinho, R O; Lima, D M A; Shiomi, H H; Siqueira, J B; Silva, H T; Lopes, P S; Guimarães, S E F; Guimarães, J D

    2014-05-01

    The objective of this study was to evaluate the effect of different cryo-protectants (glycerol, dimethylacetamide and dimethylformamide alone or combined and added to lactose-egg yolk extender) on the viability of frozen/thawed semen from the Piau breed as assessed by in vitro testing. Frozen semen samples (n=20) were used from five male swine. Five different freezing extenders, including 2% glycerol (Group 1 - G), 2% glycerol and 3% dimethylacetamide (Group 2 - GA), 2% glycerol and 3% dimethylformamide (Group 3 - GF), 5% dimethylacetamide (Group 4 - A) and 5% dimethylformamide (group 5 - F), were evaluated. To assess post-thawing sperm quality, sperm motility and morphology were evaluated. Sperm viability was determined using the hypoosmotic swelling test, supravital staining, and a fluorescent assay (carboxyfluorescein diacetate and propidium iodide). The mean total sperm motility of semen immediately after thawing was 46.2±1.3, 57.7±1.5, 53.2±2.1, 51.7±1.2, and 46.5±1.6% for groups 1-5, respectively. Groups 2 (GA) and 3 (GF) had greater motility values (P0.05). The other complementary tests (hypoosmotic swelling test and supra-vital staining) demonstrated that sperm in Groups 2 (GA), 3 (GF) and 4 (A) had the greatest viability and there were no significant differences among these three groups (P>0.05). The most effective cryo-protectant combinations likely minimized and controlled the deleterious processes that occur in the sperm cell during freezing/thawing, thus improving post-thawing sperm viability. In conclusion, the combination of amides (3%) and glycerol (2%) or dimethylacetamide (5%) alone were more efficient at cryo-protection than glycerol alone for semen freezing in the Piau swine breed. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. High survival of mouse embryos after rapid freezing and thawing inside plastic straws with 1-2 propanediol as cryoprotectant.

    Science.gov (United States)

    Renard, J P; Babinet, C

    1984-06-01

    A method for obtaining a high survival rate of frozen-thawed mouse embryos is presented. Eight-cell mouse embryos were frozen inside small plastic straws in the presence of 1-2 propanediol and stored at -196 C. After thawing, the embryos were diluted for only 5 min in a 1.0 M sucrose solution to remove the 1-2 propanediol from the cells. At high rate of thawing (is equivalent to 2500 C/min) more than 88% of the embryos survived in vitro to the blastocyst stage provided that the dilution of propanediol was performed rapidly during thawing. At a lower rate of thawing (is equivalent to 300 C/min), survival tended to be higher (94.7%) when dilution was done 5 min after thawing. When the frozen-thawed embryos were transferred to the oviducts of day 1 pseudopregnant recipients either directly after the dilution of 1-2 propanediol or after 24 or 48 hr of culture, a high proportion of them (65.9%) develop normally to viable fetuses.

  9. Effects of straw mulching on soil evaporation during the soil thawing ...

    Indian Academy of Sciences (India)

    26

    Keywords: straw mulching, soil water evaporation, soil thawing period, freezing depth, soil liquid water content. 1. Introduction. The Songnen Plain, located in northeastern China, has 594×104 ha of cultivated land area and a grain yield of 395×108 kg. It is one of the most important food production bases in China (Yan et al.

  10. Biochar can restrict N2O emissions and the risk of nitrogen leaching from an agricultural soil during the freeze-thaw period

    Directory of Open Access Journals (Sweden)

    Riitta Kettunen

    2013-12-01

    Full Text Available Freeze-thaw (FT events in soils can cause a burst of nitrous oxide (N2O and enhance N leaching during the spring-thaw event. We studied whether a soil amended with wood-derived (spruce chips biochar (10 tonnes ha-1, produced at rather low temperatures (400-450°C, could reduce the burst of N2O and the risk of N leaching from an agricultural soil after a FT event. A short-term laboratory experiment (4 weeks was conducted with 24 vegetated (Phleum pratense mesocosms (12 controls, 12 biochar-treated that had spent a dormant season in the dark at 15°C for two months after the growing season. N2O efflux to the atmosphere and ammonium (NH4+-N and nitrate (NO3-N in the percolated soil water were monitored before and after the FT event. N2O was monitored with the dark chamber method and analyzed using a gas chromatograph. We found that soil amended biochar can significantly diminish the burst of N2O after the soil FT event (by 61% just after FT event and substantially reduce the risk of NO3-N and NH4+-N leaching from the agricultural soil. Compared to the control, the decrement in concentrations of NO3-N and NH4+-N in water percolated through the biochar amended soil in the mesocosms was 58% and 22%, respectively.

  11. Cellular Biophysics During Freezing of Rat and Mouse Sperm Predicts Post-thaw Motility1

    Science.gov (United States)

    Hagiwara, Mie; Choi, Jeung Hwan; Devireddy, Ramachandra V.; Roberts, Kenneth P.; Wolkers, Willem F.; Makhlouf, Antoine; Bischof, John C.

    2009-01-01

    Though cryopreservation of mouse sperm yields good survival and motility after thawing, cryopreservation of rat sperm remains a challenge. This study was designed to evaluate the biophysics (membrane permeability) of rat in comparison to mouse to better understand the cooling rate response that contributes to cryopreservation success or failure in these two sperm types. In order to extract subzero membrane hydraulic permeability in the presence of ice, a differential scanning calorimeter (DSC) method was used. By analyzing rat and mouse sperm frozen at 5°C/min and 20°C/min, heat release signatures characteristic of each sperm type were obtained and correlated to cellular dehydration. The dehydration response was then fit to a model of cellular water transport (dehydration) by adjusting cell-specific biophysical (membrane hydraulic permeability) parameters Lpg and ELp. A “combined fit” (to 5°C/min and 20°C/min data) for rat sperm in Biggers-Whitten-Whittingham media yielded Lpg = 0.007 μm min−1 atm−1 and ELp = 17.8 kcal/mol, and in egg yolk cryopreservation media yielded Lpg = 0.005 μm min−1 atm−1 and ELp = 14.3 kcal/mol. These parameters, especially the activation energy, were found to be lower than previously published parameters for mouse sperm. In addition, the biophysical responses in mouse and rat sperm were shown to depend on the constituents of the cryopreservation media, in particular egg yolk and glycerol. Using these parameters, optimal cooling rates for cryopreservation were predicted for each sperm based on a criteria of 5%–15% normalized cell water at −30°C during freezing in cryopreservation media. These predicted rates range from 53°C/min to 70°C/min and from 28°C/min to 36°C/min in rat and mouse, respectively. These predictions were validated by comparison to experimentally determined cryopreservation outcomes, in this case based on motility. Maximum motility was obtained with freezing rates between 50°C/min and 80

  12. How to best freeze liver samples to perform the in vivo mammalian alkaline comet assay

    Directory of Open Access Journals (Sweden)

    José Manuel Enciso Gadea

    2015-06-01

    None of the different methods used was capable of giving good results, except immersing the liver samples in liquid nitrogen, followed by Jackson’s et al. (2013 thawing protocol, suggesting that the thawing process may be as critical as the freezing process. To sum up, these results highlight the importance of deepening the possibility to perform the comet assay with frozen tissue.

  13. Effect of freezing extender composition and male line on semen traits and reproductive performance in rabbits.

    Science.gov (United States)

    Viudes-de-Castro, M P; Lavara, R; Safaa, H M; Marco-Jiménez, F; Mehaisen, G M K; Vicente, J S

    2014-05-01

    This study was conducted to elucidate the effect of different freezing extenders on two lines selected for hyperprolificacy and longevity (H and LP, respectively). In extender A, dimethyl sulphoxide (Me2SO) and sucrose were used as cryoprotectants. In extenders B and C, the sucrose was replaced by 20% egg yolk, and in extender C the Me2SO was substituted by acetamide. Semen was packaged in 0.25 ml plastic straws and cooled at 5°C for 45 min, and then was frozen in liquid nitrogen vapour for 10 min before being plunged into the liquid nitrogen. Thawing was carried out by immersing the straws in a water bath at 50°C for 10 s. Frozen-thawed semen characteristics and reproductive parameters were affected by freezing. Extender C showed significantly lower post-thawing quality traits than any of the three extenders. Acrosome integrity was significantly improved when Me2SO was used as cryoprotectant. Sucrose replacement by 20% egg yolk had no effect on acrosome integrity but provided significantly lower sperm motility and viability. Freezing extender affected fertility rate, total born, number of implantation sites and gestational losses, obtaining better results when extender A was used. The acrosomal integrity after frozen-thawed process showed a significant correlation with fertility at 12th day and also at birth, indicating that an increase in acrosomal integrity leads to an increase in both fertilities (12th day and at birth). A positive correlation between motility of semen and implantation sites was found. The post-thawing quality traits of semen were not affected by the genetic line, although LP line showed higher total born and lower foetal and gestational losses. The findings of this study suggest that freezing extender composition has a significant effect on the success of rabbit sperm for preservation, and when Me2SO was used as permeable cryoprotectant sucrose provided better protection compared with egg yolk and improved reproductive traits, and, on the

  14. Monitoring Bare Soil Freeze–Thaw Process Using GPS-Interferometric Reflectometry: Simulation and Validation

    Directory of Open Access Journals (Sweden)

    Xuerui Wu

    2017-12-01

    Full Text Available Frozen soil and permafrost affect ecosystem diversity and productivity as well as global energy and water cycles. Although some space-based Radar techniques or ground-based sensors can monitor frozen soil and permafrost variations, there are some shortcomings and challenges. For the first time, we use GPS-Interferometric Reflectometry (GPS-IR to monitor and investigate the bare soil freeze–thaw process as a new remote sensing tool. The mixed-texture permittivity models are employed to calculate the frozen and thawed soil permittivities. When the soil freeze/thaw process occurs, there is an abrupt change in the soil permittivity, which will result in soil scattering variations. The corresponding theoretical simulation results from the forward GPS multipath simulator show variations of GPS multipath observables. As for the in-situ measurements, virtual bistatic radar is employed to simplify the analysis. Within the GPS-IR spatial resolution, one SNOTEL site (ID 958 and one corresponding PBO (plate boundary observatory GPS site (AB33 are used for analysis. In 2011, two representative days (frozen soil on Doy of Year (DOY 318 and thawed soil on DOY 322 show the SNR changes of phase and amplitude. The GPS site and the corresponding SNOTEL site in four different years are analyzed for comparisons. When the soil freeze/thaw process occurred and no confounding snow depth and soil moisture effects existed, it exhibited a good absolute correlation (|R| = 0.72 in 2009, |R| = 0.902 in 2012, |R| = 0.646 in 2013, and |R| = 0.7017 in 2014 with the average detrended SNR data. Our theoretical simulation and experimental results demonstrate that GPS-IR has potential for monitoring the bare soil temperature during the soil freeze–thaw process, while more test works should be done in the future. GNSS-R polarimetry is also discussed as an option for detection. More retrieval work about elevation and polarization combinations are the focus of future development.

  15. Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis

    Directory of Open Access Journals (Sweden)

    Mukul Anand

    2016-02-01

    Full Text Available Aim: The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis. Materials and Methods: A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck. Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process. Results: Significantly (p<0.01 higher value of live percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (p<0.01 higher followed by slow and rapid progression. Conclusion: This study showed that the best results for motion, vitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats.

  16. Effects of Long-Term Simulated Martian Conditions on a Freeze-Dried and Homogenized Bacterial Permafrost Community

    Science.gov (United States)

    Hansen, Aviaja A.; Jenson, Lars L.; Kristoffersen, Tommy; Mikkelsen, Karina; Merrison, Jonathan; Finster, Kai W.; Lomstein, Bente Aa.

    2009-03-01

    Indigenous bacteria and biomolecules (DNA and proteins) in a freeze-dried and homogenized Arctic permafrost were exposed to simulated martian conditions that correspond to about 80 days on the surface of Mars with respect to the accumulated UV dose. The simulation conditions included UV radiation, freeze-thaw cycles, the atmospheric gas composition, and pressure. The homogenized permafrost cores were subjected to repeated cycles of UV radiation for 3 h followed by 27 h without irradiation. The effects of the simulation conditions on the concentrations of biomolecules; numbers of viable, dead, and cultured bacteria; as well as the community structure were determined. Simulated martian conditions resulted in a significant reduction of the concentrations of DNA and amino acids in the uppermost 1.5 mm of the soil core. The total number of bacterial cells was reduced in the upper 9 mm of the soil core, while the number of viable cells was reduced in the upper 15 mm. The number of cultured aerobic bacteria was reduced in the upper 6 mm of the soil core, whereas the community structure of cultured anaerobic bacteria was relatively unaffected by the exposure conditions. As explanations for the observed changes, we propose three causes that might have been working on the biological material either individually or synergistically: (i) UV radiation, (ii) UV-generated reactive oxygen species, and (iii) freeze-thaw cycles. Currently, the production and action of reactive gases is only hypothetical and will be a central subject in future investigations. Overall, we conclude that in a stable environment (no wind-/pressure-induced mixing) biological material is efficiently shielded by a 2 cm thick layer of dust, while it is relatively rapidly destroyed in the surface layer, and that biomolecules like proteins and polynucleotides are more resistant to destruction than living biota.

  17. Association of Vitamin E with Rapid Thawing on Goat Semen

    Science.gov (United States)

    Penitente-Filho, Jurandy Mauro; Oliveira, Fabrício Albani; Jimenez, Carolina Rodriguez; Dias, Júlio César Oliveira; Oliveira, Gisele Dias; Silveira, Renata Gomes; Silveira, Camila Oliveira; Torres, Ciro Alexandre Alves

    2014-01-01

    The aim of this study was to evaluate the effects of vitamin E associated with rapid thawing on cryopreserved goat semen. Two bucks were used and eight ejaculates per animal were collected using artificial vagina. Semen was diluted with the following treatments: BIOXCELL (control), BIOXCELL + Equex (sodium lauryl sulphate) and BIOXCELL + vitamin E 100 μM. Semen was packaged into 0.25 mL straws and cooled at 5°C for 1 hour. Freezing was performed in liquid nitrogen vapor (−155°C) during 15 minutes. Then, the straws were immersed in liquid nitrogen (−196°C). Straws were thawed at 38°C/60 seconds or at 60°C/7 seconds with immediate sperm analysis. Hypoosmotic swelling test was performed adding a 20 μL aliquot of thawed semen to 1 mL of hypoosmotic solution (100 mOsm·Kg−1) followed by incubation during 60 minutes in water bath (38°C). Vitamin E did not affect any studied parameters (P > 0.05). Nevertheless, defrosting rate of 60°C/7 seconds improved sperm membrane functional integrity (P < 0.05). Current knowledge about goat semen cryopreservation is not sufficient to ensure high post-thawing recovery rates; thus, this study brings important data about using antioxidants and different thawing rates on cryopreservation process. PMID:24955428

  18. L-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm.

    Science.gov (United States)

    Fattah, A; Sharafi, M; Masoudi, R; Shahverdi, A; Esmaeili, V; Najafi, A

    2017-02-01

    Rooster semen cryopreservation is not efficient for artificial insemination in breeder flocks. L-Carnitine (LC) has been evaluated for effectiveness in cryopreservation media on the characteristics of rooster sperm after freeze-thawing. Motility characteristics, membrane functionality, abnormal morphology, apoptotic like changes, mitochondria activity and lipid peroxidation of rooster sperms were assessed after freeze-thawing with different concentrations of LC in Beltsville medium. Semen samples were collected from 12 roosters, twice a week, and diluted in the extenders that contained different concentrations of LC. Supplementation of Beltsevile with 1 and 2 mM LC was found to result in higher total motility (68.2± 1.7% and 69.1± 1.7%, respectively), progressive motility (28.4± 1.6%, 29.8± 1.6%), membrane functionality (76.2± 1.9% and 75.9± 1.9%), viability (58.2 ± 1.1%, 59.1 ± 1.1%) and lower significant of lipid peroxidation (2.53 ± 0.08 nmol/ml, 2.49 ± 0.08 nmol/ml) compared to control group containing no LC. Lower motility, progressive motility, and viability were observed in frozen-thawed sperm in extender containing 8 mM LC (35.8± 1.7%, 9.6± 1.2% and 27.1 ± 1.2%, respectively) compared to control. Morphology and mitochondrial activity were not affected by different concentrations of LC. Our results showed that supplementation of Beltsville extender with 1 and 2 mM LC significantly improved the quality of rooster sperm quality after freeze-thawing. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Effects of trehalose supplementation on cell viability and oxidative stress variables in frozen-thawed bovine calf testicular tissue.

    Science.gov (United States)

    Zhang, Xiao-Gang; Wang, Yan-Hua; Han, Cong; Hu, Shan; Wang, Li-Qiang; Hu, Jian-Hong

    2015-06-01

    Trehalose is widely used for cryopreservation of various cells and tissues. Until now, the effect of trehalose supplementation on cell viability and antioxidant enzyme activity in frozen-thawed bovine calf testicular tissue remains unexplored. The objective of the present study was to compare the effect of varying doses of trehalose in cryomedia on cell viability and key antioxidant enzymes activities in frozen-thawed bovine calf testicular tissue. Bovine calf testicular tissue samples were collected and cryopreserved in the cryomedias containing varying doses (0, 5, 10, 15, 20 and 25%; v/v) of trehalose, respectively. Cell viability, total antioxidant capacity (T-AOC) activity, catalase (CAT) activity, superoxide dismutase (SOD) activity, glutathione (GSH) content and malondialdehyde (MDA) content were measured and analyzed. The results showed that cell viability, T-AOC activity, SOD activity, CAT activity and GSH content of frozen-thawed bovine calf testicular tissue was decreased compared with that of fresh group (Pcell viability and antioxidant enzyme activity (SOD and CAT) among frozen-thawed groups (P0.05). In conclusion, the cryomedia added 15% trehalose reduced the oxidative stress and improved the cryoprotective effect of bovine calf testicular tissue. Further studies are required to obtain more concrete results on the determination of antioxidant capacity of trehalose in frozen-thawed bovine calf testicular tissue. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. On Deterioration Mechanism of Concrete Exposed to Freeze-Thaw Cycles

    Science.gov (United States)

    Trofimov, B. Ya; Kramar, L. Ya; Schuldyakov, K. V.

    2017-11-01

    At present, concrete and reinforced concrete are gaining ground in all sectors of construction including construction in the extreme north, on shelves, etc. Under harsh service conditions, the durability of reinforced concrete structures is related to concrete frost resistance. Frost resistance tests are accompanied by the accumulation of residual dilation deformations affected by temperature-humidity stresses, ice formation and other factors. Porosity is an integral part of the concrete structure which is formed as a result of cement hydration. The prevailing hypothesis of a deterioration mechanism of concrete exposed to cyclic freezing, i.e. the hypothesis of hydraulic pressure of unfrozen water in microcapillaries, does not take into account a number of phenomena that affect concrete resistance to frost aggression. The main structural element of concrete, i.e. hardened cement paste, contains various hydration products, such as crystalline, semicrystalline and gel-like products, pores and non-hydrated residues of clinker nodules. These structural elements in service can gain thermodynamic stability which leads to the concrete structure coarsening, decrease in the relaxation capacity of concrete when exposed to cycling. Additional destructive factors are leaching of portlandite, the difference in thermal dilation coefficients of hydration products, non-hydrated relicts, aggregates and ice. The main way to increase concrete frost resistance is to reduce the macrocapillary porosity of hardened cement paste and to form stable gel-like hydration products.

  1. Freezing-Enhanced Dissolution of Iron Oxides: Effects of Inorganic Acid Anions.

    Science.gov (United States)

    Jeong, Daun; Kim, Kitae; Min, Dae Wi; Choi, Wonyong

    2015-11-03

    Dissolution of iron from mineral dust particles greatly depends upon the type and amount of copresent inorganic anions. In this study, we investigated the roles of sulfate, chloride, nitrate, and perchlorate on the dissolution of maghemite and lepidocrocite in ice under both dark and UV irradiation and compared the results with those of their aqueous counterparts. After 96 h of reaction, the total dissolved iron in ice (pH 3 before freezing) was higher than that in the aqueous phase (pH 3) by 6-28 times and 10-20 times under dark and UV irradiation, respectively. Sulfuric acid was the most efficient in producing labile iron under dark condition, whereas hydrochloric acid induced the most dissolution of the total and ferrous iron in the presence of light. This ice-induced dissolution result was also confirmed with Arizona Test Dust (AZTD). In the freeze-thaw cycling test, the iron oxide samples containing chloride, nitrate, or perchlorate showed a similar extent of total dissolved iron after each cycling while the sulfate-containing sample rapidly lost its dissolution activity with repeating the cycle. This unique phenomenon observed in ice might be related to the freeze concentration of protons, iron oxides, and inorganic anions in the liquid-like ice grain boundary region. These results suggest that the ice-enhanced dissolution of iron oxides can be a potential source of bioavailable iron, and the acid anions critically influence this process.

  2. Repeated swim stress alters brain benzodiazepine receptors measured in vivo

    International Nuclear Information System (INIS)

    Weizman, R.; Weizman, A.; Kook, K.A.; Vocci, F.; Deutsch, S.I.; Paul, S.M.

    1989-01-01

    The effects of repeated swim stress on brain benzodiazepine receptors were examined in the mouse using both an in vivo and in vitro binding method. Specific in vivo binding of [ 3 H]Ro15-1788 to benzodiazepine receptors was decreased in the hippocampus, cerebral cortex, hypothalamus, midbrain and striatum after repeated swim stress (7 consecutive days of daily swim stress) when compared to nonstressed mice. In vivo benzodiazepine receptor binding was unaltered after repeated swim stress in the cerebellum and pons medulla. The stress-induced reduction in in vivo benzodiazepine receptor binding did not appear to be due to altered cerebral blood flow or to an alteration in benzodiazepine metabolism or biodistribution because there was no difference in [14C]iodoantipyrine distribution or whole brain concentrations of clonazepam after repeated swim stress. Saturation binding experiments revealed a change in both apparent maximal binding capacity and affinity after repeated swim stress. Moreover, a reduction in clonazepam's anticonvulsant potency was also observed after repeated swim stress [an increase in the ED50 dose for protection against pentylenetetrazol-induced seizures], although there was no difference in pentylenetetrazol-induced seizure threshold between the two groups. In contrast to the results obtained in vivo, no change in benzodiazepine receptor binding kinetics was observed using the in vitro binding method. These data suggest that environmental stress can alter the binding parameters of the benzodiazepine receptor and that the in vivo and in vitro binding methods can yield substantially different results

  3. Repeated swim stress alters brain benzodiazepine receptors measured in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Weizman, R.; Weizman, A.; Kook, K.A.; Vocci, F.; Deutsch, S.I.; Paul, S.M.

    1989-06-01

    The effects of repeated swim stress on brain benzodiazepine receptors were examined in the mouse using both an in vivo and in vitro binding method. Specific in vivo binding of (/sup 3/H)Ro15-1788 to benzodiazepine receptors was decreased in the hippocampus, cerebral cortex, hypothalamus, midbrain and striatum after repeated swim stress (7 consecutive days of daily swim stress) when compared to nonstressed mice. In vivo benzodiazepine receptor binding was unaltered after repeated swim stress in the cerebellum and pons medulla. The stress-induced reduction in in vivo benzodiazepine receptor binding did not appear to be due to altered cerebral blood flow or to an alteration in benzodiazepine metabolism or biodistribution because there was no difference in (14C)iodoantipyrine distribution or whole brain concentrations of clonazepam after repeated swim stress. Saturation binding experiments revealed a change in both apparent maximal binding capacity and affinity after repeated swim stress. Moreover, a reduction in clonazepam's anticonvulsant potency was also observed after repeated swim stress (an increase in the ED50 dose for protection against pentylenetetrazol-induced seizures), although there was no difference in pentylenetetrazol-induced seizure threshold between the two groups. In contrast to the results obtained in vivo, no change in benzodiazepine receptor binding kinetics was observed using the in vitro binding method. These data suggest that environmental stress can alter the binding parameters of the benzodiazepine receptor and that the in vivo and in vitro binding methods can yield substantially different results.

  4. The Effect of Freezing Stress on Percentage of Electrolytes Leakage and Survival of Flixweed (Descurainia sophia L. Seedlings

    Directory of Open Access Journals (Sweden)

    E Izadi-Darbandi

    2016-10-01

    Full Text Available Introduction Flixweed (Descurainia sophia L. is a medicinal plant from Brassicaceae family which also known as a weed for winter cereals and oil seed rape. Low temperatures are one of the most important abiotic stresses that threat Flixweed growth and productivity. Therefore it is important to recognize the freeze tolerance of Flixweed for successful planting and utilization in cold regions such as Mashhad in Khorasan Razavi Province (Iran’s north. east. Among many laboratory methods which have been developed to estimate and to evaluate plants response or their tolerance to freez ing temperatures, electrolyte leakage (EL test is widely used. This test is based on this principle that damage to the cell membranes results in enhanced leakage of solutes into the apoplastic water, hence recording the amount of leakage after stress treatments provides an estimation of tissue injury. Indeed continuing integration of plasma membrane is one important factor for survival of plants under freezing stress and any disturbance in membrane structure can lead to damage and death. So determination of LT50 point or critical temperature for electrolytes leakage and survival of plant is the most reliable, quantitative and simple methods for evaluating the cold tolerance of plants. The aim of this trial was to determine the LT50 according to the EL and SU% for Flixweed ecotypes. Materials and Methods In order to evaluate freeze tolerance in Flixweed, a factorial experiment based on completely randomized design with three replications was carried out in college of agriculture, Ferdowsi University of Mashhad. Experimental factors included five ecotypes of Flixweed (Eghlid, Sabzewar, Hamedan, Torbat-e-Jam and Neyshabour and 10 freezing temperature levels (0, -2,-4, -6, -8,-10,-12,-14,-16 and -18°C. Flixweed seeds were cultivated in pots in autumn of 2008 and were grown until 5-7 leaf stage under natural weather conditions for acclimation. Then to apply freezing

  5. Spring photosynthetic onset and net CO2 uptake in Alaska triggered by landscape thawing.

    Science.gov (United States)

    Parazoo, Nicholas C; Arneth, Almut; Pugh, Thomas A M; Smith, Ben; Steiner, Nicholas; Luus, Kristina; Commane, Roisin; Benmergui, Josh; Stofferahn, Eric; Liu, Junjie; Rödenbeck, Christian; Kawa, Randy; Euskirchen, Eugenie; Zona, Donatella; Arndt, Kyle; Oechel, Walt; Miller, Charles

    2018-04-24

    The springtime transition to regional-scale onset of photosynthesis and net ecosystem carbon uptake in boreal and tundra ecosystems are linked to the soil freeze-thaw state. We present evidence from diagnostic and inversion models constrained by satellite fluorescence and airborne CO 2 from 2012 to 2014 indicating the timing and magnitude of spring carbon uptake in Alaska correlates with landscape thaw and ecoregion. Landscape thaw in boreal forests typically occurs in late April (DOY 111 ± 7) with a 29 ± 6 day lag until photosynthetic onset. North Slope tundra thaws 3 weeks later (DOY 133 ± 5) but experiences only a 20 ± 5 day lag until photosynthetic onset. These time lag differences reflect efficient cold season adaptation in tundra shrub and the longer dehardening period for boreal evergreens. Despite the short transition from thaw to photosynthetic onset in tundra, synchrony of tundra respiration with snow melt and landscape thaw delays the transition from net carbon loss (at photosynthetic onset) to net uptake by 13 ± 7 days, thus reducing the tundra net carbon uptake period. Two global CO 2 inversions using a CASA-GFED model prior estimate earlier northern high latitude net carbon uptake compared to our regional inversion, which we attribute to (i) early photosynthetic-onset model prior bias, (ii) inverse method (scaling factor + optimization window), and (iii) sparsity of available Alaskan CO 2 observations. Another global inversion with zero prior estimates the same timing for net carbon uptake as the regional model but smaller seasonal amplitude. The analysis of Alaskan eddy covariance observations confirms regional scale findings for tundra, but indicates that photosynthesis and net carbon uptake occur up to 1 month earlier in evergreens than captured by models or CO 2 inversions, with better correlation to above-freezing air temperature than date of primary thaw. Further collection and analysis of boreal evergreen species over

  6. Canalization of freeze tolerance in an alpine grasshopper.

    Science.gov (United States)

    Hawes, Timothy C

    2015-10-01

    In the Rock and Pillar Range, New Zealand, the alpine grasshopper, Sigaus australis Hutton, survives equilibrium freezing (EF) all-year round. A comparison of freeze tolerance (FT) in grasshoppers over four austral seasons for a 1 year period finds that: (a) the majority (>70%) of the sample population of grasshoppers survive single freeze-stress throughout the year; (b) exposure to increased freeze stress (multiple freeze-stress events) does not lead to a loss of freeze tolerance; and (c) responses to increased freeze stress reveal seasonal tuning of the FT adaptation to environmental temperatures. The Rock and Pillar sample population provides a clear example of the canalization of the FT adaptation. Seasonal variability in the extent of tolerance of multiple freezing events indicates that physiology is modulated to environmental temperatures by phenotypic plasticity - i.e. the FT adaptation is permanent and adjustable. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Plasma Membrane ATPase Activity following Reversible and Irreversible Freezing Injury 1

    Science.gov (United States)

    Iswari, S.; Palta, Jiwan P.

    1989-01-01

    Plasma membrane ATPase has been proposed as a site of functional alteration during early stages of freezing injury. To test this, plasma membrane was purified from Solanum leaflets by a single step partitioning of microsomes in a dextran-polyethylene glycol two phase system. Addition of lysolecithin in the ATPase assay produced up to 10-fold increase in ATPase activity. ATPase activity was specific for ATP with a Km around 0.4 millimolar. Presence of the ATPase enzyme was identified by immunoblotting with oat ATPase antibodies. Using the phase partitioning method, plasma membrane was isolated from Solanum commersonii leaflets which had four different degrees of freezing damage, namely, slight (reversible), partial (partially reversible), substantial and total (irreversible). With slight (reversible) damage the plasma membrane ATPase specific activity increased 1.5- to 2-fold and its Km was decreased by about 3-fold, whereas the specific activity of cytochrome c reductase and cytochrome c oxidase in the microsomes were not different from the control. However, with substantial (lethal, irreversible) damage, there was a loss of membrane protein, decrease in plasma membrane ATPase specific activity and decrease in Km, while cytochrome c oxidase and cytochrome c reductase were unaffected. These results support the hypothesis that plasma membrane ATPase is altered by slight freeze-thaw stress. Images Figure 1 Figure 2 PMID:16666856

  8. Cryopreservation of human sperm: efficacy and use of a new nitrogen-free controlled rate freezer versus liquid nitrogen vapour freezing.

    Science.gov (United States)

    Creemers, E; Nijs, M; Vanheusden, E; Ombelet, W

    2011-12-01

    Preservation of spermatozoa is an important aspect of assisted reproductive medicine. The aim of this study was to investigate the efficacy and use of a recently developed liquid nitrogen and cryogen-free controlled rate freezer and this compared with the classical liquid nitrogen vapour freezing method for the cryopreservation of human spermatozoa. Ten patients entering the IVF programme donated semen samples for the study. Samples were analysed according to the World Health Organization guidelines. No significant difference in total sperm motility after freeze-thawing between the new technique and classical technique was demonstrated. The advantage of the new freezing technique is that it uses no liquid nitrogen during the freezing process, hence being safer to use and clean room compatible. Investment costs are higher for the apparatus but running costs are only 1% in comparison with classical liquid nitrogen freezing. In conclusion, post-thaw motility of samples frozen with the classical liquid nitrogen vapour technique was comparable with samples frozen with the new nitrogen-free freezing technique. This latter technique can thus be a very useful asset to the sperm cryopreservation laboratory. © 2011 Blackwell Verlag GmbH.

  9. Addition of cholesterol-loaded cyclodextrins to the thawing extender: effects on boar sperm quality.

    Science.gov (United States)

    Tomás, C; Gómez-Fernández, J; Gómez-Izquierdo, E; Mocé, E; de Mercado, E

    2014-06-01

    The aim of the present study was to evaluate the effect that the addition of cholesterol-loaded cyclodextrins (CLC) to the thawing extender has on the quality of frozen-thawed boar sperm. Pooled semen (n = 5) from three boars was used for the experiments. The semen was cryopreserved with an egg-yolk-based extender, it was diluted after thawing in Beltsville thawing solution (BTS) supplemented with different concentrations of CLC (0, 12.5, 25, 50 or 100 mg/500 × 10(6) sperm), and these samples were incubated at 37°C for 150 min. The following parameters of sperm quality were evaluated 30 and 150 min after incubation: sperm with intact plasma membrane (SIPM; %), sperm with normal acrosomal ridge (NAR; %), total motile sperm (TMS; %), progressively motile sperm (PMS; %) and kinetic parameters. Both SIPM and NAR increased (p extender was supplemented with 12.5, 25 and 50 mg CLC/500 × 10(6) sperm. Nevertheless, motility decreased (p extenders with CLC improves sperm viability and reduces acrosome damage after freezing/thawing. © 2014 Blackwell Verlag GmbH.

  10. Comparative effect of different concentrations of hydro-ethanolic extract of chamomile on freeze-thawn semen quality of rams

    Directory of Open Access Journals (Sweden)

    hossein daghighkia

    2017-04-01

    Full Text Available Oxidative stress during freeze-thawing process causes reduction in motility, viability, membrane functions and finally sperm fertility. The aim of this study was to determine the effect of Chamomile extract as natural antioxidant on quality of cryopreserved ram sperm. In this study semen was collected from five mature rams twice a week using an artificial vagina and the ejaculates were pooled equally in order to eliminate the individual effects. Different levels of extract of Chamomile (0, 50, 66.66, 100 and 200 ml/dl of diluent solution were added to diluent based tris-egg yolk. After cooling, filling and sealing of the samples, they were frozen with nitrogen vapor and immersed in liquid nitrogen and were stored until evaluation time. After thawing, results showed that addition of 66.66 ml/dl of extract increased total motility, progressive motility, curvilinear velocity, straight–line velocity, linearity, viability and plasma membrane integrity of sperm compared to the control and other treatment groups (p

  11. Impacts of post-mortem ageing prior to freezing on technological and oxidative properties of coarse ground lamb sausage in a model system

    Directory of Open Access Journals (Sweden)

    Juhui Choe

    2017-07-01

    Full Text Available Objective The objective of this study was to evaluate the effects of ageing time of lamb loins prior to freezing on technological characteristics and oxidation stability of coarse ground lamb loin sausage using in a model system. Methods Lamb loins (M. longissimus lumborum, n = 25 were aged at −1.5°C for 0, 1, 2, 3, and 8 wk and then frozen for the remaining days (a total of 30 wk. The aged/frozen/thawed lamb loins were ground, and model sausages were formulated with 75% aged/frozen/thawed lamb loin, 25% water, 1.5% sodium chloride (NaCl and 0.3% sodium tripolyphosphate. The pH and thaw/purge loss of aged/frozen/thawed lamb loins were evaluated, and protein functionality (protein solubility and emulsifying capacity, water-holding capacity and textural properties of model sausages were determined. Cooked model sausages were vacuum-packaged in a plastic bag and displayed under continuous fluorescent natural white light (3°C±1°C. Colour and lipid oxidation of the cooked model sausages were evaluated on 0 and 21 d of display storage. Results Ageing prior to freezing had no impact on pH and purge/thaw loss of lamb loins and the colour of cooked sausages (p>0.05 made from the loins. Lamb loins aged for at least 3 wk prior to freezing numerically improved total and myofibrillar protein solubilities (p>0.05 and emulsion activity index (p = 0.009 of meat batter, but decreased cooking loss (p = 0.003 and lipid oxidation (p<0.05 of model sausages. Conclusion This study suggests that post-mortem ageing of raw meat prior to freezing could improve water-holding capacity and lipid oxidative stability of sausage made from the meat.

  12. Impacts of post-mortem ageing prior to freezing on technological and oxidative properties of coarse ground lamb sausage in a model system.

    Science.gov (United States)

    Choe, Juhui; Kim, Hyun-Wook; Farouk, Mustafa M; Brad Kim, Yuan H

    2017-07-01

    The objective of this study was to evaluate the effects of ageing time of lamb loins prior to freezing on technological characteristics and oxidation stability of coarse ground lamb loin sausage using in a model system. Lamb loins ( M. longissimus lumborum , n = 25) were aged at -1.5°C for 0, 1, 2, 3, and 8 wk and then frozen for the remaining days (a total of 30 wk). The aged/frozen/thawed lamb loins were ground, and model sausages were formulated with 75% aged/frozen/thawed lamb loin, 25% water, 1.5% sodium chloride (NaCl) and 0.3% sodium tripolyphosphate. The pH and thaw/purge loss of aged/frozen/thawed lamb loins were evaluated, and protein functionality (protein solubility and emulsifying capacity), water-holding capacity and textural properties of model sausages were determined. Cooked model sausages were vacuum-packaged in a plastic bag and displayed under continuous fluorescent natural white light (3°C±1°C). Colour and lipid oxidation of the cooked model sausages were evaluated on 0 and 21 d of display storage. Ageing prior to freezing had no impact on pH and purge/thaw loss of lamb loins and the colour of cooked sausages (p>0.05) made from the loins. Lamb loins aged for at least 3 wk prior to freezing numerically improved total and myofibrillar protein solubilities (p>0.05) and emulsion activity index (p = 0.009) of meat batter, but decreased cooking loss (p = 0.003) and lipid oxidation (p<0.05) of model sausages. This study suggests that post-mortem ageing of raw meat prior to freezing could improve water-holding capacity and lipid oxidative stability of sausage made from the meat.

  13. The effects of different levels of catalase and superoxide dismutase in modified Beltsville extender on rooster post-thawed sperm quality.

    Science.gov (United States)

    Amini, Mahmood Reza; Kohram, Hamid; Zare-Shahaneh, Ahmad; Zhandi, Mahdi; Sharideh, Hossein; Nabi, Mohammad Mehdi

    2015-06-01

    Oxidative damage of sperm by means of reactive oxygen species generated by the cellular components of semen is one of the main reasons for decreased sperm motility and fertility during the freeze-thawing process. This study was conducted to determine the influence of catalase (CAT) and superoxide dismutase (SOD) on rooster sperm motility, viability and MDA level after freezing and thawing. Semen samples from 10 sexually-mature Ross 308 breeder roosters were collected and pooled, divided into nine equal parts and diluted with modified Beltsville extender containing no antioxidants (control), or supplemented with 50, 100, 200 and 300 μg/mL CAT, or 50, 100, 200 and 300 U/mL SOD. After thawing, sperm motility and motion parameters were assessed using a CASA system. Sperm viability and MDA level were assessed by eosin-nigrosin and MDA test, respectively. The results of this experiment showed that the extender supplemented with 100 and 200 μg CAT, and 50 U SOD had the highest sperm motility (Prooster semen. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Theory about the Embryo Cryo-Treatment.

    Science.gov (United States)

    Vladimirov, Iavor K; Tacheva, Desislava; Diez, Antonio

    2017-04-01

    To create hypothesis, which can give a logical explanation related to the benefits of freezing/thawing embryos. Cryopreservation is not only a technology used for storing embryos, but also a method of embryo treatment that can potentially improve the success rate in infertile couples. From the analysis of multiple results in assisted reproductive technology, which have no satisfactory explanation to date, we found evidence to support a 'therapeutic' effect of the freezing/thawing of embryos on the process of recovery of the embryo and its subsequent implantation. Freezing/thawing is a way to activate the endogenous survival and repair responses in preimplantation embryos. Several molecular mechanisms can explain the higher success rate of ET using thawed embryos compared to fresh ET in women of advanced reproductive age, the higher miscarriage rate in cases of thawed blastocyst ET compared to thawed ET at early cleavage embryo, and the higher perinatal parameters of born children after thawed ET. Embryo thawing induces a stress. Controlled stress is not necessarily detrimental, because it generates a phenomenon that is counteracted by several known biological responses aimed to repair mitochondrial damage of membrane and protein misfolding. The term for favorable biological responses to low exposures to stress is called hormesis. This thesis will summarize the role of cryopreservation in the activation of a hormetic response, preserving the mitochondrial function, improving survival, and having an impact on the process of implantation, miscarriage, and the development of pregnancy.

  15. Combinations of glycerol percent, glycerol equilibration time, and thawing rate upon freezability of bull spermatozoa in plastic straws.

    Science.gov (United States)

    Wiggin, H B; Almquist, J O

    1975-03-01

    Twelve ejaculates were used in a central composite experiment to test 15 combinations of glycerol (7, 9, 11, 13, or 15%), glycerol equilibration times (1, 2, 4, 8, or 16 h) and thawing rates (water at 35 C for 15 s, 50 C for 13 s, 65 C for 11 s, 80 C for 9 s, or 95 C for 7 s). Semen was diluted in heated skim milk-glycerol, packaged in .3-ml. Continental U.S. straws and frozen in liquid nitrogen vapor. Based on post-thaw progressive sperm motility after storage at -196 C for 9 to 11 days, estimated optima from multiple regression were 10.7% for glycerol, 2.0 h for glycerol equilibration time, and 76 C for thawing bath temperature. Only the linear effect for each variable was significant. Much faster thawing rates and shorter glycerol equilibration times than those for freezing bull spermatozoa in glass ampules should be used for maximum post-thaw sperm motility in straws.

  16. Evaluation of the Activities of Antioxidant Enzyme and Lysosomal Enzymes of the Longissimus dorsi Muscle from Hanwoo (Korean Cattle) in Various Freezing Conditions

    OpenAIRE

    Kang, Sun Moon; Kang, Geunho; Seong, Pil-Nam; Park, Beomyoung; Kim, Donghun; Cho, Soohyun

    2014-01-01

    This study was conducted to evaluate the activities of antioxidant enzyme (glutathione peroxidase (GSH-Px)) and lysosomal enzymes (alpha-glucopyranosidase (AGP) and beta-N-acetyl-glucosaminidase (BNAG)) of the longissimus dorsi (LD) muscle from Hanwoo (Korean cattle) in three freezing conditions. Following freezing at -20, -60, and -196? (liquid nitrogen), LD samples (48 h post-slaughter) were treated as follows: 1) freezing for 14 d, 2) 1 to 4 freeze-thaw cycles (2 d of freezing in each cycl...

  17. Effect of different concentration of fish oil in skim milk-egg yolk extenders on post- thawed semen qualities of Kalang swamp buffalo bull

    Directory of Open Access Journals (Sweden)

    Abdul Malik

    2018-01-01

    Full Text Available Objective: To explore the effect of fish oil at different concentrations on post-thawed semen of Kalang swamp buffalo. Methods: A total of 4 Kalang swamp buffalo bulls with 3-5 years of age and weighed about 340-360 kg were slected. Semen was regularly collected from these buffalo bulls once a week by an artificial vagina. Fish oil was supplementary at the dosages of 0 mg (control, 50 mg, 100 mg, 150 mg, and 200 mg to the extender (skim milk-egg yolk. Fresh, pre-freezing and frozen semen were thawed at 37 °C and evaluated for motility, viability, morphology, and plasma integrity of membrane. Results: The study results indicated that before freezing, supplementation of fish oil at the dose of 150 mg in the extender had significantly motility. And a significant (P<0.05 increase was observed in viability and motility of post-thawed semen at the dose of 150 mg fish oil, which was in difference with other treatment groups. Conclusions: Addition of 150 mg fish oil in the extender could be positive for the enhancement of the quality of post-thawed semen of Kalang swamp buffaloes.

  18. Thermal stress state of cryogenic HP vessels under freezing and pressurization

    International Nuclear Information System (INIS)

    Tsybenko, A.S.; Kuranov, B.A.; Chepurnoj, A.D.; Shaposhnikov, V.A.; Krishchuk, N.G.

    1986-01-01

    A mathematical model is developed for thermomechanical processes in cryogenic HP vessels under freezing either by liquid and (or) gaseous cryogen and under pressurization. Equations of nonlinear nonstationary thermal conductivity and nonisothermal thermoelastoplasticity are used for the case of the theory off low with isotropic hardening. Semiempiricaldependences of nonstationary heat exchange for gaseous medium, experimental curves of cryogenic liquid boiling, mass exchange relationships are allowed for when formulating boundary conditions. The mathematical modelis realized on the basi of the finite element method in the form of highly automated program complex TERSOD (heat resistanceof vessels), oriented for computer of the Unified System. Heat and stress-strained states for three constructions of vessels are thoroughly studied under different conditions of gaseous, liquid and combined freezing with subsequent pressurization

  19. Effect of food additives on egg yolk gelation induced by freezing.

    Science.gov (United States)

    Primacella, Monica; Fei, Tao; Acevedo, Nuria; Wang, Tong

    2018-10-15

    This study demonstrates technological advances in preventing yolk gelation during freezing and thawing. Gelation negatively affects yolk functionality in food formulation. Preventing gelation using 10% salt or sugar limits the application of the yolk. Novel food additives were tested to prevent gelation induced by freezing. Significant reduction (p freezing at -20 °C) indicates that hydrolyzed carboxymethyl cellulose (HCMC), proline, and hydrolyzed egg white and yolk (HEW and HEY) are effective gelation inhibitors. The mechanisms in which these additives prevented gelation were further studied through measuring the changes in the amount of freezable water, lipoprotein particle size, and protein surface hydrophobicity. Overall, this study provides several alternatives of gelation inhibitor that have great potentials in replacing the use of salt or sugar in commercial operation of freezing egg yolk for shelf-life extension. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. Hyaluronic acid improves frozen-thawed sperm quality and fertility potential in rooster.

    Science.gov (United States)

    Lotfi, Saied; Mehri, Morteza; Sharafi, Mohsen; Masoudi, Reza

    2017-09-01

    Beneficial effects of Hyaluronic acid (HA) has not been yet assessed for cryopreservation of rooster sperm. This study was conducted to evaluate the effects of different concentrations of HA (0, 1, 2, 4 and 8mM) in Beltsville extender on the cryopreservation of rooster sperm. Semen samples were collected from six Ross broiler breeders (24-week) using abdominal massage, then divided into five equal aliquots and cryopreserved in Beltsville extender that contained different concentrations of HA. Motion characteristics, morphology, membrane functionality, viability, acrosome integrity, lipid peroxidation and fertility potential of sperm were assessed after thawing. HA at concentration of 2mM (HA2) resulted in the highest (Prooster sperm after freeze thawing. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Cryopreservation of turkey semen: effect of breeding line and freezing method on post-thaw sperm quality, fertilization, and hatching

    Science.gov (United States)

    Cryopreservation methods for poultry semen are not reliable for germplasm preservation, especially for turkeys, where fertility rates from frozen/thawed semen are particularly low. The objective was to evaluate cryopreservation methods for effectiveness in promoting cryosurvival and post-thaw funct...

  2. Cod and rainbow trout as freeze-chilled meal elements

    DEFF Research Database (Denmark)

    Jensen, Louise Helene Søgaard; Nielsen, Jette; Jørgensen, Bo

    2010-01-01

    Meal elements' are elements of a meal, e.g. portions of pre-fried meat, sauces, frozen fish or pre-processed vegetables typically prepared industrially. The meal elements are distributed to professional satellite kitchens, where the staff can combine them into complete meals. Freeze......-chilling is a process consisting of freezing and frozen storage followed by thawing and chilled storage. Combining the two would enable the manufacturer to produce large quantities of frozen meal elements to be released into the chill chain according to demand. We have studied the influence of freeze...... days of chilled storage, and the corresponding time for rainbow trout was 10 days. After this period the sensory quality decreased and chemical indicators of spoilage were seen to increase. CONCLUSION: The consistent quality during storage and the high-quality shelf life are practically applicable...

  3. Xylem diameter changes during osmotic stress, desiccation and freezing in Pinus sylvestris and Populus tremula.

    Science.gov (United States)

    Lintunen, Anna; Lindfors, Lauri; Nikinmaa, Eero; Hölttä, Teemu

    2017-04-01

    Trees experience low apoplastic water potential frequently in most environments. Low apoplastic water potential increases the risk of embolism formation in xylem conduits and creates dehydration stress for the living cells. We studied the magnitude and rate of xylem diameter change in response to decreasing apoplastic water potential and the role of living parenchyma cells in it to better understand xylem diameter changes in different environmental conditions. We compared responses of control and heat-injured xylem of Pinus sylvestris (L.) and Populus tremula (L.) branches to decreasing apoplastic water potential created by osmotic stress, desiccation and freezing. It was shown that xylem in control branches shrank more in response to decreasing apoplastic water potential in comparison with the samples that were preheated to damage living xylem parenchyma. By manipulating the osmotic pressure of the xylem sap, we observed xylem shrinkage due to decreasing apoplastic water potential even in the absence of water tension within the conduits. These results indicate that decreasing apoplastic water potential led to withdrawal of intracellular water from the xylem parenchyma, causing tissue shrinkage. The amount of xylem shrinkage per decrease in apoplastic water potential was higher during osmotic stress or desiccation compared with freezing. During desiccation, xylem diameter shrinkage involved both dehydration-related shrinkage of xylem parenchyma and water tension-induced shrinkage of conduits, whereas dehydration-related shrinkage of xylem parenchyma was accompanied by swelling of apoplastic ice during freezing. It was also shown that the exchange of water between symplast and apoplast within xylem is clearly faster than previously reported between the phloem and the xylem. Time constant of xylem shrinkage was 40 and 2 times higher during osmotic stress than during freezing stress in P. sylvestris and P. tremula, respectively. Finally, it was concluded that the

  4. Pre-cure freezing affects proteolysis in dry-cured hams.

    Science.gov (United States)

    Bañón, S; Cayuela, J M; Granados, M V; Garrido, M D

    1999-01-01

    Several parameters (sodium chloride, moisture, intramuscular fat, total nitrogen, non-protein nitrogen, white precipitates, free tyrosine, L* a* b* values and acceptability) related with proteolysis during the curing were compared in dry-cured hams manufactured from refrigerated and frozen/thawed raw material. Pre-cure freezing increased the proteolysis levels significantly (pcured meat, although it does not significantly affect the sensory quality of the dry-cured ham.

  5. Enriching the captive elephant population genetic pool through artificial insemination with frozen-thawed semen collected in the wild.

    Science.gov (United States)

    Hildebrandt, T B; Hermes, R; Saragusty, J; Potier, R; Schwammer, H M; Balfanz, F; Vielgrader, H; Baker, B; Bartels, P; Göritz, F

    2012-10-01

    The first successful AI in an elephant was reported in 1998, using fresh semen. Since then almost 40 calves have been produced through AI in both Asian and African elephants worldwide. Following these successes, with the objective of enriching the captive population with genetic material from the wild, we evaluated the possibility of using frozen-thawed semen collected from wild bulls for AI in captivity. Semen, collected from a 36-yr-old wild African savanna elephant (Loxodonta africana) in South Africa was frozen using the directional freezing technique. This frozen-thawed semen was used for four inseminations over two consecutive days, two before and two after ovulation, in a 26-yr-old female African savanna elephant in Austria. Insemination dose of 1200 × 10(6) cells per AI with 61% motility resulted in pregnancy, which was confirmed through ultrasound examination 75, 110 and 141 days after the AI procedure. This represents the first successful AI using wild bull frozen-thawed semen in elephants. The incorporation of AI with frozen-thawed semen into the assisted reproduction toolbox opens the way to preserve and transport semen between distant individuals in captivity or, as was done in this study, between wild and captive populations, without the need to transport stressed or potentially disease-carrying animals or to remove animals from the wild. In addition, cryopreserved spermatozoa, in combination with AI, are useful methods to extend the reproductive lifespan of individuals beyond their biological lifespan and an important tool for genetic diversity management and phenotype selection in these endangered mammals. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. Construction of chitin/PVA composite hydrogels with jellyfish gel-like structure and their biocompatibility.

    Science.gov (United States)

    He, Meng; Wang, Zhenggang; Cao, Yan; Zhao, Yanteng; Duan, Bo; Chen, Yun; Xu, Min; Zhang, Lina

    2014-09-08

    High strength chitin/poly(vinyl alcohol) (PVA) composite hydrogels (RCP) were constructed by adding PVA into chitin dissolved in a NaOH/urea aqueous solution, and then by cross-linking with epichlorohydrin (ECH) and freezing-thawing process. The RCP hydrogels were characterized by field emission scanning electron microscopy, FTIR, differential scanning calorimetry, solid-state (13)C NMR, wide-angle X-ray diffraction, and compressive test. The results revealed that the repeated freezing/thawing cycles induced the bicrosslinked networks consisted of chitin and PVA crystals in the composite gels. Interestingly, a jellyfish gel-like structure occurred in the RCP75 gel with 25 wt % PVA content in which the amorphous and crystalline PVA were immobilized tightly in the chitin matrix through hydrogen bonding interaction. The freezing/thawing cycles played an important role in the formation of the layered porous PVA networks and the tight combining of PVA with the pore wall of chitin. The mechanical properties of RCP75 were much higher than the other RCP gels, and the compressive strength was 20× higher than that of pure chitin gels, as a result of broadly dispersing stress caused by the orderly multilayered networks. Furthermore, the cell culture tests indicated that the chitin/PVA composite hydrogels exhibited excellent biocompatibility and safety, showing potential applications in the field of tissue engineering.

  7. The Hydrosphere State (Hydros) Satellite Mission: An Earth System Pathfinder for Global Mapping of Soil Moisture and Land Freeze/Thaw

    Science.gov (United States)

    Entekhabi, D.; Njoku, E. G.; Spencer, M.; Kim, Y.; Smith, J.; McDonald, K. C.; vanZyl, J.; Houser, P.; Dorion, T.; Koster, R.; hide

    2004-01-01

    The Hydrosphere State Mission (Hydros) is a pathfinder mission in the National Aeronautics and Space Administration (NASA) Earth System Science Pathfinder Program (ESSP). The objective of the mission is to provide exploratory global measurements of the earth's soil moisture at 10-km resolution with two- to three-days revisit and land-surface freeze/thaw conditions at 3-km resolution with one- to two-days revisit. The mission builds on the heritage of ground-based and airborne passive and active low-frequency microwave measurements that have demonstrated and validated the effectiveness of the measurements and associated algorithms for estimating the amount and phase (frozen or thawed) of surface soil moisture. The mission data will enable advances in weather and climate prediction and in mapping processes that link the water, energy, and carbon cycles. The Hydros instrument is a combined radar and radiometer system operating at 1.26 GHz (with VV, HH, and HV polarizations) and 1.41 GHz (with H, V, and U polarizations), respectively. The radar and the radiometer share the aperture of a 6-m antenna with a look-angle of 39 with respect to nadir. The lightweight deployable mesh antenna is rotated at 14.6 rpm to provide a constant look-angle scan across a swath width of 1000 km. The wide swath provides global coverage that meet the revisit requirements. The radiometer measurements allow retrieval of soil moisture in diverse (nonforested) landscapes with a resolution of 40 km. The radar measurements allow the retrieval of soil moisture at relatively high resolution (3 km). The mission includes combined radar/radiometer data products that will use the synergy of the two sensors to deliver enhanced-quality 10-km resolution soil moisture estimates. In this paper, the science requirements and their traceability to the instrument design are outlined. A review of the underlying measurement physics and key instrument performance parameters are also presented.

  8. Cryopreservation of boar semen. II: Effect of cooling rate and duration of freezing point plateau on boar semen frozen in mini- and maxi-straws and plastic bags.

    Science.gov (United States)

    Bwanga, C O; Einarsson, S; Rodriguez-Martinez, H

    1991-01-01

    The post-thaw motility and the acrosome integrity of semen from 4 boars frozen with a programmable freezing machine, in mini (0.25 ml) and maxi (5 ml) plastic straws and in 10 x 5 cm Teflon FEP-plastic bags (0.12 mm thick, 5 ml), were compared. The freezing of the semen was monitored by way of thermo-couples placed in the straws and the bags. Three freezing programmes were used, namely A: from +5 degrees C, at a rate of 3 degrees C/min, to -6 degrees C, held for 1 min at -6 degrees C, and followed by a cooling rate of 20 degrees C/min to -100 degrees C; B: a similar curve except that there was no holding time at -6 degrees C and that the cooling rate was 30 degrees C/min, and C: from +5 degrees C to -100 degrees C, with a cooling rate of 35 degrees C/min, followed by storage in liquid N2. Despite the freezing curve assayed, both the mini-straws and the bags depicted much shorter freezing point plateaus as compared to the maxi-straws. Post-thaw sperm motility as well as the amount of normal apical ridges were equally significantly higher when semen was frozen in mini-straws or in bags than in maxi-straws. Significant differences in these post-thawing parameters were obtained between the freezing curves used. The stepwise freezing procedure A appeared as the best alternative for boar semen, considering this in vitro evaluation.

  9. DNA comet assay to identify different freezing temperatures of irradiated liver chicken

    International Nuclear Information System (INIS)

    Duarte, Renato C.; Mozeika, Michel A.; Fanaro, Gustavo B.; Villavicencio, Anna L.C.H.; Marchioni, Eric

    2009-01-01

    The cold chain is a succession of steps which maintain the food at low temperature. The thawed food never be frozen again and the best solution being to consume it quickly to avoid the microorganism growth which causes decay and nutrients damage. One of most important point is that freezing process, unlike irradiation, do not destroy microorganisms, only inactive them as long as they remain in a frozen state. The Comet Assay is an original test used to detect irradiated foods that's recognize the DNA damage and can then be used to control the overall degradation of the food and in a certain extend to evaluate the damage caused by irradiation, different forms of freeze and storage time on liver chicken cells. Different freezing temperatures were used, deep freeze -196 deg C and slow freeze -10 deg C. Samples were irradiated in a 60 Co irradiator with 1.5, 3.0 and 4.5 kGy radiation doses. Fast freezing technique induces a low percent of DNA degradation comparing to slow freezing technique. This procedure could be a good choose to chicken freezing processing. (author)

  10. DNA comet assay to identify different freezing temperatures of irradiated liver chicken

    Energy Technology Data Exchange (ETDEWEB)

    Duarte, Renato C.; Mozeika, Michel A.; Fanaro, Gustavo B.; Villavicencio, Anna L.C.H. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)], e-mail: renatocduarte@yahoo.com.br; Marchioni, Eric [Universite de Strasbourg, Illkirch (France). Faculte de Pharmacie. Lab. de Chimie Analytique et Sciences de l' Aliment

    2009-07-01

    The cold chain is a succession of steps which maintain the food at low temperature. The thawed food never be frozen again and the best solution being to consume it quickly to avoid the microorganism growth which causes decay and nutrients damage. One of most important point is that freezing process, unlike irradiation, do not destroy microorganisms, only inactive them as long as they remain in a frozen state. The Comet Assay is an original test used to detect irradiated foods that's recognize the DNA damage and can then be used to control the overall degradation of the food and in a certain extend to evaluate the damage caused by irradiation, different forms of freeze and storage time on liver chicken cells. Different freezing temperatures were used, deep freeze -196 deg C and slow freeze -10 deg C. Samples were irradiated in a {sup 60}Co irradiator with 1.5, 3.0 and 4.5 kGy radiation doses. Fast freezing technique induces a low percent of DNA degradation comparing to slow freezing technique. This procedure could be a good choose to chicken freezing processing. (author)

  11. Non-Toxic, Low-Freezing, Drop-In Replacement Heat Transfer Fluids

    Science.gov (United States)

    Cutbirth, J. Michael

    2012-01-01

    A non-toxic, non-flammable, low-freezing heat transfer fluid is being developed for drop-in replacement within current and future heat transfer loops currently using water or alcohol-based coolants. Numerous water-soluble compounds were down-selected and screened for toxicological, physical, chemical, compatibility, thermodynamic, and heat transfer properties. Two fluids were developed, one with a freezing point near 0 C, and one with a suppressed freezing point. Both fluids contain an additive package to improve material compatibility and microbial resistance. The optimized sub-zero solution had a freezing point of 30 C, and a freezing volume expansion of 10-percent of water. The toxicity of the solutions was experimentally determined as LD(50) greater than 5g/kg. The solutions were found to produce minimal corrosion with materials identified by NASA as potentially existing in secondary cooling loops. Thermal/hydrodynamic performance exceeded that of glycol-based fluids with comparable freezing points for temperatures Tf greater than 20 C. The additive package was demonstrated as a buffering agent to compensate for CO2 absorption, and to prevent microbial growth. The optimized solutions were determined to have physically/chemically stable shelf lives for freeze/thaw cycles and longterm test loop tests.

  12. Influence of immersion freezing in NaCl solutions and of frozen storage on the viscoelastic behavior of mozzarella cheese.

    Science.gov (United States)

    Ribero, G G; Rubiolo, A C; Zorrilla, S E

    2007-06-01

    The freezing of Mozzarella cheese by immersion in NaCl solutions may be an innovative procedure for the dairy industry because it combines conveniently salting and freezing processes. In this work, the influence of this type of freezing method and of the frozen storage of samples on the viscoelastic behavior of Mozzarella cheese was studied. Slabs (2 x 10 x 10 cm(3)) were immersed in 23% w/w NaCl solutions (control samples: 4 degrees C, 90 min; frozen samples: -15 degrees C, 180 min). Half of the frozen samples were immediately thawed at 4 degrees C . The other half was stored at -20 degrees C for 2 mo and then was thawed at 4 degrees C (frozen-stored samples). Samples were stored at 4 degrees C and assayed at 1, 7, 14, 20, 27, 34, and 41 d. Rheological tests were carried out in oscillatory mode (parallel-plate geometry, diameter: 20 mm, gap: 1 mm, frequency: 1 Hz). Strain sweeps were run (0.001 immersion freezing of Mozzarella cheese affects some of the studied parameters, the differences observed between frozen and frozen-stored samples with control samples were small. Therefore, it was considered that the immersion freezing might be useful for the manufacture and commercialization of Mozzarella cheese.

  13. Simple and effective methods of freezing capercaillie (Tetrao urogallus L. semen.

    Directory of Open Access Journals (Sweden)

    Artur Kowalczyk

    Full Text Available A continuous decline in the number and range of capercaillie (Tetrao urogallus L. in many European countries can be observed, mostly due to habitat destruction by human activity, unecological forestry management, and increased density of natural predators. Ex situ in vitro gene banks provide a unique opportunity to preserve the genetic material for future generations. Simple and effective cryopreservation methods for capercaillie semen are discussed. Semen was collected from seven males kept in the Capercaillie Breeding Centre at Forestry Wisła in Poland. Within five minutes after collection, ejaculates were diluted with EK diluent, then divided into two parts, and subjected to two freezing procedures: in pellets and in straws. In fresh semen, ejaculate clearness, viscosity, color and volume, as well as sperm concentration, motility and morphology, were evaluated, while in frozen-thawed semen only motility and morphology of sperm were determined. Fertilizing ability of thawed semen was examined for samples frozen in straws. Significant (P<0.05 differences between individual males were found in relation to the majority of fresh semen traits: ejaculate volume averaged 102.1 µL (varying from 49.0 to 205.0; average sperm concentration was 632.5 x 10⁶ mL⁻¹ (178.8-1257.1; percentage of live normal cells varied from 39.2 to 70.3% (58.7% on an average; percentage of motile cells ranged from 76.0 to 85.7% and motility parameters were male dependent, as well. Both cryopreservation methods had a negative effect on morphology and motility of frozen-thawed semen; however, the straw method yielded 60.7% and the pellet method 42.5% of live cells in total in thawed semen (P<0.05, while the number of live normal (intact cells was similar (22.4 and 22.2%, respectively. Egg fertility varied between 77.8 and 91.7% (average 84.4%. Both freezing procedures seem to be effective in obtaining acceptable viability and high fertilizing potency of thawed sperm and

  14. Long-term stability of acyclovir in 0.9% NaCl infusion polyolefin bags at 5±3°C after freeze-thaw treatment: a generic product versus the brand name.

    Science.gov (United States)

    Dewulf, J; Galanti, L; Godet, M; Gillet, P; Jamart, J; Hecq, J-D

    2015-03-01

    The aim of the study was to investigate the long-term stability of acyclovir 5 mg/mL (a generic product versus the brand name) in NaCl 0.9% after storage at 5±3°C and to evaluate the influence of initial freezing and microwave thawing on this stability. Five bags of Acyclovir® Hospira 5 mg/mL (A) and five bags of Zovirax® GSK 5 mg/mL (B) were prepared under aseptic conditions and stored 3 months at -20°C, then thawed and stored 30 days at 4°C. Five bags of Acyclovir® 5 mg/mL (C) and five bags of Zovirax® 5 mg/mL (D) were also prepared under aseptic conditions and stored 30 days at 5±3°C. Optic density measurement at different wavelengths, pH measurement and optic microscope observations were performed periodically during the storage. A forced degradation test with HCl 12 M and NaOH 5 M before and after heating at 100°C was also performed. The concentrations were measured by HPLC-PDA. The only one forced degradation test that yielded chromatograms with degradation products peak was the test with the acid solution heated at 100°C without interference with the native product. No significant change in pH values or optic densities were seen during the study for both products. No crystals were seen with the optic microscope during the study. Acyclovir® and Zovirax® solutions were stable for at least 21 days according to the FDA recommendations. Moreover, there was no statistical difference between regression lines of those two products and two storage conditions. Under the conditions of this study, Acyclovir® 5 mg/mL in 100 mL of NaCl 0.9% infusion remains stable at least for 21 days at 5±3°C with or without freezing at -20°C during the three previous months. There is no statistical difference between the brand name and a generic product. Acyclovir may be prepared in advanced by a centralized intravenous additive service, frozen in polyolefin bags and microwave thawed before storage under refrigeration until 21 days. Copyright © 2014 Elsevier Masson

  15. Testing a model for the critical degree of saturation at freezing of porous building materials

    DEFF Research Database (Denmark)

    Hansen, Ernst Jan De Place

    1996-01-01

    of elasticity, tensile strength, amount of freezable water, thermal expansion coefficients and parameters characterizing the pore structure and its effect on strength, modulus of elasticity and volumetric expansion. For the present, the model assumes non air-entrained homogeneous materials subjected to freeze-thaw...

  16. Effects of low voltage electrolysis and freezing on coliform content of contaminated water

    International Nuclear Information System (INIS)

    Qazi, J.I.; Saleem, F.

    2003-01-01

    A sewage sample was mixed with drinking water and subjected to low voltage (15V) electrolysis in the presence of 1% NaCl. The prepared sample was also kept in freezer with and without the presence of sodium chloride for 4-hours. Among these treatments the electrolysis proved to kill the coliforms, while the freezing reduced the bacterial content. Antibiotics sensitivity patterns revealed that certain of the coliform strains survived the freezing and thawing shocks. Nature of such surviving bacteria and need to study chemical parameters of electrolyzed water are discussed. (author)

  17. A Passive Microwave L-Band Boreal Forest Freeze/Thaw and Vegetation Phenology Study

    Science.gov (United States)

    Roy, A.; Sonnentag, O.; Pappas, C.; Mavrovic, A.; Royer, A.; Berg, A. A.; Rowlandson, T. L.; Lemay, J.; Helgason, W.; Barr, A.; Black, T. A.; Derksen, C.; Toose, P.

    2016-12-01

    The boreal forest is the second largest land biome in the world and thus plays a major role in the global and regional climate systems. The extent, timing and duration of seasonal freeze/thaw (F/T) state influences vegetation developmental stages (phenology) and, consequently, constitute an important control on how boreal forest ecosystems exchange carbon, water and energy with the atmosphere. The effective retrieval of seasonal F/T state from L-Band radiometry was demonstrated using satellite mission. However, disentangling the seasonally differing contributions from forest overstory and understory vegetation, and the soil surface to the satellite signal remains challenging. Here we present initial results from a radiometer field campaign to improve our understanding of the L-Band derived boreal forest F/T signal and vegetation phenology. Two L-Band surface-based radiometers (SBR) are installed on a micrometeorological tower at the Southern Old Black Spruce site in central Saskatchewan over the 2016-2017 F/T season. One radiometer unit is installed on the flux tower so it views forest including all overstory and understory vegetation and the moss-covered ground surface. A second radiometer unit is installed within the boreal forest overstory, viewing the understory and the ground surface. The objectives of our study are (i) to disentangle the L-Band F/T signal contribution of boreal forest overstory from the understory and ground surface, (ii) to link the L-Band F/T signal to related boreal forest structural and functional characteristics, and (iii) to investigate the use of the L-Band signal to characterize boreal forest carbon, water and energy fluxes. The SBR observations above and within the forest canopy are used to retrieve the transmissivity (γ) and the scattering albedo (ω), two parameters that describe the emission of the forest canopy though the F/T season. These two forest parameters are compared with boreal forest structural and functional

  18. Freezing effect on bread appearance evaluated by digital imaging

    Science.gov (United States)

    Zayas, Inna Y.

    1999-01-01

    In marketing channels, bread is sometimes delivered in a frozen sate for distribution. Changes occur in physical dimensions, crumb grain and appearance of slices. Ten loaves, twelve bread slices per loaf were scanned for digital image analysis and then frozen in a commercial refrigerator. The bread slices were stored for four weeks scanned again, permitted to thaw and scanned a third time. Image features were extracted, to determine shape, size and image texture of the slices. Different thresholds of grey levels were set to detect changes that occurred in crumb, images were binarized at these settings. The number of pixels falling into these gray level settings were determined for each slice. Image texture features of subimages of each slice were calculated to quantify slice crumb grain. The image features of the slice size showed shrinking of bread slices, as a results of freezing and storage, although shape of slices did not change markedly. Visible crumb texture changes occurred and these changes were depicted by changes in image texture features. Image texture features showed that slice crumb changed differently at the center of a slice compared to a peripheral area close to the crust. Image texture and slice features were sufficient for discrimination of slices before and after freezing and after thawing.

  19. the effect of freezing method on the survival of ram spermatozoa

    African Journals Online (AJOL)

    Sci.2l, 351. SALAIIiON, S., 1970. The survival of ram spermatozoa following pellet freezing below-79oC. Aust- .1. hiol..n'r. 13. 459. SALAMON, S. & BRANDON, M.R., l97l . Effect of compcition of thawing solution on sunrival of ram spernntozoa iroznn by the pellet method. Aust. J. biol. Sci. 24,355. SALAMON, S. & VISSER, D., ...

  20. Antioxidant Effects of Selenium on Seminiferous Tubules of Immature Mice Testis

    Directory of Open Access Journals (Sweden)

    Davoud Kushki

    2015-12-01

    Full Text Available Background: The freezing of immature testis tissue and then the transplant of it can be considered as a major step in fertility preservation for young boys with cancer, the survival of animal generation exposed to extinction and cloning animalistic desirable species. One of the most prevalent of damages in during the freezing-thawing process is oxidative stress. Objectives: The objective of this study was to investigate the antioxidant effects of selenium compound (Na2SeO3 on rate of seminiferous tubules injury in during of cryopreservation. Materials and Methods: In this experimental study, 8 BALB/c immature male mice (6 - 8 days old were randomly selected, and the testes removed surgically (n = 16. The testes divided into 2 groups: experimental group, control group (opposite testes. For each of the two experimental and control groups, two types of soluble (freezing solution and thawing solution were prepared. These solutions, which contain 2 mg/mL solution of Na2SeO3 and control solution, were prepared in the DMEM (Dulbecco’s modified eagle medium base medium. Of each group were 4 testes into fast freezing-thawing procedure and 4 testes were into slow freezing-thawing procedure. Then this testis for analyzing injury, after preparatory process, was stained with hematoxylin-eosin. Results: At the slow freezing-thawing procedure, seminiferous tubules injury significantly reduced in experimental group compared to control group. At the fast freezing-thawing procedure, seminiferous tubules injury significantly reduced in experimental group compared with of control group. Conclusions: It seems that Se due to its antioxidant properties, the harmful effects of freezing-thawing process reduces and protects seminiferous tubules from oxidative injury.

  1. Two-step freezing of hybridoma cells in 96-well microculture plates.

    Science.gov (United States)

    Pĕknicová, J; Kristofová, H

    1985-01-01

    Stabile hybridoma cells, colonies of hybridoma cells 14 days after fusion of immune spleen and myeloma cells, myeloma cells and fibroblasts cultured in 96-well microculture plates were frozen by the method of two-step freezing. The culture medium was aspirated, and 50 microliter of the medium containing a cryoprotectant (5% dimethyl sulphoxide) was added for 10 min at room temperature. The plates were put into microtene bags, placed at -25 degrees C in a freezer for 30 min and then stored at -100 degrees C in liquid nitrogen vapour. Plates with cells were thawed rapidly in a 50 degree C water bath. After thawing the hybrid cells were viable and continued to produce the specific antibody.

  2. Anisotropy effect of the clay soil masses on the stress-strain state of transport tunnels

    Directory of Open Access Journals (Sweden)

    Yushkov Boris Semenovich

    2014-09-01

    Full Text Available The article considers the kinds of clay soil mass anisotropy in the form of the spatial heterogeneity of properties of thawed and frozen soils, ambiguity of the frost heaving values and shrinkage in different directions. The questions of anisotropy of the clay soil properties at the positive temperatures are reported. The dependence of the heterogeneity of the physical and mechanical properties of frozen soils from the cryogenic texture, natural arrangement, different types of stratification and interbedding is considered. Indexes of the strength and strain anisotropy are noted. The accounting possibilities of the basic numerical indexes of heaving phenomena from the standpoint of anisotropy of the properties and processes inherent in the freezing through soil are analyzed by substitution in the heaving strain formula. The unevenness of thawed soil shrinkage in vertical and horizontal directions is noted during the freezing of the top layer. The unevenness of shrinkage in different directions is connected with kind of stress and cryogenic texture. Anisotropy of the frost heaving process is considered in the context of one-dimensional and non-one-dimensional problem depending on the amount of the freezing fronts and their direction. There is summarized the effect of anisotropy appearances on the stress-strain state of the transport tunnel. One can conclude that the resulting non-uniformity of heaving and shrinkage in conjunction with anisotropic properties of frozen soils, is a significant component in the complex of power factors determining the optimal design solution of a transport tunnel.

  3. Trehalose improves semen antioxidant enzymes activity, post-thaw quality, and fertility in Nili Ravi buffaloes (Bubalus bubalis).

    Science.gov (United States)

    Iqbal, Sajid; Andrabi, Syed Murtaza Hassan; Riaz, Amjad; Durrani, Aneela Zameer; Ahmad, Nasim

    2016-03-15

    Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profile during cryopreservation (after dilution, before freezing, and after thawing), (2) in vitro quality (after thawing), and (3) in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Semen samples (n = 20) from four buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of trehalose (0.0, 15, 30, 45, and 60 mM) and frozen in French straws. At post dilution, profile of sperm catalase (U/mL) was higher (P semen doses cryopreserved in extender containing 30 mM of trehalose than the control. It is concluded that addition of 30-mM trehalose in extender improves the semen antioxidant enzymes activity, post thaw quality, and fertility in Nili Ravi buffaloes. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Cost-Effectiveness of the Freeze-All Policy.

    Science.gov (United States)

    Roque, Matheus; Valle, Marcello; Guimarães, Fernando; Sampaio, Marcos; Geber, Selmo

    2015-08-01

    To evaluate the cost-effectiveness of freeze-all cycles when compared to fresh embryo transfer. This was an observational study with a cost-effectiveness analysis. The analysis consisted of 530 intracytoplasmic sperm injection (ICSI) cycles in a private center in Brazil between January 2012 and December 2013. A total of 530 intracytoplasmic sperm injection (ICSI) cycles - 351 fresh embryo transfers and 179 freeze-all cycles - with a gonadotropin-releasing hormone (GnRH) antagonist protocol and day 3 embryo transfers. The pregnancy rate was 31.1% in the fresh group and 39.7% in the freeze-all group. We performed two scenario analyses for costs. In scenario 1, we included those costs associated with the ICSI cycle (monitoring during controlled ovarian stimulation [COS], oocyte retrieval, embryo transfer, IVF laboratory, and medical costs), embryo cryopreservation of supernumerary embryos, hormone measurements during COS and endometrial priming, medication use (during COS, endometrial priming, and luteal phase support), ultrasound scan for frozen- thawed embryo transfer (FET), obstetric ultrasounds, and miscarriage. The total cost (in USD) per pregnancy was statistically lower in the freeze-all cycles (19,156.73 ± 1,732.99) when compared to the fresh cycles (23,059.72 ± 2,347.02). Even in Scenario 2, when charging all of the patients in the freeze-all group for cryopreservation (regardless of supernumerary embryos) and for FET, the fresh cycles had a statistically significant increase in treatment costs per ongoing pregnancy. The results presented in this study suggest that the freeze-all policy is a cost-effective strategy when compared to fresh embryo transfer.

  5. Freezing Injury in Onion Bulb Cells

    Science.gov (United States)

    Palta, Jiwan P.; Levitt, Jacob; Stadelmann, Eduard J.

    1977-01-01

    Onion (Allium cepa L.) bulbs were frozen to −4 and −11 C and kept frozen for up to 12 days. After slow thawing, a 2.5-cm square from a bulb scale was transferred to 25 ml deionized H2O. After shaking for standard times, measurements were made on the effusate and on the effused cells. The results obtained were as follows. Even when the scale tissue was completely infiltrated, and when up to 85% of the ions had diffused out, all of the cells were still alive, as revealed by cytoplasmic streaming and ability to plasmolyze. The osmotic concentration of the cell sap, as measured plasmolytically, decreased in parallel to the rise in conductivity of the effusate. The K+ content of the effusate, plus its assumed counterion, accounted for only 20% of the total solutes, but for 100% of the conductivity. A large part of the nonelectrolytes in the remaining 80% of the solutes was sugars. The increased cell injury and infiltration in the −11 C treatment, relative to the −4 C and control (unfrozen) treatments, were paralleled by increases in conductivity, K+ content, sugar content, and pH of the effusate. In spite of the 100% infiltration of the tissue and the large increase in conductivity of the effusate following freezing, no increase in permeability of the cells to water could be detected. The above observations may indicate that freezing or thawing involves a disruption of the active transport system before the cells reveal any injury microscopically. PMID:16660100

  6. Comparison between mechanical freezer and conventional freezing using liquid nitrogen in normozoospermia.

    Science.gov (United States)

    Rahana, A R; Ng, S P; Leong, C F; Rahimah, M D

    2011-10-01

    This study evaluated the effect of human semen cryopreservation using an ultra-low temperature technique with a mechanical freezer at -85°C as an alternative method to the conventional liquid nitrogen technique at -196°C. This was a prospective experimental study conducted in the Medically Assisted Conception unit, Department of Obstetrics and Gynaecology, National University Hospital, Malaysia from January 1, 2006 to April 30, 2007. All normozoospermic semen samples were included in the study. The concentration, motility and percentage of intact DNA of each semen sample were assessed before and after freezing and thawing on Days 7 and 30 post freezing. Sperm cryopreservation at -85°C was comparable to the conventional liquid nitrogen technique for a period of up to 30 days in a normozoospermic sample. There was no statistical difference in concentration (Day 7 p-value is 0.1, Day 30 p-value is 0.2), motility (Day 7 p-value is 0.9, Day 30 p-value is 0.5) and proportion of intact DNA (Day 7 p-value is 0.1, Day 30 p-value is 0.2) between the ultra-low temperature technique and conventional liquid nitrogen cryopreservation at Days 7 and 30 post thawing. This study clearly demonstrates that short-term storage of sperm at -85°C could be a viable alternative to conventional liquid nitrogen cryopreservation at -196°C due to their comparable post-thaw results.

  7. Intravaginal artificial insemination in bitches using frozen/thawed semen after dilution in powdered coconut water (ACP-106c).

    Science.gov (United States)

    Uchoa, D C; Silva, T F P; Mota Filho, A C; Silva, L D M

    2012-12-01

    The aim of this study was to evaluate powdered coconut water extender (ACP-106c; ACP Serviços Tecnológicos Ltda, ACP Biotecnologia, Fortaleza, Ceará, Brazil) as a diluent for freezing dog semen and the fertility after vaginal insemination of semen frozen therein. Ten ejaculates were collected from five dogs, evaluated fresh, diluted in ACP-106c, 10% egg yolk and 6% glycerol, cooled and frozen. In the first phase of the study, straws with frozen semen were thawed and immediately subjected to the same analysis as the fresh semen and, in addition, to Computer-Assisted Semen Analysis (CASA). In phase 2, 10 bitches that had been subjected to natural breeding during a preceding oestrous cycle were vaginally inseminated with thawed semen that had been re-diluted in ACP-106c. After thawing, a mean of 77% sperm motility was obtained through subjective analysis and 77.3% through CASA. Following artificial insemination, a 60% pregnancy rate was observed, resulting in a 50% parturition rate and a mean litter size of 3.4 (SEM 0.6), with 47.1% males and 52.9% females. ACP-106c can be successfully used for freezing canine semen, and vaginal deposition of such semen yields similar pregnancy rates to those reported in other studies. © 2012 Blackwell Verlag GmbH.

  8. Biogeochemical Attributes That Affect the Fate and Transport of Military Relevant Contaminants Under Freeze-thaw Conditions

    Science.gov (United States)

    LeMonte, J.; Price, C. L.; Seiter, J.; Crocker, F. H.; Douglas, T.; Chappell, M. A.

    2017-12-01

    The roles and missions that the U.S. Department of Defense (DoD) undertakes in the Arctic are being reshaped by significant changes in the operational environment as a result of rising global temperatures and increased development of the vast training ranges available in Alaska. The Arctic is warming faster than any other region on Earth resulting in changing seasonality and precipitation patterns that, in turn, are leading to alterations in above ground vegetation, permafrost stability and summer sea ice extent. Collectively, these poorly defined ecosystem changes play critical roles in affecting the transport and eventual fate of persistent military relevant contaminants through unique Arctic and Subarctic terrestrial environments. As a result, management of military contaminants in a changing Arctic represents a unique and potentially significant liability to the Army and the DoD. The United States footprint in the Arctic region falls within the state of Alaska and U.S. Army Alaska manages 10% of all active Army training lands worldwide, which cover nearly 2,500 square miles in total land area. Primary recalcitrant contaminants of concern at active training ranges and at legacy sites include energetics (i.e. RDX and 2,4-dinitrotoluene) and heavy metals (i.e. antimony and lead). Through a series of field sampling and laboratory experiments, the objectives of this work are to: 1) quantify soil biogeochemical attributes that effect the physical fate and transport of military relevant contaminants in Arctic and subarctic soils under freeze-thaw conditions with a focus on near surface processes, and 2) quantify microbial diversity in Arctic and subarctic soils and the environmental constraints on community activity while exploring the effects of amendments on community function as they relate to contaminant transformation.

  9. Influence of vertical and lateral heat transfer on permafrost thaw, peatland landscape transition, and groundwater flow

    Science.gov (United States)

    Kurylyk, Barret L.; Masaki, Masaki; Quinton, William L.; McKenzie, Jeffrey M.; Voss, Clifford I.

    2016-01-01

    Recent climate change has reduced the spatial extent and thickness of permafrost in many discontinuous permafrost regions. Rapid permafrost thaw is producing distinct landscape changes in the Taiga Plains of the Northwest Territories, Canada. As permafrost bodies underlying forested peat plateaus shrink, the landscape slowly transitions into unforested wetlands. The expansion of wetlands has enhanced the hydrologic connectivity of many watersheds via new surface and near-surface flow paths, and increased streamflow has been observed. Furthermore, the decrease in forested peat plateaus results in a net loss of boreal forest and associated ecosystems. This study investigates fundamental processes that contribute to permafrost thaw by comparing observed and simulated thaw development and landscape transition of a peat plateau-wetland complex in the Northwest Territories, Canada from 1970 to 2012. Measured climate data are first used to drive surface energy balance simulations for the wetland and peat plateau. Near-surface soil temperatures simulated in the surface energy balance model are then applied as the upper boundary condition to a three-dimensional model of subsurface water flow and coupled energy transport with freeze-thaw. Simulation results demonstrate that lateral heat transfer, which is not considered in many permafrost models, can influence permafrost thaw rates. Furthermore, the simulations indicate that landscape evolution arising from permafrost thaw acts as a positive feedback mechanism that increases the energy absorbed at the land surface and produces additional permafrost thaw. The modeling results also demonstrate that flow rates in local groundwater flow systems may be enhanced by the degradation of isolated permafrost bodies.

  10. Fertility response of artificial insemination methods in sheep with fresh and frozen-thawed semen.

    Science.gov (United States)

    Masoudi, Reza; Zare Shahneh, Ahmad; Towhidi, Armin; Kohram, Hamid; Akbarisharif, Abbas; Sharafi, Mohsen

    2017-02-01

    The aim of this study was to evaluate the fertility response of artificial insemination (AI) methods with fresh and frozen sperm in sheep. In experiment 1, one hundred and fifty fat tailed Zandi ewes were assigned into 3 equal groups and inseminated with three AI methods consisting of vaginal, laparoscopic and trans-cervical AI with fresh semen. In experiment 2, a factorial study (3 AI methods × 2 extenders) was used to analyze the effects of three AI methods and two freezing extenders containing soybean lecithin (SL) or Egg yolk (EY) on reproductive performance of 300 fat tailed Zandi ewes. Also, total motility, progressive motility, viability and lipid peroxidation of semen were evaluated after freeze-thawing in two extenders. In result, there was no significant difference among three AI methods when fresh semen was used. In experiment 2, the highest percentage of pregnancy rate, parturition rate and lambing rate were obtained in laparoscopic AI group (P semen, trans-cervical AI was more efficient than vaginal method when frozen-thawed semen was used, but its efficiency was not as high as laparoscopic method. Also, SL extender can be an efficient alternative extender to preserve ram sperm during cryopreservation procedure without adverse effects of EY. Copyright © 2016. Published by Elsevier Inc.

  11. A New Freezing Method Using Pre-Dehydration by Microwave-Vacuum Drying

    Science.gov (United States)

    Tsuruta, Takaharu; Hamidi, Nurkholis

    Partial dehydration by microwave-vacuum drying has been applied to tuna and strawberry in order to reduce cell-damages caused by the formation of large ice-crystals during freezing. The samples were subjected to microwave vacuum drying at pressure of 5 kPa and temperature less than 27°C to remove small amount of water prior to freezing. The tuna were cooled by using the freezing chamber at temperature -50°C or -150°C, while the strawberries were frozen at temperature -30°C or -80°C, respectively. The temperature transients in tuna showed that removing some water before freezing made the freezing time shorter. The observations of ice crystal clearly indicated that rapid cooling and pre-dehydration prior to freezing were effective in minimizing the size of ice crystal. It is also understood that the formation of large ice crystals has a close relation to the cell damages. After thawing, the observation of microstructure was done on the tuna and strawberry halves. The pre-dehydrated samples showed a better structure than the un-dehydrated one. It is concluded that the pre-dehydration by microwave-vacuum drying is one promising method for the cryo-preservation of foods.

  12. Microprocessor-controlled vs. "dump-freezing" platelet and lymphocyte cryopreservation: A quantitative and qualitative comparative study

    Directory of Open Access Journals (Sweden)

    Balint Bela

    2006-01-01

    Full Text Available Background/Aim. Thermodynamical and cryobiological parameters responsible for cell damages during cryopreservation (cryoinjuries have not yet been completely explained. Thus, freezing procedures should be revised, exactly optimized to obtain an enhanced structural and functional recovery of frozen- thawed cells. The aim of this study was to compare microprocessor- controlled (controlled-rate with the compensation of the released fusion heat and “dump-freezing” (uncontrolled- rate of the platelet and lymphocyte cryopreservation efficacy. Methods. Platelet quantitative recovery (post-thaw vs. unfrozen cell count, viability (using hypotonic shock response - HSR, morphological score (PMS, ultrastructural (electron microscopy properties and expression of different surface antigens were investigated. In lymphocyte setting, cell recovery and viability (using trypan blue exclusion test as well as functionality (by plant mitogens were determined. Controlled- rate freezing and uncontrolled-rate cryopreservation were combined with 6% (platelets and 10% (lymphocytes dimethyl sulfoxide (DMSO. Results. Platelet recovery and functionality were superior in the controlled-rate system. The majority of surface antigen expression was reduced in both freezing groups vs. unfrozen cells, but GP140/CD62p was significantly higher in controlled-rate vs. uncontrolled-rate setting. Controlled- rate freezing resulted with better lymphocyte recovery and viability (trypan blue-negative cell percentage. In mitogen-induced lymphocyte proliferative response no significant intergroup difference (controlled-rate vs. uncontrolled-rate were found. Conclusion. The data obtained in this study showned the dependence of cell response on the cryopreservation type. Controlled-rate freezing provided a superior platelet quantitative and functional recovery. Lymphocyte recovery and viability were better in the controlled-rate group, although only a minor intergroup difference for cell

  13. Freezing tolerance of ectomycorrhizal fungi in pure culture.

    Science.gov (United States)

    Lehto, Tarja; Brosinsky, Arlena; Heinonen-Tanski, Helvi; Repo, Tapani

    2008-10-01

    The ability to survive freezing and thawing is a key factor for the existence of life forms in large parts of the world. However, little is known about the freezing tolerance of mycorrhizal fungi and their role in the freezing tolerance of mycorrhizas. Threshold temperatures for the survival of these fungi have not been assessed experimentally. We grew isolates of Suillus luteus, Suillus variegatus, Laccaria laccata, and Hebeloma sp. in liquid culture at room temperature. Subsequently, we exposed samples to a series of temperatures between +5 degrees C and -48 degrees C. Relative electrolyte leakage (REL) and re-growth measurements were used to assess the damage. The REL test indicated that the lethal temperature for 50% of samples (LT(50)) was between -8.3 degrees C and -13.5 degrees C. However, in the re-growth experiment, all isolates resumed growth after exposure to -8 degrees C and higher temperatures. As many as 64% of L. laccata samples but only 11% in S. variegatus survived -48 degrees C. There was no growth of Hebeloma and S. luteus after exposure to -48 degrees C, but part of their samples survived -30 degrees C. The fungi tolerated lower temperatures than was expected on the basis of earlier studies on fine roots of ectomycorrhizal trees. The most likely freezing tolerance mechanism here is tolerance to apoplastic freezing and the concomitant intracellular dehydration with consequent concentrating of cryoprotectant substances in cells. Studying the properties of fungi in isolation promotes the understanding of the role of the different partners of the mycorrhizal symbiosis in the freezing tolerance.

  14. The principles of ultrasound and its application in freezing related processes of food materials: A review.

    Science.gov (United States)

    Cheng, Xinfeng; Zhang, Min; Xu, Baoguo; Adhikari, Benu; Sun, Jincai

    2015-11-01

    Ultrasonic processing is a novel and promising technology in food industry. The propagation of ultrasound in a medium generates various physical and chemical effects and these effects have been harnessed to improve the efficiency of various food processing operations. Ultrasound has also been used in food quality control as diagnostic technology. This article provides an overview of recent developments related to the application of ultrasound in low temperature and closely related processes such as freezing, thawing, freeze concentration and freeze drying. The applications of high intensity ultrasound to improve the efficiency of freezing process, to control the size and size distribution of ice crystals and to improve the quality of frozen foods have been discussed in considerable detail. The use of low intensity ultrasound in monitoring the ice content and to monitor the progress of freezing process has also been highlighted. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. A theoretical extension of the soil freezing curve paradigm

    Science.gov (United States)

    Amiri, Erfan A.; Craig, James R.; Kurylyk, Barret L.

    2018-01-01

    Numerical models of permafrost evolution in porous media typically rely upon a smooth continuous relation between pore ice saturation and sub-freezing temperature, rather than the abrupt phase change that occurs in pure media. Soil scientists have known for decades that this function, known as the soil freezing curve (SFC), is related to the soil water characteristic curve (SWCC) for unfrozen soils due to the analogous capillary and sorptive effects experienced during both soil freezing and drying. Herein we demonstrate that other factors beyond the SFC-SWCC relationship can influence the potential range over which pore water phase change occurs. In particular, we provide a theoretical extension for the functional form of the SFC based upon the presence of spatial heterogeneity in both soil thermal conductivity and the freezing point depression of water. We infer the functional form of the SFC from many abrupt-interface 1-D numerical simulations of heterogeneous systems with prescribed statistical distributions of water and soil properties. The proposed SFC paradigm extension has the appealing features that it (1) is determinable from measurable soil and water properties, (2) collapses into an abrupt phase transition for homogeneous media, (3) describes a wide range of heterogeneity within a single functional expression, and (4) replicates the observed hysteretic behavior of freeze-thaw cycles in soils.

  16. Repeated stressor exposure enhances contextual fear memory in a beta-adrenergic receptor-dependent process and increases impulsivity in a non-beta receptor-dependent fashion.

    Science.gov (United States)

    Camp, Robert M; Johnson, John D

    2015-10-15

    Memory formation is promoted by stress via the release of norepinephrine and stimulation of beta-adrenergic receptors (β-ARs). Previous data demonstrate that repeated stressor exposure increases norepinephrine turnover and β-AR signaling within the amygdala, which led to the hypothesis that some stress-induced behavioral changes are likely due to facilitated associative learning. To test this, Fischer rats were exposed to chronic mild stress for four days. On day 5, subjects (including non-stressed controls) were injected with the beta-blocker propranolol or vehicle prior to conditioning in an operant box (animals receive two mild foot shocks) or passive avoidance apparatus (animals received a foot shock upon entry into the dark chamber). Twenty-four hours later, subjects were returned to the operant box for measurement of freezing or returned to the passive avoidance apparatus for measurement of latency to enter the dark chamber. Subjects were also tested in an open field to assess context-independent anxiety-like behavior. Animals exposed to chronic stress showed significantly more freezing behavior in the operant box than did controls, and this exaggerated freezing was blocked by propranolol during the conditioning trial. There was no effect of stress on behavior in the open field. Unexpectedly, retention latency was significantly reduced in subjects exposed to chronic stress. These results indicate that chronic exposure to stress results in complex behavioral changes. While repeated stress appears to enhance the formation of fearful memories, it also results in behavioral responses that resemble impulsive behaviors that result in poor decision-making. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Inducing Heat Shock Proteins Enhances the Stemness of Frozen-Thawed Adipose Tissue-Derived Stem Cells.

    Science.gov (United States)

    Shaik, Shahensha; Hayes, Daniel; Gimble, Jeffrey; Devireddy, Ram

    2017-04-15

    Extensive research has been performed to determine the effect of freezing protocol and cryopreservation agents on the viability of adipose tissue-derived stromal/stem cells (ASCs) as well as other cells. Unfortunately, the conclusion one may draw after decades of research utilizing fundamentally similar cryopreservation techniques is that a barrier exists, which precludes full recovery. We hypothesize that agents capable of inducing a subset of heat shock proteins (HSPs) and chaperones will reduce the intrinsic barriers to the post-thaw recovery of ASCs. ASCs were exposed to 43°C for 1 h to upregulate HSPs, and the temporal HSP expression profile postheat shock was determined by performing quantitative polymerase chain reaction (PCR) and western blotting assays. The expression levels of HSP70 and HSP32 were found to be maximum at 3 h after the heat shock, whereas HSP90 and HSP27 remain unchanged. The heat shocked ASCs cryopreserved during maximal HSPs expression exhibited increased post-thaw viability than the nonheat shocked samples. Histochemical staining and quantitative reverse transcription-PCR indicated that the ASC differentiation potential was retained. Thus, suggesting that the upregulation of HSPs before a freezing insult is beneficial to ASCs and a potential alternative to the use of harmful cryoprotective agents.

  18. The effect of kinds and concentration of cryoprotectant and thawing methods on frozen semen of Arab chicken

    Directory of Open Access Journals (Sweden)

    Sofjan Iskandar

    2006-03-01

    Full Text Available The success of freezing chicken semen is the hope for preserving Indonesian native chickens. Semen from twenty Arab roosters were collected using massage technique once in a week. Cryoprotectant DMA (dimethyl acetamide or DMF (dimethyl formalmide of 7 or 9% and thawing A at temperature of 30oC for 30 seconds or in B at 5oC for 5 minutes. The volume of fresh semen was 0.3 ± 0.072 ml/ejaculate, white colour, rather thick to thick, with 2200 ± 372 millions sperms/ml and pH 6.95 ± 0.32, 4+/3+ mass movement, 80% motility, 84 ± 4.48% and abnormality of 14.75 ± 1.28%. There were not statistically significant (P>0.05 effect of interaction of treatments (kinds and concentrations of cryoprotectant, and thawing methods on motility and live-sperms. Sperm motility preserved with DMA (34.69% significantly higher than with DMF (29.84%. Sperm motility was also significantly higher (34.53% when preserved with 7% cryoprotectant than with 9% (30%. Thawing-A significantly gave higher motility (35.31% than thawing-B did (29.22%. Live-sperms of semen preserved with DMA (46.75% was significantly higher than with DMF (41.72%. Cryoprotectant concentration of 7% gave higher live-sperms (46.98% than of 9% did (41.48%. Thawing-A left live-sperms of 47.14%, which was significantly higher than thawing-B did (41.30%.

  19. AgRISTARS: Early warning and crop condition assessment. Plant cover, soil temperature, freeze, water stress, and evapotranspiration conditions

    Science.gov (United States)

    Wiegand, C. L. (Principal Investigator); Nixon, P. R.; Gausman, H. W.; Namken, L. N.; Leamer, R. W.; Richardson, A. J.

    1981-01-01

    Emissive (10.5 to 12.5 microns) and reflective (0.55 to 1.1 microns) data for ten day scenes and infrared data for six night scenes of southern Texas were analyzed for plant cover, soil temperature, freeze, water stress, and evapotranspiration. Heat capacity mapping mission radiometric temperatures were: within 2 C of dewpoint temperatures, significantly correlated with variables important in evapotranspiration, and related to freeze severity and planting depth soil temperatures.

  20. Egg freezing and egg banking: empowerment and alienation in assisted reproduction.

    Science.gov (United States)

    Robertson, John A

    2014-06-01

    With the development of rapid freezing of human oocytes, many programs have reported IVF success rates comparable to those achieved with fresh eggs and thawed frozen embryos. Egg freezing is now gaining professional and regulatory acceptance as a safe and effective technique for women who wish to avoid discarding excess embryos, who face fertility-threatening medical treatments, or who want to preserve their eggs for use when they are better situated to have a family. This article focuses on the uses of and justification for egg freezing, the path to professional acceptance, the variability in success rates, and the controversy over freezing eggs for social rather than medical reasons. It also addresses the emergence of egg banking as a separate sector in the infertility industry, the regulatory issues that it poses, and its effect on egg donation. Key here is the legal control of stored eggs by banking women and their options when they wish to dispose of those eggs. The analysis is framed around empowerment and alienation. Egg freezing is generally empowering for women, but the donation or sale of unused eggs to infertile women, egg bankers, and researchers also raises issues of alienation.

  1. Probiotic supplementation and fast freezing to improve quality attributes and oxidation stability of frozen chicken breast muscle

    Science.gov (United States)

    The objective of this study was to determine the effects of probiotic supplementation and fast freezing on quality attributes and oxidation stability of frozen/thawed chicken breast meat. Broilers were fed with a basal diet or the basal diet plus 250 ppm Sporulin (three strains of Bacillus subtilis)...

  2. Influence of Heat Shock Temperatures and Fast Freezing on Viability of Probiotic Sporeformers and the Issue of Spore Plate Count Versus True Numbers

    Directory of Open Access Journals (Sweden)

    Mojtaba Jafari

    2016-02-01

    Full Text Available Background and Objectives: The purpose of the present study was to investigate effects of various heat shock conditions and fast freezing and subsequent thawing on the viability and recovery of Bacillus coagulans and Bacillus subtilis as probiotic sporeformers, and also to compare spore plate and microscopic counts. Materials and Methods: After preparing the final suspensions of B. coagulans and Bacillus subtilis subsp. Natto spores, they were spread-plated before and after fast freezing treatment (-70°C for about 1 min. Heat shock treatments of the spores were carried out at 68oC for 15, 20, and 30 min as well as at 80oC for 10 and 15 min. Concentrations of the examined probiotic sporeformers were determined simultaneously by plate enumerations and microscopically determined counts. Student’s t-test and one-way analysis of variance (ANOVA of SPSS were used for statistical analysis of the data. Analysis of DoE results was carried out using Minitab. Results: The results presented here show that the highest recovery rates for B. coagulans (14.75 log CFU/mL and B. subtilis spores (14.80 log CFU/mL were under a heat shock condition of 68°C for 20 min in nutrient agar (p<0.05. In addition, the survival rates of B. coagulans and B. subtilis spores under the fast freezing and subsequent thawing condition were about 90% and 88%, respectively. Plate counts differed significantly from counts determined microscopically, with differences of almost 0.5 and 0.8 log for B. coagulans and B. subtilis spores, respectively (p<0.05. In addition, DoE results of the study revealed that both factors of spore count method and only freezing factor in fast freezing treatment have a significant effect on concentrations of the spores examined (p<0.05. Conclusions: Heat shock conditions, freezing and subsequent thawing circumstances, and plate counts or enumerations determined microscopically have significant influences on the viability of probiotic sporeformers and

  3. Subcellular membrane fluidity of Lactobacillus delbrueckii subsp. bulgaricus under cold and osmotic stress.

    Science.gov (United States)

    Meneghel, Julie; Passot, Stéphanie; Cenard, Stéphanie; Réfrégiers, Matthieu; Jamme, Frédéric; Fonseca, Fernanda

    2017-09-01

    Cryopreservation of lactic acid bacteria may lead to undesirable cell death and functionality losses. The membrane is the first target for cell injury and plays a key role in bacterial cryotolerance. This work aimed at investigating at a subcellular resolution the membrane fluidity of two populations of Lactobacillus delbrueckii subsp. bulgaricus when subjected to cold and osmotic stresses associated to freezing. Cells were cultivated at 42 °C in mild whey medium, and they were exposed to sucrose solutions of different osmolarities (300 and 1800 mOsm L -1 ) after harvest. Synchrotron fluorescence microscopy was used to measure membrane fluidity of cells labeled with the cytoplasmic membrane probe 1-[4 (trimethylamino) phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH). Images were acquired at 25 and 0 °C, and more than a thousand cells were individually analyzed. Results revealed that a bacterial population characterized by high membrane fluidity and a homogeneous distribution of fluidity values appeared to be positively related to freeze-thaw resistance. Furthermore, rigid domains with different anisotropy values were observed and the occurrence of these domains was more important in the freeze-sensitive bacterial population. The freeze-sensitive cells exhibited a broadening of existing highly rigid lipid domains with osmotic stress. The enlargement of domains might be ascribed to the interaction of sucrose with membrane phospholipids, leading to membrane disorganization and cell degradation.

  4. A Vivens Ex Vivo Study on the Synergistic Effect of Electrolysis and Freezing on the Cell Nucleus.

    Science.gov (United States)

    Lugnani, Franco; Zanconati, Fabrizio; Marcuzzo, Thomas; Bottin, Cristina; Mikus, Paul; Guenther, Enric; Klein, Nina; Rubinsky, Liel; Stehling, Michael K; Rubinsky, Boris

    2015-01-01

    Freezing-cryosurgery, and electrolysis-electrochemical therapy (EChT), are two important minimally invasive surgery tissue ablation technologies. Despite major advantages they also have some disadvantages. Cryosurgery cannot induce cell death at high subzero freezing temperatures and requires multiple freeze thaw cycles, while EChT requires high concentrations of electrolytic products-which makes it a lengthy procedure. Based on the observation that freezing increases the concentration of solutes (including products of electrolysis) in the frozen region and permeabilizes the cell membrane to these products, this study examines the hypothesis that there could be a synergistic effect between freezing and electrolysis in their use together for tissue ablation. Using an animal model we refer to as vivens ex vivo, which may be of value in reducing the use of animals for experiments, combined with a Hematoxylin stain of the nucleus, we show that there are clinically relevant protocols in which the cell nucleus appears intact when electrolysis and freezing are used separately but is affected by certain combinations of electrolysis and freezing.

  5. Anatomical regulation of ice nucleation and cavitation helps trees to survive freezing and drought stress

    Science.gov (United States)

    Lintunen, A.; Hölttä, T.; Kulmala, M.

    2013-01-01

    Water in the xylem, the water transport system of plants, is vulnerable to freezing and cavitation, i.e. to phase change from liquid to ice or gaseous phase. The former is a threat in cold and the latter in dry environmental conditions. Here we show that a small xylem conduit diameter, which has previously been shown to be associated with lower cavitation pressure thus making a plant more drought resistant, is also associated with a decrease in the temperature required for ice nucleation in the xylem. Thus the susceptibility of freezing and cavitation are linked together in the xylem of plants. We explain this linkage by the regulation of the sizes of the nuclei catalysing freezing and drought cavitation. Our results offer better understanding of the similarities of adaption of plants to cold and drought stress, and offer new insights into the ability of plants to adapt to the changing environment. PMID:23778457

  6. Freeze-all cycle in reproductive medicine: current perspectives.

    Science.gov (United States)

    Roque, Matheus; Valle, Marcello; Kostolias, Alessandra; Sampaio, Marcos; Geber, Selmo

    2017-02-01

    The freeze-all strategy has emerged as an alternative to fresh embryo transfer (ET) during in vitro fertilization (IVF) cycles. Although fresh ET is the norm during assisted reproductive therapies (ART), there are many concerns about the possible adverse effects of controlled ovarian stimulation (COS) over the endometrium. The supra-physiologic hormonal levels that occur during a conventional COS are associated with modifications in the peri-implantation endometrium, which may be related to a decrease in pregnancy rates and poorer obstetric and perinatal outcomes when comparing fresh to frozen-thawed embryo transfers. The main objective of this study was to assess the available literature regarding the freeze-all strategy in IVF cycles, in regards to effectiveness and safety. Although there are many potential advantages in performing a freeze-all cycle over a fresh ET, it seems that the freeze-all strategy is not designed for all IVF patients. There is a need to develop a non-invasive clinical tool to evaluate the endometrial receptivity during a fresh cycle, which enables the selection of patients that would benefit from this strategy. Today, it is reasonable to perform elective cryopreservation of all oocytes/embryos in cases with a risk of OHSS development, and in patients with supra-physiologic hormonal levels during the follicular phase of COS. It is not clear if all normal responders and poor responders may benefit from this strategy.

  7. Identification of clinical biomarkers for pre-analytical quality control of blood samples.

    Science.gov (United States)

    Kang, Hyun Ju; Jeon, Soon Young; Park, Jae-Sun; Yun, Ji Young; Kil, Han Na; Hong, Won Kyung; Lee, Mee-Hee; Kim, Jun-Woo; Jeon, Jae-Pil; Han, Bok Ghee

    2013-04-01

    Pre-analytical conditions are key factors in maintaining the high quality of biospecimens. They are necessary for accurate reproducibility of experiments in the field of biomarker discovery as well as achieving optimal specificity of laboratory tests for clinical diagnosis. In research at the National Biobank of Korea, we evaluated the impact of pre-analytical conditions on the stability of biobanked blood samples by measuring biochemical analytes commonly used in clinical laboratory tests. We measured 10 routine laboratory analytes in serum and plasma samples from healthy donors (n = 50) with a chemistry autoanalyzer (Hitachi 7600-110). The analyte measurements were made at different time courses based on delay of blood fractionation, freezing delay of fractionated serum and plasma samples, and at different cycles (0, 1, 3, 6, 9) of freeze-thawing. Statistically significant changes from the reference sample mean were determined using the repeated-measures ANOVA and the significant change limit (SCL). The serum levels of GGT and LDH were changed significantly depending on both the time interval between blood collection and fractionation and the time interval between fractionation and freezing of serum and plasma samples. The glucose level was most sensitive only to the elapsed time between blood collection and centrifugation for blood fractionation. Based on these findings, a simple formula (glucose decrease by 1.387 mg/dL per hour) was derived to estimate the length of time delay after blood collection. In addition, AST, BUN, GGT, and LDH showed sensitive responses to repeated freeze-thaw cycles of serum and plasma samples. These results suggest that GGT and LDH measurements can be used as quality control markers for certain pre-analytical conditions (eg, delayed processing or repeated freeze-thawing) of blood samples which are either directly used in the laboratory tests or stored for future research in the biobank.

  8. Thawing of Frozen Tuna Meat

    Science.gov (United States)

    Tanaka, Takeo; Nishiwaki, Kôji; Kakuda, Kitonari; Tomimatsu, Takao

    Frozen southern bluefin tuna meat discolors easily and sometimes contracts when thawed caused by thaw rigor. These phenomenon often become problematic in the transaction or handling of this kind of frozen tuna. Frozen meat blocks of southern Bluefin tuna were thawed separately by air thawing, running water thawing and microwave thawing. Changes occurring during thawing were checked for meat color by met-myoglobin ratio determination and for contract by microscopic observation. Results are as follows : (1) Discoloration scarcely occurred in the process of running water thawing (at 10°C for 50 min, or at 0°C for 6 hr). (2) No contraction was observed during thawing with running water described above and air thawing (at 18-20°C for 6 hr). (3) Discoloration and contraction seemed to be minimized, as to latently contractile blocks, when meat temperature passed through rapidly between -10°C and -5°C, and slowly (for 5-6 hr) between -5°C and -1°C. When the block was originally not contractile, discloration was minimized by rising meat temperature rapidly from -10°C to -l°C.

  9. Influence of vacuum impregnation and pulsed electric field on the freezing temperature and ice propagation rates of spinach leaves

    Science.gov (United States)

    Efforts are currently directed towards improving the quality of sensitive tissues of fruits and vegetables after freezing and thawing. One of the methods under investigation is the combination of vacuum impregnation (VI) with cryoprotectants and pulsed electric field (PEF) applied to the plant tiss...

  10. Spatial and temporal variations of thaw layer thickness and its controlling factors identified using time-lapse electrical resistivity tomography and hydro-thermal modeling

    Science.gov (United States)

    Tran, Anh Phuong; Dafflon, Baptiste; Bisht, Gautam; Hubbard, Susan S.

    2018-06-01

    Quantitative understanding of controls on thaw layer thickness (TLT) dynamics in the Arctic peninsula is essential for predictive understanding of permafrost degradation feedbacks to global warming and hydrobiochemical processes. This study jointly interprets electrical resistivity tomography (ERT) measurements and hydro-thermal numerical simulation results to assess spatiotemporal variations of TLT and to determine its controlling factors in Barrow, Alaska. Time-lapse ERT measurements along a 35-m transect were autonomously collected from 2013 to 2015 and inverted to obtain soil electrical resistivity. Based on several probe-based TLT measurements and co-located soil electrical resistivity, we estimated the electrical resistivity thresholds associated with the boundary between the thaw layer and permafrost using a grid search optimization algorithm. Then, we used the obtained thresholds to derive the TLT from all soil electrical resistivity images. The spatiotemporal analysis of the ERT-derived TLT shows that the TLT at high-centered polygons (HCPs) is smaller than that at low-centered polygons (LCPs), and that both thawing and freezing occur earlier at the HCPs compared to the LCPs. In order to provide a physical explanation for dynamics in the thaw layer, we performed 1-D hydro-thermal simulations using the community land model (CLM). Simulation results showed that air temperature and precipitation jointly govern the temporal variations of TLT, while the topsoil organic content (SOC) and polygon morphology are responsible for its spatial variations. When the topsoil SOC and its thickness increase, TLT decreases. Meanwhile, at LCPs, a thicker snow layer and saturated soil contribute to a thicker TLT and extend the time needed for TLT to freeze and thaw. This research highlights the importance of combination of measurements and numerical modeling to improve our understanding spatiotemporal variations and key controls of TLT in cold regions.

  11. Effects of Artificial Supercooling Followed by Slow Freezing on the Microstructure and Qualities of Pork Loin

    Science.gov (United States)

    2016-01-01

    This study investigated the effects of artificial supercooling followed by still air freezing (SSF) on the qualities of pork loin. The qualities of pork frozen by SSF were compared with the fresh control (CT, stored at 4℃ for 24 h), slow freezing (SAF, still air freezing) and rapid freezing (EIF, ethanol immersion freezing) treatments. Compared with no supercooling phenomena of SAF and EIF, the extent of supercooling obtained by SSF treatment was 1.4℃. Despite that SSF was conducted with the same method with SAF, application of artificial supercooling accelerated the phase transition (traverse from -0.6℃ to -5℃) from 3.07 h (SAF) to 2.23 h (SSF). The observation of a microstructure indicated that the SSF prevented tissue damage caused by ice crystallization and maintained the structural integrity. The estimated quality parameters reflected that SSF exhibited superior meat quality compared with slow freezing (SAF). SSF showed better water-holding capacity (lower thawing loss, cooking loss and expressible moisture) and tenderness than SAF, and these quality parameters of SSF were not significantly different with ultra-fast freezing treatment (EIF). Consequently, the results demonstrated that the generation of supercooling followed by conventional freezing potentially had the advantage of minimizing the quality deterioration caused by the slow freezing of meat. PMID:27857541

  12. Effects of Artificial Supercooling Followed by Slow Freezing on the Microstructure and Qualities of Pork Loin.

    Science.gov (United States)

    Kim, Yiseul; Hong, Geun-Pyo

    2016-10-31

    This study investigated the effects of artificial supercooling followed by still air freezing (SSF) on the qualities of pork loin. The qualities of pork frozen by SSF were compared with the fresh control (CT, stored at 4℃ for 24 h), slow freezing (SAF, still air freezing) and rapid freezing (EIF, ethanol immersion freezing) treatments. Compared with no supercooling phenomena of SAF and EIF, the extent of supercooling obtained by SSF treatment was 1.4℃. Despite that SSF was conducted with the same method with SAF, application of artificial supercooling accelerated the phase transition (traverse from -0.6℃ to -5℃) from 3.07 h (SAF) to 2.23 h (SSF). The observation of a microstructure indicated that the SSF prevented tissue damage caused by ice crystallization and maintained the structural integrity. The estimated quality parameters reflected that SSF exhibited superior meat quality compared with slow freezing (SAF). SSF showed better water-holding capacity (lower thawing loss, cooking loss and expressible moisture) and tenderness than SAF, and these quality parameters of SSF were not significantly different with ultra-fast freezing treatment (EIF). Consequently, the results demonstrated that the generation of supercooling followed by conventional freezing potentially had the advantage of minimizing the quality deterioration caused by the slow freezing of meat.

  13. Evaluation of motility, membrane status and DNA integrity of frozen-thawed bottlenose dolphin (Tursiops truncatus) spermatozoa after sex-sorting and recryopreservation.

    Science.gov (United States)

    Montano, G A; Kraemer, D C; Love, C C; Robeck, T R; O'Brien, J K

    2012-06-01

    Artificial insemination (AI) with sex-sorted frozen-thawed spermatozoa has led to enhanced management of ex situ bottlenose dolphin populations. Extended distance of animals from the sorting facility can be overcome by the use of frozen-thawed, sorted and recryopreserved spermatozoa. Although one bottlenose dolphin calf had been born using sexed frozen-thawed spermatozoa derived from frozen semen, a critical evaluation of in vitro sperm quality is needed to justify the routine use of such samples in AI programs. Sperm motility parameters and plasma membrane integrity were influenced by stage of the sex-sorting process, sperm type (non-sorted and sorted) and freezing method (straw and directional) (P0.05) at 24 h. The viability of sorted spermatozoa was higher (Pdolphin spermatozoa undergoing cryopreservation, sorting and recryopreservation are of adequate quality for use in AI.

  14. Fast freezing of cow embryos in French straws with an automatic program.

    Science.gov (United States)

    Massip, A; Van der Zwalmen, P; Hanzen, C; Ectors, F

    1982-09-01

    Cow embryos between day 6.5 and 9 were frozen in 1.5M DMSO in PBS at 2 degrees C/min from seeding to -25 degrees C before being plunged into liquid nitrogen directly or after 10 min at -25 degrees C. Cooling rate from 20 degrees C to -5 degrees C was 9 degrees C/min. Seeding was induced automatically at -5 degrees C by injection of liquid nitrogen vapour. Embryos were subsequently thawed by direct transfer to water at 20 degrees C (group I) or at 37 degrees C (group II). Survival was assessed by culture in vitro and by transfer. In group I, 35.7% were degenerated after thawing (compared to 35.4% in group II). Survival rate after culture in vitro for 24h was not significantly different (48.3% vs 42.8%) and hatching rate after 96h culture was quite similar (33.3% vs 34.4%). In group II, four pregnancies were obtained from 10 embryos transferred. Time at -25 degrees C did not improve the results. Automatic seeding did not impair survival. These results show that the quality of the embryo is the determinant factor for survival after freezing and that the plastic straw is the most suitable vessel for freezing, storage and transfer of embryos.

  15. The transcriptional response of microbial communities in thawing Alaskan permafrost soils

    Science.gov (United States)

    Coolen, Marco J. L.; Orsi, William D.

    2015-01-01

    Thawing of permafrost soils is expected to stimulate microbial decomposition and respiration of sequestered carbon. This could, in turn, increase atmospheric concentrations of greenhouse gasses, such as carbon dioxide and methane, and create a positive feedback to climate warming. Recent metagenomic studies suggest that permafrost has a large metabolic potential for carbon processing, including pathways for fermentation and methanogenesis. Here, we performed a pilot study using ultrahigh throughput Illumina HiSeq sequencing of reverse transcribed messenger RNA to obtain a detailed overview of active metabolic pathways and responsible organisms in up to 70 cm deep permafrost soils at a moist acidic tundra location in Arctic Alaska. The transcriptional response of the permafrost microbial community was compared before and after 11 days of thaw. In general, the transcriptional profile under frozen conditions suggests a dominance of stress responses, survival strategies, and maintenance processes, whereas upon thaw a rapid enzymatic response to decomposing soil organic matter (SOM) was observed. Bacteroidetes, Firmicutes, ascomycete fungi, and methanogens were responsible for largest transcriptional response upon thaw. Transcripts indicative of heterotrophic methanogenic pathways utilizing acetate, methanol, and methylamine were found predominantly in the permafrost table after thaw. Furthermore, transcripts involved in acetogenesis were expressed exclusively after thaw suggesting that acetogenic bacteria are a potential source of acetate for acetoclastic methanogenesis in freshly thawed permafrost. Metatranscriptomics is shown here to be a useful approach for inferring the activity of permafrost microbes that has potential to improve our understanding of permafrost SOM bioavailability and biogeochemical mechanisms contributing to greenhouse gas emissions as a result of permafrost thaw. PMID:25852660

  16. The transcriptional response of microbial communities in thawing Alaskan permafrost soils

    Directory of Open Access Journals (Sweden)

    M J L Coolen

    2015-03-01

    Full Text Available Thawing of permafrost soils is expected to stimulate microbial decomposition and respiration of sequestered carbon. This could, in turn, increase atmospheric concentrations of greenhouse gases, such as carbon dioxide and methane, and create a positive feedback to climate warming. Recent metagenomic studies suggest that permafrost has a large metabolic potential for carbon processing, including pathways for fermentation and methanogenesis. Here, we performed a pilot study using ultrahigh throughput Illumina HiSeq sequencing of reverse transcribed messenger RNA to obtain a detailed overview of active metabolic pathways and responsible organisms in up to 70 cm deep permafrost soils at a moist acidic tundra location in Arctic Alaska. The transcriptional response of the permafrost microbial community was compared before and after eleven days of thaw. In general, the transcriptional profile under frozen conditions suggests a dominance of stress responses, survival strategies, and maintenance processes, whereas upon thaw a rapid enzymatic response to decomposing soil organic matter (SOM was observed. Bacteroidetes, Firmicutes, ascomycete fungi, and methanogens were responsible for largest transcriptional response upon thaw. Transcripts indicative of heterotrophic methanogenic pathways utilizing acetate, methanol, and methylamine were found predominantly in the permafrost table after thaw. Furthermore, transcripts involved in acetogenesis were expressed exclusively after thaw suggesting that acetogenic bacteria are a potential source of acetate for acetoclastic methanogenesis in freshly thawed permafrost. Metatranscriptomics is shown here to be a useful approach for inferring the activity of permafrost microbes that has potential to improve our understanding of permafrost SOM bioavailability and biogeochemical mechanisms contributing to greenhouse gas emissions as a result of permafrost thaw.

  17. ANALYSIS OF STRESS STATE IN UPPER LAYER OF ROAD CONCRETE PAVEMENT WITH TEMPERATURE ACTION

    Directory of Open Access Journals (Sweden)

    M. K. Pshembaev

    2017-01-01

    Full Text Available While being operated auto-road pavements are subjected to intensive mechanical impacts, ultraviolet ray irradiation, freeze-thaw temperatures, freezing and thawing, drying and moistening. Due to these actions various types of pavement distresses appear on the road pavement. The most significant and dangerous type of distresses is micro-cracks on the road surface. One of the main reasons for their formation is an action of weather and climatic factors that initiate large changes in temperature of coating surface and occurrence of large temperature gradients in the upper layer. In this context while designing and operating auto-roads it is rather essential to investigate a stress state in road surface which is caused by temperature action. Purpose of the described investigations is to determine permissible temperature gradients for cement-concrete pavements that exclude formation of micro-cracks on their surface and thickness of damaged surface layer. Calculations of road pavement have been carried out at various laws for temperature distribution in its depth. A finite difference method realized in PARUS software has been used for studying a stress state of cement-concrete auto-roads. Regularities for distribution of stresses in cement-concrete pavement of auto-roads have been obtained at various surface temperatures. Permissible temperature gradients in the upper pavement layer have been determined and thickness of the layer where micro-cracks are formed has been assessed in the paper. Strength criterion based on the process of micro-crack formation and development in the concrete has been used for calculations. Risk of micro-crack formation on the auto-road pavement depends on material strength, conditions of plate fixing and temperature gradients.

  18. Investigation of stress distribution in normal and oblique partial penetration. Welded nozzles by 3-D photoelastic stress freezing method

    International Nuclear Information System (INIS)

    Miyamoto, H.; Kubo, M.; Katori, T.

    1981-01-01

    Experimental investigation by 3-D photoelasticity has been carried out to measure the stress distribution of partial penetration welded nozzles attached to the bottom head of a pressure vessel. A 3-D photoelastic stress freezing method was chosen as the most effective means of observation of the stress distribution in the vicinity of the nozzle/wall weld. The experimental model was a 1:20 scale spherical bottom head. Both an axisymmetric nozzle and an asymmetric nozzle were investigated. Epoxy resin, which is a thermosetting plastic, was used as the model material. The oblique effect was examined by comparing the stress distribution of the asymmetric nozzle with that of the axisymmetric nozzle. Furthermore, the experimental results were compared with the analytical results using 3-D finite element method (FEM). The stress distributions obtained from the frozen fringe pattern of the 3-D photoelastic model were in good agreement with those by 3-D FEM. (orig.)

  19. Freezing and low temperature photoinhibition tolerance in cultivated potato and potato hybrids

    Directory of Open Access Journals (Sweden)

    M.M. SEPPÄNEN

    2008-12-01

    Full Text Available Four Solanum tuberosum L. cultivars (Nicola, Pito, Puikula, Timo and somatic hybrids between freezing tolerant S. commersonii and freezing sensitive S. tuberosum were evaluated for their tolerance to freezing and low temperature photoinhibition. Cellular freezing tolerance was studied using ion leakage tests and the sensitivity of the photosynthetic apparatus to freezing and high light intensity stress by measuring changes in chlorophyll fluorescence (FV/FM and oxygen evolution. Exposure to high light intensities after freezing stress increased frost injury significantly in all genotypes studied. Compared with S. tuberosum cultivars, the hybrids were more tolerant both of freezing and intense light stresses. In field experiments the mechanism of frost injury varied according to the severity of night frosts. During night frosts in 1999, the temperature inside the potato canopy was significantly higher than at ground level, and did not fall below the lethal temperature for potato cultivars (from -2.5 to -3.0°C. As a result, frost injury developed slowly, indicating that damage occurred to the photosynthetic apparatus. However, as the temperature at ground level and inside the canopy fell below -4°C, cellular freezing occurred and the canopy was rapidly destroyed. This suggests that in the field visual frost damage can follow from freezing or non-freezing temperatures accompanied with high light intensity. Therefore, in an attempt to improve low temperature tolerance in potato, it is important to increase tolerance to both freezing and chilling stresses.

  20. Developmental differences in stress responding after repeated underwater trauma exposures in rats.

    Science.gov (United States)

    Altman, Daniel E; Simmons, Laurence P; Vuong, Chau T; Taylor, Rachel M; Sousa, Jason C; Marcsisin, Sean R; Zottig, Victor E; Moore, Nicole L T

    2018-05-01

    Adolescence is a distinct developmental period characterized by behavioral and physiological maturation. Rapid ongoing changes during neurodevelopment in particular present potential opportunities for stress to have lasting effects on longitudinal outcomes of behavioral and neuroendocrine function. While adult stress effects on outcomes during adulthood have been characterized, little is known about the lasting effects of adolescent repeated stressor exposure on outcomes during adolescence. We have previously reported different stress responses in adolescent rats relative to adult rats, including a blunted fear response outcome in adulthood in rats stressed during adolescence. The present study characterized the ontogeny of behavioral and neuroendocrine responses to eight underwater trauma (UWT) exposures in rats over a two week poststress time period during adolescence (P34) or adulthood (P83) relative to age-matched control groups that underwent eight swimming episodes without UWT. Repeated UWT exposures starting in adolescence, but not adulthood, resulted in adverse behavioral responses on the elevated plus maze 1 day post-stress. Corticosterone responses did not differ between UWT-exposed and controls for either age group at 1 day or at 7 days poststress, although there was an effect of age on corticosterone levels. We conclude that repeated UWT stress events have a lasting, negative behavioral effect on adolescent rats that is not observed in adult rats after the two-week exposure window. These results suggest that neurophysiological mechanisms underlying recovery from a repeated stressor are immature in adolescence relative to adulthood in rats.

  1. Effects of the technique of cryopreservation and dilution/centrifugation after thawing on the motility and vitality of spermatozoa of oligoasthenozoospermic men

    Directory of Open Access Journals (Sweden)

    Esteves Sandro C.

    2003-01-01

    Full Text Available OBJECTIVE: Comparing in human semen samples with low initial quality, the effects of 2 techniques of cryopreservation and dilution/centrifugation after thawing on the spermatic motility and vitality. MATERIALS AND METHODS: Semen samples from 15 oligo and/or asthenozoospermic individuals assisted in the infertility sector of a tertiary hospital were obtained through masturbation. The samples were divided into 2 portions of equal volume, and diluted (1:1; v/v with the cryoprotector containing glycerol (Test yolk buffer. One portion was frozen through the technique of liquid nitrogen vapor with static phases (group I - GI, while the other was frozen through a programmable biological freezer with linear speed (Planer, Kryo 10, series III (group II - GII. The following parameters were assessed before freezing and after thawing: percentage of spermatozoa with progressive motility (Prog% and percentage of live spermatozoa (Vit%. After defrosting, Prog% was assessed before and after removal of cryoprotector diluent, in different time intervals (zero, 3 h, and 24 h. The statistical analysis has been accomplished by using the non-parametric tests of Wilcoxon and Friedman. RESULTS: There was significant reduction of Prog% and Vit% from before freezing to after defrosting in both groups, I and II (p < 0.001. Values of Prog% and Vit% were not statistically different between groups, after thawing. It has been observed a significant reduction in Prog% among portions frozen with the automated technique after dilution and centrifugation for removal of cryoprotector (p = 0.006. After cryoprotector removal, Prog% has been kept unaltered, in both groups, during the first 3 hours of incubation, although being superior in group I (p = 0,04. There was a significant decrease in Prog% after 24 hours of incubation, in both groups (p < 0,01. CONCLUSION: For human semen samples with low initial quality, freezing through vapor technique or through the automated technique

  2. The use of gelatine in long-term storage (up to 48 hr) at 5°C preserves the pre-freezing and post-thawing quality of brown bear sperm.

    Science.gov (United States)

    Lopez-Urueña, E; Anel-López, L; Borragan, S; Ortega Ferrusola, C; Manrique, P; de Paz, P; Anel, L; Alvarez, M

    2016-10-01

    Sedimentation of spermatozoa occurs during long-term liquid storage and this may produce deleterious changes. Our aim was to apply gelatine supplementation during long-term pre-freezing storage of bear sperm, applying final dilution and 6% glycerol at room temperature and cool in straws. We tested four models of sperm storage using a 1:1 dilution in TTF-ULE-Bear extender (TesT-fructose-egg yolk-glycerol 6%): (i) second 1:1 dilution at room temperature (RT), cooling at 5°C in a tube and final dilution (100 × 10(6)  sperm ml(-1) ) (Standard); (ii) final dilution at RT and cooling in a tube (FD-Tube); (iii) final dilution at RT and cooling in 0.25 ml plastic straw (FD-Straw); and (iv) final dilution at RT in extender supplemented with 1.5% gelatine (Gelatine) and cooling in a 0.25 ml plastic straw. A Standard sample was stored at 5°C for 1 hr (Control); the rest of the samples (Standard, FD-Tube, FD-Straw, Gelatine) were stored for 24 or 48 hrs before freezing (100 × 10(6)  sperm ml(-1) , glycerol 6%). The quality of the samples was assessed for motility by CASA, and viability (SYBR-14/propidium iodide-PI-; VIAB), acrosomal status (PNA-FITC/PI; iACR) and apoptotic status (YO-PRO-1/PI; YOPRO-) by flow cytometry. At pre-freezing, after 48 hr, Gelatine showed significantly higher viability (for VIAB and YOPRO-) and progressiveness (PM, LIN and STR). At 48 hr, Gelatine showed similar YOPRO-, iACR, LIN, STR and ALH respect to Control. At both 24 and 48 h post-thawing, Gelatine sample had similar scores for YOPRO-, iACR, LIN, STR, WOB and VIAB (only 24 hr) when compared with Control, and lower for TM, PM, rapidPM, VAP and ALH. No differences were found among others experimental groups with respect to Control. In conclusion, gelatine could be a suitable alternative to preserve the viability and progressive motility of brown bear ejaculates during long-term pre-freezing storage at 5°C. © 2016 Blackwell Verlag GmbH.

  3. Effects of addition of sodium lauryl sulfate on frozen-thawed canine spermatozoa.

    Science.gov (United States)

    Hori, Tatsuya; Kaseki, Hanae; Fukuhara, Youko; Oba, Hiromichi; Mizutani, Tatsuji; Kawakami, Eiichi; Tsutsui, Toshihiko

    2006-10-01

    The addition of Orvus ES paste (OEP) to extender may be essential for preparing frozen dog semen. The major ingredient of OEP is sodium lauryl sulfate (SLS). In this study, we compared the effect of SLS on frozen dog semen with that of OEP. There were no significant differences between the 2-mg/ml SLS group and OEP group concerning sperm motility, viability and the percentage of viable sperm with intact acrosomes after freeze-thawing. These results suggest that the effectiveness of frozen dog semen extender containing 2 mg/ml of SLS is similar effective to that demonstrated for OEP.

  4. Repeated Predictable Stress Causes Resilience against Colitis-Induced Behavioral Changes in Mice

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    Ahmed M Hassan

    2014-11-01

    Full Text Available Inflammatory bowel disease is associated with an increased risk of mental disorders and can be exacerbated by stress. In this study which was performed with male 10-week old C57Bl/6N mice, we used dextran sulfate sodium (DSS-induced colitis to evaluate behavioral changes caused by intestinal inflammation, to assess the interaction between repeated psychological stress (water avoidance stress, WAS and colitis in modifying behavior, and to analyze neurochemical correlates of this interaction. A 7-day treatment with DSS (2 % in drinking water decreased locomotion and enhanced anxiety-like behavior in the open field test and reduced social interaction. Repeated exposure to WAS for 7 days had little influence on behavior but prevented the DSS-induced behavioral disturbances in the open field and social interaction tests. In contrast, repeated WAS did not modify colon length, colonic myeloperoxidase content and circulating proinflammatory cytokines, parameters used to assess colitis severity. DSS-induced colitis was associated with an increase in circulating neuropeptide Y (NPY, a rise in the hypothalamic expression of cyclooxygenase-2 mRNA and a decrease in the hippocampal expression of NPY mRNA, brain-derived neurotrophic factor mRNA and mineralocorticoid receptor mRNA. Repeated WAS significantly decreased the relative expression of corticotropin-releasing factor mRNA in the hippocampus. The effect of repeated WAS to blunt the DSS-evoked behavioral disturbances was associated with a rise of circulating corticosterone and an increase in the expression of hypothalamic NPY mRNA. These results show that experimental colitis leads to a particular range of behavioral alterations which can be prevented by repeated WAS, a model of predictable chronic stress, while the severity of colitis remains unabated. We conclude that the mechanisms underlying the resilience effect of repeated WAS involves hypothalamic NPY and the hypothalamic-pituitary-adrenal axis.

  5. Preparation, optimization and property of PVA-HA/PAA composite hydrogel.

    Science.gov (United States)

    Chen, Kai; Liu, Jinlong; Yang, Xuehui; Zhang, Dekun

    2017-09-01

    PVA-HA/PAA composite hydrogel is prepared by freezing-thawing, PEG dehydration and annealing method. Orthogonal design method is used to choose the optimization combination. Results showed that HA and PVA have the maximum effect on water content. PVA and freezing-thawing cycles have the maximum effect on creep resistance and stress relaxation rate of hydrogel. Annealing temperature and freezing-thawing cycles have the maximum effect on compressive elastic modulus of hydrogel. Comparing with the water content and mechanical properties of 16 kinds of combination, PVA-HA/PAA composite hydrogel with freezing-thawing cycles of 3, annealing temperature of 120°C, PVA of 16%, HA of 2%, PAA of 4% has the optimization comprehensive properties. PVA-HA/PAA composite hydrogel has a porous network structure. There are some interactions between PVA, HA and PAA in hydrogel and the properties of hydrogel are strengthened. The annealing treatment improves the crystalline and crosslinking of hydrogel. Therefore, the annealing PVA-HA/PAA composite hydrogel has good thermostability, strength and mechanical properties. It also has good lubrication property and its friction coefficient is relative low. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Boar spermatozoa cryopreservation in low glycerol/trehalose enriched freezing media improves cellular integrity.

    Science.gov (United States)

    Gutiérrez-Pérez, Oscar; Juárez-Mosqueda, María de Lourdes; Carvajal, Salvador Uribe; Ortega, María Elena Trujillo

    2009-06-01

    The use of glycerol for boar semen cryopreservation results in low fertility, possibly due to toxicity. This has led to recommend the use of solutions with less than 4% glycerol. Trehalose is a disaccharide known to stabilize proteins and biologic membranes during processes such as cryopreservation. Thus, it was decided to evaluate the cryoprotective effect of glycerol/trehalose mixtures. Effects on motility (M), viability (Vb) and acrosomal integrity (nA) were evaluated. Sperm samples were frozen in three different extenders: G4 contained 4% glycerol; T1 contained 1% glycerol plus 250 mM trehalose and T0.5 was constituted by 0.5% glycerol plus 250 mM trehalose. All extenders yielded similar post-freezing/thawing motility rates. Viability was diminished in T0.5 as compared to the others. In regard to acrosome integrity, it was twice as high (Pextender. Thus, T1 twice as many spermatozoa were alive, motile and intact, than in either T0.5 or G4, i.e. during freeze/thawing the use of T1 resulted in twice as many fertile cells as when using the other extenders. During our study, we noted that there were wide individual variations both in sperm viability and in motility.

  7. Effects of diluents and plasma on honey bee (Apis mellifera L.) drone frozen-thawed semen fertility.

    Science.gov (United States)

    Gül, Aziz; Şahinler, Nuray; Onal, Ali G; Hopkins, Brandon K; Sheppard, Walter S

    2017-10-01

    Cryopreservation is an advanced method used to protect germplasm in liquid nitrogen. Honey bees are of special interest to protect because of their pollination activity and critical role in agriculture. There has been important progress in the cryopreservation of honey bee germplasm in recent years, leading to practical recovery of genetic material for breeding purposes following freezing. However, there remains room for improvement and the goal of the present study was to evaluate the effect of different "extenders" added post-thaw on the fertilization rate of cryopreserved honey bee semen. The purpose of adding extender post-thaw was to dilute the cryoprotectant to remove chemicals after centrifugation because of potential adverse effects. The control consisted of frozen-thawed semen without the addition of an extender; treatment groups included the addition of one of the following extenders: glucose solution, fresh ram semen plasma, fresh honey bee semen plasma, extender solution. All of the above treatments and frozen-thawed control were compared to fresh semen. For each group, 15 virgin queens were instrumentally inseminated with the semen-diluent solution and introduced into nucleus colonies to determine the brood patterns of the queens. Percentages of worker brood produced in the fresh semen, frozen-thawed semen control, glucose, fresh ram semen plasma, fresh honey bee semen plasma, and extender solution supplemented groups were 98.±1.1%, 47.0 ± 0.9%, 3.0 ± 0.8%, 0.3 ± 0.1%, 48.1 ± 4.1% and 40.3 ± 2.4%, respectively. Similiarly, spermatozoa numbers in the spermathecae of the same treatment groups were 3.6 × 10 6 , 1.6 × 10 6 , 7.3 × 10 5 , 4.7 × 10 5 , 8.1 × 10 5 , and 4.6 × 10 5 spermatozoa for the same treatment, respectively. The differences in both worker brood percentage and sperm count in the spermatheca were statistically significant (P drone semen plasma group. We found a positive correlation between sperm count in

  8. Reversal of haloperidol induced motor deficits in rats exposed to repeated immobilization stress.

    Science.gov (United States)

    Shireen, Erum; Pervez, Sidra; Masroor, Maria; Ali, Wafa Binte; Rais, Qudsia; Khalil, Samira; Tariq, Anum; Haleem, Darakshan Jabeen

    2014-09-01

    Stress is defined as a non specific response of body to any physiological and psychological demand. Preclinical studies have shown that an uncontrollable stress condition produces neurochemical and behavioral deficits. The present study was conducted to test the hypothesis that a decrease in the responsiveness of somatodendritic 5-hydroxytryptamine (5-HT)-1A receptors following adaptation to stress could attenuate haloperidol induced acute parkinsonian like effect. Results showed that single exposure (2h) to immobilization stress markedly decreased food intake, growth rate and locomotor activity but these stress-induced behavioral deficits were not observed following repeated (2h/day for 5 days) exposure of immobilization stress suggesting behavioral tolerance occurs to similar stress. An important finding of present study is a reversal of haloperidol-induced motor deficits in animals exposed to repeated immobilization stress than respective control animals. It is suggested that stress induced possible desensitization of somatodendritic 5-HT-1A as well as 5-HT-2C receptors could release dopamine system from the inhibitory influence of serotonin. On the other hand, an increase in the effectiveness of postsynaptic 5-HT-1A receptors elicits a direct stimulatory influence on the activity of dopaminergic neuron and is possibly involved in the reversal of haloperidol-induced parkinsonian like symptoms in repeatedly immobilized rats.

  9. Freeze-drying of mononuclear cells derived from umbilical cord blood followed by colony formation.

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    Dity Natan

    Full Text Available BACKGROUND: We recently showed that freeze-dried cells stored for 3 years at room temperature can direct embryonic development following cloning. However, viability, as evaluated by membrane integrity of the cells after freeze-drying, was very low; and it was mainly the DNA integrity that was preserved. In the present study, we improved the cells' viability and functionality after freeze-drying. METHODOLOGY/PRINCIPAL FINDINGS: We optimized the conditions of directional freezing, i.e. interface velocity and cell concentration, and we added the antioxidant EGCG to the freezing solution. The study was performed on mononuclear cells (MNCs derived from human umbilical cord blood. After freeze-drying, we tested the viability, number of CD34(+-presenting cells and ability of the rehydrated hematopoietic stem cells to differentiate into different blood cells in culture. The viability of the MNCs after freeze-drying and rehydration with pure water was 88%-91%. The total number of CD34(+-presenting cells and the number of colonies did not change significantly when evaluated before freezing, after freeze-thawing, and after freeze-drying (5.4 x 10(4+/-4.7, 3.49 x 10(4+/-6 and 6.31 x 10(4+/-12.27 cells, respectively, and 31+/-25.15, 47+/-45.8 and 23.44+/-13.3 colonies, respectively. CONCLUSIONS: This is the first report of nucleated cells which have been dried and then rehydrated with double-distilled water remaining viable, and of hematopoietic stem cells retaining their ability to differentiate into different blood cells.

  10. Optimization of dendritic cell loading with tumor cell lysates for cancer immunotherapy.

    Science.gov (United States)

    Hatfield, Paul; Merrick, Alison E; West, Emma; O'Donnell, Dearbhaile; Selby, Peter; Vile, Richard; Melcher, Alan A

    2008-09-01

    The immune response to cancer is critically determined by the way in which tumor cells die. As necrotic, stress-associated death can be associated with activation of antitumor immunity, whole tumor cell antigen loading strategies for dendritic cell (DC)-based vaccination have commonly used freeze-thaw "necrotic" lysates as an immunogenic source of tumor-associated antigens. In this study, the effect of such lysates on the ability of DCs to mature in response to well-established maturation stimuli was examined, and methods to enhance lysate-induced DC activation explored. Freeze-thaw lysates were prepared from murine tumor cell lines and their effects on bone marrow-derived DC maturation and function examined. Unmodified freeze-thaw tumor cell lysates inhibited the toll-like receptor-induced maturation and function of bone marrow-derived DCs, preventing up-regulation of CD40, CD86, and major histocompatibility complex class II, and reducing secretion of inflammatory cytokines [interleukin (IL)-12 p70, tumor necrosis factor-alpha, and IL-6]. Although IL-10 secretion was increased by lysate-pulsed DCs, this was not responsible for the observed suppression of IL-12. Although activation of the nuclear factor-kappaB pathway remained intact, the kinase activity of phosphorylated p38 mitogen-activated protein kinase was inhibited in lysate-pulsed DCs. Lysate-induced DC suppression was partially reversed in vitro by induction of tumor cell stress before lysis, and only DCs loaded with stressed lysates afforded protection against tumor challenge in vivo. These data suggest that ex vivo freeze-thaw of tumor cells does not effectively mimic in vivo immunogenic necrosis, and advocates careful characterization and optimization of tumor cell-derived vaccine sources for cancer immunotherapy.

  11. Hysteresis analysis of graphene transistor under repeated test and gate voltage stress

    International Nuclear Information System (INIS)

    Yang Jie; Jia Kunpeng; Su Yajuan; Zhao Chao; Chen Yang

    2014-01-01

    The current transport characteristic is studied systematically based on a back-gate graphene field effect transistor, under repeated test and gate voltage stress. The interface trapped charges caused by the gate voltage sweep process screens the gate electric field, and results in the neutral point voltage shift between the forth and back sweep direction. In the repeated test process, the neutral point voltage keeps increasing with test times in both forth and back sweeps, which indicates the existence of interface trapped electrons residual and accumulation. In gate voltage stress experiment, the relative neutral point voltage significantly decreases with the reducing of stress voltage, especially in −40 V, which illustrates the driven-out phenomenon of trapped electrons under negative voltage stress. (semiconductor devices)

  12. Reduced freezing in posttraumatic stress disorder patients while watching affective pictures

    NARCIS (Netherlands)

    Fragkaki, I.; Roelofs, K.; Stins, J.F.; Jongedijk, R.A.; Hagenaars, M.A.

    2017-01-01

    Besides fight and flight responses, animals and humans may respond to threat with freezing, an adaptive response characterized by bradycardia and physical immobility. Risk assessment is enhanced during freezing to promote optimal decision-making. Indeed, healthy participants showed freezing-like

  13. Impacts of post-mortem ageing prior to freezing on technological and oxidative properties of coarse ground lamb sausage in a model system

    OpenAIRE

    Juhui Choe; Hyun-Wook Kim; Mustafa M. Farouk; Yuan H. Brad Kim

    2017-01-01

    Objective The objective of this study was to evaluate the effects of ageing time of lamb loins prior to freezing on technological characteristics and oxidation stability of coarse ground lamb loin sausage using in a model system. Methods Lamb loins (M. longissimus lumborum, n = 25) were aged at ?1.5?C for 0, 1, 2, 3, and 8 wk and then frozen for the remaining days (a total of 30 wk). The aged/frozen/thawed lamb loins were ground, and model sausages were formulated with 75% aged/frozen/thawed ...

  14. The effects of different levels of vitamin E and vitamin C in modified Beltsville extender on rooster post-thawed sperm quality.

    Science.gov (United States)

    Amini, Mahmood Reza; Kohram, Hamid; Zare Shahaneh, Ahmad; Zhandi, Mahdi; Sharideh, Hossein; Nabi, Mohammad Mehdi

    2015-12-01

    Oxidative damage of sperm by means of reactive oxygen species generated by the cellular components of semen is one of the main reason of declined motility and fertility of sperm during the freeze-thawing process. This study was conducted to determine the influence of vitamin C and vitamin E on rooster post-thawed sperm motility, viability and malondialdehyde (MDA) level. Semen samples from 10 sexually-mature Ross 308 breeder roosters were collected and pooled, divided into nine equal parts and diluted with modified Beltsville extender containing with no antioxidants (control), or containing 100 (C100), 200 (C200), 400 (C400), 800 (C800) µg/mL vitamin C, and 2 (E2), 5 (E5), 10 (E10) and 15 (E15) µg/mL vitamin E. After thawing, total and progressive sperm motility, sperm viability and semen MDA level were assessed. The results shown that C200 and E5 extenders resulted in higher total motility (p rooster spermatozoa.

  15. Negative effect of combined cysteine and glutathione in soy lecithin-based extender on post-thawed ram spermatozoa.

    Science.gov (United States)

    Zhandi, Mahdi; Sharafi, Mohsen

    2015-09-01

    This study was conducted to evaluate the effect of combined cysteine and glutathione in soy lecithin-based semen extender on post-thawed ram sperm quality. A total of 28 ejaculates were collected twice a week (from four rams) during breeding season. In each replicate, semen samples (n = 4, one ejaculate for each ram) were pooled and divided into three equal parts, and each part was diluted with one of following extender: (1) soy lecithin-based extender containing no cysteine and no glutathione (C0-G0), (2) soy lecithin-based extender containing cysteine (5 mM) and glutathione (5 mM) (C5-G5), and (3) soy lecithin-based extender containing cysteine (10 mM) and glutathione (10 mM) (C10-G10). After freeze-thawing process, motility and velocity parameters, plasma membrane integrity and functionality, mitochondrial activity, and apoptosis features of spermatozoa were evaluated. The obtained results showed that total and progressive motility, plasma membrane integrity and functionality, and live post-thawed spermatozoa was lower in C10-G10 extender compared to C0-G0 and C5-G5 extenders (P 0.05). In conclusion, it seems that high concentration of combined cysteine and glutathione in soy lecithin-based semen extender has a detrimental effect of post-thawed ram sperm quality.

  16. Permafrost soil characteristics and microbial community structure across a boreal forest watershed vary over short spatial scales and dictate community responses to thaw.

    Science.gov (United States)

    Stegen, J.; Bottos, E. M.; Kennedy, D.; Romero, E. B.; Fansler, S.; Chu, R. K.; Tfaily, M.; Jansson, J.; Bernstein, H. C.; Brown, J. M.; Markillie, L. M.

    2017-12-01

    Understanding drivers of permafrost microbial community structure and function is critical for understanding permafrost microbiology and predicting ecosystem responses to thaw; however, studies describing ecological controls on these communities are lacking. We hypothesize that permafrost communities are uniquely shaped by constraints imposed by prolonged freezing, and decoupled from the selective factors that influence non-permafrost soil communities, but that pre-thaw environmental and community characteristics will be strong determinants of community structure and function post-thaw. We characterized patterns of environmental variation and microbial community composition in sixty permafrost samples spanning landscape gradients in a boreal forest watershed, and monitored community responses to thaw. Consistent with our hypothesis, we found that, proportionally, the strongest process influencing permafrost community composition was dispersal limitation (0.36), exceeding the influence of homogenous selection (0.21) and variable selection (0.16), and that deterministic selection arose primarily from energetic constraints of the permafrost environment. Our data supported a structural equation model in which organic carbon thermodynamics and organic acid content, influenced redox conditions and total selection. Post-thaw community composition was found to be driven primarily by pre-thaw community composition, indicating a strong influence of historical conditions. Together, these results suggest that community responses to thaw may be highly varied over short distances and that changes in community structure and function are likely to be drastic, as changes to system hydrology mobilize organisms and nutrients, thereby relieving the primary constraints on the system. These findings are being integrated with metabolomic and metatranscriptomic analyses to improve understanding of how pre-thaw conditions can be used to predict microbial activity post-thaw.

  17. Freeze-thaw lysates of Plasmodium falciparum-infected red blood cells induce differentiation of functionally competent regulatory T cells from memory T cells.

    Science.gov (United States)

    Finney, Olivia C; Lawrence, Emma; Gray, Alice P; Njie, Madi; Riley, Eleanor M; Walther, Michael

    2012-07-01

    In addition to naturally occurring regulatory T (nTreg) cells derived from the thymus, functionally competent Treg cells can be induced in vitro from peripheral blood lymphocytes in response to TCR stimulation with cytokine costimulation. Using these artificial stimulation conditions, both naïve as well as memory CD4(+) T cells can be converted into induced Treg (iTreg) cells, but the cellular origin of such iTreg cells in vivo or in response to more physiologic stimulation with pathogen-derived antigens is less clear. Here, we demonstrate that a freeze/thaw lysate of Plasmodium falciparum schizont extract (PfSE) can induce functionally competent Treg cells from peripheral lymphocytes in a time- and dose-dependent manner without the addition of exogenous costimulatory factors. The PfSE-mediated induction of Treg cells required the presence of nTreg cells in the starting culture. Further experiments mixing either memory or naïve T cells with antigen presenting cells and CFSE-labeled Treg cells identified CD4(+) CD45RO(+) CD25(-) memory T cells rather than Treg cells as the primary source of PfSE-induced Treg cells. Taken together, these data suggest that in the presence of nTreg cells, PfSE induces memory T cells to convert into iTreg cells that subsequently expand alongside PfSE-induced effector T cells. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Comparative study on the freeze stability of yeast and chemical leavened steamed bread dough.

    Science.gov (United States)

    Wang, Pei; Yang, Runqiang; Gu, Zhenxin; Xu, Xueming; Jin, Zhengyu

    2017-04-15

    The present study comparatively evaluated the evolution of yeast and chemical leavened steamed bread dough (YLD/CLD) quality during freeze/thaw (FT) cycles. The steamed bread quality of CLD was more freeze-stable than that of the YLD after 3 FT cycles. Decreased yeast viability contributed to the loss of gassing power in YLD while no significant differences were observed for CLD during FT cycles. However, faster gas release rate in frozen CLD indicated gas retention loss due to the distortion of gluten network. Glutenin macropolymers (GMP) depolymerization via breakage of inter-chain disulfide (SS) bonds and conversions of α-helix and β-turn to β-sheet structures were the main indicators of gluten deterioration. Gluten network was more vulnerable in frozen YLD, resulting in detectable loss of viscoelasticity. The results suggested that supplement of chemical leavener contributed to a more freeze-tolerant gluten network besides its stable gassing power. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Modern human sperm freezing: Effect on DNA, chromatin and acrosome integrity.

    Science.gov (United States)

    Rahiminia, Tahereh; Hosseini, Akram; Anvari, Morteza; Ghasemi-Esmailabad, Saeed; Talebi, Ali Reza

    2017-08-01

    Presence of vitrification method in sperm freezing and the introduction of solid surface vitrification beside rapid freezing in vapour, opens an easy and safe way to help infertility centres. While the effects of cryopreservation on motility, morphology and viability of sperm are documented, the question of the probable alteration of sperm DNA, chromatin and acrosome integrity after freezing and thawing procedures in different methods is still controversial. Normal sample were collected according to WHO strict criteria. Sperm suspensions were mixed 1:1 with 0.5 M sucrose and divided into four equal aliquots for freezing: fresh, nitrogen direct immersion vitrification (Vit), solid surface vitrification (SSV) and in vapour (Vapour). Sperm suspensions were transferred into a 0.25 ml sterile plastic. Then straw was inserted inside the 0.5 ml straw. For thawing, the straws were immersed in a 42 °C water bath. Beside the sperm parameters, we assessed the acrosome reaction by double staining, chromatin integrity by toluidine blue (Tb) and chromomycin A3 (CMA3) and DNA integrity by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) respectively. In progressive motility, the highest rate occurred in Vit (39.9 ± 13.3). Moreover, the lowest rate of immotile sperm was in Vit (32.7 ± 16.3). In normal morphology, the group Vit was similar to the fresh, while SSV and Vapour were significantly different from the fresh. The percentage of acrosome-reacted sperms was more in Vit (81.3 ± 10.2) than the fresh group. TUNEL+ results showed that DNA fragmentation was significantly increased in Vit (p-value = 0.025). While in SSV and Vapour results were comparable to fresh. There was a significant correlation between TUNEL+ and normal morphology, TB, CMA3 and presence of intact acrosome. Sperm in Vapour was healthier in terms of DNA, chromatin and acrosome integrity. In contrast of higher motility and normal morphology; DNA, chromatin and acrosome

  20. Effects of the technique of cryopreservation and dilution/centrifugation after thawing on the motility and vitality of spermatozoa of oligoasthenozoospermic men.

    Science.gov (United States)

    Esteves, Sandro C; Spaine, Deborah M; Cedenho, Agnaldo P; Srougi, Miguel

    2003-01-01

    Comparing in human semen samples with low initial quality, the effects of 2 techniques of cryopreservation and dilution/centrifugation after thawing on the spermatic motility and vitality. Semen samples from 15 oligo and/or asthenozoospermic individuals assisted in the infertility sector of a tertiary hospital were obtained through masturbation. The samples were divided into 2 portions of equal volume, and diluted (1:1; v/v) with the cryoprotector containing glycerol (Test yolk buffer). One portion was frozen through the technique of liquid nitrogen vapor with static phases (group I - GI), while the other was frozen through a programmable biological freezer with linear speed (Planer, Kryo 10, series III) (group II - GII). The following parameters were assessed before freezing and after thawing: percentage of spermatozoa with progressive motility (Prog%) and percentage of live spermatozoa (Vit%). After defrosting, Prog% was assessed before and after removal of cryoprotector diluent, in different time intervals (zero, 3 h, and 24 h). The statistical analysis has been accomplished by using the non-parametric tests of Wilcoxon and Friedman. There was significant reduction of Prog% and Vit% from before freezing to after defrosting in both groups, I and II (p live spermatozoa with progressive motility. The effects of dilution and centrifugation to remove the cryoprotector had a negative impact only in samples frozen through the automated technique. In both techniques, progressive motility is kept constant during the first 3 hours after thawing and removal of the cryoprotector, but is drastically diminished by the end of an incubation period of 24 hours.

  1. Effect of a pre-freezing treatment with cholesterol-loaded cyclodextrins on boar sperm longevity, capacitation dynamics, ability to adhere to porcine oviductal epithelial cells in vitro and DNA fragmentation dynamics.

    Science.gov (United States)

    Tomás, C; Blanch, E; Fazeli, A; Mocé, E

    2013-01-01

    The aim of this work was to examine how a pre-freezing treatment with cholesterol-loaded cyclodextrins (CLC) affects boar sperm longevity, capacitation dynamics, ability to bind to a porcine telomerase-immortalised oviductal epithelial cell line (TERT-OPEC) in vitro and DNA integrity dynamics after freeze-thawing. Although the samples treated with CLC exhibited lower sperm quality than the control samples (P0.05) after long-term incubation (26h at 37 or 16°C). Additionally, the CLC-treated spermatozoa underwent similar capacitation and DNA fragmentation dynamics as the control spermatozoa (P>0.05). However, CLC-treated spermatozoa were better able to bind to TERT-OPEC in vitro (POPEC in vitro, which could have an effect on the establishment of the sperm reservoir in the ampullary--isthmic junction in vivo. Additionally, frozen-thawed spermatozoa can be stored at 16°C for at least 6h without a significant observable decline in sperm quality, which could be beneficial for the transport of thawed diluted doses of spermatozoa from the laboratory to the farm.

  2. Optimization of the freezing process for hematopoietic progenitor cells: effect of precooling, initial dimethyl sulfoxide concentration, freezing program, and storage in vapor-phase or liquid nitrogen on in vitro white blood cell quality.

    Science.gov (United States)

    Dijkstra-Tiekstra, Margriet J; Setroikromo, Airies C; Kraan, Marcha; Gkoumassi, Effimia; de Wildt-Eggen, Janny

    2014-12-01

    Adding dimethyl sulfoxide (DMSO) to hematopoietic progenitor cells (HPCs) causes an exothermic reaction, potentially affecting their viability. The freezing method might also influence this. The aim was to investigate the effect of 1) precooling of DMSO and plasma (D/P) and white blood cell (WBC)-enriched product, 2) DMSO concentration of D/P, 3) freezing program, and 4) storage method on WBC quality. WBC-enriched product without CD34+ cells was used instead of HPCs. This was divided into six or eight portions. D/P (20 or 50%; precooled or room temperature [RT]) was added to the WBC-enriched product (precooled or RT), resulting in 10% DMSO, while monitoring temperature. The product was frozen using controlled-rate freezing ("fast-rate" or "slow-rate") and placed in vapor-phase or liquid nitrogen. After thawing, WBC recovery and viability were determined. Temperature increased most for precooled D/P to precooled WBC-enriched product, without influence of 20 or 50% D/P, but remained for all variations below 30°C. WBC recovery for both freezing programs was more than 95%. Recovery of WBC viability was higher for slow-rate freezing compared to fast-rate freezing (74% vs. 61%; p liquid nitrogen was marginal. Based on these results, precooling is not necessary. Fifty percent D/P is preferred over 20% D/P. Slow-rate freezing is preferred over fast-rate freezing. For safety reasons storage in vapor-phase nitrogen is preferred over storage in liquid nitrogen. Additional testing using real HPCs might be necessary. © 2014 AABB.

  3. Bulletin of Materials Science | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    In the present study starch has been blended with poly(vinyl alcohol) to design macroporous architectures following a repeated freeze-thaw method. These macroporous cryogels were loaded with an antibiotic drug, ciprofloxacin hydrochloride (Cfx), and evaluated for its in vitro delivery in a completely controlled manner ...

  4. Pre-analytical factors influencing the stability of cerebrospinal fluid proteins

    DEFF Research Database (Denmark)

    Simonsen, Anja H; Bahl, Justyna M C; Danborg, Pia B

    2013-01-01

    Cerebrospinal fluid (CSF) is a potential source for new biomarkers due to its proximity to the brain. This study aimed to clarify the stability of the CSF proteome when undergoing pre-analytical factors. We investigated the effects of repeated freeze/thaw cycles, protease inhibitors and delayed s...

  5. Incipient balancing selection through adaptive loss of aquaporins in natural Saccharomyces cerevisiae populations.

    Directory of Open Access Journals (Sweden)

    Jessica L Will

    2010-04-01

    Full Text Available A major goal in evolutionary biology is to understand how adaptive evolution has influenced natural variation, but identifying loci subject to positive selection has been a challenge. Here we present the adaptive loss of a pair of paralogous genes in specific Saccharomyces cerevisiae subpopulations. We mapped natural variation in freeze-thaw tolerance to two water transporters, AQY1 and AQY2, previously implicated in freeze-thaw survival. However, whereas freeze-thaw-tolerant strains harbor functional aquaporin genes, the set of sensitive strains lost aquaporin function at least 6 independent times. Several genomic signatures at AQY1 and/or AQY2 reveal low variation surrounding these loci within strains of the same haplotype, but high variation between strain groups. This is consistent with recent adaptive loss of aquaporins in subgroups of strains, leading to incipient balancing selection. We show that, although aquaporins are critical for surviving freeze-thaw stress, loss of both genes provides a major fitness advantage on high-sugar substrates common to many strains' natural niche. Strikingly, strains with non-functional alleles have also lost the ancestral requirement for aquaporins during spore formation. Thus, the antagonistic effect of aquaporin function-providing an advantage in freeze-thaw tolerance but a fitness defect for growth in high-sugar environments-contributes to the maintenance of both functional and nonfunctional alleles in S. cerevisiae. This work also shows that gene loss through multiple missense and nonsense mutations, hallmarks of pseudogenization presumed to emerge after loss of constraint, can arise through positive selection.

  6. Incipient balancing selection through adaptive loss of aquaporins in natural Saccharomyces cerevisiae populations.

    Science.gov (United States)

    Will, Jessica L; Kim, Hyun Seok; Clarke, Jessica; Painter, John C; Fay, Justin C; Gasch, Audrey P

    2010-04-01

    A major goal in evolutionary biology is to understand how adaptive evolution has influenced natural variation, but identifying loci subject to positive selection has been a challenge. Here we present the adaptive loss of a pair of paralogous genes in specific Saccharomyces cerevisiae subpopulations. We mapped natural variation in freeze-thaw tolerance to two water transporters, AQY1 and AQY2, previously implicated in freeze-thaw survival. However, whereas freeze-thaw-tolerant strains harbor functional aquaporin genes, the set of sensitive strains lost aquaporin function at least 6 independent times. Several genomic signatures at AQY1 and/or AQY2 reveal low variation surrounding these loci within strains of the same haplotype, but high variation between strain groups. This is consistent with recent adaptive loss of aquaporins in subgroups of strains, leading to incipient balancing selection. We show that, although aquaporins are critical for surviving freeze-thaw stress, loss of both genes provides a major fitness advantage on high-sugar substrates common to many strains' natural niche. Strikingly, strains with non-functional alleles have also lost the ancestral requirement for aquaporins during spore formation. Thus, the antagonistic effect of aquaporin function-providing an advantage in freeze-thaw tolerance but a fitness defect for growth in high-sugar environments-contributes to the maintenance of both functional and nonfunctional alleles in S. cerevisiae. This work also shows that gene loss through multiple missense and nonsense mutations, hallmarks of pseudogenization presumed to emerge after loss of constraint, can arise through positive selection.

  7. Repeated restraint stress exposure during early withdrawal accelerates incubation of cue-induced cocaine craving.

    Science.gov (United States)

    Glynn, Ryan M; Rosenkranz, J Amiel; Wolf, Marina E; Caccamise, Aaron; Shroff, Freya; Smith, Alyssa B; Loweth, Jessica A

    2018-01-01

    A major challenge for treating cocaine addiction is the propensity for abstinent users to relapse. Two important triggers for relapse are cues associated with prior drug use and stressful life events. To study their interaction in promoting relapse during abstinence, we used the incubation model of craving and relapse in which cue-induced drug seeking progressively intensifies ('incubates') during withdrawal from extended-access cocaine self-administration. We tested rats for cue-induced cocaine seeking on withdrawal day (WD) 1. Rats were then subjected to repeated restraint stress or control conditions (seven sessions held between WD6 and WD14). All rats were tested again for cue-induced cocaine seeking on WD15, 1 day after the last stress or control session. Although controls showed a time-dependent increase in cue-induced cocaine seeking (incubation), rats exposed to repeated stress in early withdrawal exhibited a more robust increase in seeking behavior between WD1 and WD15. In separate stressed and control rats, equivalent cocaine seeking was observed on WD48. These results indicate that repeated stress in early withdrawal accelerates incubation of cocaine craving, although craving plateaus at the same level were observed in controls. However, 1 month after the WD48 test, rats subjected to repeated stress in early withdrawal showed enhanced cue-induced cocaine seeking following acute (24 hours) food deprivation stress. Together, these data indicate that chronic stress exposure enhances the initial rate of incubation of craving during early withdrawal, resulting in increased vulnerability to cue-induced relapse during this period, and may lead to a persistent increase in vulnerability to the relapse-promoting effects of stress. © 2016 Society for the Study of Addiction.

  8. Effects of Repeated Stress on Age-Dependent GABAergic Regulation of the Lateral Nucleus of the Amygdala.

    Science.gov (United States)

    Zhang, Wei; Rosenkranz, J Amiel

    2016-08-01

    The adolescent age is associated with lability of mood and emotion. The onset of depression and anxiety disorders peaks during adolescence and there are differences in symptomology during adolescence. This points to differences in the adolescent neural circuitry that underlies mood and emotion, such as the amygdala. The human adolescent amygdala is more responsive to evocative stimuli, hinting to less local inhibitory regulation of the amygdala, but this has not been explored in adolescents. The amygdala, including the lateral nucleus (LAT) of the basolateral amygdala complex, is sensitive to stress. The amygdala undergoes maturational processes during adolescence, and therefore may be more vulnerable to harmful effects of stress during this time period. However, little is known about the effects of stress on the LAT during adolescence. GABAergic inhibition is a key regulator of LAT activity. Therefore, the purpose of this study was to test whether there are differences in the local GABAergic regulation of the rat adolescent LAT, and differences in its sensitivity to repeated stress. We found that LAT projection neurons are subjected to weaker GABAergic inhibition during adolescence. Repeated stress reduced in vivo endogenous and exogenous GABAergic inhibition of LAT projection neurons in adolescent rats. Furthermore, repeated stress decreased measures of presynaptic GABA function and interneuron activity in adolescent rats. In contrast, repeated stress enhanced glutamatergic drive of LAT projection neurons in adult rats. These results demonstrate age differences in GABAergic regulation of the LAT, and age differences in the mechanism for the effects of repeated stress on LAT neuron activity. These findings provide a substrate for increased mood lability in adolescents, and provide a substrate by which adolescent repeated stress can induce distinct behavioral outcomes and psychiatric symptoms.

  9. Effect of Controlled Ice Nucleation on Stability of Lactate Dehydrogenase During Freeze-Drying.

    Science.gov (United States)

    Fang, Rui; Tanaka, Kazunari; Mudhivarthi, Vamsi; Bogner, Robin H; Pikal, Michael J

    2018-03-01

    Several controlled ice nucleation techniques have been developed to increase the efficiency of the freeze-drying process as well as to improve the quality of pharmaceutical products. Owing to the reduction in ice surface area, these techniques have the potential to reduce the degradation of proteins labile during freezing. The objective of this study was to evaluate the effect of ice nucleation temperature on the in-process stability of lactate dehydrogenase (LDH). LDH in potassium phosphate buffer was nucleated at -4°C, -8°C, and -12°C using ControLyo™ or allowed to nucleate spontaneously. Both the enzymatic activity and tetramer recovery after freeze-thawing linearly correlated with product ice nucleation temperature (n = 24). Controlled nucleation also significantly improved batch homogeneity as reflected by reduced inter-vial variation in activity and tetramer recovery. With the correlation established in the laboratory, the degradation of protein in manufacturing arising from ice nucleation temperature differences can be quantitatively predicted. The results show that controlled nucleation reduced the degradation of LDH during the freezing process, but this does not necessarily translate to vastly superior stability during the entire freeze-drying process. The capability of improving batch homogeneity provides potential advantages in scaling-up from lab to manufacturing scale. Copyright © 2018 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  10. Validity of a portable urine refractometer: the effects of sample freezing.

    Science.gov (United States)

    Sparks, S Andy; Close, Graeme L

    2013-01-01

    The use of portable urine osmometers is widespread, but no studies have assessed the validity of this measurement technique. Furthermore, it is unclear what effect freezing has on osmolality. One-hundred participants of mean (±SD) age 25.1 ± 7.6 years, height 1.77 ± 0.1 m and weight 77.1 ± 10.8 kg provided single urine samples that were analysed using freeze point depression (FPD) and refractometry (RI). Samples were then frozen at -80°C (n = 81) and thawed prior to re-analysis. Differences between methods and freezing were determined using Wilcoxon's signed rank test. Relationships between measurements were assessed using intraclass correlation coefficients (ICC) and typical error of estimate (TE). Osmolality was lower (P = 0.001) using RI (634.2 ± 339.8 mOsm · kgH2O(-1)) compared with FPD (656.7 ± 334.1 mOsm · kgH2O(-1)) but the TE was trivial (0.17). Freezing significantly reduced mean osmolality using FPD (656.7 ± 341.1 to 606.5 ± 333.4 mOsm · kgH2O(-1); P < 0.001), but samples were still highly related following freezing (ICC, r = 0.979, P < 0.001, CI = 0.993-0.997; TE = 0.15; and r=0.995, P < 0.001, CI = 0.967-0.986; TE = 0.07 for RI and FPD respectively). Despite mean differences between methods and as a result of freezing, such differences are physiologically trivial. Therefore, the use of RI appears to be a valid measurement tool to determine urine osmolality.

  11. Effects of Technological Parameters and Fishing Ground on Quality Attributes of Thawed, Chilled Cod Fillets Stored in Modified Atmosphere Packaging

    DEFF Research Database (Denmark)

    Bøknæs, Niels; Østerberg, Carsten; Sørensen, Rie

    2001-01-01

    . The parameters investigated were: (1) packaging in modified atmosphere during frozen storage, (2)frozen storage period and temperature, (3),fishing ground and chill storage temperature, together with (4) the addition of trimethylamine oxide (TMAO) and sodium chloride (NaCl) to cod fillets before freezing......Effects were studied of various technological parameters and fishing ground on quality attributes of thawed, chilled cod fillets stored in modified atmosphere packaging Frozen fillets of Baltic Sea and Barents Sea cod, representing two commercial fishing grounds, were used as raw material...... of Baltic Sea cod. Therefore, addition of trimethylamine oxide and NaCl to Baltic Sea cod fillets was evaluated and shown to protect P, phosphoreum against fro::en storage inactivation and this explained the observed differences in growth of the spoilage bacteria and trimethylamine production between thawed...

  12. Semen quality before cryopreservation and after thawing in 543 patients with testicular cancer

    Science.gov (United States)

    MacKenna, Antonio; Crosby, Javier; Huidobro, Cristián; Correa, Eduardo; Duque, Gonzalo

    2017-01-01

    Objective The main objective of this study was to assess semen characteristics of patients with testicular cancer before cryopreservation and after thawing, to evaluate the consequences of this technique on sperm quality in patients with testicular cancer. Methods Five hundred eighty-nine samples from 543 patients with testicular cancer were cryopreserved between 1995 and 2015, one aliquot per patient was used for a thawing test to assess the impact of cryopreservation on sperm motility; semen analysis was performed before cryo preservation and after thawing, the result interpretation was carried out using the 2010 World Health Organization (WHO) Laboratory Manual, and consent forms were signed by the patients for freezing and when sperm was used for reproductive purposes. Results Hypospermia was observed in 28.7% of samples, the median sperm concentration was 18 million/mL with 35% oligozoospermia; twenty-two patients (4.1%) had azoospermia and 12.7% had severe oligozoospermia, the median sperm count was 31.3 million and 261 semen samples (44.3%) were normal in all parameters according to the WHO; total motile sperm count before cryopreservation and after thawing was 12 (0-412.2) and 7 (0-303.9) million sperm, respectively (p < 0.00001, 95% CI 5.48-14.91), which represents a 32% reduction; concerning the utilization of cryopreserved semen samples, only twelve patients (2.2%) used their frozen sperm for reproductive purposes. Conclusions An impairment in semen quality was found in almost half of the samples from patients with testicular cancer, only few patients had azoospermia or severe oligozoospermia; sperm cryopreservation significantly reduces sperm motility and total motile sperm count and very few patients use their frozen sperm for reproductive purposes. PMID:28333030

  13. Digital laminography assessment of the damage in concrete exposed to freezing temperatures

    KAUST Repository

    Wakimoto, Kentaro

    2008-10-01

    The research explores the possibility of using digital laminography as a non-destructive inspection X-ray method to image the damage existing in concrete exposed to low temperatures. Freezing-thawing and scaling tests were performed and digital laminography was used to determine the degree of damage existing inside the concrete samples. First, digital laminography was performed on the concrete sample and then a visual inspection was done by slicing the sample after it was vacuum-impregnated with epoxy in order to compare the differences in crack width. © 2008 Elsevier Ltd. All rights reserved.

  14. Digital laminography assessment of the damage in concrete exposed to freezing temperatures

    KAUST Repository

    Wakimoto, Kentaro; Blunt, Joshua; Carlos, Cruz; Monteiro, Paulo J.M.; Ostertag, Claudia P.; Albert, Richard

    2008-01-01

    The research explores the possibility of using digital laminography as a non-destructive inspection X-ray method to image the damage existing in concrete exposed to low temperatures. Freezing-thawing and scaling tests were performed and digital laminography was used to determine the degree of damage existing inside the concrete samples. First, digital laminography was performed on the concrete sample and then a visual inspection was done by slicing the sample after it was vacuum-impregnated with epoxy in order to compare the differences in crack width. © 2008 Elsevier Ltd. All rights reserved.

  15. Cathepsin activities and membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling.

    Science.gov (United States)

    Zhang, T; Rawson, D M; Tosti, L; Carnevali, O

    2008-04-01

    This study investigated enzymatic activity of cathepsins and the membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling. Stage III oocytes (>0.5mm), obtained through dissection of anaesthetised female fish and desegregation of ovarian cumulus, were exposed to 2M methanol or 2M DMSO (both prepared in Hank's medium) for 30min at 22 degrees C before being loaded into 0.5ml plastic straws and placed into a programmable cooler. After controlled slow freezing, samples were plunged into liquid nitrogen (LN) and held for at least 10min, and thawed by immersing straws into a 27 degrees C water bath for 10s. Thawed oocytes were washed twice in Hank's medium. Cathepsin activity and membrane integrity of oocytes were assessed both after cryoprotectant treatment at 22 degrees C and after freezing in LN. Cathepsin B and L colorimetric analyses were performed using substrates Z-Arg-ArgNNap and Z-Phe-Arg-4MbetaNA-HCl, respectively, and 2-naphthylamine and 4-methoxy-2-naphthylamine were used as standards. Cathepsin D activity was performed by analysing the level of hydrolytic action on haemoglobin. Oocytes membrane integrity was assessed using 0.2% Trypan blue staining for 5min. Analysis of cathepsin activities showed that whilst the activity of cathepsin B and D was not affected by 2M DMSO treatment, their activity was lowered when treated with 2M methanol. Following freezing to -196 degrees C, the activity of all cathepsins (B, D and L) was significantly decreased in both 2M DMSO and 2M methanol. Trypan blue staining showed that 63.0+/-11.3% and 72.7+/-5.2% oocytes membrane stayed intact after DMSO and methanol treatment for 30min at 22 degrees C, respectively, whilst 14.9+/-2.6% and 1.4+/-0.8% stayed intact after freezing in DMSO and methanol to -196 degrees C. The results indicate that cryoprotectant treatment and freezing modified the activities of lysosomal enzymes involved in oocyte maturation and yolk

  16. Infant attachment predicts bodily freezing in adolescence: evidence from a prospective longitudinal study

    Directory of Open Access Journals (Sweden)

    Hannah C. M. Niermann

    2015-10-01

    Full Text Available Early life-stress, particularly maternal deprivation, is associated with long-lasting deviations in animals’ freezing responses. Given the relevance of freezing for stress-coping, translational research is needed to examine the relation between insecure infant-parent attachment and bodily freezing-like behavior in humans. Therefore, we investigated threat-related reductions in body sway (indicative of freezing-like behavior in 14-year-old adolescents (N=79, for whom attachment security was earlier assessed in infancy. As expected, insecure (versus secure attachment was associated with less body sway for angry versus neutral faces. This effect remained when controlling for intermediate life-events. These results suggest that the long-lasting effects of early negative caregiving experiences on the human stress and threat systems extend to the primary defensive reaction of freezing. Additionally, we replicated earlier work in adults, by observing a significant correlation (in adolescents assessed as securely attached between subjective state anxiety and reduced body sway in response to angry versus neutral faces. Together, this research opens venues to start exploring the role of freezing in the development of human psychopathology.

  17. Improved recovery of post-thaw motility and vitality of human spermatozoa cryopreserved in the presence of dithiothreitol.

    Science.gov (United States)

    Rao, B; David, G

    1984-10-01

    Semen was collected in the laboratory from nine healthy donors. The concentrations and the percentages of live and motile spermatozoa in all semen samples were within the normal range. Each sample was diluted with citrate-egg yolk-glycerol medium with and without 5 mM dithiothreitol (DTT). Samples were frozen in liquid nitrogen vapor (-70 degrees C) for 7 min and subsequently stored in liquid nitrogen. The effect of DTT in cryopreservation of sperm was determined by comparing percentage of motile and live spermatozoa between controls and DTT-treated post-thaw samples. Percentage of motile spermatozoa was determined by two techniques, laser Doppler velocimetry (LDV) and light microscopy. The percentage of live spermatozoa was measured by microscopic evaluation after staining with eosin-nigrosin. It was shown that the addition of DTT to the freezing medium significantly improved the recovery of motile and live spermatozoa in the post-thaw samples. The mean motility recovery, as measured by LDV, was 44.9% in the controls as compared to 73.9% in the DTT-treated samples. Similarly the mean recovery of live spermatozoa in the controls and DTT-treated samples was 66.5 and 86.6%, respectively. Based on these results, a new hypothesis implicating lipid peroxidation in cryoinjury is proposed. It is also suggested that the use of DTT in the freezing medium may offer an advantage over the commonly used techniques of human sperm cryopreservation.

  18. Detection of lipid peroxidation in frozen-thawed avian spermatozoa using C(11)-BODIPY(581/591).

    Science.gov (United States)

    Partyka, Agnieszka; Lukaszewicz, Ewa; Niżański, Wojciech; Twardoń, Jan

    2011-06-01

    The aim of this study was to perform flow cytometric analysis of C11-BODIPY581/591 oxidation in fowl and geese sperm as a marker for membrane lipid peroxidation (LPO) and to establish if the cryopreservation process would make sperm membranes more susceptible to oxidative stress. The experiment was carried out on 10 meat type line Flex roosters and 10 White Koluda® geese. The semen was collected two times a week, by dorso-abdominal massage method and pooled from 10 individuals of each species. Fowl semen samples were subjected to cryopreservation using the "pellet" method and Dimethylacetamide (DMA) as a cryoprotectant. Geese semen samples were cryopreserved in plastic straws in a programmable freezing unit with Dimethyloformamide (DMF) as the cryoprotectant. A fluorescent lipid probe C11-BODIPY581/591 provided with two double bonds that are oxidized during their contact with ROS, was used for the purpose of the assessment of the LPO in freshly diluted semen samples and frozen-thawed semen samples. This probe changes its color according to its state (non peroxidized: red; peroxidized: green). Flow cytometric analysis was used to monitor these changes. The White Koluda® geese fresh semen had a higher level of LPO than the Flex fresh semen (P > 0.01). The cryopreservation of fowl semen significantly (P > 0.01) increased the percentage of live and dead spermatozoa with lipid peroxidation. In frozen-thawed semen of White Koluda® geese the percentage of live spermatozoa with LPO significantly decreased (P > 0.05) whereas significantly (P > 0.01) higher level of dead cells with LPO was observed. There were significant differences between the two studied species. After thawing, the percentage of live and dead spermatozoa with lipid peroxidation was higher in fowl semen than in geese semen (P > 0.01). In conclusion, our data clearly indicate the existence of species specific differences in susceptibility of spermatozoa to the oxidation of PUFAs in the cell membranes

  19. Transcriptional and metabolomic analysis of Ascophyllum nodosum mediated freezing tolerance in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Nair Prasanth

    2012-11-01

    Full Text Available Abstract Background We have previously shown that lipophilic components (LPC of the brown seaweed Ascophyllum nodosum (ANE improved freezing tolerance in Arabidopsis thaliana. However, the mechanism(s of this induced freezing stress tolerance is largely unknown. Here, we investigated LPC induced changes in the transcriptome and metabolome of A. thaliana undergoing freezing stress. Results Gene expression studies revealed that the accumulation of proline was mediated by an increase in the expression of the proline synthesis genes P5CS1 and P5CS2 and a marginal reduction in the expression of the proline dehydrogenase (ProDH gene. Moreover, LPC application significantly increased the concentration of total soluble sugars in the cytosol in response to freezing stress. Arabidopsis sfr4 mutant plants, defective in the accumulation of free sugars, treated with LPC, exhibited freezing sensitivity similar to that of untreated controls. The 1H NMR metabolite profile of LPC-treated Arabidopsis plants exposed to freezing stress revealed a spectrum dominated by chemical shifts (δ representing soluble sugars, sugar alcohols, organic acids and lipophilic components like fatty acids, as compared to control plants. Additionally, 2D NMR spectra suggested an increase in the degree of unsaturation of fatty acids in LPC treated plants under freezing stress. These results were supported by global transcriptome analysis. Transcriptome analysis revealed that LPC treatment altered the expression of 1113 genes (5% in comparison with untreated plants. A total of 463 genes (2% were up regulated while 650 genes (3% were down regulated. Conclusion Taken together, the results of the experiments presented in this paper provide evidence to support LPC mediated freezing tolerance enhancement through a combination of the priming of plants for the increased accumulation of osmoprotectants and alteration of cellular fatty acid composition.

  20. Mapping the sensitivity of citrus crops to freeze stress using a geographical information system in Ramsar, Iran

    Directory of Open Access Journals (Sweden)

    Hasan Zabihi

    2016-12-01

    Full Text Available Citrus, a cold-sensitive plant, often suffers from low temperature, which seriously affects citrus productivity. Environmental constraint factors have mixed impacts on horticulture that differ among the areas, periods and crops. This study presents a statistical analysis to investigate the freeze stress (FS conditions and morphometry, especially altitude and minimum temperature on citriculture at a regional scale. Based on the temperature isolines map and topography, this paper highlights the impact of altitude and minimum temperature on the citrus crop production using geographic information system (GIS techniques, statistical analysis and climatic data in Ramsar, Iran over a period of 30 years from 1980 to 2010. This study shows that the suitability varies in relation to the critical temperature and concludes that both minimum temperature and altitude have significant negative impact on citrus crop production. Climate change, in particular, occurring cold fronts in recent years during the citrus harvest time have been complicating this issue and increased the importance of freezes stress (FS. The results highlight that citrus crop orchards were more strongly affected by the minimum temperature, and along with the elevation ranges theses are major challenging factors.

  1. Context and strain-dependent behavioral response to stress

    Directory of Open Access Journals (Sweden)

    Baum Amber E

    2008-06-01

    Full Text Available Abstract Background This study posed the question whether strain differences in stress-reactivity lead to differential behavioral responses in two different tests of anxiety. Strain differences in anxiety-measures are known, but strain differences in the behavioral responses to acute prior stress are not well characterized. Methods We studied male Fisher 344 (F344 and Wistar Kyoto (WKY rats basally and immediately after one hour restraint stress. To distinguish between the effects of novelty and prior stress, we also investigated behavior after repeated exposure to the test chamber. Two behavioral tests were explored; the elevated plus maze (EPM and the open field (OFT, both of which are thought to measure activity, exploration and anxiety-like behaviors. Additionally, rearing, a voluntary behavior, and grooming, a relatively automatic, stress-responsive stereotyped behavior were measured in both tests. Results Prior exposure to the test environment increased anxiety-related measures regardless of prior stress, reflecting context-dependent learning process in both tests and strains. Activity decreased in response to repeated testing in both tests and both strains, but prior stress decreased activity only in the OFT which was reversed by repeated testing. Prior stress decreased anxiety-related measures in the EPM, only in F344s, while in the OFT, stress led to increased freezing mainly in WKYs. Conclusion Data suggest that differences in stressfulness of these tests predict the behavior of the two strains of animals according to their stress-reactivity and coping style, but that repeated testing can overcome some of these differences.

  2. Fertility and flow cytometric evaluations of frozen-thawed rooster semen in cryopreservation medium containing low-density lipoprotein.

    Science.gov (United States)

    Shahverdi, A; Sharafi, M; Gourabi, H; Yekta, A Amiri; Esmaeili, V; Sharbatoghli, M; Janzamin, E; Hajnasrollahi, M; Mostafayi, F

    2015-01-01

    Frozen-thawed rooster semen is not reliable for use in artificial insemination in commercial stocks. Low-density lipoprotein (LDL) has been assessed for effectiveness as a cryoprotectant in the extender to improve the quality of frozen-thawed rooster semen. Although LDL has been evaluated in a few studies in other species for semen cryopreservation, so far no study has been conducted to examine this cryoprotectant for cryopreservation of fowl semen. Thus, this study aims to analyze the effects of different concentrations of LDL (0%, 2%, 4%, 6%, and 8%) in a Beltsville extender for cryopreservation of rooster spermatozoa. In experiment 1, motion parameters, membrane integrity, acrosome integrity, apoptosis status, and mitochondria activity were assessed after freeze-thawing. The highest quality frozen-thawed semen was selected to be used for evaluation of the fertility rate in experiment 2. Semen was collected from six roosters, twice weekly, then extended in a Beltsville extender that contained different concentrations of LDL as follows: 0% (control), 1% (Beltsville plus 1% LDL [BLDL1]), 2% (BLDL2), 4% (BLDL4), 6% (BLDL6), and 8% (BLDL8). Supplementation of the Beltsville extender with 4% LDL produced the most significant percentage of motility (43.1 ± 1.3), membrane integrity (59.4 ± 2.1),mitochondria activity (49.1 ± 1.19), and viable spermatozoa (45 ± 2.28) compared with the control treatment with the results of 22.7 ± 1.3 (motility), 38.4 ± 2.1 (membrane integrity), 40.25 ± 1.19 (mitochondrial activity), and 37.8 ± 2.28 (viability). In experiment 2, a significantly higher percentage of fertility rate was observed for frozen-thawed semen in the extender supplemented with 4% LDL (49.5 ± 1.6) compared with the control (29.2 ± 2.9). Progressive motility and acrosome integrity were not affected by LDL levels in the extenders. The results revealed that supplementation of the Beltsville extender with 4% LDL resulted in higher quality of frozen-thawed rooster

  3. Evaluation of green tea extract as a glazing material for shrimp frozen by cryogenic freezing.

    Science.gov (United States)

    Sundararajan, Srijanani; Prudente, Alfredo; Bankston, J David; King, Joan M; Wilson, Paul; Sathivel, Subramaniam

    2011-09-01

    Solutions of green tea (Camellia sinensis) extract (GTE) in distilled water were evaluated as a glazing material for shrimp frozen by cryogenic freezing. Total of 2%, 3%, and/or 5% GTE solutions (2GTE, 3GTE, 5GTE) were used for glazing. Distilled water glazed (GDW) and nonglazed shrimp (NG) served as controls. The GTE was characterized by measuring color, pH, (o) Brix, total phenols, and % antiradical activity. Individual catechins were identified by HPLC. The freezing time, freezing rate, and energy removal rate for freezing shrimp by cryogenic freezing process were estimated. The frozen shrimp samples were stored in a freezer at -21 °C for 180 d. Samples were analyzed for pH, moisture content, glazing yield, thaw yield, color, cutting force, and thiobarbituric acid reactive substances (TBARS) after 1, 30, 90, and 180 d. The HPLC analysis of GTE revealed the presence of catechins and their isomers and the total polyphenol content was 148.10 ± 2.49 g/L. The freezing time (min) and energy removal rate (J/s) were 48.67 ± 2.3 and 836.67 ± 78.95, respectively. Glazed samples had higher moisture content compared to NG shrimp after 180 d storage. GTE was effective in controlling the lipid oxidation in shrimp. Glazing with GTE affected a* and b* color values, but had no significant effect on the L* values of shrimp. © 2011 Institute of Food Technologists®

  4. Effect of cyclodextrin-loaded cholesterol conjugates on plasma membrane viability of Piau swine breed frozen/thawed spermatozoa.

    Science.gov (United States)

    Pinho, R O; Lima, D M A; Shiomi, H H; Siqueira, J B; Silveira, C O; Faria, V R; Lopes, P S; Guimarães, S E F; Guimarães, J D

    2016-08-01

    The objective of this study was to investigate the effect of cyclodextrin-loaded cholesterol conjugates addition to freezing extenders on plasma membrane viability of frozen-thawed spermatozoa of the Piau swine breed. Twenty semen samples were used from five males. The freezing extender was based on lactose-egg yolk extender, added to 2% glycerol, 3% dimethylacetamide. The addition of cyclodextrin-loaded cholesterol conjugates was performed after centrifugation, when semen was diluted with the cooling extender. Four groups were subjected to the following treatment: without addition (group 1); 1.5 mg of cyclodextrin-loaded cholesterol/120 × 10(6) sperm (group 2); 1.5 mg of cyclodextrin-loaded cholestanol/120 × 10(6) sperm (group 3); 1.5 mg of cyclodextrin-loaded desmosterol/120 × 10(6) sperm (group 4). To check post-thawing sperm quality sperm motility and sperm morphology evaluation were used. Additionally, to check sperm viability the hypoosmotic swelling test, supravital staining, and fluorescent assay were used. The mean values recorded for total sperm motility of semen immediately after thawing were 54.5 ± 5.8, 55.5 ± 5.3, 53.7 ± 6.7, and 52.5 ± 6.6% respectively for groups one to four, without difference between themselves (p > 0.05). Regarding fluorescent assay the results were 28.3 ± 13.2, 26.9 ± 12.2, 22.2 ± 11.4, and 32.0 ± 15.3% respectively for groups one to four, also without difference between groups (p > 0,05). Similarly, complementary tests for evaluating the integrity and functionality of the plasma membrane showed no difference between treatments (p > 0.05). In conclusion, use of cyclodextrin-loaded cholesterol conjugates added to the plasma membrane of sperm did not demonstrate any additive effect on increasing and/or maintaining sperm motility. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Reduced freezing in posttraumatic stress disorder patients while watching affective pictures

    NARCIS (Netherlands)

    Fragkaki, Iro; Roelofs, Karin; Stins, John; Jongedijk, Ruud A.; Hagenaars, Muriel A.

    2017-01-01

    Besides fight and flight responses, animals and humans may respond to threat with freezing, a response characterized by bradycardia and physical immobility. Risk assessment is proposed to be enhanced during freezing to promote optimal decision making. Indeed, healthy participants showed

  6. Reduced freezing in posttraumatic stress disorder patients while watching affective pictures

    NARCIS (Netherlands)

    Fragkaki, Iro; Roelofs, Karin; Stins, John; Jongedijk, Ruud A.; Hagenaars, Muriel A.

    Besides fight and flight responses, animals and humans may respond to threat with freezing, a response characterized by bradycardia and physical immobility. Risk assessment is proposed to be enhanced during freezing to promote optimal decision making. Indeed, healthy participants showed

  7. Effects of Oxidized Glutathione, Bovine Serum Albumin, Cysteine and Lycopene on the Quality of Frozen-Thawed Ram Semen

    Directory of Open Access Journals (Sweden)

    O. Uysal

    2007-01-01

    Full Text Available Free radicals are known to be involved in lipid peroxidation as well as DNA and sperm membrane damages that may lead to decreased sperm motility or cell death. The balance between free radical production and their detoxification may be an important factor in sperm survival and function before, during and after cryopreservation. The aim of this study was to determine the effects of the addition of the antioxidants of oxidized glutathione (GSSG, bovine serum albumin (BSA, cysteine and lycopene to freezing media on the post-thawing sperm characteristics, including motility, morphology, acrosome integrity, viability and membrane integrity. A total number of 42 ejaculates were collected using the artifi cial vagina from 4 Akkaraman rams and 10 replicates of the ejaculates were diluted with a Tris-based extender containing additives and no additives as control. GSSG (5 mM, BSA (20 mg/ml, cysteine (10 mM and lycopene (800 μg showed more positive effects than other concentrations of the supplements and controls in protecting sperm characteristics after the freezing-thawing process (P < 0.001. Many aspects of sperm protection, e.g. sperm motility, viability and membrane stabilisation of the sperm cells during relative cryopreservation, are the key factors in determining the preservation of sperm function. The results of this study provide a new approach to the cryopreservation of sperm from rams and related breeds, and thereby contribute to the improvement of these breeds for the world sheep industry.

  8. Preparation and properties of GO-PVA composite hydrogel with oriented structure

    Science.gov (United States)

    Liu, Huanqing; Zhang, Gongzheng; Li, Huanjun

    2017-03-01

    We fabricated GO-PVA composite hydrogels with oriented structure by directional freezing and repeated freeze-thawing, which owned superior mechanical property and thermostability than PVA hydrogel. Due to physical interactions such as hydrogen bonding between surface of GO and PVA chains, GO-PVA composite hydrogel possessed higher crosslinking density and smaller pore size and can resist higher temperature and stronger force from outside than PVA hydrogel. These unique properties will endow GO-PVA hydrogel with greater potential application in biomedical materials.

  9. Isolation of baker's yeast mutants with proline accumulation that showed enhanced tolerance to baking-associated stresses.

    Science.gov (United States)

    Tsolmonbaatar, Ariunzaya; Hashida, Keisuke; Sugimoto, Yukiko; Watanabe, Daisuke; Furukawa, Shuhei; Takagi, Hiroshi

    2016-12-05

    During bread-making processes, yeast cells are exposed to baking-associated stresses such as freeze-thaw, air-drying, and high-sucrose concentrations. Previously, we reported that self-cloning diploid baker's yeast strains that accumulate proline retained higher-level fermentation abilities in both frozen and sweet doughs than the wild-type strain. Although self-cloning yeasts do not have to be treated as genetically modified yeasts, the conventional methods for breeding baker's yeasts are more acceptable to consumers than the use of self-cloning yeasts. In this study, we isolated mutants resistant to the proline analogue azetidine-2-carboxylate (AZC) derived from diploid baker's yeast of Saccharomyces cerevisiae. Some of the mutants accumulated a greater amount of intracellular proline, and among them, 5 mutants showed higher cell viability than that observed in the parent wild-type strain under freezing or high-sucrose stress conditions. Two of them carried novel mutations in the PRO1 gene encoding the Pro247Ser or Glu415Lys variant of γ-glutamyl kinase (GK), which is a key enzyme in proline biosynthesis in S. cerevisiae. Interestingly, we found that these mutations resulted in AZC resistance of yeast cells and desensitization to proline feedback inhibition of GK, leading to intracellular proline accumulation. Moreover, baker's yeast cells expressing the PRO1 P247S and PRO1 E415K gene were more tolerant to freezing stress than cells expressing the wild-type PRO1 gene. The approach described here could be a practical method for the breeding of proline-accumulating baker's yeasts with higher tolerance to baking-associated stresses. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Repeated Exposure to Conditioned Fear Stress Increases Anxiety and Delays Sleep Recovery Following Exposure to an Acute Traumatic Stressor

    OpenAIRE

    Greenwood, Benjamin N.; Thompson, Robert S.; Opp, Mark R.; Fleshner, Monika

    2014-01-01

    Repeated stressor exposure can sensitize physiological responses to novel stressors and facilitate the development of stress-related psychiatric disorders including anxiety. Disruptions in diurnal rhythms of sleep–wake behavior accompany stress-related psychiatric disorders and could contribute to their development. Complex stressors that include fear-eliciting stimuli can be a component of repeated stress experienced by human beings, but whether exposure to repeated fear can prime the develo...

  11. Freezing of bentonite. Experimental studies and theoretical considerations

    Energy Technology Data Exchange (ETDEWEB)

    Birgersson, Martin; Karnland, Ola; Nilsson, Ulf (Clay Technology AB, Lund (Sweden))

    2010-01-15

    recorded continuously. The samples have been varied with respect to bentonite type (e.g. calcium or sodium dominated), smectite content and density. The experimental results basically confirm all predictions of the developed theory. In particular: - A weak swelling pressure response above 0 deg C. - A strong and positive swelling pressure response below the freezing point of the external aqueous reservoir, in the order of 1.2 MPa/deg C. - The swelling pressure is completely lost at a specific temperature T{sub C}. The value of T{sub C} is determined solely by the value of the swelling pressure at 0 deg C. - Ice formation (i.e. freezing) occurs in the bentonite only below T{sub C}. - The freezing/thawing is completely reversible. The success of the single pore-type model to describe the process together with the observation that no pressure peaks was observed as the 0 deg C level was passed suggests that water saturated bentonite do contain a negligible amount of larger pores (> 50 nm) since these should freeze at temperatures close to 0 deg C with a resulting pressure increase. From a safety assessment point of view it can be concluded that freezing of the buffer will not occur during the repository lifetime as the reference density corresponds to a T{sub C} below -5 deg C and the lowest predicted temperature at repository depth is approximately -2 deg C. The possibility that the backfilled parts of the repository will freeze during its lifetime cannot be excluded as the backfill has a higher freezing temperature and will also be exposed to lower temperatures in vertically extended structures (ramps and shafts). The possible freezing of the backfill will not impose a problem however as freezing/thawing has been shown to be a reversible process. Part of the bentonite in the borehole seals will also freeze because of its location closer to ground level. The possibility of forming ice lenses by transporting water from lower unfrozen parts of the surrounding rock via the

  12. Freeze Tolerance of Seed-Producing Turf Bermudagrasses.

    Science.gov (United States)

    Anderson, Jeffrey A.; Taliaferro, Charles M.

    2002-01-01

    Bermudagrass, Cynodon dactylon (L.) Pers., suffers periodic severe winter-kill throughout much of its area of use in the contiguous USA. A research goal is to increase freeze tolerance in cultivars to lessen the risk of such damage. An identified research need is for Cynodon germplasm resources to be characterized for freeze tolerance and hybridization potential. Accordingly, the objective of this research was to characterize the relative freeze tolerance of selected fertile bermudagrass plants. Nine tetraploid (2n = 4x = 36) C. dactylon and two triploid (2n = 3x = 27) hybrid (C. dactylon x C. transvaalensis Burtt Davy) clonal plants (standards) were evaluated in two experiments. Plants were propagated clonally and established in Cone-tainers (Ray Leach Cone-tainer Nursery, Canby, OR) for about 10 wk. Acclimation took place for 4 wk in controlled environment chambers at 8/2 degrees C (day/night) temperatures with a 10-h photoperiod. Following acclimation, Cone-tainers were placed into a freeze chamber and cooled rapidly to -2 degrees C, induced to freeze with ice chips, then held overnight at -2 degrees C. The freeze chamber was then programmed to cool linearly at 1 degrees C per hour. For each cultivar, three Cone-tainers were removed at each test temperature. Following thawing, Cone-tainers were transferred to a greenhouse and regrowth was evaluated visually. Nonlinear regression was used to estimate T(mid), which corresponded to the midpoint of the sigmoidal response curve of survival vs temperature. Within experiment one, Tifgreen (T(mid) = -7.2 degrees C) was significantly less cold hardy than Quickstand (-9.0 degrees C), A-12204 (-9.2 degrees C), Midiron (-9.9 degrees C), and A-12195 (-10.5 degrees C). A-12195 was significantly hardier than all genotypes except Midiron. In the second experiment, Arizona Common (-6.6 degrees C), Tifgreen (-7.1 degrees C), and A-12205 (-7.1 degrees C) were less hardy than A-9959 (-8.7 degrees C), A-12156 (-8.9 degrees C), A

  13. Combining reduced glutathione and ascorbic acid has supplementary beneficial effects on boar sperm cryotolerance.

    Science.gov (United States)

    Giaretta, Elisa; Estrada, Efrén; Bucci, Diego; Spinaci, Marcella; Rodríguez-Gil, Joan E; Yeste, Marc

    2015-02-01

    The main aim of this work was to evaluate how supplementing freezing and thawing media with reduced glutathione (GSH) and l-ascorbic acid (AA) affected the quality parameters of frozen-thawed boar spermatozoa. With this purpose, semen samples of 12 ejaculates coming from 12 boars were used. Each ejaculate was split into seven aliquots to which 5 mM of GSH and 100 μM of AA were added separately or together at two different steps of freeze-thawing. Various sperm parameters (levels of free cysteine residues in sperm nucleoproteins, sperm viability, acrosome membrane integrity, intracellular peroxide and superoxide levels [ROS], and total and progressive motility) were evaluated before freezing and at 30 and 240 minutes after thawing. Both GSH and AA significantly improved boar sperm cryotolerance when they were separately added to freezing and thawing media. However, the highest improvement was recorded when both freezing and thawing media were supplemented with 5 mM of GSH plus 100 μM of AA. This improvement was observed in sperm viability and acrosome integrity, sperm motility, and nucleoprotein structure. Although ROS levels were not much increased by freeze-thawing procedures, the addition of GSH and AA to both freezing and thawing extenders significantly decreased intracellular peroxide levels and had no impact on superoxide levels. According to our results, we can conclude that supplementation of freezing and thawing media with both GSH and AA has a combined, beneficial effect on frozen-thawed boar sperm, which is greater than that obtained with the separate addition of either GSH or AA. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Bupleurum falcatum prevents depression and anxiety-like behaviors in rats exposed to repeated restraint stress.

    Science.gov (United States)

    Lee, Bombi; Yun, Hye-Yeon; Shim, Insop; Lee, Hyejung; Hahm, Dae-Hyun

    2012-03-01

    Previous studies have demonstrated that repeated restraint stress in rodents produces increases in depression and anxietylike behaviors and alters the expression of corticotrophinreleasing factor (CRF) in the hypothalamus. The current study focused on the impact of Bupleurum falcatum (BF) extract administration on repeated restraint stress-induced behavioral responses using the forced swimming test (FST) and elevated plus maze (EPM) test. Immunohistochemical examinations of tyrosine hydroxylase (TH) expression in rat brain were also conducted. Male rats received daily doses of 20, 50, or 100 mg/kg (i.p.) BF extract for 15 days, 30 min prior to restraint stress (4 h/day). Hypothalamicpituitary- adrenal axis activation in response to repeated restraint stress was confirmed base on serum corticosterone levels and CRF expression in the hypothalamus. Animals that were pre-treated with BF extract displayed significantly reduced immobility in the FST and increased open-arm exploration in the EPM test in comparison with controls. BF also blocked the increase in TH expression in the locus coeruleus of treated rats that experienced restraint stress. Together, these results demonstrate that BF extract administration prior to restraint stress significantly reduces depression and anxiety-like behaviors, possibly through central adrenergic mechanisms, and they suggest a role for BF extract in the treatment of depression and anxiety disorders.

  15. A fresh look at the freeze-all protocol: a SWOT analysis.

    Science.gov (United States)

    Blockeel, Christophe; Drakopoulos, Panagiotis; Santos-Ribeiro, Samuel; Polyzos, Nikolaos P; Tournaye, Herman

    2016-03-01

    The 'freeze-all' strategy with the segmentation of IVF treatment, namely with the use of a GnRH antagonist protocol, GnRH agonist triggering, the elective cryopreservation of all embryos by vitrification and a frozen-thawed embryo transfer in a subsequent cycle, has become more popular. However, the approach still encounters drawbacks. In this opinion paper, a SWOT (strengths, weaknesses, opportunities and threats) analysis sheds light on the different aspects of this strategy. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. Protein crowding in solution, frozen and freeze-dried states: small-angle neutron and X-ray scattering study of lysozyme/sorbitol/water systems

    Science.gov (United States)

    Krueger, Susan; Khodadadi, Sheila; Clark, Nicholas; McAuley, Arnold; Cristiglio, Viviana; Theyencheri, Narayanan; Curtis, Joseph; Shalaev, Evgenyi

    2015-03-01

    For effective preservation, proteins are often stored as frozen solutions or in glassy states using a freeze-drying process. However, aggregation is often observed after freeze-thaw or reconstitution of freeze-dried powder and the stability of the protein is no longer assured. In this study, small-angle neutron and X-ray scattering (SANS and SAXS) have been used to investigate changes in protein-protein interaction distances of a model protein/cryoprotectant system of lysozyme/sorbitol/water, under representative pharmaceutical processing conditions. The results demonstrate the utility of SAXS and SANS methods to monitor protein crowding at different stages of freezing and drying. The SANS measurements of solution samples showed at least one protein interaction peak corresponding to an interaction distance of ~ 90 Å. In the frozen state, two protein interaction peaks were observed by SANS with corresponding interaction distances at 40 Å as well as 90 Å. On the other hand, both SAXS and SANS data for freeze-dried samples showed three peaks, suggesting interaction distances ranging from ~ 15 Å to 170 Å. Possible interpretations of these interaction peaks will be discussed, as well as the role of sorbitol as a cryoprotectant during the freezing and drying process.

  17. DESCRIPTION OF ULTRASTRUCTURAL DAMAGES IN FROZEN-THAWED CANINE SPERMATOZOA DESCRIÇÃO DE DANOS ULTRAESTRUTURAIS EM ESPERMATOZOIDES CANINOS CONGELADOS-DESCONGELADOS

    Directory of Open Access Journals (Sweden)

    Viviane Helena Chirinéa

    2009-07-01

    Full Text Available In spite of the advances in transmission electronic microscopy (TEM, there are few studies presenting a systematic description of the canine spermatozoa and they are only focused on sperm cell heads. The goal of the present study was to describe ultrastructural appearance of canine fresh and frozen-thawed spermatozoa, focusing on damages induced by freezing-thawing in different sperm regions. Ten ejaculates from five proven stud dogs (two ejaculates/dog were collected, evaluated, extended in Tris-egg yolk-glycerol, frozen and stored in liquid nitrogen, and thawed six weeks later.  Samples were evaluated for progressive motility, morphology, and for ultrastructural analysis by TEM. Concerning to TEM, the most striking differences between fresh and frozen-thawed samples were observed over the mid-piece since the fresh spermatozoa showed a well preserved mid-piece. However, the frozen-thawed spermatozoa mid-piece showed signs of damage such as mitochondrial vacuolization. In conclusion, freezing-thawing processes using a Tris-egg yolk extender induce ultrastructural damages in head and mid-piece of canine sperm, affecting the mitochondrial ultrastructure besides.

    KEY WORDS: Cryopreservation, dog, semen, ultrastructure.
    Apesar dos avanços na microscopia eletrônica de transmissão (MET, existem poucos estudos apresentando uma descrição sistemática do espermatozoide canino, os quais estão focados apenas na cabeça espermática. Objetivou-se, com o presente estudo, descrever a aparência ultraestrutural do espermatozoide canino fresco e congelado-descongelado, enfocando os danos induzidos pela congelação e descongelação em diferentes regiões espermáticas. Dez ejaculados obtidos de cinco cães (dois ejaculados por cão foram coletados, avaliados e diluídos em Tris-gema-glicerol, congelados e armazenados em nitrogênio líquido, e descongelados seis semanas após. Avaliaram-se as amostras quanto à motilidade progressiva

  18. Normal freezing of ideal ternary systems of the pseudobinary type

    Science.gov (United States)

    Li, C. H.

    1972-01-01

    Perfect liquid mixing but no solid diffusion is assumed in normal freezing. In addition, the molar compositions of the freezing solid and remaining liquid, respectively, follow the solidus and liquidus curves of the constitutional diagram. For the linear case, in which both the liquidus and solidus are perfectly straight lines, the normal freezing equation giving the fraction solidified at each melt temperature and the solute concentration profile in the frozen solid was determined as early as 1902, and has since been repeatedly published. Corresponding equations for quadratic, cubic or higher-degree liquidus and solidus lines have also been obtained. The equation of normal freezing for ideal ternary liquid solutions solidified into ideal solid solutions of the pseudobinary type is given. Sample computations with the use of this new equation were made and are given for the Ga-Al-As system.

  19. Fator do plasma seminal associado à integridade de membrana de espermatozóides suínos pós-descongelamento Seminal plasma factor associated to post-thawing swine sperm membrane integrity

    Directory of Open Access Journals (Sweden)

    I. Bianchi

    2008-04-01

    Full Text Available Neste estudo, identificaram-se polipeptídeos associados à integridade da membrana plasmática (IMP de espermatozóides suínos após o processo de congelamento/descongelamento. Por meio do perfil protéico do plasma seminal em SDS-PAGE, observou-se a presença de nove bandas polipeptídicas com pesos moleculares que variaram de 11,97 a 122,52kDa. Detectou-se que uma banda de 26,58kDa esteve associada à baixa IMP (Polypeptides associate to membrane integrity (MI of swine spermatozoa submitted to freezing and thawing were identified. The protein profile of seminal plasma analyzed by SDS-PAGE allowed the identification of nine polypeptide bands with molecular weight ranging from 11.97 to 122.52kDa. One 26.58kDa band was associated with reduced MI (<55%. No associations among other bands and MI were observed. The 26.58kDa factor is associated with reduction of membrane integrity of swine spermatozoa after freezing and thawing.

  20. Repeated social stress leads to contrasting patterns of structural plasticity in the amygdala and hippocampus.

    Science.gov (United States)

    Patel, D; Anilkumar, S; Chattarji, S; Buwalda, B

    2018-03-23

    Previous studies have demonstrated that repeated immobilization and restraint stress cause contrasting patterns of dendritic reorganization as well as alterations in spine density in amygdalar and hippocampal neurons. Whether social and ethologically relevant stressors can induce similar patterns of morphological plasticity remains largely unexplored. Hence, we assessed the effects of repeated social defeat stress on neuronal morphology in basolateral amygdala (BLA), hippocampal CA1 and infralimbic medial prefrontal cortex (mPFC). Male Wistar rats experienced social defeat stress on 5 consecutive days during confrontation in the resident-intruder paradigm with larger and aggressive Wild-type Groningen rats. This resulted in clear social avoidance behavior one day after the last confrontation. To assess the morphological consequences of repeated social defeat, 2 weeks after the last defeat, animals were sacrificed and brains were stained using a Golgi-Cox procedure. Morphometric analyses revealed that, compared to controls, defeated Wistar rats showed apical dendritic decrease in spine density on CA1 but not BLA. Sholl analysis demonstrated a significant dendritic atrophy of CA1 basal dendrites in defeated animals. In contrast, basal dendrites of BLA pyramidal neurons exhibited enhanced dendritic arborization in defeated animals. Social stress failed to induce lasting structural changes in mPFC neurons. Our findings demonstrate for the first time that social defeat stress elicits divergent patterns of structural plasticity in the hippocampus versus amygdala, similar to what has previously been reported with repeated physical stressors. Therefore, brain region specific variations may be a universal feature of stress-induced plasticity that is shared by both physical and social stressors. Copyright © 2018 Elsevier B.V. All rights reserved.