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Sample records for bk060042 cell freeze

  1. Freezing mammalian cells for production of biopharmaceuticals.

    Science.gov (United States)

    Seth, Gargi

    2012-03-01

    Cryopreservation techniques utilize very low temperatures to preserve the structure and function of living cells. Various strategies have been developed for freezing mammalian cells of biological and medical significance. This paper highlights the importance and application of cryopreservation for recombinant mammalian cells used in the biopharmaceutical industry to produce high-value protein therapeutics. It is a primer that aims to give insight into the basic principles of cell freezing for the benefit of biopharmaceutical researchers with limited or no prior experience in cryobiology. For the more familiar researchers, key cell banking parameters such as the cell density and hold conditions have been reviewed to possibly help optimize their specific cell freezing protocols. It is important to understand the mechanisms underlying the freezing of complex and sensitive cellular entities as we implement best practices around the techniques and strategies used for cryopreservation. PMID:22226818

  2. PEM Fuel Cell Freeze Durability and Cold Start Project

    Energy Technology Data Exchange (ETDEWEB)

    Patterson, T.; O' Neill, Jonathan

    2008-01-02

    UTC has taken advantage of the unique water management opportunities inherent in micro-porous bipolar-plates to improve the cold-start performance of its polymer electrolyte fuel cells (PEFC). Diagnostic experiments were used to determine the limiting factors in micro-porous plate PEFC freeze performance and the causes of any performance decay. Alternative cell materials were evaluated for their freeze performance. Freeze-thaw cycling was also performed to determine micro-porous plate PEFC survivability. Data from these experiments has formed the basis for continuing development of advanced materials capable of supporting DOE's cold-start and durability objectives.

  3. Determination of cell volume during equilibrium freezing process

    Institute of Scientific and Technical Information of China (English)

    ZHAO Gang; GAO Dayong; HE Liqun; WANG Peitao; DING Weiping; XIE Xiaojian; LIU Zhong; ZHANG Haifeng; SHU Zhiquan; LUO Dawei

    2003-01-01

    A new type electronic particle counter (EPC, MultisizerTM 3, Beckman Coulter Inc., USA) was used to determine the volumes of human red blood cells (RBCs) in NaCl solutions of different osmolalities. The thermodynamics model describing cell response during freezing process was used to simulate the volume change of RBC in 0.9% NaCl solution during equilibrium freezing process. It was assumed that the effect of temperature on cell volume can be neglected compared to that of osmolality, then by using the phase diagram for the binary system sodium chloride/water, the osmolalities of the NaCl solution under different sub-zero temperatures can be obtained (converted from mass concentration), then the calculated values of RBC volumes can be validated by the experiments.

  4. 21 CFR 864.9225 - Cell-freezing apparatus and reagents for in vitro diagnostic use.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Cell-freezing apparatus and reagents for in vitro diagnostic use. 864.9225 Section 864.9225 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH... Establishments That Manufacture Blood and Blood Products § 864.9225 Cell-freezing apparatus and reagents for...

  5. Intracellular trehalose improves the survival of human red blood cells by freeze-drying

    Institute of Scientific and Technical Information of China (English)

    HE Hui; LIU Baolin; HUA Zezhao; LI Chuan; WU Zhengzheng

    2007-01-01

    Freeze-drying of human red blood cells has a potential important application for blood transfusion.The aim of this study was to investigate the effects ofintracellular trehalose on the survival of red blood cells after freeze-drying and rehydration.Fresh red blood cells were incubated in trehalose solutions of various concentrations at 37℃ for 7 h following freeze-drying.Polyvinylpyrrolidone,Trehalose,sodium citrate,and human serum albumin were used as extracellular protective agents for the freeze-drying of red blood cells.The results indicated that the intracellular trehalose concentration was increased with increasing concentration of extracellular trehalose solution,and the maximum concen tration of intracellular trehalose reached 35 mmol/L.The viability of freeze-dried red blood cells increased with the increment of intracellular trehalose concentration.

  6. Roles of the plasma membrane and the cell wall in the responses of plant cells to freezing.

    Science.gov (United States)

    Yamada, Tomoyoshi; Kuroda, Katsushi; Jitsuyama, Yutaka; Takezawa, Daisuke; Arakawa, Keita; Fujikawa, Seizo

    2002-09-01

    In an effort to clarify the responses of a wide range of plant cells to freezing, we examined the responses to freezing of the cells of chilling-sensitive and chilling-resistant tropical and subtropical plants. Among the cells of the plants that we examined, those of African violet ( Saintpaulia grotei Engl.) leaves were most chilling-sensitive, those of hypocotyls in mungbean [ Vigna radiata (L.) R. Wilcz.] seedlings were moderately chilling-sensitive, and those of orchid [ Paphiopedilum insigne (Wallich ex Lindl.) Pfitz.] leaves were chilling-resistant, when all were chilled at -2 degrees C. By contrast, all these plant cells were freezing-sensitive and suffered extensive damage when they were frozen at -2 degrees C. Cryo-scanning electron microscopy (Cryo-SEM) confirmed that, upon chilling at -2 degrees C, both chilling-sensitive and chilling-resistant plant cells were supercooled. Upon freezing at -2 degrees C, by contrast, intracellular freezing occurred in Saintpaulia leaf cells, frost plasmolysis followed by intracellular freezing occurred in mungbean seedling cells, and extracellular freezing (cytorrhysis) occurred in orchid leaf cells. We postulate that chilling-related destabilization of membranes might result in the loss of the ability of the plasma membrane to act as a barrier against the propagation of extracellular ice in chilling-sensitive plant cells. We also examined the role of cell walls in the response to freezing using cells in which the plasma membrane had been disrupted by repeated freezing and thawing. In chilling-sensitive Saintpaulia and mungbean cells, the cells with a disrupted plasma membrane responded to freezing at -2 degrees C by intracellular freezing. By contrast, in chilling-resistant orchid cells, as well as in other cells of chilling-resistant and freezing-resistant plant tissues, including leaves of orchard grass ( Dactylis glomerata L.), leaves of Arabidopsis thaliana (L.) Heynh. and cortical tissues of mulberry ( Morus

  7. An improved model for nucleation-limited ice formation in living cells during freezing.

    Directory of Open Access Journals (Sweden)

    Jingru Yi

    Full Text Available Ice formation in living cells is a lethal event during freezing and its characterization is important to the development of optimal protocols for not only cryopreservation but also cryotherapy applications. Although the model for probability of ice formation (PIF in cells developed by Toner et al. has been widely used to predict nucleation-limited intracellular ice formation (IIF, our data of freezing Hela cells suggest that this model could give misleading prediction of PIF when the maximum PIF in cells during freezing is less than 1 (PIF ranges from 0 to 1. We introduce a new model to overcome this problem by incorporating a critical cell volume to modify the Toner's original model. We further reveal that this critical cell volume is dependent on the mechanisms of ice nucleation in cells during freezing, i.e., surface-catalyzed nucleation (SCN and volume-catalyzed nucleation (VCN. Taken together, the improved PIF model may be valuable for better understanding of the mechanisms of ice nucleation in cells during freezing and more accurate prediction of PIF for cryopreservation and cryotherapy applications.

  8. An improved model for nucleation-limited ice formation in living cells during freezing.

    Science.gov (United States)

    Yi, Jingru; Liang, Xin M; Zhao, Gang; He, Xiaoming

    2014-01-01

    Ice formation in living cells is a lethal event during freezing and its characterization is important to the development of optimal protocols for not only cryopreservation but also cryotherapy applications. Although the model for probability of ice formation (PIF) in cells developed by Toner et al. has been widely used to predict nucleation-limited intracellular ice formation (IIF), our data of freezing Hela cells suggest that this model could give misleading prediction of PIF when the maximum PIF in cells during freezing is less than 1 (PIF ranges from 0 to 1). We introduce a new model to overcome this problem by incorporating a critical cell volume to modify the Toner's original model. We further reveal that this critical cell volume is dependent on the mechanisms of ice nucleation in cells during freezing, i.e., surface-catalyzed nucleation (SCN) and volume-catalyzed nucleation (VCN). Taken together, the improved PIF model may be valuable for better understanding of the mechanisms of ice nucleation in cells during freezing and more accurate prediction of PIF for cryopreservation and cryotherapy applications. PMID:24852166

  9. Cryopreservation of Endothelial Cells in Various Cryoprotective Agents and Media - Vitrification versus Slow Freezing Methods.

    Directory of Open Access Journals (Sweden)

    Achim von Bomhard

    Full Text Available Vitrification of endothelial cells (MHECT-5 has not previously been compared with controlled slow freezing methods under standardized conditions. To identify the best cryopreservation technique, we evaluated vitrification and standardized controlled-rate -1°C/minute cell freezing in a -80°C freezer and tested four cryoprotective agents (CPA, namely dimethyl sulfoxide (DMSO, ethylene glycol (EG, propylene glycol (PG, and glycerol (GLY, and two media, namely Dulbecco's modified Eagle medium Ham's F-12 (DMEMand K+-modified TiProtec (K+TiP, which is a high-potassium-containing medium. Numbers of viable cells in proliferation were evaluated by the CellTiter 96® AQueous One Solution Cell Proliferation Assay (Promega Corporation, Mannheim, Germany. To detect the exact frozen cell number per cryo vial, DNA content was measured by using Hoechst 33258 dye prior to analysis. Thus, results could be evaluated unconstrained by absolute cell number. Thawed cells were cultured in 25 cm2 cell culture flasks to confluence and examined daily by phase contrast imaging. With regard to cell recovery immediately after thawing, DMSO was the most suitable CPA combined with K+TiP in vitrification (99 ±0.5% and with DMEM in slow freezing (92 ±1.6%. The most viable cells in proliferation after three days of culture were obtained in cells vitrificated by using GLY with K+TiP (308 ±34% and PG with DMEM in slow freezing (280 ±27%.

  10. Freezing resistance in Patagonian woody shrubs: the role of cell wall elasticity and stem vessel size.

    Science.gov (United States)

    Zhang, Yong-Jiang; Bucci, Sandra J; Arias, Nadia S; Scholz, Fabian G; Hao, Guang-You; Cao, Kun-Fang; Goldstein, Guillermo

    2016-08-01

    Freezing resistance through avoidance or tolerance of extracellular ice nucleation is important for plant survival in habitats with frequent subzero temperatures. However, the role of cell walls in leaf freezing resistance and the coordination between leaf and stem physiological processes under subzero temperatures are not well understood. We studied leaf and stem responses to freezing temperatures, leaf and stem supercooling, leaf bulk elastic modulus and stem xylem vessel size of six Patagonian shrub species from two sites (plateau and low elevation sites) with different elevation and minimum temperatures. Ice seeding was initiated in the stem and quickly spread to leaves, but two species from the plateau site had barriers against rapid spread of ice. Shrubs with xylem vessels smaller in diameter had greater stem supercooling capacity, i.e., ice nucleated at lower subzero temperatures. Only one species with the lowest ice nucleation temperature among all species studied exhibited freezing avoidance by substantial supercooling, while the rest were able to tolerate extracellular freezing from -11.3 to -20 °C. Leaves of species with more rigid cell walls (higher bulk elastic modulus) could survive freezing to lower subzero temperatures, suggesting that rigid cell walls potentially reduce the degree of physical injury to cell membranes during the extracellular freezing and/or thaw processes. In conclusion, our results reveal the temporal-spatial ice spreading pattern (from stem to leaves) in Patagonian shrubs, and indicate the role of xylem vessel size in determining supercooling capacity and the role of cell wall elasticity in determining leaf tolerance of extracellular ice formation. PMID:27217529

  11. FREEZE PROFILE AND HEAT BALANCE CALCULATION OF THE 160kA DRAINED CELL

    Institute of Scientific and Technical Information of China (English)

    X.P.Li; J.Li; Y.Q.Lai; H.Q.Zhao; Y.X.Liu

    2004-01-01

    A 2D full cell thermo-electric model of 160kA drained cell was set up using finite element code to calculate its freeze profile,then the drained cell model was modified according to the freeze profile computed and its heat balance was calculated.Compared with that of a 160kA conventional Hall-Heroult cell(H-H cell),though the melts volume of the drained cell reduced greatly,the whole heat loss from it didn't drop down apparently,and an analysis was presented in the paper.On the other hand,the anodecathode distance(ACD)of a drained cell was much less than that of a H-H cell,so the voltage drop on it and heat produced decreased too,steps should be taken to keep a workable heat balance on a drained cell.

  12. 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. PMID:27364314

  13. Studies on Freezing Injury in Plant Cells : II. Protein and Lipid Changes in the Plasma Membranes of Jerusalem Artichoke Tubers during a Lethal Freezing in Vivo.

    Science.gov (United States)

    Uemura, M; Yoshida, S

    1986-01-01

    Plasma membranes were isolated from both unfrozen and frozen tissues of Jerusalem artichoke tubers (Helianthus tuberosus L.) in high purity utilizing an aqueous two-polymer phase partition system. Although the recovery of the plasma membranes was decreased significantly by freezing of tissues even at the nonlethal temperature (-5 degrees C), the isolated plasma membrane samples were considered to be representative of the plasma membranes in situ. Freezing of the tissues at sublethal temperatures resulted in marked changes in the chemical composition of the plasma membrane. Those are losses of sterols and phosphatidylethanolamine from the plasma membranes, and a change of specific proteins with relatively high molecular weights into low molecular weight peptides. These specific proteins were designated as frost susceptible proteins. The properties of the plasma membrane ATPase seem to be not affected so much by the in vivo freezing of cells. However, inhibition of the plasma membrane ATPase by N,N'-dicyclohexylcarbodiimide (DCCD) was relatively low before and after freezing in vivo at the nonlethal temperature at -5 degrees C, but was markedly enhanced by freezing in vivo at sublethal temperatures below -10 degrees C. From the results, it is assumed either that the enzyme molecule was partially modified, especially at the presumed DCCD binding sites or that the DCCD had become more accessible to the enzyme as a result of increased permeability of the plasma membranes. These observed changes are discussed in connection with the mechanism of cell injury. PMID:16664579

  14. Insufficiency of copper ion homeostasis causes freeze-thaw injury of yeast cells as revealed by indirect gene expression analysis.

    Science.gov (United States)

    Takahashi, Shunsuke; Ando, Akira; Takagi, Hiroshi; Shima, Jun

    2009-11-01

    Saccharomyces cerevisiae is exposed to freeze-thaw stress in commercial processes, including frozen dough baking. Cell viability and fermentation activity after a freeze-thaw cycle were dramatically decreased due to freeze-thaw injury. Because this type of injury involves complex phenomena, the injury mechanisms are not fully understood. We examined freeze-thaw injury by indirect gene expression analysis during postthaw incubation after freeze-thaw treatment using DNA microarray profiling. The results showed that genes involved in the homeostasis of metal ions were frequently contained in genes that were upregulated, depending on the freezing period. We assessed the phenotype of deletion mutants of the metal ion homeostasis genes that exhibited freezing period-dependent upregulation and found that the strains with deletion of the MAC1 and CTR1 genes involved in copper ion homeostasis exhibited freeze-thaw sensitivity, suggesting that copper ion homeostasis is required for freeze-thaw tolerance. We found that supplementation with copper ions during postthaw incubation increased intracellular superoxide dismutase activity and intracellular levels of reactive oxygen species were decreased. Moreover, cell viability was increased by supplementation with copper ions. These results suggest that insufficiency of copper ion homeostasis may be one of the causes of freeze-thaw injury. PMID:19749072

  15. Effect of uncontrolled freezing on biological characteristics of human dental pulp stem cells.

    Science.gov (United States)

    Kumar, Ajay; Bhattacharyya, Shalmoli; Rattan, Vidya

    2015-12-01

    Human dental pulp stem cells (hDPSCs) hold great promise as a source of adult stem cells for utilization in regenerative medicine. Successful storage and post thaw recovery of DPSCs without loss of function is a key issue for future clinical application. Most of the cryopreservation methods use controlled rate freezing and vapor phase nitrogen to store stem cells. But these methods are both expensive and laborious. In this study, we isolated DPSCs from a patient undergoing impacted mandibular third molar extraction. We adopted eight different methods of cryopreservation at -80 °C for long term storage of the DPSC aliquots. Various parameters like proliferation, cell death, cell cycle, retention of stemness markers and differentiation potential were studied post cryopreservation period of 1 year. We observed successful recovery of stem cells in every method and a significant difference in proliferation potential and cell death between samples stored by different methods. However, post thaw, all cells retained their stemness markers. All DPSCs stored by different methods were able to differentiate into osteoblast like cells, adipocytes and neural cells. Based on these parameters we concluded that uncontrolled freezing at a temperature of -80 °C is as effective as controlled freezing using ethanol vessels and other cryopreservation methods. To the best of our knowledge, our study provides the first proof of concept that long term storage in uncontrolled freezing of cells at -80 °C in 10 % DMSO does not affect the revival capacity of hDPSCs. This implies that DPSCs may be used successfully for tissue engineering and cell based therapeutics even after long term, uncontrolled cryopreservation.

  16. Freezing tolerance of sea urchin embryonic cells: Differentiation commitment and cytoskeletal disturbances in culture.

    Science.gov (United States)

    Odintsova, Nelly A; Ageenko, Natalya V; Kipryushina, Yulia O; Maiorova, Mariia A; Boroda, Andrey V

    2015-08-01

    This study focuses on the freezing tolerance of sea urchin embryonic cells. To significantly reduce the loss of physiological activity of these cells that occurs after cryopreservation and to study the effects of ultra-low temperatures on sea urchin embryonic cells, we tested the ability of the cells to differentiate into spiculogenic or pigment directions in culture, including an evaluation of the expression of some genes involved in pigment differentiation. A morphological analysis of cytoskeletal disturbances after freezing in a combination of penetrating (dimethyl sulfoxide and ethylene glycol) and non-penetrating (trehalose and polyvinylpyrrolidone) cryoprotectants revealed that the distribution pattern of filamentous actin and tubulin was similar to that in the control cultures. In contrast, very rare spreading cells and a small number of cells with filamentous actin and tubulin were detected after freezing in the presence of only non-penetrating cryoprotectants. The largest number of pigment cells was found in cultures frozen with trehalose or trehalose and dimethyl sulfoxide. The ability to induce the spicule formation was lost in the cells frozen only with non-penetrating cryoprotectants, while it was maximal in cultures frozen in a cryoprotective mixture containing both non-penetrating and penetrating cryoprotectants (particularly, when ethylene glycol was present). Using different markers for cell state assessment, an effective cryopreservation protocol for sea urchin cells was developed: three-step freezing with a low cooling rate (1-2°C/min) and a combination of non-penetrating and penetrating cryoprotectants made it possible to obtain a high level of cell viability (up to 65-80%). PMID:26049089

  17. GMP cryopreservation of large volumes of cells for regenerative medicine: active control of the freezing process.

    Science.gov (United States)

    Massie, Isobel; Selden, Clare; Hodgson, Humphrey; Fuller, Barry; Gibbons, Stephanie; Morris, G John

    2014-09-01

    Cryopreservation protocols are increasingly required in regenerative medicine applications but must deliver functional products at clinical scale and comply with Good Manufacturing Process (GMP). While GMP cryopreservation is achievable on a small scale using a Stirling cryocooler-based controlled rate freezer (CRF) (EF600), successful large-scale GMP cryopreservation is more challenging due to heat transfer issues and control of ice nucleation, both complex events that impact success. We have developed a large-scale cryocooler-based CRF (VIA Freeze) that can process larger volumes and have evaluated it using alginate-encapsulated liver cell (HepG2) spheroids (ELS). It is anticipated that ELS will comprise the cellular component of a bioartificial liver and will be required in volumes of ∼2 L for clinical use. Sample temperatures and Stirling cryocooler power consumption was recorded throughout cooling runs for both small (500 μL) and large (200 mL) volume samples. ELS recoveries were assessed using viability (FDA/PI staining with image analysis), cell number (nuclei count), and function (protein secretion), along with cryoscanning electron microscopy and freeze substitution techniques to identify possible injury mechanisms. Slow cooling profiles were successfully applied to samples in both the EF600 and the VIA Freeze, and a number of cooling and warming profiles were evaluated. An optimized cooling protocol with a nonlinear cooling profile from ice nucleation to -60°C was implemented in both the EF600 and VIA Freeze. In the VIA Freeze the nucleation of ice is detected by the control software, allowing both noninvasive detection of the nucleation event for quality control purposes and the potential to modify the cooling profile following ice nucleation in an active manner. When processing 200 mL of ELS in the VIA Freeze-viabilities at 93.4% ± 7.4%, viable cell numbers at 14.3 ± 1.7 million nuclei/mL alginate, and protein secretion at 10.5 ± 1.7

  18. Comparison of conventional and directional freezing for the cryopreservation of human umbilical vein endothelial cells

    Institute of Scientific and Technical Information of China (English)

    Bing; Qi; Qing-Shan; Ji; Guang-Hui; Hou; Liu; Li; Xian-Fen; Cao; Jing; Wu

    2014-01-01

    AIM:To compare conventional slow equilibrium cooling and directional freezing(DF) by gauze package for cryopreservation of human umbilical vein endothelial cells(HUVECs).METHODS:HUVECs were randomly assigned to conventional freezing(CF) and DF by gauze package group. The two groups of HUVECs were incubated with a freezing liquid consisting of 10% dimethylsulfoxide(DMSO), 60% fetal bovine serum(FBS) and 30%Dulbecco’s modified Eagle’s medium(DMEM) and then put into cryopreserved tubes. CF group, slow equilibrium cooling was performed with the following program:precool in 4℃ for 30 min,-20℃ for 1h, and then immersion in-80℃ refrigerator. DF group, the tubes were packaged with gauze and then directional freezing in-80℃ refrigerator straightly. One month later, the vitality of HUVECs were calculated between two groups.RESULTS:There was no significant difference in the survival rate and growth curve between CF and DF groups. The DF group was significantly better than CFgroup in adherent rates, morphological changes and proliferative ability.CONCLUSION:In the conventional cryopreserved method, cells are slow equilibrium cooling by steps(4℃,-20℃ and finally-80℃), which is a complicated and time-consuming process. But the improved DF by gauze package method is better than conventional method, for which is convenient and easy to operate.

  19. Performance and durability of PEM fuel cells operated at sub-freezing temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Mukundan, Rangachary [Los Alamos National Laboratory; Davey, John R [Los Alamos National Laboratory; Lujan, Roger W [Los Alamos National Laboratory; Spendelow, Jacob S [Los Alamos National Laboratory

    2008-01-01

    The durability of polymer electrolyte membrane (PEM) fuel cells operated at sub-freezing temperatures has received increasing attention in recent years. The Department of Energy's PEM fuel cell stack technical targets for the year 2010 include unassisted start-up from -40 {sup o}C and startup from -20 {sup o}C ambient in as low as 30 seconds with < 5 MJ energy consumption. Moreover, the sub-freezing operations should not have any impact on acieving other technical targets including 5000 hours durability. The effect of MEA preparation on the performance of single-PEM fuel cells operated at sub-freezing temperatures is presented. The cell performance and durability are dependent on the MEA and are probably influenced by the porosity of the catalyst layers. When a cell is operated isothermally at -10 {sup o}C in constant current mode, the voltage gradually decreases over time and eventually drops to zero. AC impedance analysis indicated that the rate of voltage loss is initially due to an increase in the charge transfer resistance and is gradual. After a period, the rate of decay accelerates rapidly due to mass transport limitations at the catalyst and/or gas diffusion layers. The high frequency resistance also increases over time during the isothermal operation at sub-freezing temperatures and was a function of the initial membrane water content. LANL prepared MEAs showed very little loss in the catalyst surface area with multiple sub-freezing operations, whereas the commercial MEAs exhibited significant loss in cathode surface area with the anode being unaffected. These results indicate that catalyst layer ice formation is influenced strongly by the MEA and is responsible for the long-term degradation of fuel cells operated at sub-freezing temperatures. This ice formation was monitored using neutron radiography and was found to be concentrated near cell edges at the flow field turns. The water distribution also indicated that ice may be forming mainly in the

  20. Fundamental aspects of the freezing of cells, with emphasis on mammalian ova and embryos

    Energy Technology Data Exchange (ETDEWEB)

    Mazur, P.

    1980-01-01

    The problem in cryobiology is how to cool cells to -196/sup 0/C and return them to normal temperatures without killing them. One important factor is the presence of a protective additive like glycerol or dimethyl sulfoxide. Mammalian cells rarely survive freezing to below -40/sup 0/C in its absence. In the presence of an additive, survival is critically dependent on the cooling rate. Supraoptimal rates and suboptimal rates are both damaging. Death at supraoptimal rates is the result of the formation of intracellular ice and its recrystallization during warming. Death at suboptimal rates is a consequence of the major alterations in aqueous solutions produced by ice formation. The chief effects are a major reduction in the fraction of the solution remaining unfrozen at a given temperature and a major increase in the solute concentration of that fraction. The introduction of molar concentrations of additive greatly reduces both the fraction frozen and the concentration of electrolytes in the unfrozen channels and in the cell interior. Usually, freezing either kills cells outright or it results in survivors that retain full capacity to function. But there is the possibility that in some cases survivors may in fact be impaired genetically or physiologically. All evidence indicates that genetic damage does not occur. But there are clear examples in which freezing does induce nonlethal physiological damage. (ERB)

  1. Visualization of Fuel Cell Water Transport and Performance Characterization under Freezing Conditions

    Energy Technology Data Exchange (ETDEWEB)

    Kandlikar, Satish G. [Rochester Inst. of Technology, Rochester, NY (United States); Lu, Zijie [Rochester Inst. of Technology, Rochester, NY (United States); Rao, Navalgund [Rochester Inst. of Technology, Rochester, NY (United States); Sergi, Jacqueline [Rochester Inst. of Technology, Rochester, NY (United States); Rath, Cody [Rochester Inst. of Technology, Rochester, NY (United States); McDade, Christopher [Rochester Inst. of Technology, Rochester, NY (United States); Trabold, Thomas [General Motors, Honeoye Falls, NY (United States); Owejan, Jon [General Motors, Honeoye Falls, NY (United States); Gagliardo, Jeffrey [General Motors, Honeoye Falls, NY (United States); Allen, Jeffrey [Michigan Technological Univ., Houghton, MI (United States); Yassar, Reza S. [Michigan Technological Univ., Houghton, MI (United States); Medici, Ezequiel [Michigan Technological Univ., Houghton, MI (United States); Herescu, Alexandru [Michigan Technological Univ., Houghton, MI (United States)

    2010-05-30

    In this program, Rochester Institute of Technology (RIT), General Motors (GM) and Michigan Technological University (MTU) have focused on fundamental studies that address water transport, accumulation and mitigation processes in the gas diffusion layer and flow field channels of the bipolar plate. These studies have been conducted with a particular emphasis on understanding the key transport phenomena which control fuel cell operation under freezing conditions.

  2. The effect of magnetic field during freezing and thawing of rat bone marrow-derived mesenchymal stem cells.

    Science.gov (United States)

    Shikata, H; Kaku, M; Kojima, S-I; Sumi, H; Kojima, S-T; Yamamoto, T; Yashima, Y; Kawata, T; Tanne, K; Tanimoto, K

    2016-08-01

    Previous studies showed that a programmed freezer with magnetic field can maintain a high survival rate of mesenchymal stem cells (MSCs). The purpose of this study was to evaluate the influences of magnetic field during freezing and thawing on the survival of MSCs isolated from rat bone marrow. The cells were frozen by a normal programmed freezer or a programmed freezer with magnetic field (CAS-LAB1) and cryopreserved for 7 days at -150 °C. Then, the cells were thawed in the presence or absence of magnetic field. Immediately after thawing, the number of surviving or viable cells was counted. The cell proliferation was examined after 1-week culture. Cryopreserved MSCs which were frozen by a normal freezer or a CAS freezer were transplanted into bone defects artificially made in calvaria of 4-week-old rats. Non-cryopreserved MSCs were used as a control. The rats were sacrificed at 8, 16, or 24 weeks after transplantation and the bone regeneration area was measured. Proliferation rates of MSCs after 1 week were significantly higher in the CAS-freezing-thawing group than in the CAS-freezing group. The extent of new bone formation in the CAS-freezing-thawing group tended to be larger than in CAS-freezing group 24 weeks after transplantation. These results suggest that a magnetic field enhances cell survival during thawing as well as freezing. PMID:27346603

  3. Subcellular imaging of freeze-fractured cell cultures by TOF-SIMS and Laser-SNMS

    Science.gov (United States)

    Fartmann, M.; Dambach, S.; Kriegeskotte, C.; Lipinsky, D.; Wiesmann, H. P.; Wittig, A.; Sauerwein, W.; Arlinghaus, H. F.

    2003-01-01

    We have examined atomic and molecular distributions in freeze-fractured freeze-dried primary osteoblasts and cancer cells using time-of-flight secondary ion mass spectrometry (TOF-SIMS) and non-resonant laser secondary neutral mass spectrometry (NR-Laser-SNMS). A pulsed Ga primary ion beam with a diameter of approximately 200 nm was employed to bombard the sample. Ion-induced electron-images were used to identify individual cells. High resolution elemental and molecular images were obtained from cell cultures. From these data the K/Na ratio was determined. It shows a higher K-concentration inside individual cells demonstrating that the chemical and structural integrity of living cells were preserved by the applied preparation technique. Consecutive presputtering of the sample with different primary ion dose densities was used to move the analysis plane toward the inside of the cell. It can be concluded that TOF-SIMS and Laser-SNMS are well suited for imaging trace element and molecule concentrations in biological samples.

  4. The effect of freeze-thaw cycles on gene expression levels in lymphoblastoid cell lines.

    Directory of Open Access Journals (Sweden)

    Minal Çalışkan

    Full Text Available Epstein-Barr virus (EBV transformed lymphoblastoid cell lines (LCLs are a widely used renewable resource for functional genomic studies in humans. The ability to accumulate multidimensional data pertaining to the same individual cell lines, from complete genomic sequences to detailed gene regulatory profiles, further enhances the utility of LCLs as a model system. However, the extent to which LCLs are a faithful model system is relatively unknown. We have previously shown that gene expression profiles of newly established LCLs maintain a strong individual component. Here, we extend our study to investigate the effect of freeze-thaw cycles on gene expression patterns in mature LCLs, especially in the context of inter-individual variation in gene expression. We report a profound difference in the gene expression profiles of newly established and mature LCLs. Once newly established LCLs undergo a freeze-thaw cycle, the individual specific gene expression signatures become much less pronounced as the gene expression levels in LCLs from different individuals converge to a more uniform profile, which reflects a mature transformed B cell phenotype. We found that previously identified eQTLs are enriched among the relatively few genes whose regulations in mature LCLs maintain marked individual signatures. We thus conclude that while insight drawn from gene regulatory studies in mature LCLs may generally not be affected by the artificial nature of the LCL model system, many aspects of primary B cell biology cannot be observed and studied in mature LCL cultures.

  5. Ultra-high temperature isothermal furnace liners (IFLS) for copper freeze point cells

    Science.gov (United States)

    Dussinger, P. M.; Tavener, J. P.

    2013-09-01

    Primary Laboratories use large fixed-point cells in deep calibration furnaces utilizing heat pipes to achieve temperature uniformity. This combination of furnace, heat pipe, and cell gives the smallest of uncertainties. The heat pipe, also known as an isothermal furnace liner (IFL), has typically been manufactured with Alloy 600/601 as the envelope material since the introduction of high temperature IFLs over 40 years ago. Alloy 600/601 is a widely available high temperature material, which is compatible with Cesium, Potassium, and Sodium and has adequate oxidation resistance and reasonable high temperature strength. Advanced Cooling Technologies, Inc. (ACT) Alloy 600/Sodium IFLs are rated to 1100°C for approximately 1000 hours of operation (based on creep strength). Laboratories interested in performing calibrations and studies around the copper freezing point (1084.62°C) were frustrated by the 1000 hours at 1100°C limitation and the fact that expensive freeze-point cells were getting stuck and/or crushed inside the IFL. Because of this growing frustration/need, ACT developed an Ultra High Temperature IFL to take advantage of the exceptional high temperature strength properties of Haynes 230.

  6. A freeze-dried graphene counter electrode enhances the performance of dye-sensitized solar cells

    Energy Technology Data Exchange (ETDEWEB)

    Hung, Kai-Hsiang; Wang, Hong-Wen, E-mail: hongwen@cycu.edu.tw

    2014-01-01

    A flexible graphene/polyimide (PI) counter electrode without a fluorine-doped tin oxide (FTO) layer has been fabricated for dye-sensitized solar cell (DSSCs) applications. The flexible counter electrode consists of polyimide double-sided tape as a substrate beneath a graphene film acting as the conductive and catalytic layer. Chemically reduced graphene oxide (rGO) on the PI electrode (rGO-PI) shows comparable catalytic activity to that of the reference sputtered platinum/FTO counter electrodes (Sputter-Pt/FTO). A DSSC with a freeze-dried rGO-PI (FD-rGO-PI) counter electrode shows an overall conversion efficiency (η) of 5.45%, while that of the conventional Sputter-Pt/FTO electrode is 5.52%. The DSSC with a thermally dried rGO-PI (Gel-rGO-PI) counter electrode (not freeze-dried) exhibits a smooth morphology and much poorer performance (η = 1.61%). Field emission scanning electron microscopy, electrochemical impedance spectroscopy, and cyclic voltammetry measurements demonstrate that the FD-rGO-PI electrode possesses a porous structure, numerous edges, minimum charge-transfer resistance and a higher electrocatalytic activity toward the I{sub 3}{sup −}/I{sup −} redox couple than that of the Gel-rGO-PI electrode. The high electrocatalytic activity, facile preparation procedure, absence of FTO, and material flexibility render the FD-rGO-PI electrode an ideal alternative to conventional DSSC counter electrodes. - Highlights: • Highly rough and conductive graphene-based counter electrode is synthesized. • The characteristics of graphene surface by freeze drying are different. • The graphene counter electrode exhibits comparable performance to that of sputtered Pt one.

  7. Meat Freezing Theories and Novel Freezing Technologies%肉类冷冻理论与冷冻新技术

    Institute of Scientific and Technical Information of China (English)

    金文刚

    2008-01-01

    Main meat freezing theories,including transition theory,glass transformation theory and ice crystallization theory,were introduced.Some novel freezing technologies such as high pressure freezing,ultrasonic freezing,ice nucleus bacteria freezing protein,cell alive system freezing and decompression freezing were also generally reviewed.

  8. The effect of initial tonicity on freeze/thaw injury to human red cells suspended in solutions of sodium chloride.

    Science.gov (United States)

    Pegg, D E; Diaper, M P

    1991-02-01

    Human red blood cells, suspended in solutions of sodium chloride, have been frozen to temperatures between -2 and -14 degrees C and thawed, and the extent of hemolysis was measured. In parallel experiments, red cells were exposed to similar cycles of change in the composition of the suspending solution, but by dialysis at 21 degrees C. The tonicity of the saline in which the cells were initially suspended was varied between 0.6x isotonic and 4x isotonic; some samples from each experimental treatment were returned to isotonic saline before hemolysis was measured. It was found that the tonicity of the saline used to suspend the cells for the main body of the experiment affected the amount of hemolysis measured: raising the tonicity from 0.6x to 1x to 2x reduced hemolysis, both in the freezing and in the dialysis experiments, whereas raising the tonicity further to 4x reversed that trend. There was little difference between the freeze/thaw and the dialysis treatments for the cells suspended in 1x or 2x saline, whether or not the cells were returned to isotonic conditions. However, the cells suspended in 0.6x saline showed greater damage from freezing and thawing than from the comparable change in the composition of the solution, whether or not they were returned to isotonic conditions. Cells that were suspended in 4x saline and exposed to changes in salt concentration by dialysis showed less hemolysis when they were assayed in the 4x solution than cells that had received the comparable freezing/thaw treatment, but when the experiment included a return to isotonicity, the two treatments gave similar results. Returning the cells to isotonic saline had a negligible affect on the cells in 0.6x and 1x saline, but caused considerable hemolysis in the 2x and 4x samples, more so after dialysis than after freezing and thawing. We conclude that cells suspended in 0.6x and 4x saline behave differently from cells suspended in 1x and 2x saline and hence that cells suspended in a

  9. High throughput cryopreservation of cells by rapid freezing of sub-μl drops using inkjet printing--cryoprinting.

    Science.gov (United States)

    Dou, Rui; Saunders, Rachel E; Mohamet, Lisa; Ward, Christopher M; Derby, Brian

    2015-09-01

    We have successfully used inkjet printing to cryopreserve 3T3 mouse fibroblast cells and human neuroprogenitor cells (NPCs) derived from human embryonic stem cells (hESCs). Sessile drops of volume 114 nl were formed by printing cell suspensions containing dimethyl sulphoxide (DMSO) as a cryoprotection agent (CPA) at rates in the region 100 Hz-20 kHz, from individual droplets of 380 pl. After printing and a freeze/thaw cycle (with a minimum 24 hours hold period at liquid N2 temperature), 3T3 cells showed an average viability of >90% with CPA concentration polyimide substrate. The viability of 3T3 cells after 2 months storage at liquid nitrogen temperature was slightly reduced compared to the cells held for 24 hours but there was no significant further deterioration after 4 or 6 months storage. The viability of NPCs after an identical freeze/thaw cycle were only 55% but this is comparable with conventional cryopreservation methods that use much higher CPA concentrations. A parallel series of experiments printing cells onto substrates held at 195 K or directly into liquid N2 showed considerable variation in cell survival rate with drop deposition rate. Cell suspensions required higher levels of CPA than when printing followed by freezing. At low deposition rates a combination of DMSO and polyethylene glycol (PEG) was needed to allow cell viability after freezing. These results show that inkjet printing provides a practical high throughput method for the cryopreservation of cells with lower CPA concentrations than are required for current low volume cryopreservation methods. PMID:26190571

  10. Biotransport and intracellular ice formation phenomena in freezing human embryonic kidney cells (HEK293T).

    Science.gov (United States)

    Xu, Yunpeng; Zhao, Gang; Zhou, Xiaoming; Ding, Weiping; Shu, Zhiquan; Gao, Dayong

    2014-04-01

    The objective of this study is to determine the cryobiological characteristics of human embryonic kidney (HEK293T) cells. The cell membrane hydraulic conductivity (L(pg)) and the activation energy of water transport (E(Lp)) were determined in the absence/presence of cryoprotectant agent (CPA), while the nucleation rate kinetic and thermodynamic parameters (Ωo(SCN) and κo(SCN)) were determined in the absence of CPA. Since dehydration and intracellular ice formation (IIF) are two factors that may cause damage to cells during the freezing process, systematical freezing experiments were carried out at different cooling rates (5, 10, 15, 20, 30, and 60°C/min) under the commercial available cryomicroscopy (FDCS 196, Linkham, Waterfield, UK) to further explore the cryoinjury mechanism for HEK293T cells. By simultaneously fitting the water transport equation to the experimentally measured volumetric shrinkage data at 5, 10, and 15°C/min, the "combined best fit" membrane permeability parameters for HEK293T cells in both phosphate buffer saline (PBS) and CPA media (0.75M Me2SO in PBS) are determined. They are L(pg)=2.85×10(-14)m/s/Pa (0.17μm/min/atm), E(Lp)=142.91kJ/mol (34.13kcal/mol) (R(2)=0.990), and L(pg)[cpa]=2.73±0.44×10(-14)m/s/Pa (0.16±0.03μm/min/atm), E(Lp)[cpa]=152.52±27.69kJ/mol (36.42±6.61kcal/mol) (R(2)=0.993), respectively. An optimal cooling rate B(opt) (the highest cooling rate without IIF) was determined to be 14.24°C/min in the absence of CPA. Additionally, the ice nucleation parameters (Ωo(SCN) and κo(SCN)) were averaged to be 1.31±0.11×10(8)m(-2)s(-1) and 7.67±2.55×10(9)K(5) for the cooling rates 20, 30, and 60°C/min. PMID:24582893

  11. Behavioral profile and dorsal hippocampal cells in carioca high-conditioned freezing rats.

    Science.gov (United States)

    Dias, Gisele Pereira; Bevilaqua, Mário Cesar do Nascimento; Silveira, Anna Claudia Domingos; Landeira-Fernandez, Jesus; Gardino, Patrícia Franca

    2009-12-28

    Selection for contextual fear conditioning is an important behavioral paradigm for studying the role of genetic variables and their interaction with the surrounding environment in the etiology and development of anxiety disorders. Recently, a new line of animals selectively bred for high levels of freezing in response to contextual cues previously associated with footshock was developed from a Wistar population. The purpose of the present study was to evaluate the emotional and cognitive aspects of this new line of animals, which has been named Carioca High-Freezing (CHF). For the characterization of anxious behavior, CHF and control animals were tested in the elevated plus-maze (EPM) and the social interaction test. CHF animals were significantly more anxious than control rats in terms of both the number of entries into EPM open arms and the percentage of time spent in these arms. The time spent in social interaction behavior was also significantly decreased. No statistical differences were found in locomotor activity, as measured by both the number of entries into the closed arms of the EPM and the number of crossings into the social interaction test arena. No differences between CHF and control groups were found in the depression forced swimming test, suggesting that the anxiety trait selected in the CHF line did not interact with affective disorders traits such as those for depression. Cognitive aspects of the CHF rats were evaluated in the object recognition task. Results from this test indicated no difference between the two groups. The present study also encompassed histological analysis of the dorsal hippocampus from CHF and control animals. Results revealed an absence of qualitative and quantitative differences between these two groups of animals in cells located in the dentate gyrus, CA1, and CA3 areas. Therefore, future studies are required to further investigate the possible neural mechanisms involved in the origin and development of the anxious

  12. Incorporation of [14C]-palmitate into lipids of Brassica cells during the induction of freezing tolerance

    International Nuclear Information System (INIS)

    Changes in plasma membrane lipid composition have been causally related to increased freezing tolerance. Studies of lipid metabolism during ABA induction of freezing tolerance in Brassica napus suspension cultures were undertaken. Cells were labeled with [14C]-palmitate four days after transfer to fresh medium (control) or medium containing ABA (which increases freezing tolerance). At times between one and 20 hrs after labeling, ABA-treated cells incorporated almost twice the amount of label as controls cells. Approximately 80% of the radioactivity was associated with neutral lipids in ABA-treated cells and controls. Incorporation of label into total cellular polar lipids was 4.9 x 105 dpm/mg protein for control cells and 1 x 106 dpm/mg protein for cells transferred to medium containing ABA. Analysis of lipids following alkaline hydrolysis indicated that incorporation of [14C]-palmitate into glucosylceramide of ABA-treated cells was less than 60% of control values when expressed relative to that of the total polar lipids. Incorporation into ceramides was also depressed in ABA-treated cells

  13. Ice formation in PEM fuel cells operated isothermally at sub-freezing temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Mukundan, Rangachary [Los Alamos National Laboratory; Luhan, Roger W [Los Alamos National Laboratory; Davey, John R [Los Alamos National Laboratory; Spendelow, Jacob S [Los Alamos National Laboratory; Borup, Rodney L [Los Alamos National Laboratory; Hussey, Daniel S [NIST; Jacobson, David L [NIST; Arif, Muhammad [NIST

    2009-01-01

    The effect of MEA and GDL structure and composition on the performance of single-PEM fuel cells operated isothermally at subfreezing temperatures is presented. The cell performance and durability are not only dependent on the MEA/GDL materials used but also on their interfaces. When a cell is operated isothermally at sub-freezing temperatures in constant current mode, the water formation due to the current density initially hydrates the membrane/ionomer and then forms ice in the catalyst layer/GDL. An increase in high frequency resistance was also observed in certain MEAs where there is a possibility of ice formation between the catalyst layer and GDL leading to a loss in contact area. The total water/ice holding capacity for any MEA was lower at lower temperatures and higher current densities. The durability of MEAs subjected to multiple isothermal starts was better for LANL prepared MEAs as compared to commercial MEAs, and cloth GDLs when compared to paper GDLs. The ice formation was monitored using high-resolution neutron radiography and was found to be concentrated near the cathode catalyst layer. However, there was significant ice formation in the GDLs especially at the higher temperature ({approx} -10 C) and lower current density (0.02 A/cm{sup 2}) operations. These results are consistent with the longer-term durability observations that show more severe degradation at the lower temperatures.

  14. Small molecule ice recrystallization inhibitors enable freezing of human red blood cells with reduced glycerol concentrations.

    Science.gov (United States)

    Capicciotti, Chantelle J; Kurach, Jayme D R; Turner, Tracey R; Mancini, Ross S; Acker, Jason P; Ben, Robert N

    2015-01-01

    In North America, red blood cells (RBCs) are cryopreserved in a clinical setting using high glycerol concentrations (40% w/v) with slow cooling rates (~1°C/min) prior to storage at -80°C, while European protocols use reduced glycerol concentrations with rapid freezing rates. After thawing and prior to transfusion, glycerol must be removed to avoid intravascular hemolysis. This is a time consuming process requiring specialized equipment. Small molecule ice recrystallization inhibitors (IRIs) such as β-PMP-Glc and β-pBrPh-Glc have the ability to prevent ice recrystallization, a process that contributes to cellular injury and decreased cell viability after cryopreservation. Herein, we report that addition of 110 mM β-PMP-Glc or 30 mM β-pBrPh-Glc to a 15% glycerol solution increases post-thaw RBC integrity by 30-50% using slow cooling rates and emphasize the potential of small molecule IRIs for the preservation of cells.

  15. A Simple and Efficient Method of Slow Freezing for Human Embryonic Stem Cells and Induced Pluripotent Stem Cells.

    Science.gov (United States)

    Imaizumi, Keitaro; Iha, Momoe; Nishishita, Naoki; Kawamata, Shin; Nishikawa, Shinichi; Akuta, Teruo

    2016-01-01

    Protocols available for the cryopreservation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells are very inefficient and laborious compared to those for the cryopreservation of murine ES/iPS cells or other general cell lines. While the vitrification method may be adequate when working with small numbers of human ES/iPS cells, it requires special skills and is unsuitable when working with large cell numbers. Here, we describe a simple and efficient method for the cryopreservation of hES/hiPS cells that is based on a conventional slow freezing method that uses a combination of Pronase/EDTA for Stem™ and CP-5E™ [final concentrations: 6 % hydroxyethyl starch, 5 % DMSO, and 5 % ethylene glycol in saline]. CP-5E™ is highly effective for the cryopreservation of small cell clumps produced by hES/hiPS colony detachment in the presence of Pronase and EDTA (Pronase/EDTA for Stem™, a formulation containing multiple digestive enzymes from Streptomyces griseus). This novel method would be quite useful for large-scale hES/iPS cell banking for use in clinical applications.

  16. Pore architecture and cell viability on freeze dried 3D recombinant human collagen-peptide (RHC)–chitosan scaffolds

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Jing; Zhou, Aimei; Deng, Aipeng [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Yang [Faculty of Engineering, University of Nottingham, Nottingham NG7 2RD (United Kingdom); Gao, Lihu; Zhong, Zhaocai [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Yang, Shulin, E-mail: yshulin@njust.edu.cn [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China)

    2015-04-01

    Pore architecture of 3D scaffolds used in tissue engineering plays a critical role in the maintenance of cell survival, proliferation and further promotion of tissue regeneration. We investigated the pore size and structure, porosity, swelling as well as cell viability of a series of recombinant human collagen-peptide–chitosan (RHCC) scaffolds fabricated by lyophilization. In this paper, freezing regime containing a final temperature of freezing (T{sub f}) and cooling rates was applied to obtain scaffolds with pore size ranging from 100 μm to 120 μm. Other protocols of RHC/chitosan suspension concentration and ratio modification were studied to produce more homogenous and appropriate structural scaffolds. The mean pore size decreased along with the decline of T{sub f} at a slow cooling rate of 0.7 °C/min; a more rapid cooling rate under 5 °C/min resulted to a smaller pore size and more homogenous microstructure. High concentration could reduce pore size and lead to thick well of scaffold, while improved the ratio of RHC, lamellar and fiber structure coexisted with cellular pores. Human umbilical vein endothelial cells (HUVECs) were seeded on these manufactured scaffolds, the cell viability represented a negative correlation to the pore size. This study provides an alternative method to fabricate 3D RHC–chitosan scaffolds with appropriate pores for potential tissue engineering. - Highlights: • Fabrication of recombinant human collagen-chitosan scaffolds by freezing drying • Influence of freeze drying protocols on lyophilized scaffolds • Pore size, microstructure, porosity, swelling and cell viability were compared. • The optimized porous scaffold is suitable for cell (HUVEC) seeding.

  17. Pore architecture and cell viability on freeze dried 3D recombinant human collagen-peptide (RHC)–chitosan scaffolds

    International Nuclear Information System (INIS)

    Pore architecture of 3D scaffolds used in tissue engineering plays a critical role in the maintenance of cell survival, proliferation and further promotion of tissue regeneration. We investigated the pore size and structure, porosity, swelling as well as cell viability of a series of recombinant human collagen-peptide–chitosan (RHCC) scaffolds fabricated by lyophilization. In this paper, freezing regime containing a final temperature of freezing (Tf) and cooling rates was applied to obtain scaffolds with pore size ranging from 100 μm to 120 μm. Other protocols of RHC/chitosan suspension concentration and ratio modification were studied to produce more homogenous and appropriate structural scaffolds. The mean pore size decreased along with the decline of Tf at a slow cooling rate of 0.7 °C/min; a more rapid cooling rate under 5 °C/min resulted to a smaller pore size and more homogenous microstructure. High concentration could reduce pore size and lead to thick well of scaffold, while improved the ratio of RHC, lamellar and fiber structure coexisted with cellular pores. Human umbilical vein endothelial cells (HUVECs) were seeded on these manufactured scaffolds, the cell viability represented a negative correlation to the pore size. This study provides an alternative method to fabricate 3D RHC–chitosan scaffolds with appropriate pores for potential tissue engineering. - Highlights: • Fabrication of recombinant human collagen-chitosan scaffolds by freezing drying • Influence of freeze drying protocols on lyophilized scaffolds • Pore size, microstructure, porosity, swelling and cell viability were compared. • The optimized porous scaffold is suitable for cell (HUVEC) seeding

  18. Small molecule ice recrystallization inhibitors mitigate red blood cell lysis during freezing, transient warming and thawing

    Science.gov (United States)

    Briard, Jennie G.; Poisson, Jessica S.; Turner, Tracey R.; Capicciotti, Chantelle J.; Acker, Jason P.; Ben, Robert N.

    2016-03-01

    During cryopreservation, ice recrystallization is a major cause of cellular damage. Conventional cryoprotectants such as dimethyl sulfoxide (DMSO) and glycerol function by a number of different mechanisms but do not mitigate or control ice recrystallization at concentrations utilized in cryopreservation procedures. In North America, cryopreservation of human red blood cells (RBCs) utilizes high concentrations of glycerol. RBC units frozen under these conditions must be subjected to a time-consuming deglycerolization process after thawing in order to remove the glycerol to <1% prior to transfusion thus limiting the use of frozen RBC units in emergency situations. We have identified several low molecular mass ice recrystallization inhibitors (IRIs) that are effective cryoprotectants for human RBCs, resulting in 70–80% intact RBCs using only 15% glycerol and slow freezing rates. These compounds are capable of reducing the average ice crystal size of extracellular ice relative to a 15% glycerol control validating the positive correlation between a reduction in ice crystal size and increased post-thaw recovery of RBCs. The most potent IRI from this study is also capable of protecting frozen RBCs against the large temperature fluctuations associated with transient warming.

  19. Small molecule ice recrystallization inhibitors mitigate red blood cell lysis during freezing, transient warming and thawing.

    Science.gov (United States)

    Briard, Jennie G; Poisson, Jessica S; Turner, Tracey R; Capicciotti, Chantelle J; Acker, Jason P; Ben, Robert N

    2016-01-01

    During cryopreservation, ice recrystallization is a major cause of cellular damage. Conventional cryoprotectants such as dimethyl sulfoxide (DMSO) and glycerol function by a number of different mechanisms but do not mitigate or control ice recrystallization at concentrations utilized in cryopreservation procedures. In North America, cryopreservation of human red blood cells (RBCs) utilizes high concentrations of glycerol. RBC units frozen under these conditions must be subjected to a time-consuming deglycerolization process after thawing in order to remove the glycerol to recrystallization inhibitors (IRIs) that are effective cryoprotectants for human RBCs, resulting in 70-80% intact RBCs using only 15% glycerol and slow freezing rates. These compounds are capable of reducing the average ice crystal size of extracellular ice relative to a 15% glycerol control validating the positive correlation between a reduction in ice crystal size and increased post-thaw recovery of RBCs. The most potent IRI from this study is also capable of protecting frozen RBCs against the large temperature fluctuations associated with transient warming.

  20. A freeze-fracture study on the developing satellite cells of spinal ganglia in the chick embryo

    Directory of Open Access Journals (Sweden)

    Claudio A. Ferraz de Carvalho

    1988-03-01

    Full Text Available A freeze-fracture analysis of the satellite cells of spinal ganglia of the chick embryo was performed in 8 successive stages of development, from the 5th incubation day to hatching. The characteristic laminar disposition of the cells were first observed on the 7th day. Tight junctions were found at the 20th incubation day. Small groups or irregular aggregates of particles, but not gap junctions, were described on the 7th and 8th days. Pinocytotic vesicles were pointed out in the different stages considered.

  1. [Effect of inert gas xenon on the functional state of nucleated cells of peripheral blood during freezing].

    Science.gov (United States)

    Laptev, D S; Polezhaeva, T V; Zaitseva, O O; Khudyakov, A N; Utemov, S V; Knyazev, M G; Kostyaev, A A

    2015-01-01

    A new method of preservation of nucleated cells in the electric refrigerator with xenon. After slow freezing and storage is even one day at -80 °C persists for more than 60% leukocytes. Cell membranes are resistant to the vital dye. In 85% of granulocytes stored baseline lysosomal-cationic protein, reduced lipid peroxidation and antioxidant activity. Cryopreservation of biological objects in inert gases is a promising direction in the practice of medicine and can be an alternative to the traditional method using liquid nitrogen. PMID:26027341

  2. Freezing avoidance by supercooling in Olea europaea cultivars: the role of apoplastic water, solute content and cell wall rigidity.

    Science.gov (United States)

    Arias, Nadia S; Bucci, Sandra J; Scholz, Fabian G; Goldstein, Guillermo

    2015-10-01

    Plants can avoid freezing damage by preventing extracellular ice formation below the equilibrium freezing temperature (supercooling). We used Olea europaea cultivars to assess which traits contribute to avoid ice nucleation at sub-zero temperatures. Seasonal leaf water relations, non-structural carbohydrates, nitrogen and tissue damage and ice nucleation temperatures in different plant parts were determined in five cultivars growing in the Patagonian cold desert. Ice seeding in roots occurred at higher temperatures than in stems and leaves. Leaves of cold acclimated cultivars supercooled down to -13 °C, substantially lower than the minimum air temperatures observed in the study site. During winter, leaf ice nucleation and leaf freezing damage (LT50 ) occurred at similar temperatures, typical of plant tissues that supercool. Higher leaf density and cell wall rigidity were observed during winter, consistent with a substantial acclimation to sub-zero temperatures. Larger supercooling capacity and lower LT50 were observed in cold-acclimated cultivars with higher osmotically active solute content, higher tissue elastic adjustments and lower apoplastic water. Irreversible leaf damage was only observed in laboratory experiments at very low temperatures, but not in the field. A comparative analysis of closely related plants avoids phylogenetic independence bias in a comparative study of adaptations to survive low temperatures. PMID:25737264

  3. Freezing avoidance by supercooling in Olea europaea cultivars: the role of apoplastic water, solute content and cell wall rigidity.

    Science.gov (United States)

    Arias, Nadia S; Bucci, Sandra J; Scholz, Fabian G; Goldstein, Guillermo

    2015-10-01

    Plants can avoid freezing damage by preventing extracellular ice formation below the equilibrium freezing temperature (supercooling). We used Olea europaea cultivars to assess which traits contribute to avoid ice nucleation at sub-zero temperatures. Seasonal leaf water relations, non-structural carbohydrates, nitrogen and tissue damage and ice nucleation temperatures in different plant parts were determined in five cultivars growing in the Patagonian cold desert. Ice seeding in roots occurred at higher temperatures than in stems and leaves. Leaves of cold acclimated cultivars supercooled down to -13 °C, substantially lower than the minimum air temperatures observed in the study site. During winter, leaf ice nucleation and leaf freezing damage (LT50 ) occurred at similar temperatures, typical of plant tissues that supercool. Higher leaf density and cell wall rigidity were observed during winter, consistent with a substantial acclimation to sub-zero temperatures. Larger supercooling capacity and lower LT50 were observed in cold-acclimated cultivars with higher osmotically active solute content, higher tissue elastic adjustments and lower apoplastic water. Irreversible leaf damage was only observed in laboratory experiments at very low temperatures, but not in the field. A comparative analysis of closely related plants avoids phylogenetic independence bias in a comparative study of adaptations to survive low temperatures.

  4. Role of intracellular freezing in the death of cells cooled at supraoptimal rates. [Preservation of erythrocytes, bone marrow cells, and yeasts by freezing

    Energy Technology Data Exchange (ETDEWEB)

    Mazur, P.

    1976-01-01

    Cooling velocity is one of the major factors that determines whether viable cells can be frozen to temperatures that permit indefinite storage. Cooling either too slowly or too rapidly tends to be damaging. Optimum cooling rates are reported for mouse marrow stem cells, yeast, and human red cells.

  5. A study on ice crystal formation behavior at intracellular freezing of plant cells using a high-speed camera.

    Science.gov (United States)

    Ninagawa, Takako; Eguchi, Akemi; Kawamura, Yukio; Konishi, Tadashi; Narumi, Akira

    2016-08-01

    Intracellular ice crystal formation (IIF) causes several problems to cryopreservation, and it is the key to developing improved cryopreservation techniques that can ensure the long-term preservation of living tissues. Therefore, the ability to capture clear intracellular freezing images is important for understanding both the occurrence and the IIF behavior. The authors developed a new cryomicroscopic system that was equipped with a high-speed camera for this study and successfully used this to capture clearer images of the IIF process in the epidermal tissues of strawberry geranium (Saxifraga stolonifera Curtis) leaves. This system was then used to examine patterns in the location and formation of intracellular ice crystals and to evaluate the degree of cell deformation because of ice crystals inside the cell and the growing rate and grain size of intracellular ice crystals at various cooling rates. The results showed that an increase in cooling rate influenced the formation pattern of intracellular ice crystals but had less of an effect on their location. Moreover, it reduced the degree of supercooling at the onset of intracellular freezing and the degree of cell deformation; the characteristic grain size of intracellular ice crystals was also reduced, but the growing rate of intracellular ice crystals was increased. Thus, the high-speed camera images could expose these changes in IIF behaviors with an increase in the cooling rate, and these are believed to have been caused by an increase in the degree of supercooling.

  6. Generalized structural theory of freezing

    International Nuclear Information System (INIS)

    The first-principles order parameter theory of freezing, proposed in an earlier work, has been successful in yielding quantitative agreement with known freezing parameters for monoatomic liquids forming solids with one atom per unit cell. A generalization of this theory is presented here to include the effects of a basis set of many atoms per unit cell. The basic equations get modified by the 'density structure factors' fsub(i) which arise from the density variations within the unit cell. Calculations are presented for the important case of monoatomic liquids freezing into hexagonal close packed solids. It is concluded that all freezing transitions can be described by using structural correlations in the liquid instead of the pair potential; and that the three body correlations are important in deciding the type of solid formed after freezing. (author)

  7. Deficiency in the glycerol channel Fps1p confers increased freeze tolerance to yeast cells: application of the fps1delta mutant to frozen dough technology.

    Science.gov (United States)

    Izawa, Shingo; Ikeda, Kayo; Maeta, Kazuhiro; Inoue, Yoshiharu

    2004-12-01

    Intracellular glycerol content affects the freeze-thaw stress tolerance of Saccharomyces cerevisiae. We have recently reported that intracellular-glycerol-enriched cells cultured in glycerol medium acquire tolerance to freeze stress and retain high leavening ability even in dough after frozen storage [Izawa et al. (2004) Appl Microbiol Biotechnol http://dx.doi.org/10.1007/s00253-004-1624-4]. A deletion mutant of the FPS1 gene, which encodes a glycerol channel, accumulates glycerol inside the cell without an exogenous supply of glycerol into the medium. We found that the fps1delta cells acquired tolerance to freeze stress and retained high leavening ability in dough after frozen storage for 7 days. These results suggest that the fps1delta mutant is a useful strain for developing better frozen-dough with a commercial advantage. PMID:15278313

  8. Freezing of Fresh Wharton's Jelly From Human Umbilical Cords Yields High Post-Thaw Mesenchymal Stem Cell Numbers for Cell-Based Therapies.

    Science.gov (United States)

    Fong, Chui-Yee; Subramanian, Arjunan; Biswas, Arijit; Bongso, Ariff

    2016-04-01

    Some cord blood banks freeze entire pieces of UC (mixed cord, MC) which after post-thaw yields mixed heterogeneous populations of mesenchymal stem cells (MSCs) from all its microanatomical compartments. Freezing of such entire tissues results in sub-optimal post-thaw cell recovery because of poor cryoprotectant diffusion and intracellular ice-formation, heat and water transport issues, and damage to intercellular junctions. To develop a simple method of harvesting pure homogeneous MSCs for cord blood banks, we compared the post-thaw behavior of three groups of frozen UC tissues: (i) freshly harvested WJ without cell separation; (ii) MSCs isolated from WJ (WJSC); and (iii) MC, WJ, and WJSC produced high post-thaw cell survival rates (93.52 ± 6.12% to 90.83 ± 4.51%) and epithelioid monolayers within 24 h in primary culture whereas post-thaw MC explants showed slow growth with mixed epithelioid and fibroblastic cell outgrowths after several days. Viability and proliferation rates of post-thawed WJ and hWJSC were significantly greater than MC. Post-thaw WJ and WJSC produced significantly greater CD24(+) and CD108(+) fluorescence intensities and significantly lower CD40(+) contaminants. Post-thaw WJ and WJSC produced significantly lesser annexin-V-positive and sub-G1 cells and greater degrees of osteogenic and chondrogenic differentiation compared to MC. qRT-PCR analysis of post-thaw MC showed significant decreases in anti-apoptotic gene expression (SURVIVIN, BCL2) and increases in pro-apoptotic (BAX) and cell cycle regulator genes (P53, P21, ROCK 1) compared to WJ and WJSC. We conclude that freezing of fresh WJ is a simple and reliable method of generating large numbers of clinically utilizable MSCs for cell-based therapies. PMID:26365815

  9. In Vitro Study on Lethal Effect of Human Choroidal Melanoma OCM-1 Cell Line by Repeating-70℃Freeze Thawing%In Vitro Study on Lethal Effect of Human Choroidal Melanoma OCM-1 Cell Line by Repeating-70℃ Freeze Thawing

    Institute of Scientific and Technical Information of China (English)

    Bin Liu; Yongping Li; Bo Zhang; Wenxin Zhang

    2007-01-01

    Objective: To investigate the effects of repeating -70℃ freeze thawing on human choroidal melanoma cell line OCM-1.Methods: OCM-1 cells were frozen by repeating -70℃ freeze thawing with various durations and frequencies. Then the inhibit rate of cells was examined by MTT essay.The cell viability was measured by monoclonal formation assay. We also used the HE staining, immunohistochemistry staining and the laser-scanning confocal microscopy (LSCM) to investigate the morphological changes of the cells.Results: The growth of OCM-1 cells was inhibited by repeating -70℃ freeze thawing in time-dependent and frequency-dependent manners (P < 0.01). Different morphous including necrosis and apoptosis of the cells could be observed after -70℃ freeze thawing by the LSCM.Conclusion: Repeating -70℃ freeze thawing can not only kill cells directly and induce considerable cells to apoptosis, but also inhibit the growth of the survivals. The kill and wound ratio of the cells disposal with different times and frequencies present variance. And the distinction when treated with different frequencies during the same time is much more significant than different times with the same frequency, which guide clinical workers to choose repeating cryotherapy with short term method instead of single cryotherapy with long term in choroidal melanoma treatment.

  10. Intracellular glycerol influences resistance to freeze stress in Saccharomyces cerevisiae: analysis of a quadruple mutant in glycerol dehydrogenase genes and glycerol-enriched cells.

    Science.gov (United States)

    Izawa, Shingo; Sato, Machiko; Yokoigawa, Kumio; Inoue, Yoshiharu

    2004-11-01

    Glycerol is well known as a cryoprotectant similar to trehalose. However, there is little information about the effects of intracellular glycerol on the freeze-thaw stress tolerance of yeast. Through analysis of a quadruple-knockout mutant of glycerol dehydrogenase genes (ara1 Delta gcy1 Delta gre3 Delta ypr1 Delta) in Saccharomyces cerevisiae, we revealed that the decrease in glycerol dehydrogenase activity led to increased levels of intracellular glycerol. We also found that this mutant showed higher tolerance to freeze stress than wild type strain W303-1A. Furthermore, we demonstrated that intracellular-glycerol-enriched cells cultured in glycerol medium acquire tolerance to freeze stress and retain high leavening ability in dough even after frozen storage for 7 days. These results suggest the possibility of using intracellular-glycerol-enriched cells to develop better frozen dough. PMID:15127164

  11. Freezing osteoblast cells attached to hydroxyapatite discs and glass coverslips: Mechanisms of damage

    Institute of Scientific and Technical Information of China (English)

    McGRATH; John

    2007-01-01

    Damage mechanisms for osteoblast cells (OBs) attached to hydroxyapatite (HA) discs and glass coverslips were comprehensively investigated. Cell-cell, cell-matrix interaction altered the cryobiological properties of cells. Attached cells were sub- ject to more severe mechanical damage than isolated cells because attached cells had larger contacting area with ice and the three dimensional movements of iso- lated cells made them more flexible than attached cells that could only deform in one dimension. Results showed that the viability of attached OB cells decreased significantly compared with the viability of isolated OB cells under the same cryo- preservation procedure. Extracellular ice, differential thermal contraction, and mechanical stresses were the major damaging factors for OB cells attached to HA discs and glass coverslips.

  12. Freezing osteoblast cells attached to hydroxyapatite discs and glass coverslips: Mechanisms of damage

    Institute of Scientific and Technical Information of China (English)

    LIU BaoLin; McGRATH John

    2007-01-01

    Damage mechanisms for osteoblast cells (OBs) attached to hydroxyapatite (HA)discs and glass coverslips were comprehensively investigated. Cell-cell, cell-matrix interaction altered the cryobiological properties of cells. Attached cells were subject to more severe mechanical damage than isolated cells because attached cells had larger contacting area with ice and the three dimensional movements of isolated cells made them more flexible than attached cells that could only deform in one dimension. Results showed that the viability of attached OB cells decreased significantly compared with the viability of isolated OB cells under the same cryopreservation procedure. Extracellular ice, differential thermal contraction, and mechanical stresses were the major damaging factors for OB cells attached to HA discs and glass coverslips.

  13. Evaluation of Methylcellulose and Dimethyl Sulfoxide as the Cryoprotectants in a Serum-Free Freezing Media for Cryopreservation of Adipose-Derived Adult Stem Cells

    OpenAIRE

    Thirumala, Sreedhar; Gimble, Jeffrey M.; Devireddy, Ram V.

    2010-01-01

    Developing effective techniques for the cryopreservation of human adipose-derived adult stem cells (ASCs) could increase the usefulness of these cells in tissue engineering and regenerative medicine. To this end, we investigated the post-freeze/thaw viability and apoptotic behavior of Passage 1 (P1) adult stem cells (ASCs) in 11 different media: (i) the traditional media containing Dulbecco’s modified Eagle’s medium (DMEM) with 80% fetal calf serum (FCS) and 10% dimethyl sulfoxide (DMSO), (ii...

  14. Freezing injury in potato leaves.

    Science.gov (United States)

    Sukumaran, N P; Weiser, C J

    1972-11-01

    Time-temperature profiles of freezing leaves from frost-resistant (Solanum acaule Bitt.) and frost-susceptible (Solanum tuberosum L. subsp. tuberosum Hawkes) types of potatoes did not reveal any major differences. The pattern of change in resistance of leaves to low voltage, low frequency current during freezing was different in the frost-resistant and susceptible leaves. In tissue sections from both types of leaves, cells freeze extracellularly at cooling velocities lower than 5 C per minute. Cells from leaves of resistant plants showed a higher osmotic pressure but not a higher water permeability than those from susceptible plants. The extent of injury caused by even very slow freezing was greater than that caused by equivalent isopiestic desiccation, particularly in susceptible leaves. The higher osmotic pressure in cells of leaves from resistant plants can account for the greater desiccation resistance but not for the frost resistance observed. PMID:16658217

  15. Anhydrobiosis and Freezing-Tolerance

    DEFF Research Database (Denmark)

    McGill, Lorraine; Shannon, Adam; Pisani, Davide;

    2015-01-01

    Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode...... Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth...

  16. Freezing and Food Safety

    Science.gov (United States)

    ... freeze well. Examples are mayonnaise, cream sauce and lettuce. Raw meat and poultry maintain their quality longer ... illness. [ Top of Page ] Does Freezing Destroy Bacteria & Parasites? Freezing to 0 °F inactivates any microbes — bacteria, ...

  17. Fusion of liposomes with the plasma membrane of epithelial cells: Fate of incorporated lipids as followed by freeze fracture and autoradiography of plastic sections

    OpenAIRE

    Knoll, G.; Burger, K.N.J.; Bron, R.; van Meer, G.; Verkleij, A. J.

    1988-01-01

    The fusion of liposomes with the plasma membrane of influenza virus- infected monolayers of an epithelial cell line, Madin-Darby canine kidney cells (van Meer et al., 1985. Biochemistry. 24:3593-3602), has been analyzed by morphological techniques. The distribution of liposomal lipids over the apical and basolateral plasma membrane domains after fusion was assessed by autoradiography of liposomal [3H]dipalmitoylphosphatidylcholine after rapid freezing or chemical fixation and further processi...

  18. Robotic milking and milk quality: effects on bacterial counts, somatic cell counts, freezing point and free fatty acids

    Directory of Open Access Journals (Sweden)

    Yvonne van der Vorst

    2010-01-01

    Full Text Available Changes in milk quality after the introduction of automatic milking systems (AM-systems on dairy farms in TheNetherlands, Germany and Denmark were examined and the data were compared with milk quality results of farms withconventional milking technology. After introduction, a small, but significant increase in total bacterial count, somatic cellcount, freezing point and free fatty acids was observed. The highest levels for total plate count and cell count are foundin the first six months after introduction. After this period the milk quality slightly improves to a more stable level.Risk factors related with milk quality concern general farm characteristics, animal health, AM-system, cleaning and cooling,housing, management skills of the farmer and the hygiene on the farm. Total plate count was significantly relatedto milk yield of the herd, cleaning of the area around the AM-system and the overall hygiene on the farm. Bulk milksomatic cell count appeared to be significantly related to milk yield of the herd and the number of milkings before replacementof the liners. An increased milking frequency is not the only explanation of increased free fatty acid levels. Technicalfactors related to free fatty acids mainly concerned the air inlet in the teat cups, bubbling (excessive air inlet and a toolong post run time of the milk pump. However, several questions regarding the causes of increased free fatty acid levelsremained unclear.

  19. Freezing injury in onion bulb cells: I. Evaluation of the conductivity method and analysis of ion and sugar efflux from injured cells.

    Science.gov (United States)

    Palta, J P; Levitt, J; Stadelmann, E J

    1977-09-01

    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 H(2)O. 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

  20. Neutron Computed Tomography of Freeze/thaw Phenomena in Polymer Electrolyte Fuel Cells

    Energy Technology Data Exchange (ETDEWEB)

    Matthew M. Mech; Jack Brenizer; Kenan Unlu; A.K. Heller

    2008-12-12

    This report summarizes the final year's progress of the three-year NEER program. The overall objectives of this program were to 1) design and construct a sophisticated hight-resolution neutron computed tomography (NCT) facility, 2) develop novel and sophisticated liquid water and ice quantification analysis software for computed tomography, and 3) apply the advanced software and NCT capability to study liquid and ice distribution in polymer electrolyte fuel cells (PEFCs) under cold-start conditions. These objectives have been accomplished by the research team, enabling a new capability for advanced 3D image quantification with neutron imaging for fuel cell and other applications. The NCT water quantification methodology and software will greatly add to the capabilities of the neutron imaging community, and the quantified liquid water and ice distribution provided by its application to PEFCs will enhance understanding and guide design in the fuel cell community.

  1. Effects of the ionizing radiations, freezing and thawing duration on chicken liver cells quality

    Science.gov (United States)

    Duarte, R. C.; Araújo, M. M.; Salum, D. C.; Marchioni, E.; Villavicencio, A. L. C. H.

    2009-07-01

    All food storage processes modify the food. Irradiation reduces and could stop cell division, avoid infestation, reduce contamination and delay food decomposition. The cold chain is a succession of steps which maintains the food at low temperature. Defrosted food shall never be frozen again, the best way being to consume it quickly then avoiding multiplication and acceleration of microbial growth, which causes decay and nutrients damage. The Comet Assay indicates DNA damage and can then be used to control the overall quality of the food and in a certain extent to evaluate the damage caused by irradiation and storage on liver chicken cells. In this work, different thawing temperatures and radiation doses were checked to establish a "DNA damage index" by using the Comet Assay. Samples were irradiated in a 60Co irradiator with 1.5, 3.0 and 4.5 kGy radiation doses. Our results showed that no intact cells were detected in frozen samples: however, irradiated liver samples in natura showed some intact cells depending on the applied radiation doses.

  2. Supercooling as a viable non-freezing cell preservation method of rat hepatocytes.

    Directory of Open Access Journals (Sweden)

    O Berk Usta

    Full Text Available Supercooling preservation holds the potential to drastically extend the preservation time of organs, tissues and engineered tissue products, and fragile cell types that do not lend themselves well to cryopreservation or vitrification. Here, we investigate the effects of supercooling preservation (SCP at -4(oC on primary rat hepatocytes stored in cryovials and compare its success (high viability and good functional characteristics to that of static cold storage (CS at +4(oC and cryopreservation. We consider two prominent preservation solutions a Hypothermosol (HTS-FRS and b University of Wisconsin solution (UW and a range of preservation temperatures (-4 to -10 (oC. We find that there exists an optimum temperature (-4(oC for SCP of rat hepatocytes which yields the highest viability; at this temperature HTS-FRS significantly outperforms UW solution in terms of viability and functional characteristics (secretions and enzymatic activity in suspension and plate culture. With the HTS-FRS solution we show that the cells can be stored for up to a week with high viability (~56%; moreover we also show that the preservation can be performed in large batches (50 million cells with equal or better viability and no loss of functionality as compared to smaller batches (1.5 million cells performed in cryovials.

  3. Temperature-sensitivity and cell biocompatibility of freeze-dried nanocomposite hydrogels incorporated with biodegradable PHBV

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Qingsong, E-mail: zqs8011@163.com; Chen, Li, E-mail: chenlis@tjpu.edu.cn; Dong, Youyu; Lu, Si

    2013-04-01

    The structure, morphology, thermal behaviors and cytotoxicity of novel hydrogels, composed of poly(N-isopropylacrylamide)(PNIPAM) and biodegradable polyester poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) under nanoclay hectorite “Laponite XLG” severed as physical cross-linker, were characterized by X-ray diffraction, scanning electron microscopy, gravimetric method, differential scanning calorimetry, and cell culture experiments. It was found that, due to the introduction of hydrophobic PHBV, the homogeneity of interior pore in the pure PNIPAM nanocomposite hydrogel was disrupted, the transparency and swelling degree gradually decreased. Although the weight ratio between PHBV and NIPAM increased from 5 to 40 wt.%, the volume phase transition temperature (VPTTs) of hydrogel were not altered compared with the pure PNIPAM nanocomposite hydrogel. No matter what PHBV content, the PHBV/PNIPAM/Hectorite hydrogels always exhibit good stimuli-responsibility. In addition, human hepatoma cells(HepG2) adhesion and spreading on the surface of PHBV-based hydrogels was greatly improved than that of pure PNIPAM nanocomposite hydrogel at 37 °C due to the introduction of PHBV. Highlights: ► Thermo-responsive and cell biocompatible hydrogels incorporated PHBV was synthesized. ► The introduction of PHBV decreases the transparency of nanocomposite hydrogel. ► The introduction of PHBV has a little shift on VPTTs of nanocomposite hydrogel. ► The HepG2 cells could adhere and spread on the surface of PHBV-based hydrogels. ► Cell sheet could be detached simultaneously from the surface of hydrogels.

  4. Active control of the nucleation temperature enhances freezing survival of multipotent mesenchymal stromal cells.

    Science.gov (United States)

    Lauterboeck, L; Hofmann, N; Mueller, T; Glasmacher, B

    2015-12-01

    Cryopreservation is a technique that has been extensively used for storage of multipotent mesenchymal stromal cells (MSCs) in regenerative medicine. Therefore, improving current cryopreservation procedures in terms of increasing cell viability and functionality is important. In this study, we optimized the cryopreservation protocol of MSCs derived from the common marmoset Callithrix jacchus (cj), which can be used as a non-human primate model in various pathological and transplantation studies and have a great potential for regenerative medicine. We have investigated the effect of the active control of the nucleation temperature using induced nucleation at a broad range of temperatures and two different dimethylsulfoxide concentrations (Me2SO, 5% (v/v) and 10%, (v/v)) to evaluate the overall effect on the viability, metabolic activity and recovery of cells after thawing. Survival rate and metabolic activity displayed an optimum when ice formation was induced at -10 °C. Cryomicroscopy studies indicated differences in ice crystal morphologies as well as differences in intracellular ice formation with different nucleation temperatures. High subzero nucleation temperatures resulted in larger extracellular ice crystals and cellular dehydration, whereas low subzero nucleation temperatures resulted in smaller ice crystals and intracellular ice formation. PMID:26499840

  5. Trehalose levels and survival ratio of freeze-tolerant versus freeze-sensitive yeasts.

    Science.gov (United States)

    Hino, A; Mihara, K; Nakashima, K; Takano, H

    1990-05-01

    Five freeze-tolerant yeast strains suitable for frozen dough were compared with ordinary commercial bakers' yeast. Kluyveromyces thermotolerans FRI 501 cells showed high survival ability after freezing when their resting cells were fermented for 0 to 180 min in modified liquid medium, and they grew to log and stationary phases. Among the freeze-tolerant strains of Saccharomyces cerevisiae, FRI 413 and FRI 869 showed higher surviving and trehalose-accumulating abilities than other S. cerevisiae strains, but were affected by a prolonged prefermentation period and by growth phases. The freeze tolerance of the yeasts was, to some extent, associated with the basal amount of intracellular trehalose after rapid degradation at the onset of the prefermentation period. In the freeze-sensitive yeasts, the degree of hydrolysis of trehalose may thus be affected by the kind of saccharide, unlike in freeze-tolerant yeasts. PMID:2339891

  6. Improved method for freeze-fracture radioautography of tissues and cells, as applied to duodenal epithelium and thymic lymphocytes

    International Nuclear Information System (INIS)

    An improved method has been devised for the localization of radioactive substances to either one of the leaflets of cellular membranes. After tissue specimens are freeze-fractured and covered with a platinum-carbon replica, they are freeze-dried to allow coating with radioautographic emulsion at room temperature. After exposure at 4 degrees C and development, the emulsion is protected by layers of carbon and grease before the tissue underlying the replica is dissolved in sodium hypochlorite. The grease is removed in Freon 14 and the replica with its emulsion cover is mounted on a specimen grid for electron microscopic examination. The accuracy of radioactivity localization was demonstrated using 3H-thymidine-labeled liver by finding silver grains over the same sites after freeze-fracture as after thin section radioautography. Tests with 3H-methacrylate revealed that the interposition of a platinum-carbon replica decreased the radioautographic reaction by over 80%; hence, the need for long exposure. Only 67% of the silver grains came from radiation sources located beyond the upper 0.05 micron of the specimen and, therefore, the emulsion could be affected by radiation sources located not only within membrane leaflets but also in nearby cytoplasm. Thus, when 3H-fucose was injected into rats to locate newly formed glycoproteins within intestinal epithelium membranes, some of the silver grains found over E and P faces might be produced by radiation coming from the adjacent cytoplasm. To localize label within membrane leaflets in the absence of radiation sources in the cytoplasm, lymphocyte suspensions were incubated with 3H-concanavalin A at 0 degrees C. The plasmalemma radioactivity was then restricted to the two membrane leaflets, with 87-93% of the silver grains on the E leaflet and 7-13% on the P leaflet

  7. Functional compartmentation of the Golgi apparatus of plant cells : immunocytochemical analysis of high-pressure frozen- and freeze-substituted sycamore maple suspension culture cells.

    Science.gov (United States)

    Zhang, G F; Staehelin, L A

    1992-07-01

    The Golgi apparatus of plant cells is engaged in both the processing of glycoproteins and the synthesis of complex polysaccharides. To investigate the compartmentalization of these functions within individual Golgi stacks, we have analyzed the ultrastructure and the immunolabeling patterns of high-pressure frozen and freeze-substituted suspension-cultured sycamore maple (Acer pseudoplatanus L.) cells. As a result of the improved structural preservation, three morphological types of Golgi cisternae, designated cis, medial, and trans, as well as the trans Golgi network, could be identified. The number of cis cisternae per Golgi stack was found to be fairly constant at approximately 1, whereas the number of medial and trans cisternae per stack was variable and accounted for the varying number of cisternae (3-10) among the many Golgi stacks examined. By using a battery of seven antibodies whose specific sugar epitopes on secreted polysaccharides and glycoproteins are known, we have been able to determine in which types of cisternae specific sugars are added to N-linked glycans, and to xyloglucan and polygalacturonic acid/rhamnogalacturonan-I, two complex polysaccharides. The findings are as follows. The beta-1,4-linked d-glucosyl backbone of xyloglucan is synthesized in trans cisternae, and the terminal fucosyl residues on the trisaccharide side chains of xyloglucan are partly added in the trans cisternae, and partly in the trans Golgi network. In contrast, the polygalacturonic/rhamnogalacturonan-I backbone is assembled in cis and medial cisternae, methylesterification of the carboxyl groups of the galacturonic acid residues in the polygalacturonic acid domains occurs mostly in medial cisternae, and arabinose-containing side chains of the polygalacturonic acid domains are added to the nascent polygalacturonic acid/rhamnogalacturonan-I molecules in the trans cisternae. Double labeling experiments demonstrate that xyloglucan and polygalacturonic acid

  8. Surface freezing of water.

    Science.gov (United States)

    Pérez-Díaz, J L; Álvarez-Valenzuela, M A; Rodríguez-Celis, F

    2016-01-01

    Freezing, melting, evaporation and condensation of water are essential ingredients for climate and eventually life on Earth. In the present work, we show how surface freezing of supercooled water in an open container is conditioned and triggered-exclusively-by humidity in air. Additionally, a change of phase is demonstrated to be triggered on the water surface forming surface ice crystals prior to freezing of bulk. The symmetry of the surface crystal, as well as the freezing point, depend on humidity, presenting at least three different types of surface crystals. Humidity triggers surface freezing as soon as it overpasses a defined value for a given temperature, generating a plurality of nucleation nodes. An evidence of simultaneous nucleation of surface ice crystals is also provided. PMID:27330895

  9. Freeze drying apparatus

    Science.gov (United States)

    Coppa, Nicholas V.; Stewart, Paul; Renzi, Ernesto

    2001-01-01

    The present invention provides methods and apparatus for freeze drying in which a solution, which can be a radioactive salt dissolved within an acid, is frozen into a solid on vertical plates provided within a freeze drying chamber. The solid is sublimated into vapor and condensed in a cold condenser positioned above the freeze drying chamber and connected thereto by a conduit. The vertical positioning of the cold condenser relative to the freeze dryer helps to help prevent substances such as radioactive materials separated from the solution from contaminating the cold condenser. Additionally, the system can be charged with an inert gas to produce a down rush of gas into the freeze drying chamber to also help prevent such substances from contaminating the cold condenser.

  10. Freeze drying method

    Science.gov (United States)

    Coppa, Nicholas V.; Stewart, Paul; Renzi, Ernesto

    1999-01-01

    The present invention provides methods and apparatus for freeze drying in which a solution, which can be a radioactive salt dissolved within an acid, is frozen into a solid on vertical plates provided within a freeze drying chamber. The solid is sublimated into vapor and condensed in a cold condenser positioned above the freeze drying chamber and connected thereto by a conduit. The vertical positioning of the cold condenser relative to the freeze dryer helps to help prevent substances such as radioactive materials separated from the solution from contaminating the cold condenser. Additionally, the system can be charged with an inert gas to produce a down rush of gas into the freeze drying chamber to also help prevent such substances from contaminating the cold condenser.

  11. Aquaporin expression correlates with freeze tolerance in baker's yeast, and overexpression improves freeze tolerance in industrial strains.

    Science.gov (United States)

    Tanghe, An; Van Dijck, Patrick; Dumortier, Françoise; Teunissen, Aloys; Hohmann, Stefan; Thevelein, Johan M

    2002-12-01

    Little information is available about the precise mechanisms and determinants of freeze resistance in baker's yeast, Saccharomyces cerevisiae. Genomewide gene expression analysis and Northern analysis of different freeze-resistant and freeze-sensitive strains have now revealed a correlation between freeze resistance and the aquaporin genes AQY1 and AQY2. Deletion of these genes in a laboratory strain rendered yeast cells more sensitive to freezing, while overexpression of the respective genes, as well as heterologous expression of the human aquaporin gene hAQP1, improved freeze tolerance. These findings support a role for plasma membrane water transport activity in determination of freeze tolerance in yeast. This appears to be the first clear physiological function identified for microbial aquaporins. We suggest that a rapid, osmotically driven efflux of water during the freezing process reduces intracellular ice crystal formation and resulting cell damage. Aquaporin overexpression also improved maintenance of the viability of industrial yeast strains, both in cell suspensions and in small doughs stored frozen or submitted to freeze-thaw cycles. Furthermore, an aquaporin overexpression transformant could be selected based on its improved freeze-thaw resistance without the need for a selectable marker gene. Since aquaporin overexpression does not seem to affect the growth and fermentation characteristics of yeast, these results open new perspectives for the successful development of freeze-resistant baker's yeast strains for use in frozen dough applications. PMID:12450819

  12. Does Freeze-Thawing Influence the Effects of Platelet Concentrates? An In Vitro Study on Human Adipose-Derived Stem Cells.

    Science.gov (United States)

    Ceci, Caterina; Niada, Stefania; Del Fabbro, Massimo; Lolato, Alessandra; Taschieri, Silvio; Giannasi, Chiara; Brini, Anna Teresa

    2016-03-01

    Human adipose-derived stem cells (hASCs) have been proposed as a possible therapy for tissue regeneration in aesthetic, plastic, and reconstructive surgery. Today, platelet concentrates are used in a wide range of disciplines, but their storage has become a controversial aspect. The purpose of this in vitro study was to evaluate the effect of plasma rich in growth factors (PRGF), after a freeze-thawing cycle, on the proliferation and biological activity of progenitor cells involved in soft tissue healing. Different formulations of activated PRGF were added to hASCs cultured in serum-free medium. Cell proliferation was assessed by MTT test and cell count up to 7 and 12-day incubation. Osteo-differentiation ability of hASCs was also tested after 7 and 14-day incubation by alkaline phosphatase assay. The effects of 4 PRGF preparations (fresh/frozen and with/without platelets) were compared with corresponding formulations of plasma poor in growth factors and with standard medium. hASCs cultured in the presence of platelet concentrates increased proliferation rate with respect to cells grown in standard medium without significant differences among all the tested plasma formulations on cell viability up to 12 days of culture. PRGF activity is preserved after cryopreservation and platelet-rich preparations promoted osteo-differentiation of hASCs at day 7. In conclusion, PRGF supports the proliferation and the differentiation of progenitor cells in vitro also when applied after cryopreservation. Platelet concentrates, either alone or in combination with mesenchymal stem cells, might be a valuable tool in the field of tissue regeneration. PMID:26872279

  13. Prevention and therapy of squamous cell carcinoma of the rodent esophagus using freeze-dried black raspberries

    Institute of Scientific and Technical Information of China (English)

    Gary D STONER; Robeena M AZIZ

    2007-01-01

    Aim: This study was conducted to determine if short-term treatment of N-nitrosomethylbenzylamine (NMBA)-induced tumors in the rat esophagus with dietary freeze-dried black raspberries (FBR) would result in tumor regression and enhanced survival of the animals. Methods: Four-week-old male Fisher-344 ratswere administered an AIN-76A control diet and injected subcutaneously with 0.5 mg/kg NMBA once per week for 15 weeks. At 19 weeks, when rats had an average of 5-6 tumors (papillomas) per esophagus, they were given a control diet contain-ing either 5%, 10%, or 20% FBR. After 7 weeks of berry treatment, all surviving rats were killed and tumor incidence, number and volume were determined. Results:Esophageal tumor incidences, numbers and volumes in NMBA-treated rats were not influenced by any of the berry treatments. There were progressive increases in the survival of NMBA-treated rats fed 5%-20% FBR diets; however, these increases were not significant. Conclusion: FBR at 5%, 10%, and 20% of the diet had no effect on the development of NMBA-induced tumors in the rat esophagus or on animal survival when administered for 7 weeks beginning at the papilloma stage of tumor development. Thus, FBR appear to have no therapeutic value in the treatment of esophageal tumors. In contrast, dietary FBR are highly effectivein preventing the development of NMBA-induced esophageal tumors in rats when administered before and during NMBA treatment or shortly after NMBA treat-ment when the esophagi contain preneoplastic (dysplastic) lesions of varyingdegrees of severity.

  14. Caffeine-induced Ca2+ transients and exocytosis in Paramecium cells. A correlated Ca2+ imaging and quenched-flow/freeze-fracture analysis.

    Science.gov (United States)

    Klauke, N; Plattner, H

    1998-01-01

    Caffeine causes a [Ca2+]i increase in the cortex of Paramecium cells, followed by spillover with considerable attenuation, into central cell regions. From [Ca2+]resti approximately 50 to 80 nm, [Ca2+]acti rises within /=2 sec. Chelation of Ca2+o considerably attenuated [Ca2+]i increase. Therefore, caffeine may primarily mobilize cortical Ca2+ pools, superimposed by Ca2+ influx and spillover (particularly in tl cells with empty trichocyst docking sites). In nd cells, caffeine caused trichocyst contents to decondense internally (Ca2+-dependent stretching, normally occurring only after membrane fusion). With 7S cells this usually occurred only to a small extent, but with increasing frequency as [Ca2+]i signals were reduced by [Ca2+]o chelation. In this case, quenched-flow and ultrathin section or freeze-fracture analysis revealed dispersal of membrane components (without fusion) subsequent to internal contents decondensation, opposite to normal membrane fusion when a full [Ca2+]i signal was generated by caffeine stimulation (with Ca2+i and Ca2+o available). We conclude the following. (i) Caffeine can mobilize Ca2+ from cortical stores independent of the presence of Ca2+o. (ii) To yield adequate signals for normal exocytosis, Ca2+ release and Ca2+ influx both have to occur during caffeine stimulation. (iii) Insufficient [Ca2+]i increase entails caffeine-mediated access of Ca2+ to the secretory contents, thus causing their decondensation before membrane fusion can occur. (iv) Trichocyst decondensation in turn gives a signal for an unusual dissociation of docking/fusion components at the cell membrane. These observations imply different threshold [Ca2+]i-values for membrane fusion and contents discharge.

  15. Freeze-thaw lysates of Plasmodium falciparum-infected red blood cells induce differentiation of functionally competent regulatory T cells from memory T cells

    OpenAIRE

    Finney, Olivia C; Lawrence, Emma; Gray, Alice P; Njie, Madi; Riley, Eleanor M.; Walther, Michael

    2012-01-01

    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-derive...

  16. Evaluation of the ability of primary selective enrichment to resuscitate heat-injured and freeze-injured Listeria monocytogenes cells.

    OpenAIRE

    Budu-Amoako, E; Toora, S; Ablett, R F; J. Smith

    1992-01-01

    Resuscitation rates of injured Listeria monocytogenes on conventional selective Listeria enrichment broth and nonselective Trypticase soy broth containing 0.6% yeast extract were compared. Cells were heated to 60 degrees C for 5 min or frozen at -20 degrees C for 7 days. Inoculation of Trypticase soy broth-yeast extract with the stressed cells resulted in growth that was superior to that in Listeria enrichment broth. Injured cells were fully recovered at 6 to 8 h.

  17. Mycobacterium bovis bacillus Calmette-Guérin killed by extended freeze-drying targets plasmacytoid dendritic cells to regulate lung inflammation.

    Science.gov (United States)

    Lagranderie, Micheline; Abolhassani, Mohammad; Vanoirbeek, Jeroen A J; Lima, Carla; Balazuc, Anne-Marie; Vargaftig, B Boris; Marchal, Gilles

    2010-01-15

    We have previously shown that bacillus Calmette-Guérin (BCG) inactivated by extended freeze-drying (EFD) reduces airway hyperresponsiveness, whereas live and heat-killed BCG fail to do so. However, the cells involved in the protective effect and the signaling and transcriptional networks that could reprogram T cell commitment after EFD BCG treatment remained to be elucidated. We investigated whether EFD BCG targets plasmacytoid dendritic cells (pDCs) potentially involved in the polarization of regulatory T cells (Tregs) and the transcriptional factors that regulate allergic inflammation. OVA-sensitized mice were s.c. injected with EFD, live, or heat-killed BCG. We analyzed after the injection of the various BCG preparations: 1) pDCs recruited in the draining lymph nodes (day 4); 2) transcription factors involved in inflammation and T cell commitment in spleen and lungs after OVA challenge (day 28). Airway hyperresponsiveness and transcription factors were determined after in vivo depletion of pDCs or Tregs in EFD BCG-treated and OVA-challenged mice. EFD BCG reduced inflammation via the recruitment of pDCs polarizing the differentiation of naive CD4+ T lymphocytes into Tregs. In vivo, pDC or Treg depletion at the time of EFD BCG treatment abrogated the protection against inflammation. EFD BCG treatment upregulated Forkhead-winged helix transcription factor (Treg signature) and downregulated GATA-3 and RORgammat (Th2 and Th17 signatures) more efficiently than live and heat-killed BCG. Moreover, only EFD BCG enhanced peroxisome proliferator-activated receptor gamma expression and blocked NF-kappaB activation, cyclooxygenase expression, and p38 MAPK phosphorylation. EFD BCG reduced allergic inflammation by recruiting pDCs that promoted Tregs; EFD BCG acted as a peroxisome proliferator-activated receptor gamma agonist and thus could be used in asthma and other inflammatory diseases. PMID:20007537

  18. Ultrasound-Assisted Freezing

    Science.gov (United States)

    Delgado, A. E.; Sun, Da-Wen

    Freezing is a well-known preservation method widely used in the food industry. The advantages of freezing are to a certain degree counterbalanced by the risk of damage caused by the formation and size of ice crystals. Over recent years new approaches have been developed to improve and control the crystallization process, and among these approaches sonocrystallization has proved to be very useful, since it can enhance both the nucleation rate and the crystal growth rate. Although ultrasound has been successfully used for many years in the evaluation of various aspects of foods and in medical applications, the use of power ultrasound to directly improve processes and products is less popular in food manufacturing. Foodstuffs are very complex materials, and research is needed in order to define the specific sound parameters that aid the freezing process and that can later be used for the scale-up and production of commercial frozen food products.

  19. Design of a blood-freezing system for leukemia research

    Science.gov (United States)

    Williams, T. E.; Cygnarowicz, T. A.

    1978-01-01

    Leukemia research involves the use of cryogenic freezing and storage equipment. In a program being carried out at the National Cancer Institute (NCI), bone marrow (white blood cells) was frozen using a standard cryogenic biological freezer. With this system, it is difficult to maintain the desired rate of freezing and repeatability from sample to sample. A freezing system was developed that satisfies the requirements for a repeatable, constant freezing rate. The system was delivered to NIC and is now operational. This report describes the design of the major subsystems, the analyses, the operating procedure, and final system test results.

  20. Aquaporin-mediated improvement of freeze tolerance of Saccharomyces cerevisiae is restricted to rapid freezing conditions.

    Science.gov (United States)

    Tanghe, An; Van Dijck, Patrick; Colavizza, Didier; Thevelein, Johan M

    2004-06-01

    Previous observations that aquaporin overexpression increases the freeze tolerance of baker's yeast (Saccharomyces cerevisiae) without negatively affecting the growth or fermentation characteristics held promise for the development of commercial baker's yeast strains used in frozen dough applications. In this study we found that overexpression of the aquaporin-encoding genes AQY1-1 and AQY2-1 improves the freeze tolerance of industrial strain AT25, but only in small doughs under laboratory conditions and not in large doughs under industrial conditions. We found that the difference in the freezing rate is apparently responsible for the difference in the results. We tested six different cooling rates and found that at high cooling rates aquaporin overexpression significantly improved the survival of yeast cells, while at low cooling rates there was no significant effect. Differences in the cultivation conditions and in the thawing rate did not influence the freeze tolerance under the conditions tested. Survival after freezing is determined mainly by two factors, cellular dehydration and intracellular ice crystal formation, which depend in an inverse manner on the cooling velocity. In accordance with this so-called two-factor hypothesis of freezing injury, we suggest that water permeability is limiting, and therefore that aquaporin function is advantageous, only under rapid freezing conditions. If this hypothesis is correct, then aquaporin overexpression is not expected to affect the leavening capacity of yeast cells in large, industrial frozen doughs, which do not freeze rapidly. Our results imply that aquaporin-overexpressing strains have less potential for use in frozen doughs than originally thought. PMID:15184134

  1. Examination of the Blood Groups and Biologal Activity of the Freeze-drying Red Blood Cells%冻干前后红细胞血型及生物活性指标的检测

    Institute of Scientific and Technical Information of China (English)

    吴学忠; 刘忠; 吕蓉; 李敏; 李素萍; 於娟; 赵丹

    2011-01-01

    目的 检测冻干前、后红细胞血型抗原及其生物学活性,了解冻干前、后红细胞血型抗原及其生物学活性的变化情况.方法红细胞血型抗原的检测采用血型血清学方法,2,3-DPG和ATP用ELISA法.结果在ABO、Rh、MNSs、Kell、Duffy、P血型抗原中,冷冻前、冻干后红细胞血型抗原一致;在Lewis血型抗原中,冷冻前、冻干后红细胞血型抗原有变化.冷冻前、冻干后红细胞2,3-DPG和ATP含量均无明显变化.结论冷冻前、冻干后红细胞Lewis血型系统中的Lea和Leb血型抗原变化有明显差异;冷冻前、冻干后红细胞2,3-DPG和ATP含量变化无统计学意义.%Objective To detect the blood group antigens, the levels of 2,3-DPG and ATP of red blood cells (RBCs) before and after freeze-drying. Also the changes of blood group antigens and biological activity of RBCs before and after RBCs freeze-drying was observed. Methods The blood group antigens of RBCs were detected by the blood group serology, the levels of 2,3-DPG and ATP were detected by ELISA. Results The blood group antigens of RBCs were the same as before and after freeze-drying about ABO, Rh, MNSs, Kell, Duffy and P blood group systems. But the blood group antigens of RBCs of Lewis blood group systems were different before and after freeze-drying. The levels of 2,3-DPG and ATP of RBCs before and after freeze-drying didn't change significantly. Conclusion The Lea and Leb blood group antigens of RBCs before and after freeze-drying of Lewis blood group systems have changed significantly. The levels of 2,3-DPG and ATP of RBCs before and after freeze-drying havn't chenged significantly.

  2. Theoretical prediction of 'optimal' freezing programmes

    NARCIS (Netherlands)

    Woelders, H.; Chaveiro, A.

    2004-01-01

    We have developed a quantitative description of the osmotic behaviour of cells during freezing without a presupposed value of the cooling rate. Instead, at all times the intracellular supercooling is maximised provided that it does not exceed a predetermined value 'p' (e.g., 2°C). This should preclu

  3. Freeze-drying of lactic acid bacteria.

    Science.gov (United States)

    Fonseca, Fernanda; Cenard, Stéphanie; Passot, Stéphanie

    2015-01-01

    Lactic acid bacteria are of great importance for the food and biotechnology industry. They are widely used as starters for manufacturing food (e.g., yogurt, cheese, fermented meats, and vegetables) and probiotic products, as well as for green chemistry applications. Freeze-drying or lyophilization is a convenient method for preservation of bacteria. By reducing water activity to values below 0.2, it allows long-term storage and low-cost distribution at suprazero temperatures, while minimizing losses in viability and functionality. Stabilization of bacteria via freeze-drying starts with the addition of a protectant solution to the bacterial suspension. Freeze-drying includes three steps, namely, (1) freezing of the concentrated and protected cell suspension, (2) primary drying to remove ice by sublimation, and (3) secondary drying to remove unfrozen water by desorption. In this chapter we describe a method for freeze-drying of lactic acid bacteria at a pilot scale, thus allowing control of the process parameters for maximal survival and functionality recovery.

  4. Air-Cooled Stack Freeze Tolerance Freeze Failure Modes and Freeze Tolerance Strategies for GenDriveTM Material Handling Application Systems and Stacks Final Scientific Report

    Energy Technology Data Exchange (ETDEWEB)

    Hancock, David, W.

    2012-02-14

    Air-cooled stack technology offers the potential for a simpler system architecture (versus liquid-cooled) for applications below 4 kilowatts. The combined cooling and cathode air allows for a reduction in part count and hence a lower cost solution. However, efficient heat rejection challenges escalate as power and ambient temperature increase. For applications in ambient temperatures below freezing, the air-cooled approach has additional challenges associated with not overcooling the fuel cell stack. The focus of this project was freeze tolerance while maintaining all other stack and system requirements. Through this project, Plug Power advanced the state of the art in technology for air-cooled PEM fuel cell stacks and related GenDrive material handling application fuel cell systems. This was accomplished through a collaborative work plan to improve freeze tolerance and mitigate freeze-thaw effect failure modes within innovative material handling equipment fuel cell systems designed for use in freezer forklift applications. Freeze tolerance remains an area where additional research and understanding can help fuel cells to become commercially viable. This project evaluated both stack level and system level solutions to improve fuel cell stack freeze tolerance. At this time, the most cost effective solutions are at the system level. The freeze mitigation strategies developed over the course of this project could be used to drive fuel cell commercialization. The fuel cell system studied in this project was Plug Power's commercially available GenDrive platform providing battery replacement for equipment in the material handling industry. The fuel cell stacks were Ballard's commercially available FCvelocity 9SSL (9SSL) liquid-cooled PEM fuel cell stack and FCvelocity 1020ACS (Mk1020) air-cooled PEM fuel cell stack.

  5. Freezing-induced deformation of biomaterials in cryomedicine

    Science.gov (United States)

    Ozcelikkale, Altug

    Cryomedicine utilizes low temperature treatments of biological proteins, cells and tissues for cryopreservation, materials processing and cryotherapy. Lack of proper understanding of cryodamage that occurs during these applications remains to be the primary bottleneck for development of successful tissue cryopreservation and cryosurgery procedures. An engineering approach based on a view of biological systems as functional biomaterials can help identify, predict and control the primary cryodamage mechanisms by developing an understanding of underlying freezing-induced biophysical processes. In particular, freezing constitutes the main structural/mechanical origin of cryodamage and results in significant deformation of biomaterials at multiple length scales. Understanding of these freezing-induced deformation processes and their effects on post-thaw biomaterial functionality is currently lacking but will be critical to engineer improved cryomedicine procedures. This dissertation addresses this problem by presenting three separate but related studies of freezing-induced deformation at multiple length scales including nanometer-scale protein fibrils, single cells and whole tissues. A combination of rigorous experimentation and computational modeling is used to characterize post-thaw biomaterial structure and properties, predict biomaterial behavior and assess its post-thaw biological functionality. Firstly, freezing-induced damage on hierarchical extracellular matrix structure of collagen is investigated at molecular, fibril and matrix levels. Results indicate to a specific kind of fibril damage due to freezing-induced expansion of intrafibrillar fluid. This is followed by a study of freezing-induced cell and tissue deformation coupled to osmotically driven cellular water transport. Computational and semi empirical modeling of these processes indicate that intracellular deformation of the cell during freezing is heterogeneous and can interfere with cellular water

  6. Evaluation of Sericin as a Fetal Bovine Serum-Replacing Cryoprotectant During Freezing of Human Mesenchymal Stromal Cells and Human Osteoblast-Like Cells

    OpenAIRE

    Verdanova, Martina; Pytlik, Robert; Kalbacova, Marie Hubalek

    2014-01-01

    A reliable, cryoprotective, xeno-free medium suitable for different cell types is highly desirable in regenerative medicine. There is danger of infection or allergic reaction with the use of fetal bovine serum (FBS), making it problematic for medical applications. The aim of the present study was to develop an FBS-free cryoprotective medium for human mesenchymal stromal cells (hMSCs; primary cells) and immortalized human osteoblasts (SAOS-2 cell line). Furthermore, we endeavored to eliminate ...

  7. Freeze-Tolerant Condensers

    Science.gov (United States)

    Crowley, Christopher J.; Elkouhk, Nabil

    2004-01-01

    Two condensers designed for use in dissipating heat carried by working fluids feature two-phase, self-adjusting configurations such that their working lengths automatically vary to suit their input power levels and/or heat-sink temperatures. A key advantage of these condensers is that they can function even if the temperatures of their heat sinks fall below the freezing temperatures of their working fluids and the fluids freeze. The condensers can even be restarted from the frozen condition. The top part of the figure depicts the layout of the first condenser. A two-phase (liquid and vapor) condenser/vapor tube is thermally connected to a heat sink typically, a radiatively or convectively cooled metal panel. A single-phase (liquid) condensate-return tube (return artery) is also thermally connected to the heat sink. At intervals along their lengths, the condenser/vapor tube and the return artery are interconnected through porous plugs. This condenser configuration affords tolerance of freezing, variable effective thermal conductance (such that the return temperature remains nearly constant, independently of the ultimate sink temperature), and overall pressure drop smaller than it would be without the porous interconnections. An additional benefit of this configuration is that the condenser can be made to recover from the completely frozen condition either without using heaters, or else with the help of heaters much smaller than would otherwise be needed. The second condenser affords the same advantages and is based on a similar principle, but it has a different configuration that affords improved flow of working fluid, simplified construction, reduced weight, and faster recovery from a frozen condition.

  8. Influence of freezing and thawing cycles on mechanical properties of closed-cell expanded perlite cemented soil%冻融循环对闭孔珍珠岩水泥土力学性能的影响

    Institute of Scientific and Technical Information of China (English)

    侯宇慧; 申向东

    2013-01-01

    Due to poor frost resistance of cement-soil,in permafrost and seasonal frozen soil area,the application and promotion of the soil cement is subject to a certain limit,how to improve the strength and durability of cemented soil in repeated freeze-thaw conditions to ensure the service life of the engineering is the key for further promotion and application of soil cement material in cold regions.By adding closed-cell expanded perlite in cemented soil,it came to the strength that cemented soil in different closed-cell expanded perlite under freezing and thawing cycles,analyzed the effect of freezing and thawing cycles on closed-cell expanded perlite and the changes before and after freezing and thawing cycles closed-cell expanded perlite cemented soil.The cemented soil adding closed-cell expanded perlite are preliminary analyzed.%由于水泥土抗冻性能较差,在多年冻土和季节性冻土地区,水泥土的应用和推广受到了一定的限制,如何提高反复冻融条件下水泥土的强度和耐久性,保证工程的使用寿命,是水泥土材料在寒冷地区进一步推广应用的关键.通过在水泥土中加入闭孔珍珠岩,得出水泥土在不同闭孔珍珠岩掺量下冻融循环后的强度值,分析了冻融循环次数对闭孔珍珠岩水泥土性能的影响及冻融循环前后闭孔珍珠岩水泥土强度变化,对掺入闭孔珍珠岩的水泥土做了初步的机理分析.

  9. Performance Characteristics of an Isothermal Freeze Valve

    Energy Technology Data Exchange (ETDEWEB)

    Hailey, A.E.

    2001-08-22

    This document discusses performance characteristics of an isothermal freeze valve. A freeze valve has been specified for draining the DWPF melter at the end of its lifetime. Two freeze valve designs have been evaluated on the Small Cylindrical Melter-2 (SCM-2). In order to size the DWPF freeze valve, the basic principles governing freeze valve behavior need to be identified and understood.

  10. Studies on Freezing RAM Semen in Absence of Glycerol.

    Science.gov (United States)

    Abdelnaby, Abdelhady Abdelhakeam

    1988-12-01

    Glycerol is widely used as a major cryoprotective agent for freezing spermatozoa of almost all species. However, it reduces fertility of sheep inseminated cervically compared with intrauterine insemination. Studies were conducted to develop a method and procedure for freezing ram semen in the absence of glycerol. Post -thaw survival of ram spermatozoa frozen in the absence of glycerol was affected by time and temperature after collection and before dilution and time after dilution and before freezing. Increase in time at 5^ circC before or after dilution and before freezing increased both post-thaw motility and number of cells passing through Sephadex filter. A cold dilution method was developed. Slow cooling of fresh ram semen and diluting at 5^circ C 2-3 hr. after collection, then freezing 1 hr. after dilution improved both post-thaw motility and number of cells passing through Sephadex filter compared with immediate dilution at 30-37^circC after collection and freezing 3-4 hr. later (P < 0.05). An extender was developed to freeze ram semen in the absence of glycerol. An increase in post-thaw motility was obtained when semen was extended in TES titrated with Tris to pH 7.0 (TEST) and osmotic pressure of 375-400 mOsm/kg, containing 25-30% (v/v) egg yolk and 10% (v/v) maltose. A special device (boat) for freezing was constructed to insure the same height of the sample above LN _2 and thus the same freezing rate from freeze to freeze. Freezing of semen in 0.25cc straws at 5-10 cm above LN_2 (73.8 to 49.5 ^circC/min) yielded higher post-thaw motility than the rates resulted from freezing at 15 cm above LN_2 or 1 cm above LN _2. Faster Thawing in 37^ circC water for 30 sec. (7.8^ circC/sec.) increased post-thaw motility compared with slower thawing in 5 or 20^circ C water (P < 0.05). A lambing rate of 52.2% was obtained in one fertility trial conducted with ram semen frozen without glycerol and 17.1% in a second trial. One injection (IM) of 15 mg PGF_{2alpha}/ewe for

  11. Understanding Slag Freeze Linings

    Science.gov (United States)

    Fallah-Mehrjardi, Ata; Hayes, Peter C.; Jak, Evgueni

    2014-09-01

    Slag freeze linings, the formation of protective deposit layers on the inner walls of furnaces and reactors, are increasingly used in industrial pyrometallurgical processes to ensure that furnace integrity is maintained in these aggressive, high-temperature environments. Most previous studies of freeze-linings have analyzed the formation of slag deposits based solely on heat transfer considerations. These thermal models have assumed that the interface between the stationary frozen layer and the agitated molten bath at steady-state deposit thickness consists of the primary phase, which stays in contact with the bulk liquid at the liquidus temperature. Recent experimental studies, however, have clearly demonstrated that the temperature of the deposit/liquid bath interface can be lower than the liquidus temperature of the bulk liquid. A conceptual framework has been proposed to explain the observations and the factors influencing the microstructure and the temperature of the interface at steady-state conditions. The observations are consistent with a dynamic steady state that is a balance between (I) the rate of nucleation and growth of solids on detached crystals in a subliquidus layer as this fluid material moves toward the stagnant deposit interface and (II) the dissolution of these detached crystals as they are transported away from the interface by turbulent eddies. It is argued that the assumption that the interface temperature is the liquidus of the bulk material represents only a limiting condition, and that the interface temperature can be between T liquidus and T solidus depending on the process conditions and bath chemistry. These findings have implications for the modeling approach and boundary conditions required to accurately describe these systems. They also indicate the opportunity to integrate considerations of heat and mass flows with the selection of melt chemistries in the design of future high temperature industrial reactors.

  12. Nonenzymatic cryogenic isolation of therapeutic cells: novel approach for enzyme-free isolation of pancreatic islets using in situ cryopreservation of islets and concurrent selective freeze destruction of acinar tissue.

    Science.gov (United States)

    Taylor, Michael J; Baicu, Simona C

    2014-01-01

    Cell-based therapies, which all involve processes for procurement and reimplantation of living cells, currently rely upon expensive, inconsistent, and even toxic enzyme digestion processes. A prime example is the preparation of isolated pancreatic islets for the treatment of type 1 diabetes by transplantation. To avoid the inherent pitfalls of these enzymatic methods, we have conceptualized an alternative approach based on the hypothesis that cryobiological techniques can be used for differential freeze destruction of the pancreas (Px) to release islets that are selectively cryopreserved in situ. Pancreata were procured from juvenile pigs using approved procedures. The concept of cryoisolation is based on differential processing of the pancreas in five stages: 1) infiltrating islets in situ preferentially with a cryoprotectant (CPA) cocktail via antegrade perfusion of the major arteries; 2) retrograde ductal infusion of water to distend the acinar; 3) freezing the entire Px solid to dithizone for identification of intact islets and with Syto 13/PI for fluorescence viability testing and glucose-stimulated insulin release assessment. As predicted, the cryoisolate contained small fragments of residual tissue comprising an amorphous mass of acinar tissue with largely intact and viable (>90%) embedded islets. Islets were typically larger (range 50-500 µm diameter) than their counterparts isolated from juvenile pigs using conventional enzyme digestion techniques. Functionally, the islets from replicate cryoisolates responded to a glucose challenge with a mean stimulation index = 3.3 ± 0.7. An enzyme-free method of islet isolation relying on in situ cryopreservation of islets with simultaneous freeze destruction of acinar tissue is feasible and proposed as a new and novel method that avoids the problems associated with conventional collagenase digestion methods. PMID:23992741

  13. Influence of dough freezing on Saccharomyces cerevisiae metabolism

    OpenAIRE

    Pejin Dušanka J.; Došanović Irena S.; Popov Stevan D.; Suturović Zvonimir J.; Ranković Jovana A.; Dodić Siniša N.; Dodić Jelena M.; Vučurović Vesna M.

    2007-01-01

    The need to freeze dough is increasing in bakery production. Frozen dough can be stored for a long time without quality change. The capacity of bakery production can be increased in this way, and in the same time, the night shifts can be decreased. Yeast cells can be damaged by freezing process resulting in poor technological quality of dough after defrostation (longer fermentation of dough). The influence of frozen storage time of dough on survival percentage of Saccharomyces cerevisiae was ...

  14. 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. PMID:23036662

  15. Fundamentals of freeze-drying.

    Science.gov (United States)

    Nail, Steven L; Jiang, Shan; Chongprasert, Suchart; Knopp, Shawn A

    2002-01-01

    Given the increasing importance of reducing development time for new pharmaceutical products, formulation and process development scientists must continually look for ways to "work smarter, not harder." Within the product development arena, this means reducing the amount of trial and error empiricism in arriving at a formulation and identification of processing conditions which will result in a quality final dosage form. Characterization of the freezing behavior of the intended formulation is necessary for developing processing conditions which will result in the shortest drying time while maintaining all critical quality attributes of the freeze-dried product. Analysis of frozen systems was discussed in detail, particularly with respect to the glass transition as the physical event underlying collapse during freeze-drying, eutectic mixture formation, and crystallization events upon warming of frozen systems. Experiments to determine how freezing and freeze-drying behavior is affected by changes in the composition of the formulation are often useful in establishing the "robustness" of a formulation. It is not uncommon for seemingly subtle changes in composition of the formulation, such as a change in formulation pH, buffer salt, drug concentration, or an additional excipient, to result in striking differences in freezing and freeze-drying behavior. With regard to selecting a formulation, it is wise to keep the formulation as simple as possible. If a buffer is needed, a minimum concentration should be used. The same principle applies to added salts: If used at all, the concentration should be kept to a minimum. For many proteins a combination of an amorphous excipient, such as a disaccharide, and a crystallizing excipient, such as glycine, will result in a suitable combination of chemical stability and physical stability of the freeze-dried solid. Concepts of heat and mass transfer are valuable in rational design of processing conditions. Heat transfer by conduction

  16. Effect of freezing on rabbit cultured chondrocytes

    Directory of Open Access Journals (Sweden)

    R.R Filgueiras

    2011-02-01

    Full Text Available This work evaluated the effect of freezing on chondrocytes maintained in culture, aiming the establishment of a cell bank for future application as heterologous implant. Chondrocytes extracted from joint cartilage of nine healthy New Zealand White rabbits were cultivated and frozen with the cryoprotector 5% dimethylsulfoxide for six months. Phenotypic and scanning electron microscopy analyses were carried out to identify morphological and functional differences between fresh and thawed cells. After enzymatic digestion, a total of 4.8x10(5cells per rabbit were obtained. Fresh chondrocytes showed a high mitotic rate and abundant matrix was present up to 60 days of culture. Loss of phenotypic stability was notable in the thawed chondrocytes, with a low labeling of proteoglycans and weak immunostaining of type II collagen. The present study showed important loss of chondrocyte viability under the freezing conditions. For future in vivo studies of heterologous implant, these results suggests that a high number of cells should be implanted in the host site in order to achieve an adequate number of viable cells. Furthermore, the chondrocytes should be implanted after two weeks of culture, when the highest viability rate is found

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

    Science.gov (United States)

    Nah, Gyoungju; Lee, Moonsub; Kim, Do-Soon; Rayburn, A Lane; Voigt, Thomas; Lee, D K

    2016-01-01

    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. PMID:27032112

  18. Freeze-fracture study of Trichomonas vaginalis

    Directory of Open Access Journals (Sweden)

    Marlene Benchimol

    1990-12-01

    Full Text Available The freeze-fracture technique was used to analyse the organization of the plasma membrane, as well as membranes of cytoplasmic organelles, of the pathogenic protozoan Trichomonas vaginalis. Rosettes formed by 4 to 14 intramembranous particles were seen on the fracture faces of the membrane lining the anterior flagella as well as in fracture faces of the plasma membrane enclosing the anterior region of the protozoan and in cytoplasmic organelles. Special organization of the membrane particles were also seen in the region of association of the recurrent flagellum to the cell body.

  19. Freeze-in through portals

    CERN Document Server

    Blennow, Mattias; Zaldivar, Bryan

    2014-01-01

    The popular freeze-out paradigm for Dark Matter (DM) production, relies on DM-baryon couplings of the order of the weak interactions. However, different search strategies for DM have failed to provide a conclusive evidence of such (non-gravitational) interactions, while greatly reducing the parameter space of many representative models. This motivates the study of alternative mechanisms for DM genesis. In the freeze-in framework, the DM is slowly populated from the thermal bath while never reaching equilibrium. In this work, we analyse in detail the possibility of producing a frozen-in DM via a mediator particle which acts as a portal. We give analytical estimates of different freeze-in regimes and support them with full numerical analyses, taking into account the proper distribution functions of bath particles. Finally, we constrain the parameter space of generic models by requiring agreement with DM relic abundance observations.

  20. Freeze Protection in Gas Holders

    DEFF Research Database (Denmark)

    Hjorth, Poul G.; Duursma, Gail

    In cold weather, the water seals of gasholders need protection from freez- ing to avoid compromising the seal. These holders have a large reservoir of “tank water” at the base which is below ground. At present freeze- protection is achieved by external heating of the seal water which...... is in a slotted channel called a cup. Electrical heating or circulation of heated tank water to the cup are examples of systems presently used. The tank water has a large thermal capacity and National Grid wishes to inves- tigate whether circulation of the tank water without external heating could provide...... sufficient energy input to avoid freezing. Only tanks in which the tank water is below ground are investigated in the report. The soil temperature under the reservoir at depth of 10m and lower is almost constant....

  1. 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. PMID:26268795

  2. Factors influencing the outcome of embryo freezing and Ihawing program

    Institute of Scientific and Technical Information of China (English)

    叶英辉; 金帆; 徐晨明; 邢兰凤

    2002-01-01

    Objective: To investigate the factors that might influence the sucess of an ernbryo freezing and thawing program.Method: The relationship between the pregnancy rate in 73 cycles of embryo freezing and thewing program and the following factors was analyzed;matermal age,E2 level at the time of HCG trigger,embryo storage time,number of thawed embryos transferred,presence of sponsoring embryos and intact embryos.And the suvival rate of thawed embryos with different morphology,cell stage and storage time was evaluated.Result:Transfer with three of more than three thawed embryos resulted in pragnancy rates of 38.5% and 35.7%,respectively.compared with 5.3% for transfer of fewer than three embryos.The presence of sponsoring embryos and intact embryos significantly incresses pregnancy rate in embryo freezing and thawing program .No other factor examined had any effect on pregnancy outcome.The survival rate of good morphology embryos was higher than poor ones,but was not influenced by cell stage and storage time.Conclusion:Embryo morphology before freezing , number of thawed embryos transferred and the presence of intact embryos are important to the outcome of embryo freezing and thawing program.

  3. Factors influencing the outcome of embryo freezing and thawing program

    Institute of Scientific and Technical Information of China (English)

    叶英辉; 金帆; 徐晨明; 邢兰凤

    2002-01-01

    Objective: To investigate the factors that might influence the succ ess of an embryo freezing and thawing program. Method: The relationship betwee n the pregnancy rate in 73 cycles of embryo freezing and thawing program and the following factors was analyzed: maternal age, E2 level at the time of HCG trigg er, embryo storage time, number of thawed embryos transferred, presence of spons oring embryos and intact embryos. And the survival rate of thawed embryos with d ifferent morphology, cell stage and storage time was evaluated. Result: Tra nsfer with three or more than three thawed embryos resulted in pregnancy rates o f 38.5% and 35.7%, respectively, compared with 5.3% for transfer of fewer th an t hree embryos. The presence of sponsoring embryos and intact embryos significantl y increases pregnancy rate in embryo freezing and thawing program. No other fact or examined had any effect on pregnancy outcome. The survival rate of good morph ology embryos was higher than poor ones, but was not influenced by cell stage an d storage time. Conclusion: Embryo morphology before freezing, number of thaw ed embryos transferred and the presence of intact embryos are important to the o utcome of embryo freezing and thawing program.

  4. Ice adhesions in relation to freeze stress.

    Science.gov (United States)

    Olien, C R; Smith, M N

    1977-10-01

    In freezing, competitive interaction between ice and hydrophilic plant substances causes an energy of adhesion to develop through the interstitial liquid. The thermodynamic basis for the adhesion energy is discussed, with estimates of the energies involved. In this research, effects of adhesion energy were observed microscopically in conjunction with energies of crystallization and frost desiccation. The complex character of ice in intact crown tissue of winter barley (Hordeum vulgare L.) and the problems of sectioning frozen tissue without producing artifacts led to an alternative study of single barley cells in a mesh of ice and cell wall polymers. Adhesions between ice, cell wall polymers, and the plasmalemma form a complexly interacting system in which the pattern of crystallization is a major factor in determination of stress and injury. PMID:16660124

  5. Response of Cytoskeleton of Murine Osteoblast Cultures to Two-step Freezing

    Institute of Scientific and Technical Information of China (English)

    Bao-Lin LIU; John McGRATH

    2005-01-01

    Understanding the ultrastructural response of cells to the freezing process is important for designing cryopreservation strategies for cells and tissues. The cellular structures of attached cells are targets of cryopreservation-induced damage. Specific fluorescence staining was used to assess the status of the actin filaments (F-actin) of murine osteoblasts attached to hydroxyapatite discs and plastic coverslips for a two-step freezing process. The F-actin of dead cells was depolymerized and distorted in the freezing process,whereas that of live cells had little change. The results suggest that the cytoskeleton may support the robustness of cells during cryopreservation. The present study helps to investigate the damage mechanism of attached cells during the freezing process.

  6. Time dependence of immersion freezing

    Directory of Open Access Journals (Sweden)

    A. Welti

    2012-05-01

    Full Text Available The time dependence of immersion freezing was studied for temperatures between 236 K and 243 K. Droplets with single immersed, size-selected 400 nm and 800 nm kaolinite particles were produced at 300 K, cooled down to supercooled temperatures typical for mixed-phase cloud conditions, and the fraction of frozen droplets with increasing residence time was detected. To simulate the conditions of immersion freezing in mixed-phase clouds we used the Zurich Ice Nucleation Chamber (ZINC and its vertical extension, the Immersion Mode Cooling chAmber (IMCA. We observed that the frozen fraction of droplets increased with increasing residence time in the chamber. This suggests that there is a time dependence of immersion freezing and supports the importance of a stochastic component in the ice nucleation process. The rate at which droplets freeze was observed to decrease towards higher temperatures and smaller particle sizes. Comparison of the laboratory data with four different ice nucleation models, three based on classical nucleation theory with different representations of the particle surface properties and one singular, suggest that the classical, stochastic approach combined with a distribution of contact angles is able to reproduce the ice nucleation observed in these experiments most accurately. Using the models to calculate the increase in frozen fraction at typical mixed-phase cloud temperatures over an extended period of time, yields an equivalent effect of −1 K temperature shift and an increase in time scale by a factor of ~10.

  7. A Mathematical Model for Freeze-Drying

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Based on the experiments on freeze-drying carrot and potato slabs, the effects of some parameters, such as heating temperature and pressure on the freeze-drying process are examined. A simple model of freeze-drying is established to predict drying time and the mass variations of materials during the drying. The experimental results agree well with those calculated by the model.

  8. Asymmetric Melting and Freezing Kinetics in Silicon.

    OpenAIRE

    Aziz, Michael; Tsao, Jeff Y.; Thompson, Michael O.; Peercy, Paul S.

    1986-01-01

    We report measurements of the melting velocity of amorphous Si relative to that of (100) crystalline Si. These measurements permit the first severe experimental test of theories describing highly nonequilibrium freezing and melting. The results indicate that freezing in Si is inherently slower than melting; this asymmetry can be interpreted in terms of an entropy-related reduction in the freezing rate.

  9. 9 CFR 590.534 - Freezing facilities.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Freezing facilities. 590.534 Section 590.534 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG..., and Facility Requirements § 590.534 Freezing facilities. (a) Freezing rooms, either on or off...

  10. 9 CFR 590.536 - Freezing operations.

    Science.gov (United States)

    2010-01-01

    ... 9 Animals and Animal Products 2 2010-01-01 2010-01-01 false Freezing operations. 590.536 Section 590.536 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE EGG..., and Facility Requirements § 590.536 Freezing operations. (a) Freezing rooms shall be kept clean...

  11. 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.

  12. Fusion of liposomes with the plasma membrane of epithelial cells: Fate of incorporated lipids as followed by freeze fracture and autoradiography of plastic sections

    NARCIS (Netherlands)

    Knoll, G.; Burger, K.N.J.; Bron, R.; van Meer, G.; Verkleij, A.J.

    1988-01-01

    The fusion of liposomes with the plasma membrane of influenza virus-infected monolayers of an epithelial cell line, Madin-Darby canine kidney cells (van Meer et al., 1985. Biochemistry, 24: 3593-3602), has been analyzed by morphological techniques. The distribution of liposomal lipids over the apica

  13. Freeze Technology for Nuclear Applications - 13590

    Energy Technology Data Exchange (ETDEWEB)

    Rostmark, Susanne C.; Knutsson, Sven [Lulea University of Technology (Sweden); Lindberg, Maria [Studsvik Nuclear AB, 611 82 Nykoeping (Sweden)

    2013-07-01

    Freezing of soil materials is a complicated process of a number of physical processes: - freezing of pore water in a thermal gradient, - cryogenic suction causing water migration and - ice formation expanding pores inducing frost heave. Structural changes due to increase of effective stress during freezing also take place. The over consolidation gives a powerful dewatering/drying effect and the freeze process causes separation of contaminates. Artificial ground freezing (AGF is a well established technique first practiced in south Wales, as early as 1862. AGF is mostly used to stabilize tunnels and excavations. During the last ten years underwater applications of freeze technologies based on the AGF have been explored in Sweden. The technology can, and has been, used in many different steps in a remediation action. Freeze Sampling where undisturbed samples are removed in both soft and hard sediment/sludge, Freeze Dredging; retrieval of sediment with good precision and minimal redistribution, and Freeze Drying; volume reduction of contaminated sludge/sediment. The application of these technologies in a nuclear or radioactive environment provides several advantages. Sampling by freezing gives for example an advantage of an undisturbed sample taken at a specified depth, salvaging objects by freezing or removal of sludges is other applications of this, for the nuclear industry, novel technology. (authors)

  14. Advances of cell injury mechanism by freezing stress during processing of probiotic ice cream%冰淇淋中益生菌冷冻胁迫致菌体损伤机制研究进展

    Institute of Scientific and Technical Information of China (English)

    张健; 郑义; 朱金锦; 刘心怡; 杨贞耐

    2015-01-01

    冰淇淋是优良的益生菌载体,但其加工中的老化、凝冻和硬化等阶段性降温至冷冻的过程,会造成益生菌生理损伤,导致益生菌活力下降,功能性变差。近年,有关冰淇淋中冷冻胁迫致益生菌的损伤,冰淇淋中益生菌活性的保护,以及组学技术推动下的菌株损伤相关代谢研究取得了一定进展,为研究影响冰淇淋中益生菌存活能力、菌体细胞膜特性及代谢酶活性,明确冰淇淋基质中益生菌冷冻胁迫损伤的分子机制,建立冰淇淋加工和贮藏过程中益生菌的保护策略提供了可能,也为未来改善益生菌冰淇淋的功效和品质提供科学依据。%Ice cream is a favorable probiotic carrier. However, the viability and functions of probiotic strains tend to be lowered in ice cream processing, including aging, freezing and hardening. Recently, progress of researches about the probiotics injury, protection and their metabolic changes in ice cream based on omics techniques have been made, which provides the possibility to specify survival ability, membrane properties, and key molecules of metabolic enzyme activities of probiotics in ice cream. These studies also facilitate to uncover the molecular mechanism of cell injury caused by freezing stress during the processing of probiotic ice cream and establish protective strategies for probiotics during processing and storage of ice cream, and provide a scientific basis for the improvement of functionality and quality of such product.

  15. Contact freezing: a review of experimental studies

    Directory of Open Access Journals (Sweden)

    L. A. Ladino Moreno

    2013-10-01

    Full Text Available This manuscript compiles both theoretical and experimental information on contact freezing with the aim to better understand this potentially important but still not well quantified heterogeneous freezing mode. There is no complete theory that describes contact freezing and how the energy barrier has to be overcome to nucleate an ice crystal by contact freezing. Experiments on contact freezing conducted using the cold plate technique indicate that it can initiate ice formation at warmer temperatures than immersion freezing. Additionally, a qualitative difference in the freezing temperatures between contact and immersion freezing has been found using different instrumentation and different ice nuclei. There is a lack of data on collision rates in most of the reported data, which inhibits a quantitative calculation of the freezing efficiencies. Thus, new or modified instrumentation to study contact nucleation in the laboratory and in the field are needed to identify the conditions at which contact nucleation could occur in the atmosphere. Important questions concerning contact freezing and its potential role for ice cloud formation and climate are also summarized.

  16. Differential freezing resistance and photoprotection in C3 and C4 eudicots and grasses.

    Science.gov (United States)

    Liu, Mei-Zhen; Osborne, Colin P

    2013-05-01

    Globally, C4 plants dominate hot, open environments, but this general pattern is underpinned by important differences in the biogeography of C4 lineages. In particular, the species richness of C4 Poaceae (grasses) increases strongly with increasing temperature, whereas that of the major C4 eudicot group Chenopodiaceae correlates positively with aridity. Freezing tolerance is a crucial determinant of biogeographical relationships with temperature and is mediated by photodamage and cellular disruption by desiccation, but little is known about differences between C4 families. This study hypothesized that there is a greater risk of freezing damage via these mechanisms in C4 Poaceae than Chenopodiaceae, that freezing protection differs between the taxonomic groups, and that freezing tolerance of species is linked to arid habitat preference. Chlorophyll fluorescence, water relations, and freezing injury were compared in four C3 and six C4 species of Poaceae and Chenopodiaceae from the same Mongolian flora. Contrary to expectations, freezing-induced leaf mortality and photodamage were lower in Poaceae than Chenopodiaceae species, and unrelated to photosynthetic pathway. The freezing resistance of Poaceae species resulted from constitutive protection and cold acclimation and an ability to protect the photosynthetic apparatus from photodamage. Freezing protection was associated with low osmotic potential and low tissue elasticity, and freezing damage was accompanied by electrolyte leakage, consistent with cell-membrane disruption by ice. Both Chenopodiaceae and Poaceae had the potential to develop cold acclimation and withstand freezing during the growing season, which conflicted with the hypothesis. Instead, freezing tolerance was more closely associated with life history and ecological preference in these Mongolian species. PMID:23599273

  17. Characteristics of sugar surfactants in stabilizing proteins during freeze-thawing and freeze-drying.

    Science.gov (United States)

    Imamura, Koreyoshi; Murai, Katsuyuki; Korehisa, Tamayo; Shimizu, Noriyuki; Yamahira, Ryo; Matsuura, Tsutashi; Tada, Hiroko; Imanaka, Hiroyuki; Ishida, Naoyuki; Nakanishi, Kazuhiro

    2014-06-01

    Sugar surfactants with different alkyl chain lengths and sugar head groups were compared for their protein-stabilizing effect during freeze-thawing and freeze-drying. Six enzymes, different in terms of tolerance against inactivation because of freeze-thawing and freeze-drying, were used as model proteins. The enzyme activities that remained after freeze-thawing and freeze-drying in the presence of a sugar surfactant were measured for different types and concentrations of sugar surfactants. Sugar surfactants stabilized all of the tested enzymes both during freeze-thawing and freeze-drying, and a one or two order higher amount of added sugar surfactant was required for achieving protein stabilization during freeze-drying than for the cryoprotection. The comprehensive comparison showed that the C10-C12 esters of sucrose or trehalose were the most effective through the freeze-drying process: the remaining enzyme activities after freeze-thawing and freeze-drying increased at the sugar ester concentrations of 1-10 and 10-100 μM, respectively, and increased to a greater extent than for the other surfactants at higher concentrations. Results also indicate that, when a decent amount of sugar was also added, the protein-stabilizing effect of a small amount of sugar ester through the freeze-drying process could be enhanced.

  18. Freezing of testicular tissue as a minced suspension preserves sperm quality better than whole-biopsy freezing when glycerol is used as cryoprotectant.

    Science.gov (United States)

    Crabbé, E; Verheyen, G; Tournaye, H; Van Steirteghem, A

    1999-02-01

    Frozen-thawed testicular spermatozoa have been used successfully for ICSI, especially in cases of obstructive azoospermia with normal spermatogenesis. Fewer attempts, however, have been made to check whether these rather immature spermatozoa, in a different environment with several other cell types present, have cryobiological requirements other than those of ejaculated spermatozoa. This is the reason why the freezing protocols and cryoprotectants (glycerol) used for freezing testicular tissue are based on experience with semen freezing. This study aimed to assess whether cryosurvival and/or motility was influenced by freezing of testicular tissue either as an intact biopsy or as a shredded tissue suspension, when glycerol was used as cryoprotectant. Freezing of testicular tissue as a suspension preserved motility (type B + C) significantly better than freezing of whole biopsies (9.2% vs. 4.0%). Similar observations have been made for vitality (39.3% vs. 25.4%). Centrifugation on 50% Percoll in order to remove the cryoprotectant resulted in a huge loss of spermatozoa (or late spermatids) and should therefore be especially avoided in cases of testicular failure. On the basis of these observations, mincing of the testicular biopsies before freezing may be advocated. Testicular spermatozoa seem to be better preserved when frozen in suspension, at least when slowly permeating glycerol is used as a cryoprotectant. PMID:10068943

  19. 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)

  20. Atmospheric freeze drying assisted by power ultrasound

    OpenAIRE

    Santacatalina Bonet, Juan Vicente; Carcel Carrión, Juan Andrés; García Pérez, José Vicente; Mulet Pons, Antonio; Simal, S.

    2012-01-01

    [EN] Atmospheric freeze drying (AFD) is considered an alternative to vacuum freeze drying to keep the quality of fresh product. AFD allows continuous drying reducing fix and operating costs, but presents, as main disadvantage, a long drying time required. The application of power ultrasound (US) can accelerate AFD process. The main objective of the present study was to evaluate the application of power ultrasound to improve atmospheric freeze drying of carrot. For that pur...

  1. Fundamental Technical Elements of Freeze-fracture/Freeze-etch in Biological Electron Microscopy

    Science.gov (United States)

    Freeze-fracture/freeze-etch describes a process whereby specimens, typically biological or nanomaterial in nature, are frozen, fractured, and replicated to generate a carbon/platinum "cast" intended for examination by transmission electron microscopy. Specimens are subjected to u...

  2. Issues in Freeze Drying of Aqueous Solutions

    Institute of Scientific and Technical Information of China (English)

    王维; 陈墨; 陈国华

    2012-01-01

    Freeze drying or lyophilization of aqueous solutions is widely used in pharmaceutical industry. The in-creased importance Of the process is gaining a worldwide interest of research. A growing body of literature has demonstrated that the scientific approach can result in improved product quality with minimum trial and error em-piricism. Formulation and process development need a systematical understanding of the physical chemistry of freezing and freeze drying, material science and mechanisms of heat and mass transfer. This paper presents an overview on freeze ding of aqueous solutions based on publications in the past few decades. The important issuesof the process are analyzed.

  3. When hot water freezes before cold

    CERN Document Server

    Katz, J I

    2006-01-01

    I suggest that the origin of the Mpemba effect (the freezing of hot water before cold) is freezing-point depression by solutes, either gaseous or solid, whose solubility decreases with increasing temperature so that they are removed when water is heated. They are concentrated ahead of the freezing front by zone refining in water that has not been heated, reduce the temperature of the freezing front, and thereby reduce the temperature gradient and heat flux, slowing the progress of the front. I present a simple calculation of this effect, and suggest experiments to test this hypothesis.

  4. Effect of freezing and thawing on UMTRA covers

    International Nuclear Information System (INIS)

    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

  5. Improvement of parameters of freezing medium and freezing protocol for bull sperm using two osmotic supports.

    NARCIS (Netherlands)

    Chaveiro, A.; Machado, A.L.; Frijters, A.; Engel, B.; Woelders, H.

    2006-01-01

    The aim of this study was to improve the freezing protocol of bull sperm, by investigating the influence on sperm viability after freeze/thawing of different freezing medium components, as well as the effect of cooling rates in the different stages of the cooling protocol, in single factor experimen

  6. Comparison of three-dimensional printing and vacuum freeze-dried techniques for fabricating composite scaffolds.

    Science.gov (United States)

    Sun, Kai; Li, Ruixin; Jiang, Wenxue; Sun, Yufu; Li, Hui

    2016-09-01

    In this study, the performances of different preparation methods of the scaffolds were analyzed for chondrocyte tissue engineering. Silk fibroin/collagen (SF/C) was fabricated using a vacuum freeze-dried technique and by 3D printing. The porosity, water absorption expansion rates, mechanical properties, and pore sizes of the resulting materials were evaluated. The proliferation and metabolism of the cells was detected at different time points using an MTT assay. Cell morphologies and distributions were observed by histological analysis and scanning electron microscopy (SEM). The porosity, water absorption expansion rate, and Young's modulus of the material obtained via 3D printing were significantly higher than those obtained by the freeze-dried method, while the pore size did not differ significantly between the two methods. MTT assay results showed that the metabolism of cells seeded on the 3D printed scaffolds was more viable than the metabolism on the freeze-dried material. H&E staining of the scaffolds revealed that the number of cells in the 3D printed scaffold was higher in comparison to a similar measurement on the freeze-dried material. Consequently, stem cells grew well inside the 3D printed scaffolds, as measured by SEM, while the internal structure of the freeze-dried scaffold was disordered. Compared with the freeze-dried technique, the 3D printed scaffold exhibited better overall performance and was more suitable for cartilage tissue engineering. PMID:27404126

  7. Mechanisms of deterioration of nutrients. [of freeze dried foods

    Science.gov (United States)

    Karel, M.; Flink, J. M.

    1976-01-01

    Methods which produce freeze dried foods of improved quality were examined with emphasis on storage stability. Specific topics discussed include: microstructure of freeze dried systems, investigation of structural changes in freeze dried systems, artificial food matrices, osmotic preconcentration to yield improved quality freeze dried fruits, and storage stability of osmotically preconcentrated freeze dried fruits.

  8. -80℃非程控冷冻保存人外周血干细胞的效果观察%Cryopreservation of human peripheral blood stem cells by-80℃non rate-controlled freezing method

    Institute of Scientific and Technical Information of China (English)

    裴仁治; 岑东; 王奇军; 刘旭辉; 罗建平; 石兆玲

    2008-01-01

    Objective To investigate the cryopreservation of human peripheraI blood stern cells (PBSC)by-80℃non rate-controlled freezing method.Methods Forty-one samples of PBSC were obtained from 39 malignant tumor patients and 2 nornlal donors.PBSC in all malignant tumor patients were mobilized with special cherootherapy protocols combined with granulocyte colony-stimulating factor or granulocyte/monocyte colony-stimulating factor before collection.The obtained PBSC were mixed with same volume of cryoprotectant composed of 12 g/L hydroxyethyl starch,10% dimethylsulfoxide and 80 g/L human serunl albumin,and then eryopreserved in-80℃deep freeze refrigerator directly.Trypan blue viability and recovery rate of nucleated cells(NC),mononuclear cells(MNC),CD34+cells(CD34+),colony-forming unit-granulocyte macrophages (CFU-GM),burst forming unit-erythroid(BFU-E) were detected before and after cryopreservation in different periods.Results Having been cryopreserved from 1 month to 10 years in-80℃.there were no statistically significant differences in trypan blue viability,NC and MNC recovery rate(P>0.05).AIthough recovery rate of CD34+, CFU-M and BFU-E was decreased obviously after cryopreservation for 5-10 years(P<0.05),10-year recovery rate was 89.6%,85.1%and 83.7%,respectively.Thirty-four patients subject to corresponding pretreatment were transfused with-80℃ cryopreserved PBSC which had been stored for 13 to 35 days(mean 19 days)and hematological reconstitution was obtained successfully in 11 to 27 days (mean 14.7 days).Conclusion -80℃non rate-controlled freezing method is suitable to human PBSC long-term cryopreservation with the cryoproteetant composed of 60 g/L hydroxyethyl starch,5%dimethylsulfoxide and 40 g/L human serum albumin,and its cryopreservation effect is ideal%目的 探讨-80℃非程控冷冻保存人外周血干细胞(PBSC)的效果.方法 41份PBSC标本,39份采自恶性肿瘤患者,2份采自正常供者.对于恶性肿瘤患者,干细胞采集前

  9. Anhydrobiosis and freezing-tolerance: adaptations that facilitate the establishment of Panagrolaimus nematodes in polar habitats.

    Science.gov (United States)

    McGill, Lorraine M; Shannon, Adam J; Pisani, Davide; Félix, Marie-Anne; Ramløv, Hans; Dix, Ilona; Wharton, David A; Burnell, Ann M

    2015-01-01

    Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode isolated from Ross Island Antarctica, can survive intracellular ice formation when fully hydrated. A capacity to survive freezing while fully hydrated has also been observed in some other Antarctic nematodes. We experimentally determined the anhydrobiotic and freezing-tolerance phenotypes of 24 Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth of ice crystals. We show that P. davidi belongs to a clade of anhydrobiotic and freezing-tolerant panagrolaimids containing strains from temperate and continental regions and that P. superbus, an early colonizer at Surtsey island, Iceland after its volcanic formation, is closely related to a species from Pennsylvania, USA. Ancestral state reconstructions show that anhydrobiosis evolved deep in the phylogeny of Panagrolaimus. The early-diverging Panagrolaimus lineages are strongly anhydrobiotic but weakly freezing-tolerant, suggesting that freezing tolerance is most likely a derived trait. The common ancestors of the davidi and the superbus clades were anhydrobiotic and also possessed robust freezing tolerance, along with a capacity to inhibit the growth and recrystallization of ice crystals. Unlike other endemic Antarctic nematodes, the life history traits of P. davidi do not show evidence of an evolved response to polar conditions. Thus we suggest that the colonization of Antarctica by P. davidi and of Surtsey by P. superbus may be examples of recent "ecological fitting

  10. Anhydrobiosis and freezing-tolerance: adaptations that facilitate the establishment of Panagrolaimus nematodes in polar habitats.

    Directory of Open Access Journals (Sweden)

    Lorraine M McGill

    Full Text Available Anhydrobiotic animals can survive the loss of both free and bound water from their cells. While in this state they are also resistant to freezing. This physiology adapts anhydrobiotes to harsh environments and it aids their dispersal. Panagrolaimus davidi, a bacterial feeding anhydrobiotic nematode isolated from Ross Island Antarctica, can survive intracellular ice formation when fully hydrated. A capacity to survive freezing while fully hydrated has also been observed in some other Antarctic nematodes. We experimentally determined the anhydrobiotic and freezing-tolerance phenotypes of 24 Panagrolaimus strains from tropical, temperate, continental and polar habitats and we analysed their phylogenetic relationships. We found that several other Panagrolaimus isolates can also survive freezing when fully hydrated and that tissue extracts from these freezing-tolerant nematodes can inhibit the growth of ice crystals. We show that P. davidi belongs to a clade of anhydrobiotic and freezing-tolerant panagrolaimids containing strains from temperate and continental regions and that P. superbus, an early colonizer at Surtsey island, Iceland after its volcanic formation, is closely related to a species from Pennsylvania, USA. Ancestral state reconstructions show that anhydrobiosis evolved deep in the phylogeny of Panagrolaimus. The early-diverging Panagrolaimus lineages are strongly anhydrobiotic but weakly freezing-tolerant, suggesting that freezing tolerance is most likely a derived trait. The common ancestors of the davidi and the superbus clades were anhydrobiotic and also possessed robust freezing tolerance, along with a capacity to inhibit the growth and recrystallization of ice crystals. Unlike other endemic Antarctic nematodes, the life history traits of P. davidi do not show evidence of an evolved response to polar conditions. Thus we suggest that the colonization of Antarctica by P. davidi and of Surtsey by P. superbus may be examples of recent

  11. Viability of Two Freeze-dried Strains of Bifidobacterium Preparations at Various Temperatures during Prolonged Storage

    Institute of Scientific and Technical Information of China (English)

    DU Peng; DU Li-hui; HUO Gui-cheng

    2005-01-01

    Viability of bifidobacteria in freeze-dried probiotic products at various temperatures during prolonged storage was assessed. Bifidobacterium longum and Bifidobacterium infantis were freeze-dried. The freeze-dried preparations were stored at -18,4, and 20℃. Cell counts were enumerated using BS agar at 37℃ for 48 h under anaerobic conditions at 0, 45 and 120 days. Storage at 20℃ showed the greatest decline in the viability of bifidobacteria, whereas that at -18℃ showed the least decrease.

  12. Cooling and Freezing Behaviors of Aqueous Sodium Chloride Solution in a Closed Rectangular Container

    Science.gov (United States)

    Narumi, Akira; Kashiwagi, Takao; Nakane, Ichirou

    This paper investigates cooling and freezing behaviors of NaCl aqueous solution in a rectangular container equipped with horizontal partitions of micro porous film in order to determine the mechanisms of heat and mass transfer through cell wall for the purpose of freezing food. For comparison, experiments were performed using partitions of copper plate, no partition, and water. These processes were visualized and measured using real-time laser holographic interferometry. It was found that there was very little difference in the cooling process due to partitions, but that there were significant differences in freezing process when NaCl aqueous solution is used.

  13. Lipids differentially degraded during tissue freezing and thawing

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    @@ Plants cope with freezing and thawing by altering the lipid composition of their cell membranes. Such cellular responses go through three phases Successful test flight of an airship Researchers with the Balloon Aircraft Research Center (BARC) of the Academy of Opto-electronics, CAS, succeeded in their first test flight of an aeroboat with a flight altitude up to 1,000 meters and an effective payload of 20 kilograms in Shandong on 25 December, 2007.

  14. Freeze conditioning agents ease winter railcar unloading

    Energy Technology Data Exchange (ETDEWEB)

    Olson, G.E.

    1982-02-01

    A US midwest utility's freeze control programme is described. All coal is treated with a glycol-based freeze control agent. Some rail wagons were treated with a side release agent which stops coal sticking to the metal wagon. The use of calcium chloride or heat to thaw frozen coal is also discussed.

  15. Updating freeze: Aligning animal and human research

    NARCIS (Netherlands)

    Hagenaars, M.A.; Oitzl, M.S.; Roelofs, K.

    2014-01-01

    Freezing is widely used as the main outcome measure for fear in animal studies. Freezing is also getting attention more frequently in human stress research, as it is considered to play an important role in the development of psychopathology. Human models on defense behavior are largely based on anim

  16. Repeatability and randomness in heterogeneous freezing nucleation

    Directory of Open Access Journals (Sweden)

    G. Vali

    2008-08-01

    Full Text Available This study is aimed at clarifying the relative importance of the specific character of the nuclei and of the duration of supercooling in heterogeneous freezing nucleation by immersed impurities. Laboratory experiments were carried out in which sets of water drops underwent multiple cycles of freezing and melting. The drops contained suspended particles of mixtures of materials; the resulting freezing temperatures ranged from −6°C to −24°C. Rank correlation coefficients between observed freezing temperatures of the drops in successive runs were >0.9 with very high statistical significance, and thus provide strong support for the modified singular model of heterogeneous immersion freezing nucleation. For given drops, changes in freezing temperatures between cycles were relatively small (<1°C for the majority of the events. These frequent small fluctuations in freezing temperatures are interpreted as reflections of the random nature of embryo growth and are associated with a nucleation rate that is a function of a temperature difference from the characteristic temperatures of nuclei. About a sixth of the changes were larger, up to ±5°C, and exhibited some systematic patterns. These are thought to arise from alterations of the nuclei, some being permanent and some transitory. The results are used to suggest ways of describing ice initiation in cloud models that account for both the temperature and the time dependence of freezing nucleation.

  17. Repeatability and randomness in heterogeneous freezing nucleation

    Directory of Open Access Journals (Sweden)

    G. Vali

    2008-02-01

    Full Text Available This study is aimed at clarifying the relative importance of the specific character of the nuclei and of the duration of supercooling in heterogeneous freezing nucleation by immersed impurities. Laboratory experiments were carried out in which sets of water drops underwent multiple cycles of freezing and melting. The drops contained suspended particles of mixtures of materials; the resulting freezing temperatures ranged from −6°C to −24°C. Rank correlation coefficients between observed freezing temperatures of the drops in successive runs were >0.9 with very high statistical significance, and thus provide strong support for the modified singular model of heterogeneous immersion freezing nucleation. For given drops, changes in freezing temperatures between cycles were relatively small (<1°C for the majority of the events. These frequent small fluctuations in freezing temperatures are interpreted as reflections of the random nature of embryo growth and are associated with a nucleation rate that is a function of a temperature difference from the characteristic temperatures of nuclei. About a sixth of the changes were larger, up to ±5°C, and exhibited some systematic patterns. These are thought to arise from alterations of the nuclei, some being permanent and some transitory. The results are used to suggest ways of describing ice initiation in cloud models that account for both the temperature and the time dependence of freezing nucleation.

  18. Bioinspired Design: Magnetic Freeze Casting

    Science.gov (United States)

    Porter, Michael Martin

    Nature is the ultimate experimental scientist, having billions of years of evolution to design, test, and adapt a variety of multifunctional systems for a plethora of diverse applications. Next-generation materials that draw inspiration from the structure-property-function relationships of natural biological materials have led to many high-performance structural materials with hybrid, hierarchical architectures that fit form to function. In this dissertation, a novel materials processing method, magnetic freeze casting, is introduced to develop porous scaffolds and hybrid composites with micro-architectures that emulate bone, abalone nacre, and other hard biological materials. This method uses ice as a template to form ceramic-based materials with continuously, interconnected microstructures and magnetic fields to control the alignment of these structures in multiple directions. The resulting materials have anisotropic properties with enhanced mechanical performance that have potential applications as bone implants or lightweight structural composites, among others.

  19. Thermodynamics of freezing and melting.

    Science.gov (United States)

    Pedersen, Ulf R; Costigliola, Lorenzo; Bailey, Nicholas P; Schrøder, Thomas B; Dyre, Jeppe C

    2016-01-01

    Although the freezing of liquids and melting of crystals are fundamental for many areas of the sciences, even simple properties like the temperature-pressure relation along the melting line cannot be predicted today. Here we present a theory in which properties of the coexisting crystal and liquid phases at a single thermodynamic state point provide the basis for calculating the pressure, density and entropy of fusion as functions of temperature along the melting line, as well as the variation along this line of the reduced crystalline vibrational mean-square displacement (the Lindemann ratio), and the liquid's diffusion constant and viscosity. The framework developed, which applies for the sizable class of systems characterized by hidden scale invariance, is validated by computer simulations of the standard 12-6 Lennard-Jones system. PMID:27530064

  20. Freeze concentration of lime juice

    Directory of Open Access Journals (Sweden)

    Ampawan Tansakul

    2008-11-01

    Full Text Available The main objective of this research was to study the effects of processing conditions, i.e. cooling medium temperature (-6, -12 and -18C and scraper blade rotational speed (50, 100 and 150 rpm on the freeze concentration of lime juice. The initial soluble solid content of lime juice was 7.6 Brix. Results showed that soluble solid content of lime juice increased as cooling medium temperature decreased while scraper blade rotational speed increased. It was also found that the processing condition with -18˚C cooling medium temperature and 150 rpm rotational speed of the scraper blade was the best among all studied conditions, although the loss of the soluble solids with ice crystals during ice separation was relatively high at 35%.

  1. Impregnation of leather during "freeze-drying"

    DEFF Research Database (Denmark)

    Storch, Mikkel; Vestergaard Poulsen Sommer, Dorte; Hovmand, Ida;

    2016-01-01

    Freeze-drying is a recognized method for the preservation of waterlogged objects. Naturally, freeze-drying has also been used for waterlogged archaeological leather often after treatment with Na2.EDTA and impregnation with PEG; but the treated leather sometimes suffers from “excessive drying......” becoming too stiff and brittle. The aim of this study was to examine the effect of a conventional freeze-drying method against an alternative freeze-drying method that preserves the natural moisture content of the leather. Both new and archaeological waterlogged leather were included in the study...... and the leather samples were treated in one of four ways: pre-treatment with Na2EDTA, impregnation with PEG 400, pre-treatment with Na2EDTA followed by impregnation with PEG 400 as well as no treatment. After the treatments, the leather samples were freeze-dried either by the conventional or by the alternative...

  2. Food freezing with simultaneous surface dehydration: approximate prediction of freezing time

    Energy Technology Data Exchange (ETDEWEB)

    Campanone, Laura A.; Salvadori, Viviana O.; Mascheroni, Rodolfo H. [Centro de Investigacion Desarollo en Criotecnologia de Alimentos (CIDCA), Facultad de Ciencias Exactas, La Plata (Argentina); MODIAL, Facultad de Ingenieria, La Plata (Argentina)

    2005-03-01

    Freezing of unpackaged foods induces mass transfer in the form of surface ice sublimation, which in turn modifies heat transfer conditions. At present there are no simplified methods for predicting freezing times when surface dehydration occurs. This paper uses a previously developed model for the simulation of simultaneous heat and mass transfer during food freezing and storage to generate a complete set of predicted freezing times when dehydration occurs. Based on these data a simplified analytical method for the prediction of freezing time during freezing of unpackaged frozen foods was developed. The method accounts for product characteristics (shape, size and composition) and operating conditions (initial and refrigerant temperature, heat transfer coefficient, relative humidity). The prediction equation is very simple and results of its use - simulating usual freezing conditions for different products - shows very good accuracy when tested against the previously cited numerical model and all the available experimental data. (Author)

  3. Influence of dough freezing on Saccharomyces cerevisiae metabolism

    Directory of Open Access Journals (Sweden)

    Pejin Dušanka J.

    2007-01-01

    Full Text Available The need to freeze dough is increasing in bakery production. Frozen dough can be stored for a long time without quality change. The capacity of bakery production can be increased in this way, and in the same time, the night shifts can be decreased. Yeast cells can be damaged by freezing process resulting in poor technological quality of dough after defrostation (longer fermentation of dough. The influence of frozen storage time of dough on survival percentage of Saccharomyces cerevisiae was investigated. Dough samples were taken after 1, 7, 14 and 28 days of frozen storage at -20°C. After defrosting, at room temperature, samples were taken from the surface and the middle part of dough (under aseptic conditions, and the percentage of living S. cerevisiae cells was determined. During frozen storage of dough, the number of living S. cerevisiae decreased. After 28 days of frozen storage, the percentage of live cells on the surface and inside the dough was 53,1% and 54,95%, respectively. The addition of k-carragenan to dough increased the percentage of living cells in the middle part of dough up to 64,63%. Pure cultures, isolated from survived S. cerevisia cells in frozen dough by agar plates method (Koch's method, were multiplied in optimal liquid medium for yeasts. The content of cytochromes in S. cerevisiae cells was determined by spectrophotometric method. The obtained results showed that the content of cytochromes in survived S. cerevisiae cells was not affected by dough freezing process. Growth rate and fermentative activity (Einchor's method were determined in multiplied cells.

  4. The relationship of freeze tolerance with intracellular compounds in baker's yeasts.

    Science.gov (United States)

    Shi, Xiaojian; Miao, Yelian; Chen, Jie Yu; Chen, Jun; Li, Wenli; He, Xun; Wang, Jining

    2014-03-01

    Freeze-tolerant baker's yeasts are required for the processing of frozen doughs. The present study was carried out to investigate the cell survival rate after frozen storage and the change of fermentability in dough due to frozen storage, and to discuss quantitatively the relationship of freeze tolerance with intracellular trehalose, amino acids, and glycerol, using six types of baker's yeasts as the test materials. The experimental results showed that the fermentability of yeast cells in frozen dough was strongly correlated with the cell survival rate. The baker's yeast with a higher level of cell survival rate had a larger increase in the total intracellular compound content after frozen storage, and the cell survival rate increased linearly with increasing total intracellular compound content in frozen yeast cells. Trehalose was a primary compound affecting freeze tolerance, followed by glutamic acid, arginine, proline, asparagic acid, and glycerol. The basic information provided by the present study is useful for exploring the freeze-tolerance mechanisms of baker's yeast cells, breeding better freeze-tolerant baker's yeast strains, and developing more effective cryoprotectants. PMID:24482281

  5. Lipid composition of commercial bakers' yeasts having different freeze-tolerance in frozen dough.

    Science.gov (United States)

    Murakami, Y; Yokoigawa, K; Kawai, F; Kawai, H

    1996-11-01

    The lipid composition of some commercial bakers' yeasts having different freeze-sensitivity in frozen dough was investigated to clarify the correlation between their lipid composition and freeze-tolerance. The total lipid content including neutral lipid, free fatty acid, sterol, and phospholipid ranged between 23.0 to 32.2 mg/100 mg protein of the yeasts tested. Phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidylserine were the main phospholipids found in all yeast strains, but no distinct difference in these components between freeze-tolerant and freeze-sensitive strains was observed. Palmitoleic (C16:1), oleic (C18:1), palmitic (16:0), and stearic (C18:0) acids were the major fatty acids present in total lipid and phospholipid, and unsaturation indices of fatty acid in these lipid components were almost equal by the strains. The molar ratios of sterol to phospholipid of freeze-sensitive strains were higher than those of freeze-tolerant strains. The difference in the sterol-phospholipid ratio that influences the fluidity of plasma membranes in yeast cells was supposed to reflect the difference in freeze-sensitivity of bakers' yeast. PMID:8987866

  6. Development of Three-Layer Simulation Model for Freezing Process of Food Solution Systems

    Science.gov (United States)

    Kaminishi, Koji; Araki, Tetsuya; Shirakashi, Ryo; Ueno, Shigeaki; Sagara, Yasuyuki

    A numerical model has been developed for simulating freezing phenomena of food solution systems. The cell model was simplified to apply to food solution systems, incorporating with the existence of 3 parts such as unfrozen, frozen and moving boundary layers. Moreover, the moving rate of freezing front model was also introduced and calculated by using the variable space network method proposed by Murray and Landis (1957). To demonstrate the validity of the model, it was applied to the freezing processes of coffee solutions. Since the model required the phase diagram of the material to be frozen, the initial freezing temperatures of 1-55 % coffee solutions were measured by the DSC method. The effective thermal conductivity for coffee solutions was determined as a function of temperature and solute concentration by using the Maxwell - Eucken model. One-dimensional freezing process of 10 % coffee solution was simulated based on its phase diagram and thermo-physical properties. The results were good agreement with the experimental data and then showed that the model could accurately describe the change in the location of the freezing front and the distributions of temperature as well as ice fraction during a freezing process.

  7. Pulmonary delivery of an inulin-stabilized influenza subunit vaccine prepared by spray-freeze drying induces systemic, mucosal humoral as well as cell-mediated immune responses in BALB/c mice

    NARCIS (Netherlands)

    Amorij, J-P.; Saluja, V.; Petersen, A.H.; Hinrichs, W.L.J.; Huckriede, A.; Frijlink, H.W.

    2007-01-01

    In this study pulmonary vaccination with a new influenza subunit vaccine powder was evaluated. Vaccine powder was produced by spray-freeze drying (SFD) using the oligosaccharide inulin as stabilizer. Immune responses after pulmonary vaccination of BALB/c mice with vaccine powder were determined and

  8. Exploring the Nature of Contact Freezing

    Science.gov (United States)

    Kiselev, A. A.; Hoffmann, N.; Duft, D.; Leisner, T.

    2012-12-01

    The freezing of supercooled water droplets upon contact with aerosol particles (contact nucleation of ice) is the least understood mechanism of ice formation in atmospheric clouds. Although experimental evidences suggest that some aerosols can be better IN in the contact than in the immersion mode (that is, triggering ice nucleation at higher temperature), no final explanation of this phenomena currently exists. On the other hand, the contact freezing is believed to be responsible for the enhanced rate of secondary ice formation occasionally observed in LIDAR measurements in the cold mixed phase clouds. Recently we have been able to show that the freezing of supercooled droplets electrodynamically levitated in the laminar flow containing mineral dust particles (kaolinite) is a process solely governed by a rate of collisions between the supercooled droplet and the aerosol particles. We have shown that the probability of droplet freezing on a single contact with aerosol particle may differ over an order of magnitude for kaolinite particles having different genesis and morphology. In this presentation we extend the study of contact nucleation of ice and compare the IN efficiency measured for DMA-selected kaolinite, illite and hematite particles. We show that the freezing probability increases towards unity as the temperature decreases and discuss the functional form of this temperature dependence. We explore the size dependence of the contact freezing probability and show that it scales with the surface area of the particles, thus resembling the immersion freezing behavior. However, for all minerals investigated so far, the contact freezing has been shown to dominate over immersion freezing on the short experimental time scales. Finally, based on the combined ESEM and electron microprobe analysis, we discuss the significance of particle morphology and variability of chemical composition on its IN efficiency in contact mode.

  9. Effect of the technology of high cell density cultivation and vacuum freeze-drying on cryotolerance property of lactic acid bacteria%高密度发酵和真空冷冻干燥工艺对乳酸菌抗冷冻性的影响

    Institute of Scientific and Technical Information of China (English)

    刘彩虹; 邵玉宇; 任艳; 孟和毕力格; 张和平

    2013-01-01

    经真空冷冻干燥得到的乳酸菌发酵剂存活率和后期的低温贮藏稳定性与诸多因素相关.本文综述了制备乳酸菌发酵剂过程中高密度发酵和真空冷冻干燥工艺的不同对乳酸菌抗冷冻性的影响,其中高密度发酵过程中的培养基组分、培养温度、发酵恒定pH、中和剂的选择、菌体收获时期和发酵结束后处理以及真空冷冻干燥过程中保护剂的添加、预冷冻处理等是影响乳酸菌抗冷冻性的重要因素.通过对这些相关因素的综述分析,为提高乳酸菌发酵剂的冻干存活率和后期的低温贮藏稳定性提供新的思路,且应用抗冷冻性强、活力高的乳酸菌发酵剂对有效提高乳制品的质量和企业的经济效益意义重大.%The survival rate and low temperature stability of lactic acid bacterial starter obtained by vacuum freeze-drying are governed by several factors. In this paper, the influence of the technology of high cell density cultivation and vacuum freeze-drying on cryotolerance of lactic acid bacteria for use as starters was analyzed. During fermentation, the following factors had a significant effect on the cryosurvival of lactic acid bacteria: culture medium, temperature control, pH stat, the neutralizer used, the harvesting stage of the cell crop, and post-fermentation handling of the concentrated cells. Factors affecting cell viability subjected to lyophilization include the following: cryoprotectants used, conditions used in initial freezing of the cell concentrate, and during vacuum freeze-drying. A good understanding of these factors will provide a reliable technology for preserving high cell density starter. The use of starter bacteria with high cryotolerance and viability can improve the quality of fermented milk products and boost economic benefits to the dairy industry.

  10. A new application of the SFDA-staining method to assessment of the freezing tolerance in leaves of alpine plants

    OpenAIRE

    Yamori,Wataru/Kogami,Hiroyuki/Masuzawa,Takehiro

    2006-01-01

    For the first time, this study used 5- (6-) sulfofluorescein diacetate (SFDA), a fluorescent product in plant cells converted by esterase activity to fluorescein-5- (and 6-) sulfonic acid (FSA), to assess the freezing tolerance of leaf cells. We were able to readily distinguish living and dead cells, and detect differences in freezing tolerance among five alpine plants using the SFDA-staining method. We also compared this method with two conventional methods, the electrolyte leakage test and ...

  11. Harnessing the energy accompanying freezing

    Energy Technology Data Exchange (ETDEWEB)

    Akyurt, M., E-mail: makyurt@kau.edu.s [Departments of Mechanical Engineering, King Abdulaziz University, Jeddah 21589 (Saudi Arabia); Tuerkmen, N. [Departments of Mechanical Engineering, King Abdulaziz University, Jeddah 21589 (Saudi Arabia)

    2011-05-15

    Research highlights: {yields} Ice pressurization allows the burst and leak testing of practically all tubular materials. {yields} The assembly can be made fully portable for maintenance operations without the use of liquid CO{sub 2} and N{sub 2}. {yields} Ice pressurization can work where conventional interference fitting, axial pressing and heat treatment fail. {yields} Uniform pressures can be developed in ice pressurization as opposed to Herzian distributions under plungers. -- Abstract: The progression of freezing of water inside a pipe is reviewed, with special emphasis on bursting. The process of pressure rise in confined bodies of water is discussed. The development of a method utilizing liquid carbon dioxide and liquid nitrogen, for the development of pressures inside closed containers is summarized. Then a novel method, utilizing mechanical refrigeration, is explained for the generation of high pressures. An experimental setup for the latter technique is described and results of experiments are summarized. A number of ways of utilizing the ice-pressurization technique are presented. Certain characteristics and advantages of ice-pressurization are enumerated as regards to burst and leak testing. It is noted that a number of other techniques such as shrink fitting, embossing and compaction of powders also seem to be particularly suitable. It is concluded that, with the advent of the portable and novel chilling apparatus, new vistas are approachable for undertaking maintenance operations in hospitals, power plants, nuclear facilities, and other systems that require uninterrupted operation.

  12. Induction of Freeze-sensitive Mutants from a Freeze-tolerant Yeast, Torulaspora delbrueckii.

    Science.gov (United States)

    Murakami, Y; Hahn, Y S; Yokoigawa, K; Endo, K; Kawai, H

    1994-01-01

    Freeze-sensitive strains of yeast were induced from a freeze-tolerant yeast Torulaspora delbrueckii by incubation with ethyl-methane sulfonate as a mutagen. A maximum ratio of mutation was attained by the incubation at 30°C for 75min. One-hundred and fifty strains of freeze-sensitive yeast were selected by plating-culture for the first screening. The freeze-tolerance ratio of each strain was examined based on the fermentative activity before and after freezing in liquid medium and dough. Strain 60B3 showed the highest freeze-sensitivity in a pre-fermented frozen dough (pre-fermented at 30°C for 2h, and frozen at -20°C for 7 days) among eight strains finally selected. PMID:27315725

  13. The influence of freezing and tissue porosity on the material properties of vegetable tissues

    CERN Document Server

    Ralfs, J D

    2002-01-01

    Tissue porosity and fluid flow have been shown to be important parameters affecting the mechanical and sensorial behaviour of edible plant tissues. The quantity of fluid and the manner with which it was released on compression of the plant tissue were also important regarding the sensory perception and a good indication of any structural damage resulting from freezing, for example. Potato, carrot and Chinese water chestnut were used to study the effects freezing has on model plant tissues. Mechanical and structural measurements of the plant tissue were correlated with sensory analysis. Conventional freezing was shown to cause severe structural damage predominantly in the form of cavities between or through cells, resulting in decreases in mechanical strength and stiffness, and samples that were perceived in the mouth as 'soft' and 'wet'. The location and size of the cavities formed from ice crystals, depended on the particular plant tissue being frozen, the processing it was subjected to prior to freezing, th...

  14. 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.

  15. Freezing phenomena in ice-water systems

    Energy Technology Data Exchange (ETDEWEB)

    Akyurt, M.; Zaki, G.; Habeebullah, B. [Fakieh Center for Applied Research, Makkah Al-Mukarramah (Saudi Arabia); King Abdulaziz University, Jeddah (Saudi Arabia). Dept. of Mechanical Engineering

    2002-09-01

    The characteristics of solidification and melting are reviewed. The properties of water and ice and the phase diagram of water are discussed with special emphasis on ice density. A concise account of the freezing process and the Stefan problem is presented. To this end, the four stages of freezing are identified, supercooling, nucleation and the formation of dendritic ice, the growth of concentric rings of solid ice at 0{sup o}C and the final cooling of the solid ice are treated in some detail. The subject of bursting of pipes is given particular emphasis. Attention is drawn to a common misconception on pipe bursting and to misleading relationships for the computation of freezing time for ice blockage. Several current applications of melting and freezing systems are outlined. (author)

  16. The Influence of Freezing Drizzle on Wire Icing during Freezing Fog Events

    Institute of Scientific and Technical Information of China (English)

    ZHOU Yue; NIU Shengjie; L(U) Jingjing

    2013-01-01

    Both direct and indirect effects of freezing drizzle on ice accretion were analyzed for ten freezing drizzle events during a comprehensive ice thickness,fog,and precipitation observation campaign carried out during the winter of 2008 and 2009 at Enshi Radar Station (30°17′N,109°16′E),Hubei Province,China.The growth rate of ice thickness was 0.85 mm h-1 during the freezing drizzle period,while the rate was only 0.4 mm h-1 without sleet and freezing drizzle.The rain intensity,liquid water content (LWC),and diameter of freezing drizzle stayed at low values.The development of microphysical properties of fog was suppressed in the freezing drizzle period.A threshold diameter (Dc) was proposed to estimate the influence of freezing drizzle on different size ranges of fog droplets.Fog droplets with a diameter less than Dc would be affected slightly by freezing drizzle,while larger fog droplets would be affected significantly.Dc had a correlation with the average rain intensity,with a correlation coefficient of 0.78.The relationships among the microphysical properties of fog droplets were all positive when the effect of freezing drizzle was weak,while they became poor positive correlations,or even negative correlations during freezing drizzle period.The direct contribution of freezing drizzle to ice thickness was about 14.5%.Considering both the direct and indirect effects,we suggest that freezing drizzle could act as a “catalyst” causing serious icing conditions.

  17. Characterization of Caulobacter crescentus response to low temperature and identification of genes involved in freezing resistance.

    Science.gov (United States)

    Mazzon, Ricardo R; Lang, Elza A S; Braz, Vânia S; Marques, Marilis V

    2008-11-01

    Free-living bacteria must respond to a wide range of temperature changes, and have developed specific mechanisms to survive in extreme environments. In this work we describe a remarkable resistance of mesophilic bacterium Caulobacter crescentus to several cycles of freezing at -80 degrees C, which was able to grow at low temperatures. Exponentially growing cells and late stationary-phase cells presented higher freezing resistance at both -20 and -80 degrees C than early stationary-phase cells. Cryotolerance was observed when log-phase cultures grown at 30 degrees C were preincubated at 5, 15 or 20 degrees C before freezing at -20 degrees C. A transposon library was screened to identify mutants sensitive to freezing at -80 degrees C and three strains presenting <10% survival were isolated. Identification of genes disrupted in each mutant showed that they encoded an AddA family DNA helicase, a DEAD/DEAH box RNA helicase and a putative RND (resistance, nodulation, cell division) efflux system component. These strains showed longer generation times than wild-type cells when growing at 15 degrees C, with the RNA helicase mutant presenting a severe growth defect. These analyses suggest that the singular intrinsic resistance to freezing of C. crescentus is in fact a consequence of several independent traits, especially the maintenance of a proper degree of supercoiling of nucleic acids.

  18. High pressure freezing/freeze substitution fixation improves the ultrastructural assessment of Wolbachia endosymbiont-filarial nematode host interaction.

    Directory of Open Access Journals (Sweden)

    Kerstin Fischer

    Full Text Available BACKGROUND: Wolbachia α-proteobacteria are essential for growth, reproduction and survival for many filarial nematode parasites of medical and veterinary importance. Endobacteria were discovered in filarial parasites by transmission electron microscopy in the 1970's using chemically fixed specimens. Despite improvements of fixation and electron microscopy techniques during the last decades, methods to study the Wolbachia/filaria interaction on the ultrastructural level remained unchanged and the mechanisms for exchange of materials and for motility of endobacteria are not known. METHODOLOGY/PRINCIPAL FINDING: We used high pressure freezing/freeze substitution to improve fixation of Brugia malayi and its endosymbiont, and this led to improved visualization of different morphological forms of Wolbachia. The three concentric, bilayer membranes that surround the endobacterial cytoplasm were well preserved. Vesicles with identical membrane structures were identified close to the endobacteria, and multiple bacteria were sometimes enclosed within a single outer membrane. Immunogold electron microscopy using a monoclonal antibody directed against Wolbachia surface protein-1 labeled the membranes that enclose Wolbachia and Wolbachia-associated vesicles. High densities of Wolbachia were observed in the lateral chords of L4 larvae, immature, and mature adult worms. Extracellular Wolbachia were sometimes present in the pseudocoelomic cavity near the developing female reproductive organs. Wolbachia-associated actin tails were not observed. Wolbachia motility may be explained by their residence within vacuoles, as they may co-opt the host cell's secretory pathway to move within and between cells. CONCLUSIONS/SIGNIFICANCE: High pressure freezing/freeze substitution significantly improved the preservation of filarial tissues for electron microscopy to reveal membranes and sub cellular structures that could be crucial for exchange of materials between Wolbachia

  19. Positive impact of sucrose supplementation during slow freezing of cat ovarian tissues on cellular viability, follicle morphology, and DNA integrity.

    Science.gov (United States)

    Tanpradit, Nae; Comizzoli, Pierre; Srisuwatanasagul, Sayamon; Chatdarong, Kaywalee

    2015-06-01

    The objectives of the study were to (1) examine and optimize the impact of sucrose during slow freezing and (2) compare the results of two freezing methods (slow freezing and vitrification) on cellular viability (germinal and stromal cells), follicle morphology, DNA integrity, and gap junction protein expression (connexin 43 [Cx 43]). Different sucrose supplementations (0, 0.1, and 0.3 M) in standard freezing medium were compared before and after slow freezing. Ovarian tissue slow frozen using 0.1- (4.0 ± 0.4) or 0.3-M sucrose (3.9 ± 0.5) yielded better follicular viability (number of positive follicles per 0.0625 mm(2)) than the group without sucrose (1.9 ± 0.2; P sucrose-treated groups (0.1 M, 47.4% and 0.3 M, 43.5%) than the group without sucrose (0 M, 33.8%; P sucrose groups (0.1 M, 1.2% and 0.3 M, 1.9%) than the group without sucrose (7.7%; P sucrose concentrations. In terms of the freezing methods used, vitrified ovarian tissues had fewer viable follicles (3.2 ± 0.6) than the slow-freezing method (4.6 ± 0.6; P sucrose supplementation and slow-freezing method on the follicular viability, follicular histologic appearances of follicles, and apoptosis of the follicles and stromal cells in cat ovarian tissues.

  20. Improvement of tolerance to freeze-thaw stress of baker's yeast by cultivation with soy peptides.

    Science.gov (United States)

    Izawa, Shingo; Ikeda, Kayo; Takahashi, Nobuyuki; Inoue, Yoshiharu

    2007-06-01

    The tolerance to freeze-thaw stress of yeast cells is critical for frozen-dough technology in the baking industry. In this study, we examined the effects of soy peptides on the freeze-thaw stress tolerance of yeast cells. We found that the cells cultured with soy peptides acquired improved tolerance to freeze-thaw stress and retained high leavening ability in dough after frozen storage for 7 days. The final quality of bread regarding its volume and texture was also improved by using yeast cells cultured with soy peptides. These findings promote the utilization of soy peptides as ingredients of culture media to improve the quality of baker's yeast. PMID:17505771

  1. 冻干无佐剂Veto细胞狂犬病疫苗免疫应答动态观察%A dynamic observation on the immune response to the freeze-dried without adjuvant Veto cell rabies vaccine

    Institute of Scientific and Technical Information of China (English)

    杨书宏; 陈胤忠; 陈万庚; 姜仁杰; 沈进进; 吴巨飞; 何飞; 王立成

    2009-01-01

    目的 评价国产冻干无佐剂Vero细胞狂犬病疫苗的免疫原性.方法 选择既往无明确狂犬病疫苗接种史和犬伤史、符合研究方案制定的入选标准和排除标准为研究对象,对暴露于狂犬病的患者采用常规5针注射.观察对象于首针接种前、首针接种后7、14、28、45 d,全程后6个月采集血样检测抗狂犬病中和抗体.结果 符合入选标准和排除标准的观察对象90名常规接种5针冻干无佐剂Vero细胞狂犬病疫苗.观察对象接种前狂犬病抗体均为阴性,接种首针后7 d狂犬病抗体阳转率为12.22%,接种首针后14 d阳转率达到100%,接种首针后28、45 d和全程后6个月的阳性率均为100%.接种首针后7 d抗狂犬病病毒综合抗体的几何抗体平均滴度(GMT)仅为0.27 IU/ml,接种首针后14 d狂犬病抗体的GlVlT达到2.52 IU/ml,较首针接种后7 d增长9.33倍.接种首针后28、45 d狂犬病抗体的GMT分别达到4.43、7.08 IU/ml,较首针后14 d分别增长1.76倍、2.81倍.全程接种后6个月狂犬病抗体的GMT仍达到8.41 IU/ml.结论 国产冻干无佐剂Vero细胞狂犬病疫苗具有良好的免疫原性,6个月内再被暴露于狂犬病动物者可以不需要接种狂犬病疫苗.%Objective The purpose of the present study was to evaluate the immunogenicity of the freeze-dried without adjuvant Vero cell rabies vaccine made in China. Methods A conventional vaccination procedure that included 5 injections of vaccine was applied to 90 patients who had exposed to rabies and were without history of rabies vaccination and dog bite injury, selected according to the requirements for recruitment and exclusion in the project. Blood samples were acquired before the first vaccination, 7, 14, 28 and45 days after the first vaccination and 180 days after the whole procedure of vaccination, respectively, for determination of anti-rabies neutralizing antibody. Results No anti-rabies neutralizing antibody was detected before the

  2. A case study on stress preconditioning of a Lactobacillus strain prior to freeze-drying.

    Science.gov (United States)

    Bergenholtz, Åsa Schoug; Wessman, Per; Wuttke, Anne; Håkansson, Sebastian

    2012-06-01

    Freeze-drying of bacterial cells with retained viability and activity after storage requires appropriate formulation, i.e. mixing of physiologically adapted cell populations with suitable protective agents, and control of the freeze-drying process. Product manufacturing may alter the clinical effects of probiotics and it is essential to identify and understand possible factor co-dependencies during manufacturing. The physical solid-state behavior of the formulation and the freeze-drying parameters are critical for bacterial survival and thus process optimization is important, independent of strain. However, the maximum yield achievable is also strain-specific and strain survival is governed by e.g. medium, cell type, physiological state, excipients used, and process. The use of preferred compatible solutes for cross-protection of Lactobacilli during industrial manufacturing may be a natural step to introduce robustness, but knowledge is lacking on how compatible solutes, such as betaine, influence formulation properties and cell survival. This study characterized betaine formulations, with and without sucrose, and tested these with the model lactic acid bacteria Lactobacillus coryniformis Si3. Betaine alone did not act as a lyo-protectant and thus betaine import prior to freeze-drying should be avoided. Differences in protective agents were analyzed by calorimetry, which proved to be a suitable tool for evaluating the characteristics of the freeze-dried end products.

  3. Effects of freezing on soil temperature, freezing front propagation and moisture redistribution in peat: laboratory investigations

    Directory of Open Access Journals (Sweden)

    R. M. Nagare

    2012-02-01

    Full Text Available There are not many studies that report water movement in freezing peat. Soil column studies under controlled laboratory settings can help isolate and understand the effects of different factors controlling freezing of the active layer in organic covered permafrost terrain. In this study, four peat Mesocosms were subjected to temperature gradients by bringing the Mesocosm tops in contact with sub-zero air temperature while maintaining a continuously frozen layer at the bottom (proxy permafrost. Soil water movement towards the freezing front (from warmer to colder regions was inferred from soil freezing curves, liquid water content time series and from the total water content of frozen core samples collected at the end of freezing cycle. A substantial amount of water, enough to raise the upper surface of frozen saturated soil within 15 cm of the soil surface at the end of freezing period appeared to have moved upwards during freezing. Diffusion under moisture gradients and effects of temperature on soil matric potential, at least in the initial period, appear to drive such movement as seen from analysis of freezing curves. Freezing front (separation front between soil zones containing and free of ice propagation is controlled by latent heat for a long time during freezing. A simple conceptual model describing freezing of an organic active layer initially resembling a variable moisture landscape is proposed based upon the results of this study. The results of this study will help in understanding, and ultimately forecasting, the hydrologic response of wetland-dominated terrain underlain by discontinuous permafrost.

  4. The role of antioxidant system in freezing acclimation-induced freezing resistance of Populus suaveolens cuttings

    Institute of Scientific and Technical Information of China (English)

    Luo Lei; Lin Shan-zhi; Zheng Hui-quan; Lei Yang; Zhang Qian; Zhang Zhi-yi

    2007-01-01

    We investigated the changes in the contents of H2O2, malonaldehyde (MDA) and endogenous antioxidants, the activities of protective enzymes and some critical enzymes involved in the ascorbate-glutathione (ASA-GSH) cycle as well as freezing resistance(expressed as LT50) and correlations mentioned above, in detail using Populus suaveolens cuttings. The purpose was to explore the physiological mechanism of the enhancement of freezing resistance induced by freezing acclimation at -20℃, and to elucidate the physiological mechanisms by which trees adapt to freezing. The results showed that freezing acclimation enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), monodehydroascorbate reductase (MDAR), ascorbate peroxidase(APX), dehydroascorbate reductase (DHAR) and glutathione reductase (GR). And it increased the contents of reduced ascorbate(ASA), reduced glutathione (GSH), dehydroascorbate (DHA) and oxidized glutathione (GSSG). However, H2O2 and MDA contents and LT50 of cuttings were decreased. LT50 in cuttings was found to be closely correlated to the levels of SOD, POD, CAT, APX,DHAR, MDAR, GR, H2O2, MDA, ASA, GSH, DHA and GSSG during freezing acclimation. This suggested that the enhancement of freezing resistance of cuttings induced by freezing acclimation may relate to the distinct increase for the levels of SOD, POD, CAT,APX, DHAR, MDAR,GR,ASA, GSH, DHA, and GSSG. In addition, the observed levels of APX, DHAR, MDAR, GR, ASA, DHA,GSH and GSSG were higher than those of SOD, POD and CAT during freezing acclimation. It indicated that a higher capacity of the ASA-GSH cycle is required for H2O2 detoxification, and growth and development of cuttings. Based on the obtained results, it can be concluded that the ASA-GSH cycle plays an important role in enhancement of freezing resistance of P. suaveolens cuttings during freezing acclimation.

  5. Monolayer freeze-fracture autoradiography: quantitative analysis of the transmembrane distribution of radioiodinated concanavalin A

    International Nuclear Information System (INIS)

    The technique of monolayer freeze-fracture autoradiography (MONOFARG) has been developed and the principles, quantitation, and application of the method are described. Cell monolayers attached to polylysine-treated glass were freeze-fractured, shadowed, and coated with dry, Parlodion-supported Ilford L4 photographic emulsion at room temperature. Quantitative aspects of MONOFARG were examined using radioiodinated test systems. Background was routinely -4 grains/μm2/day, the highest overall efficiency was between 25% and 45%, and grain density and efficiency were dependent on radiation dose for iodine-125 and D-19 development. Corrected grain densities were linearly proportional to iodine-125 concentration. The method was applied to an examination of the transmembrane distribution of radioiodinated and fluoresceinated concanavalin A (125I-FITC-Con-A). Human erythrocytes were labeled, column-purified, freeze-dried or freeze-fractured, autoradiographed, and examined by electron microscopy. The number of silver grains per square micrometer of unsplit single membrane was essentially identical to that of split extracellular membrane halves. These data demonstrate that 125I-FITC-Con-A partitions exclusively with the extracellular half of the membrane upon freeze-fracturing and can be used as a quantitative marker for the fraction of extracellular split membrane halves. This method should be able to provide new information about certain transmembrane properties of biological membrane molecules and probes, as well as about the process of freeze-fracture per se

  6. Amplitude Manipulation Evokes Upper Limb Freezing during Handwriting in Patients with Parkinson’s Disease with Freezing of Gait

    OpenAIRE

    Elke Heremans; Evelien Nackaerts; Griet Vervoort; Sarah Vercruysse; Sanne Broeder; Carolien Strouwen; Stephan P Swinnen; Alice Nieuwboer

    2015-01-01

    Background Recent studies show that besides freezing of gait (FOG), many people with Parkinson's disease (PD) also suffer from freezing in the upper limbs (FOUL). Up to now, it is unclear which task constraints provoke and explain upper limb freezing. Objective To investigate whether upper limb freezing and other kinematic abnormalities during writing are provoked by (i) gradual changes in amplitude or by (ii) sustained amplitude generation in patients with and without freezing of gait. Metho...

  7. Dynamics of leaf gas exchange, chlorophyll fluorescence and stem diameter changes during freezing and thawing of Scots pine seedlings.

    Science.gov (United States)

    Lindfors, Lauri; Hölttä, Teemu; Lintunen, Anna; Porcar-Castell, Albert; Nikinmaa, Eero; Juurola, Eija

    2015-12-01

    Boreal trees experience repeated freeze-thaw cycles annually. While freezing has been extensively studied in trees, the dynamic responses occurring during the freezing and thawing remain poorly understood. At freezing and thawing, rapid changes take place in the water relations of living cells in needles and in stem. While freezing is mostly limited to extracellular spaces, living cells dehydrate, shrink and their osmotic concentration increases. We studied how the freezing-thawing dynamics reflected on leaf gas exchange, chlorophyll fluorescence and xylem and living bark diameter changes of Scots pine (Pinus sylvestris L.) saplings in controlled experiments. Photosynthetic rate quickly declined following ice nucleation and extracellular freezing in xylem and needles, almost parallel to a rapid shrinking of xylem diameter, while that of living bark followed with a slightly longer delay. While xylem and living bark diameters responded well to decreasing temperature and water potential of ice, the relationship was less consistent in the case of increasing temperature. Xylem showed strong temporal swelling at thawing suggesting water movement from bark. After thawing xylem diameter recovered to a pre-freezing level but living bark remained shrunk. We found that freezing affected photosynthesis at multiple levels. The distinct dynamics of photosynthetic rate and stomatal conductance reveals that the decreased photosynthetic rate reflects impaired dark reactions rather than stomatal closure. Freezing also inhibited the capacity of the light reactions to dissipate excess energy as heat, via non-photochemical quenching, whereas photochemical quenching of excitation energy decreased gradually with temperature in agreement with the gas exchange data. PMID:26423334

  8. An Investigation of Freezing of Supercooled Water on Anti-Freeze Protein Modified Surfaces

    Institute of Scientific and Technical Information of China (English)

    Thibaut V J Charpentier; Anne Neville; Paul Millner; Rob Hewson; Ardian Morina

    2013-01-01

    This work investigates how functionalization ofaluminium surfaces with natural type Ⅲ Anti-Freeze Protein (AFP) affects the mechanism of heterogeneous ice nucleation.First the bulk ice nucleation properties of distilled water and aqueous solution of AFP were evaluated by differential scanning calorimetry.Then the modified surface was characterized by Secondary Ions Mass Spectroscopy (SIMS),Fourier Transform InfraRed (FTIR) spectroscopy and contact angle measurement.Freezing experiments were then conducted in which water droplets underwent a slow controlled cooling.This study shows that compared to uncoated aluminium,the anti-freeze proteins functionalized surfaces exhibit a higher and narrower range of freezing temperature.It was found that these proteins that keep living organisms from freezing in cold environment act in the opposite way once immobilized on surfaces by promoting ice nucleation.Some suggestions regarding the mechanism of action of the observed phenomena were proposed based on the Classical Nucleation Theory (CNT).

  9. Effect of protective agents and previous acclimation on ethanol resistance of frozen and freeze-dried Lactobacillus plantarum strains.

    Science.gov (United States)

    Bravo-Ferrada, Bárbara Mercedes; Brizuela, Natalia; Gerbino, Esteban; Gómez-Zavaglia, Andrea; Semorile, Liliana; Tymczyszyn, E Elizabeth

    2015-12-01

    The aim of this work was to study the protective effect of sucrose, trehalose and glutamate during freezing and freeze-drying of three oenological Lactobacillus plantarum strains previously acclimated in the presence of ethanol. The efficiency of protective agents was assessed by analyses of membrane integrity and bacterial cultivability in a synthetic wine after the preservation processes. No significant differences in the cultivability, with respect to the controls cells, were observed after freezing at -80 °C and -20 °C, and pre-acclimated cells were more resistant to freeze-drying than non-acclimated ones. The results of multiparametric flow cytometry showed a significant level of membrane damage after freeze-drying in two of the three strains. The cultivability was determined after incubation in wine-like medium containing 13 or 14% v/v ethanol at 21 °C for 24 h and the results were interpreted using principal component analysis (PCA). Acclimation was the most important factor for preservation, increasing the bacterial resistance to ethanol after freezing and freeze-drying. Freeze-drying was the most drastic method of preservation, followed by freezing at -20 °C. The increase of ethanol concentration from 6 to 10% v/v in the acclimation medium improved the recovery of two of the three strains. In turn, the increase of ethanol content in the synthetic wine led to a dramatic decrease of viable cells in the three strains investigated. The results of this study indicate that a successful inoculation of dehydrated L. plantarum in wine depends not only on the use of protective agents, but also on the cell acclimation process prior to preservation, and on the ethanol content of wine.

  10. Female Fertility: Is it Safe to "Freeze?"

    Directory of Open Access Journals (Sweden)

    Lu Zhang

    2015-01-01

    Full Text Available Objective: To evaluate the safety and risk of cryopreservation in female fertility preservation. Data sources: The data analyzed in this review were the English articles from 1980 to 2013 from journal databases, primarily PubMed and Google scholar. The criteria used in the literature search show as following: (1 human; embryo; cryopreservation/freezing/vitrification, (2 human; oocyte/immature oocyte; cryopreservation/ freezing/vitrification, (3 human; ovarian tissue transplantation; cryopreservation/freezing/vitrification, (4 human; aneuploidy/DNA damage/epigenetic; cryopreservation/freezing/vitrification, and (5 human; fertility preservation; maternal age. Study selection: The risk ratios based on survival rate, maturation rate, fertilization rate, cleavage rate, implantation rate, pregnancy rate, and clinical risk rate were acquired from relevant meta-analysis studies. These studies included randomized controlled trials or studies with one of the primary outcome measures covering cryopreservation of human mature oocytes, embryos, and ovarian tissues within the last 7 years (from 2006 to 2013, since the pregnancy rates of oocyte vitrification were significantly increased due to the improved techniques. The data involving immature oocyte cryopreservation obtained from individual studies was also reviewed by the authors. Results: Vitrifications of mature oocytes and embryos obtained better clinical outcomes and did not increase the risks of DNA damage, spindle configuration, embryonic aneuploidy, and genomic imprinting as compared with fresh and slow-freezing procedures, respectively. Conclusions: Both embryo and oocyte vitrifications are safe applications in female fertility preservation.

  11. Identification and classification of genes required for tolerance to freeze-thaw stress revealed by genome-wide screening of Saccharomyces cerevisiae deletion strains.

    Science.gov (United States)

    Ando, Akira; Nakamura, Toshihide; Murata, Yoshinori; Takagi, Hiroshi; Shima, Jun

    2007-03-01

    Yeasts used in bread making are exposed to freeze-thaw stress during frozen-dough baking. To clarify the genes required for freeze-thaw tolerance, genome-wide screening was performed using the complete deletion strain collection of diploid Saccharomyces cerevisiae. The screening identified 58 gene deletions that conferred freeze-thaw sensitivity. These genes were then classified based on their cellular function and on the localization of their products. The results showed that the genes required for freeze-thaw tolerance were frequently involved in vacuole functions and cell wall biogenesis. The highest numbers of gene products were components of vacuolar H(+)-ATPase. Next, the cross-sensitivity of the freeze-thaw-sensitive mutants to oxidative stress and to cell wall stress was studied; both of these are environmental stresses closely related to freeze-thaw stress. The results showed that defects in the functions of vacuolar H(+)-ATPase conferred sensitivity to oxidative stress and to cell wall stress. In contrast, defects in gene products involved in cell wall assembly conferred sensitivity to cell wall stress but not to oxidative stress. Our results suggest the presence of at least two different mechanisms of freeze-thaw injury: oxidative stress generated during the freeze-thaw process, and defects in cell wall assembly. PMID:16989656

  12. 冷冻-解冻循环及气体吹扫对质子交换膜燃料电池的影响%Effects of Freeze/Thaw Cycles and Gas Purging Method on Polymer Electrolyte Membrane Fuel Cells

    Institute of Scientific and Technical Information of China (English)

    张生生; 俞红梅; 朱红; 侯俊波; 衣宝廉; 明平文

    2006-01-01

    At subzero temperature, the startup capability and performance of polymer electrolyte membrane fuel cell (PEMFC) deteriorates markedly. The object of this work is to study the degradation mechanism of key components of PEMFC-membrane-electrode assembly (MEA) and seek feasible measures to avoid degradation. The effect of freeze/thaw cycles on the structure of MEA is investigated based on porosity and SEM measurement. The performance of a single cell was also tested before and after repetitious freeze/thaw cycles. The experimental results indicated that the performance of a PEMFC decreased along with the total operating time as well as the pore size distribution shifting and micro configuration changing. However, when the redundant water had been removed by gas purging, the performance of the PEMFC stack was almost resumed when it experienced again the same subzero temperature test. These results show that it is necessary to remove the water in PEMFCs to maintain stable performance under subzero temperature and gas purging is proved to be the effective operation.

  13. Immersion freezing of biological particles at LACIS

    Science.gov (United States)

    Clauss, T.; Hartmann, S.; Temkiv, T. S.; Augustin, S.; Gosewinkel Karlson, U.; Sahyoun, M. M.; Niedermeier, D.; Wex, H.; Voigtländer, J.; Raddatz, M.; Stratmann, F.

    2012-04-01

    Biological particles, especially bacteria being ubiquitous in the atmosphere, belong to the most efficient ice nuclei (IN) (Möhler, 2008) and hence might have a large impact on weather and climate. In this study, the immersion freezing behavior of different size segregated biological particles is investigated at the laminar flow tube LACIS (Leipzig Aerosol Cloud Interaction Simulator, Hartmann et al., 2011). For these experiments, SNOMAX and outer membrane vesicles (OMV) are used as IN. SNOMAX industrially produced from Pseudomonas-syringae bacteria, which are very ice nucleation active, can be seen as a proxy for ice nucleating bacteria in general. On the surface of these bacteria, ice nucleating proteins that initiate the freezing are situated (Maki et al., 1974). Additionally, it has been found that some ice nucleating bacteria strains have the ability to produce OMV, i.e., strangulated parts of the bacterial cell consisting of the same membrane material (Phelps et al., 1986). These OMV might contain the same ice nucleating proteins on their surface and thus might be able to nucleate ice as well. The OMV used in our experiments were extracted from bacteria cultivated from rain samples collected in Denmark from 30 m height. In our experiments, the biological particles are suspended in air via atomization, size selected by means of a Differential Mobility Particle Sizer, and then fed into LACIS. In LACIS, well defined droplets are produced by activating the biological particles to cloud droplets, so that each droplet contains only one biological particle. By decreasing the temperature in LACIS, these droplets are frozen. To determine the ice fraction, i.e., the fraction of frozen droplets to all particles, the liquid and frozen droplets are distinguished by means of a newly self-built optical device, which is positioned under LACIS, using the depolarization of light scattered by a single particle. The ice fractions are measured as a function of temperature and

  14. Freeze tolerance of soil chytrids from temperate climates in Australia.

    Science.gov (United States)

    Gleason, Frank H; Letcher, Peter M; McGee, Peter A

    2008-08-01

    Very little is known about the capacity of soil chytrids to withstand freezing in the field. Tolerance to freezing was tested in 21 chytrids isolated from cropping and undisturbed soils in temperate Australia. Samples of thalli grown on peptone-yeast-glucose (PYG) agar were incubated for seven days at -15 degrees C. Recovery of growth after thawing and transferring to fresh medium at 20 degrees C indicated survival. All isolates in the Blastocladiales and Spizellomycetales survived freezing in all tests. All isolates in the Chytridiales also survived freezing in some tests. None of the isolates in the Rhizophydiales survived freezing in any of the tests. However, some isolates in the Rhizophydiales recovered growth after freezing if they were grown on PYG agar supplemented with either 1% sodium chloride or 1% glycerol prior to freezing. After freezing, the morphology of the thalli of all isolates was observed under LM. In those isolates that recovered growth after transfer to fresh media, mature zoosporangia were observed in the monocentric isolates and resistant sporangia or resting spores in the polycentric isolates. Encysted zoospores in some monocentric isolates also survived freezing. In some of the experiments the freezing and thawing process caused visible structural damage to the thalli. The production of zoospores after freezing and thawing was also used as an indicator of freeze tolerance. The chytrids in this study responded differently to freezing. These data add significantly to our limited knowledge of freeze tolerance in chytrids but leave many questions unanswered. PMID:18550351

  15. Simple improvement in freeze-tolerance of bakers' yeast with poly-gamma-glutamate.

    Science.gov (United States)

    Yokoigawa, Kumio; Sato, Machiko; Soda, Kenji

    2006-09-01

    We examined the effect of poly-gamma-glutamate (PGA) on the freeze-tolerance of four types of commercial bakers' yeast (freeze-tolerant, osmotic-tolerant, low-temperature-sensitive, and ordinary bakers' yeasts). The survival ratio of ordinary bakers' yeast cells frozen at -30 degrees C for 3 d in a medium (0.5% yeast extract, 0.5% peptone, and 2% glucose: YPD medium) was improved by adding more than 1% PGA to the medium; the survival ratio increased from about 10% to more than 70%. All PGA preparations, which differed in average molecular mass (50, 2,000, 4,000, 6,000, 8,000, and 10,000 kDa), showed a similar cryoprotecive effect on the cells. Similar results were also obtained with other types of bakers' yeast, sake yeast and beer yeast. When the four types of bakers' yeast cell were frozen at -30 degrees C for 3 d in dough supplemented with more than 1% PGA, the cells (after freezing and thawing) showed higher leavening ability than those frozen in dough without PGA, irrespective of the molecular mass of PGA. Thus, PGA appears to protect bakers' yeast from lethal freeze injury, leading to a high leavening ability after freezing and thawing. PGA did not decrease the original leavening ability of the bakers' yeast, and was not decomposed by the yeast cells. PGA suppressed the decrease in leavening ability during a prolonged fermentation time, probably because PGA adsorbed inhibitory metabolites accumulated in the dough. PGA could prove useful for improving the freeze-tolerance of bakers' yeast by its addition to dough. PMID:17046536

  16. Heat transfer coefficient of cryotop during freezing.

    Science.gov (United States)

    Li, W J; Zhou, X L; Wang, H S; Liu, B L; Dai, J J

    2013-01-01

    Cryotop is an efficient vitrification method for cryopreservation of oocytes. It has been widely used owing to its simple operation and high freezing rate. Recently, the heat transfer performance of cryotop was studied by numerical simulation in several studies. However, the range of heat transfer coefficient in the simulation is uncertain. In this study, the heat transfer coefficient for cryotop during freezing process was analyzed. The cooling rates of 40 percent ethylene glycol (EG) droplet in cryotop during freezing were measured by ultra-fast measurement system and calculated by numerical simulation at different value of heat transfer coefficient. Compared with the results obtained by two methods, the range of the heat transfer coefficient necessary for the numerical simulation of cryotop was determined, which is between 9000 W/(m(2)·K) and 10000 W/(m (2)·K).

  17. Freezing precipitation in Russia and the Ukraine

    Directory of Open Access Journals (Sweden)

    A. A. Zavyalova

    2007-04-01

    Full Text Available Conditions for freezing precipitation (FP, including freezing rain (FR and freezing drizzle (FZ for 8 airports in Russia and 4 in the Ukraine are studied on the basis of 10 to 20-year series of surface observations, radiosonde and objective analysis data. Statistical characteristics are presented of the FP episode durations and of occurrence frequency dependences on surface air temperature, wind direction and speed and cloud base height. From the radiosonde data, it is found that the "classical mechanism" of FP generation (for which, stratification of "warm nose" type in the cloud layer is necessary is not frequent: most of FP cases are associated with "all cold" conditions in the lower 3-km layer, that is, with negative temperatures in and below the clouds.

  18. Hatchling turtles survive freezing during winter hibernation.

    OpenAIRE

    Storey, K. B.; Storey, J M; Brooks, S. P.; Churchill, T A; Brooks, R. J.

    1988-01-01

    Hatchlings of the painted turtle (Chrysemys picta marginata) are unique as the only reptile and highest vertebrate life form known to tolerate the natural freezing of extracellular body fluids during winter hibernation. Turtles survived frequent exposures to temperatures as low as -6 degrees C to -8 degrees C in their shallow terrestrial nests over the 1987-1988 winter. Hatchlings collected in April 1988 had a mean supercooling point of -3.28 +/- 0.24 degrees C and survived 24 hr of freezing ...

  19. Reproducing Black's experiments: freezing point depression and supercooling of water

    International Nuclear Information System (INIS)

    We carried out two historical experiments referred to by Joseph Black, one on freezing mixtures of salted water with ice and another on freezing supercooled pure water by a small disturbance. The results confirm thermodynamical predictions for the depression of the freezing point of salted water and for the latent heat of freezing of supercooled water respectively, which came after Black. The depression of the freezing point can hardly be fitted in the framework of the caloric theory of heat, which was taken for granted by Black, and the instantaneous freezing of supercooled water also poses some difficulties for that theory. (author)

  20. Cryoprotectant Production in Freeze-Tolerant Wood Frogs Is Augmented by Multiple Freeze-Thaw Cycles.

    Science.gov (United States)

    Larson, Don J; Barnes, Brian M

    2016-01-01

    Ice nucleation across the skin of wood frogs (Lithobates sylvaticus) rapidly induces endogenous production of glucose, a cryoprotectant necessary for freeze tolerance. In laboratory studies of freeze tolerance, wood frogs are cooled slowly, often at -0.05°C h(-1), to facilitate high cryoprotectant production and survival. Under natural conditions in Alaska, however, wood frogs accumulate maximal tissue glucose concentrations while cooling at much faster rates, -0.35° to -1.6°C h(-1), and in addition undergo multiple successive freeze-thaw cycles before remaining frozen for the winter. We examined whether simulating these ecologically relevant cooling rates and repeated freeze-thaw events in captive wood frogs results in the high glucose concentrations found in naturally frozen wood frogs. We found that over successive freezing and thawing events, glucose concentrations increased stepwise in all measured tissues. Short thawing periods did not result in a statistically significant decline of glucose concentrations. Wood frogs that experienced three freeze-thaw events had fresh weight glucose concentrations that approached values found in tissues of wood frogs frozen in natural conditions. Laboratory wood frogs survive frozen for 2 mo, while wood frogs frozen under natural conditions survive frozen for up to 7 mo at temperatures below -18°C. We hypothesize that repeated freeze-thaw cycles with rapid cooling and warming rates allow for greater survival in Alaskan wood frogs through enhanced cryoprotectant production. PMID:27327184

  1. Farinose flavonoids are associated with high freezing tolerance in fairy primrose (Primula malacoides) plants.

    Science.gov (United States)

    Isshiki, Ryutaro; Galis, Ivan; Tanakamaru, Shigemi

    2014-02-01

    The deposition of surface (farinose) flavonoids on aerial parts of some Primula species is a well-documented but poorly understood phenomenon. Here, we show that flavonoid deposition on the leaves and winter buds may contribute strongly to preventing freezing damage in these plants. The ice nucleation temperature of fairy primrose (Primula malacoides) leaves covered with natural flavone was approximately 6 °C lower compared to those that had their flavone artificially removed. Additionally, farinose flavonoids on the leaves reduced subsequent electrolyte leakage (EL) from the cells exposed to freezing temperatures. Interestingly, exogenous application of flavone at 4 mg/g fresh weight to P. malacoides leaves, which had the original flavone mechanically removed, restored freezing tolerance, and diminished EL from the cells to pretreatment values. Our results suggest that farinose flavonoids may function as mediators of freezing tolerance in P. malacoides, and exogenous application of flavone could be used to reduce freezing damage during sudden but predictable frost events in other plant species.

  2. Building the Method to Determine the Rate of Freezing Water in Penaeus monodon of the Freezing Process

    Directory of Open Access Journals (Sweden)

    Nguyen Tan Dzung

    2012-10-01

    Full Text Available The method of determination the rate of freezing water in Penaeus monodon of freezing process was established on base the equation of energy balance in warming up process Penaeus monodon after freezing to determine specific heat of Penaeus monodon. The result obtained was built the mathematical model (19 to determine the rate of freezing water according to the freezing temperature of Penaeus monodon. The results indicated that when water was completely frozen (ω = 1 or 100%, the optimal freezing temperature of Penaeus monodon was-22.00°C.

  3. Mass production of shaped particles through vortex ring freezing

    Science.gov (United States)

    An, Duo; Warning, Alex; Yancey, Kenneth G.; Chang, Chun-Ti; Kern, Vanessa R.; Datta, Ashim K.; Steen, Paul H.; Luo, Dan; Ma, Minglin

    2016-08-01

    A vortex ring is a torus-shaped fluidic vortex. During its formation, the fluid experiences a rich variety of intriguing geometrical intermediates from spherical to toroidal. Here we show that these constantly changing intermediates can be `frozen' at controlled time points into particles with various unusual and unprecedented shapes. These novel vortex ring-derived particles, are mass-produced by employing a simple and inexpensive electrospraying technique, with their sizes well controlled from hundreds of microns to millimetres. Guided further by theoretical analyses and a laminar multiphase fluid flow simulation, we show that this freezing approach is applicable to a broad range of materials from organic polysaccharides to inorganic nanoparticles. We demonstrate the unique advantages of these vortex ring-derived particles in several applications including cell encapsulation, three-dimensional cell culture, and cell-free protein production. Moreover, compartmentalization and ordered-structures composed of these novel particles are all achieved, creating opportunities to engineer more sophisticated hierarchical materials.

  4. Effect of tumor antigen released during photodynamic therapy or freezing-thawing on the maturation phenotypes of dendritic cells%光动力治疗和冻融对肿瘤细胞抗原释放及树突细胞成熟表型的影响

    Institute of Scientific and Technical Information of China (English)

    江珊; 夏育民; 程丹; 程鸿

    2011-01-01

    目的 探讨光动力疗法(PDT)和冻融导致的肿瘤细胞抗原暴露,对体外树突细胞(DC)成熟表型的影响.方法 对体外培养的4T1乳腺癌细胞分别予以PDT和反复冻融,检测4T1细胞的坏死与凋亡,收获上清液并与未成熟DC(imDC)或成熟DC(mDC)共同孵育,采用流式细胞法和激光共聚焦显微镜分析DC免疫表型和形态学特征.结果 经两种机制处理的4T1细胞都以坏死为主,但PDT较冻融诱导更高的凋亡率(34.7%±9.6%比16.8%±5.1%,P<0.05);与上清液孵育后,PDT组imDC的CD80、CD86与I-A/I-E阳性率分别为22.4%±4.6%、22.5%±5.6%、24.3%±6.3%,冻融组依次为17.3%±3.3%、18.0%±4.7%、20.7%±3.1%,均高于空白对照组(14.2%±3.0%、13.6%±3.3%、15.5%±2.1%,P<0.05),且PDT组升高更明显,而mDC的免疫表型变化差异无统计学意义(P>0.05).结论 与冻融相比较,PDT导致更高的细胞凋亡率,更易诱导imDC表达成熟表型.%Objective To investigate the effects of tumor antigen on the maturation phenotypes of dendritic cells through different treatments of antigenic exposure. Methods The necrosis and apoptosis of 4T1 mammary carcinoma cells were detected after an in vitro treatment of photodynamic therapy (PDT) or freezing-thawing. The supernatant of 4T1 cells was harvested and added into the culture of immature or mature dendritic cells (DCs). The immune phenotypes and morphological features of DCs were analyzed by flow cytometry and confocal laser scanning microscopy respectively. Results Necrosis predominated after these treatments. But the apoptotic ratio was higher in PDT cells than that in freezing-thawing cells (34. 7% ±9. 6% vs 16. 8% ± 5. 1%, P < 0.05 ). After co-culturing with different supernatants, the immature DCs of PDT group showed more significant increases of positive rates of CD80 (22. 4% ±4. 6% ),CD86 (22. 5% ± 5. 6% ) and I-A/I-E (24. 3% ± 6. 3% ) versus the immature DCs of freezing-thawing group (17.3% ±3.3%, 18.0% ±4.7% and

  5. Freeze-thaw induced gelation of alginates.

    Science.gov (United States)

    Zhao, Ying; Shen, Wei; Chen, Zhigang; Wu, Tao

    2016-09-01

    Adding divalent ions or lowering pH below the pKa values of alginate monomers are common ways in preparing alginate gels. Herein a new way of preparing alginate gels using freeze-thaw technique is described. Solvent crystallization during freezing drove the polymers to associate into certain structures that became the junction zones of hydrogels after thawing. It enabled the preparation of alginate gels at pH 4.0 and 3.5, two pH at which the gel could not be formed previously. At pH 3.0 where alginate gel could be formed initially, applying freeze-thaw treatment increased the gel storage modulus almost 100 times. The formation of hydrogels and the resulting gel properties, such as dynamic moduli and gel syneresis were influenced by the pH values, number of freeze-thaw cycles, alginate concentrations, and ionic strengths. The obtained hydrogels were soft and demonstrated a melting behavior upon storage, which may find novel applications in the biomedical industry.

  6. Scaling-Up Eutectic Freeze Crystallization

    NARCIS (Netherlands)

    Genceli, F.E.

    2008-01-01

    A novel crystallization technology, Eutectic Freeze Crystallization (EFC) has been investigated and further developed in this thesis work. EFC operates around the eutectic temperature and composition of aqueous solutions and can be used for recovery of (valuable) dissolved salts (and/or or acids) an

  7. Freezing in the Antarctic limpet, Nacella concinna.

    Science.gov (United States)

    Hawes, T C; Worland, M R; Bale, J S

    2010-08-01

    The process of organismal freezing in the Antarctic limpet, Nacella concinna, is complicated by molluscan biology. Internal ice formation is, in particular, mediated by two factors: (a) the provision of an inoculative target for ice formation in the exposed mucus-secreting foot; and (b) osmoconformity to the marine environment. With regard to the first, direct observations of the independent freezing of pedal mucus support the hypothesis that internal ice formation is delayed by the mucal film. As to the second, ice nucleation parametrics of organismal tissue (head, midgut, gonad, foot) and mucus in both inter- and subtidal populations were characterized by high melting points (range=-4.61 to -6.29 degrees C), with only c.50% of a given sample osmotically active. At this stage it would be premature to ascribe a cryo-adaptive function to the mucus as the protective effects are more readily attributed to the physical properties of the secretion (i.e. viscosity) and their corresponding effects on the rate of heat transfer. As it is difficult to thermally distinguish between the freezing of mucus and the rest of the animal, the question as to whether it is tolerant of internal as well as external ice formation remains problematic, although it may be well suited to the osmotic stresses of organismal freezing. PMID:20599885

  8. Freeze-thaw induced gelation of alginates.

    Science.gov (United States)

    Zhao, Ying; Shen, Wei; Chen, Zhigang; Wu, Tao

    2016-09-01

    Adding divalent ions or lowering pH below the pKa values of alginate monomers are common ways in preparing alginate gels. Herein a new way of preparing alginate gels using freeze-thaw technique is described. Solvent crystallization during freezing drove the polymers to associate into certain structures that became the junction zones of hydrogels after thawing. It enabled the preparation of alginate gels at pH 4.0 and 3.5, two pH at which the gel could not be formed previously. At pH 3.0 where alginate gel could be formed initially, applying freeze-thaw treatment increased the gel storage modulus almost 100 times. The formation of hydrogels and the resulting gel properties, such as dynamic moduli and gel syneresis were influenced by the pH values, number of freeze-thaw cycles, alginate concentrations, and ionic strengths. The obtained hydrogels were soft and demonstrated a melting behavior upon storage, which may find novel applications in the biomedical industry. PMID:27185114

  9. Liquid carbon: structure near the freezing line

    NARCIS (Netherlands)

    Ghiringhelli, L.M.; Los, J.H.; Meijer, E.J.; Fasolino, A.; Frenkel, D.

    2005-01-01

    We present a detailed analysis of the structure of liquid carbon near the freezing line. The results are obtained by molecular simulation using a recently developed state-of-the-art bond order potential. We find that along the melting line the liquid is predominantly threefold coordinated up to pres

  10. Anomalous freezing behavior of nanoscale liposomes

    DEFF Research Database (Denmark)

    Spangler, E. J.; Kumar, P. B. S.; Laradji, M.

    2012-01-01

    The effect of the finite size of one-component liposomes on their phase behavior is investigated via simulations of an implicit-solvent model of self-assembled lipid bilayers. We found that the high curvature of nanoscale liposomes has a significant effect on their freezing behavior. While...

  11. Osteogenesis and vascularization of bone marrow mesenchymal stem cells transfected by pcDNA3/hVEGF165 combined with freeze-dried cancellous bone in vivo%脂质体介导pcDNA3/hVEGF165转染骨髓基质干细胞复合冻干骨的体内成骨和血管化

    Institute of Scientific and Technical Information of China (English)

    张鹏; 董玲; 杨连甲

    2011-01-01

    BACKGROUND: Previous studies have shown that vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) can improve the implant livability and growth.OBJECTIVE: To study the effect of freeze-dried cancellous bone on the osteogenesis and vascularization of bone marrow stem cells transfected with pcDNA3/hVEGF165 in vivo.METHODS: The rabbit bone marrow stem cells, which were transfected with pcDNA3/hVEGF165 by liposome mediated method and then adhered to freeze-dried cancellous bone, were implanted in the muscle pouches of rabbits. The rabbits were divided into three groups: freeze-dried cancellous bone group (A group), freeze-dried cancellous bone combined with bone marrow stem cells group (B group), freeze-dried cancellous bone combined with bone marrow stem cells transfected with VEGF group (C group).RESULTS AND CONCLUSION: At the 8th week after implantation, it was found, compared with A group and B group, C group grew a large number of osteoblasts, osteoclasts and cartilage, and the number of vessels in C group was more than that in A or B group. The osteogenesis of the freeze-dried cancellous bone combined with bone marrow stem cells transfected with VEGF using pcDNA3/hVEGF165 by liposome mediated method is better than the freeze-dried cancellous bone or freeze-dried cancellous bone combined with bone marrow stem cells.%背景:以往的研究表明血管内皮生长因子、碱性成纤维细胞生长因子可以促进移植物的存活和体内生长.目的:观察脂质体介导的pcDNA3/hVEGF165转染骨髓基质干细胞后复合冻干松质骨在体内的成骨和血管化效果.方法:取同种异体新西兰大白兔的耾骨和股骨制备冻干骨,用脂质体将血管内皮生长因子转染入体外培养扩增新西兰大白兔骨髓基质干细胞中,使其附着于同种异体冻干松质骨.将新西兰大白兔分为3组,于兔竖脊肌分别植入单纯冻干骨、单纯骨髓间充质干细胞复合冻干骨组、转染

  12. Freezing characteristics and texture variation after freezing and thawing of four fruit types

    Directory of Open Access Journals (Sweden)

    Arpassorn Sirijariyawat

    2012-11-01

    Full Text Available One major problem with frozen fruits is a loss of texture. Therefore this study investigated the effects of the freezingprocess on the freezing profiles, texture, and drip loss of apple, mango, cantaloupe, and pineapple fruit samples. All frozenthawedfruits varied in these three properties because of diversity in the fresh fruits. Mango had the highest total solublesolids content and the lowest freezing point, whereas pineapple showed the highest freezing rate. The highest firmness andcrunchy texture were found in fresh apple, and these properties were absent in the other fresh fruits. The firmness of allfrozen fruits significantly decreased by different percentages as compared to those of the fresh fruits. The drip loss of eachfruit type was also significantly different with apple samples having the highest firmness decrease and drip loss. This studyshows that freezing characteristics and frozen fruit properties depend on type of fruit.

  13. Female Fertility: Is it Safe to "Freeze?"

    Institute of Scientific and Technical Information of China (English)

    Lu Zhang; Li-Ying Yan; Xu Zhi; Jie Yan; Jie Qiao

    2015-01-01

    Objective:To evaluate the safety and risk of cryopreservation in female fertility preservation.Data sources:The data analyzed in this review were the English articles from 1980 to 2013 from journal databases,primarily PubMed and Google scholar.The criteria used in the literature search show as following:(1) human; embryo; cryopreservation/freezing/vitrification,(2) human; oocyte/immature oocyte; cryopreservation/freezing/vitrification,(3) human; ovarian tissue transplantation; cryopreservation/ freezing/vitrification,(4) human; aneuploidy/DNA damage/epigenetic; cryopreservation/freezing/vitrification,and (5) human; fertility preservation; maternal age.Study selection:The risk ratios based on survival rate,maturation rate,fertilization rate,cleavage rate,implantation rate,pregnancy rate,and clinical risk rate were acquired from relevant meta-analysis studies.These studies included randomized controlled trials or studies with one of the primary outcome measures covering cryopreservation of human mature oocytes,embryos,and ovarian tissues within the last 7 years (from 2006 to 2013,since the pregnancy rates of oocyte vitrification were significantly increased due to the improved techniques).The data involving immature oocyte cryopreservation obtained from individual studies was also reviewed by the authors.Results:Vitrifications of mature oocytes and embryos obtained better clinical outcomes and did not increase the risks of DNA damage,spindle configuration,embryonic aneuploidy,and genomic imprinting as compared with fresh and slow-freezing procedures,respectively.Conclusions:Both embryo and oocyte vitrifications are safe applications in female fertility preservation.

  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; Derksen, Chris; Royer, Alain; Belair, Stephane; Houser, Paul; McDonald, Kyle; Entin, Jared; Lewis, Kristen

    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. 7 CFR 58.620 - Freezing and packaging rooms.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Freezing and packaging rooms. 58.620 Section 58.620 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards....620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall...

  16. COMPREHENSIVE DESIGN METHOD OF FREEZE WALL AND ITS APPLICATION

    Institute of Scientific and Technical Information of China (English)

    CHENXiangsheng

    1995-01-01

    Artificially ground freezing (AGF) is one of the main methods to establish temporary support for shaft sinking in unstable water bearing strata. Domke(1915) formula based on frozen soil strength has widely been used for designing freeze wall thickness. However, it can not ensure the stability of freeze wall, nor guarantee the safety of shaft construction as frozen depth increase in unstable water bearing strata. F.A.Auld (1985, 1988) presented a design method of freeze wall, which is on the basis of strength and stability, together with deformation of freeze wall.He combined deformation of freeze wall, lining and deformation of freeze tube to set up a comprehensive design method for freeze wall. This paper, according to the practice in China, describes a comprehensive design method for deep freeze wall, considering influence of excavation rate of advance, unsupported length of freeze wall and the sump state on inward deformation of freeze wall, and the allowable pipe deformation caused by inward deformation of freeze wall. Finally, successful application of this method to the large scale coal mine-Jining No.2 Mine in Shandong Province of China, is presented.It saved much investment compared with F.A.Auld's design for the same mine on behalf of Shell Coal International.

  17. Genetics of winter wheat response to two freezing treatments

    Science.gov (United States)

    The inheritance of the ability of winter wheat plants to survive two kinds of freezing stress was investigated in a five-parent diallel cross. Plants were acclimated at +4°C for 5 wks and frozen with or without a –3°C, 16-hour pre-freezing (PF) period prior to freezing to damaging temperatures. The ...

  18. Ribosome binding sites visualized on freeze-fractured membranes of the rough endoplasmic reticulum

    OpenAIRE

    1980-01-01

    Freeze-fracture micrographs of cells of the green alga Micrasterias denticulata stabilized by ultrarapid freezing reveal imprints of polysomes on the rough endoplasmic reticulum membranes. The imprints appear as broad, spiral ridges on the P faces and as corresponding wide grooves on the E faces of the membranes. Distinct 110-A particles with a spacing of 270 +/- 45 A are associated with the P-face ridges. Where imprints of individual ribosomes can be discerned, it is seen that there is a 1:1...

  19. Comparing contact and immersion freezing from continuous flow diffusion chambers

    Science.gov (United States)

    Nagare, Baban; Marcolli, Claudia; Welti, André; Stetzer, Olaf; Lohmann, Ulrike

    2016-07-01

    Ice nucleating particles (INPs) in the atmosphere are responsible for glaciating cloud droplets between 237 and 273 K. Different mechanisms of heterogeneous ice nucleation can compete under mixed-phase cloud conditions. Contact freezing is considered relevant because higher ice nucleation temperatures than for immersion freezing for the same INPs were observed. It has limitations because its efficiency depends on the number of collisions between cloud droplets and INPs. To date, direct comparisons of contact and immersion freezing with the same INP, for similar residence times and concentrations, are lacking. This study compares immersion and contact freezing efficiencies of three different INPs. The contact freezing data were obtained with the ETH CoLlision Ice Nucleation CHamber (CLINCH) using 80 µm diameter droplets, which can interact with INPs for residence times of 2 and 4 s in the chamber. The contact freezing efficiency was calculated by estimating the number of collisions between droplets and particles. Theoretical formulations of collision efficiencies gave too high freezing efficiencies for all investigated INPs, namely AgI particles with 200 nm electrical mobility diameter, 400 and 800 nm diameter Arizona Test Dust (ATD) and kaolinite particles. Comparison of freezing efficiencies by contact and immersion freezing is therefore limited by the accuracy of collision efficiencies. The concentration of particles was 1000 cm-3 for ATD and kaolinite and 500, 1000, 2000 and 5000 cm-3 for AgI. For concentrations nucleation process that is enhanced compared to immersion freezing due to the position of the INP on the droplet, and we discriminate it from collisional contact freezing, which assumes an enhancement due to the collision of the particle with the droplet. For best comparison with contact freezing results, immersion freezing experiments of the same INPs were performed with the continuous flow diffusion chamber Immersion Mode Cooling chAmber-Zurich Ice

  20. Potential role of salicylic acid in modulating diacylglycerol homeostasis in response to freezing temperatures in Arabidopsis.

    Science.gov (United States)

    Tan, Wei-Juan; Xiao, Shi; Chen, Qin-Fang

    2015-01-01

    In our recent article in Molecular Plant, we reported that 3 lipase-like defense regulators SENESCENCE-ASSOCIATED GENE101 (SAG101), ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1) and PHYTOALEXIN DEFICIENT4 (PAD4) are involved in the regulation of freezing tolerance in Arabidopsis. The transcripts of SAG101, EDS1 and PAD4 were inducible by cold stress and their knockout or knockdown mutants exhibited enhanced chilling and freezing tolerance in comparison to the wild type. The freezing tolerance phenotype showed in the sag101, eds1 and pad4 mutants was correlated with the transcriptional upregulation of C-REPEAT/DRE BINDING FACTORs (CBFs) and their regulons as well as increased levels of proline. Upon cold exposure, the sag101, eds1 and pad4 mutants showed ameliorated cell death and accumulation of hydrogen peroxide, which were highly induced by freezing stress in the wild-type leaves. Moreover, the contents of salicylic acid (SA) and diacylglycerol (DAG) were significantly decreased in the sag101, eds1 and pad4 mutants compared to the wild type. Taken together, our results suggest that the SAG101, EDS1 and PAD4 are negative regulators in the freezing response and function, at least in part, by modulating the homeostasis of SA and DAG in Arabidopsis.

  1. MICROSTRUCTURE MODIFICATIONS INDUCED IN SPRUCE WOOD BY FREEZING

    Directory of Open Access Journals (Sweden)

    Maria Bernadett SZMUTKU

    2011-12-01

    Full Text Available Scanning Electron Microscopy (SEM is amodern, non-invasive method for objective andspecialized image analysis of anatomical materialfeatures at microscopic level. Referring to wood, itoffers the possibility to view in 3D a bunch ofneighboring cells, in all three grain directions.This allows the imaging of modifications thatmight appear in the structure of the wood cellmembrane (e.g. micro-fissures caused by differentfactors, including temperature variations. This paperpresents the results of the SEM analysis performedon European spruce (Picea abies samples, cut fromboards which were subjected to freezing and thawingunder different conditions of temperature variationand time of exposure.The main aim of this research was to reveal theconditions which determine the occurrence of microfissuresin the cell wall and consequently lead tostrength losses in wood.

  2. Freezing Tolerance of Bulb Scales of Lily Cultivars : Effects of Freezing and Storage Duration and Partial Dehydration

    NARCIS (Netherlands)

    Bonnier, Frans J.M.; Jansen, Ritsert C.; Tuyl, Jaap M. van

    1997-01-01

    Effects of freezing duration, previous storage duration of bulbs at -2 °C, and partial dehydration of scales on freezing tolerance of lily (Lilium hybrids) scales were studied for a series of cultivars. Freezing tolerance of scales was estimated by measuring ion leakage and recording scale bulblet r

  3. High ice nucleation activity located in blueberry stem bark is linked to primary freeze initiation and adaptive freezing behaviour of the bark.

    Science.gov (United States)

    Kishimoto, Tadashi; Yamazaki, Hideyuki; Saruwatari, Atsushi; Murakawa, Hiroki; Sekozawa, Yoshihiko; Kuchitsu, Kazuyuki; Price, William S; Ishikawa, Masaya

    2014-01-01

    Controlled ice nucleation is an important mechanism in cold-hardy plant tissues for avoiding excessive supercooling of the protoplasm, for inducing extracellular freezing and/or for accommodating ice crystals in specific tissues. To understand its nature, it is necessary to characterize the ice nucleation activity (INA), defined as the ability of a tissue to induce heterogeneous ice nucleation. Few studies have addressed the precise localization of INA in wintering plant tissues in respect of its function. For this purpose, we recently revised a test tube INA assay and examined INA in various tissues of over 600 species. Extremely high levels of INA (-1 to -4 °C) in two wintering blueberry cultivars of contrasting freezing tolerance were found. Their INA was much greater than in other cold-hardy species and was found to be evenly distributed along the stems of the current year's growth. Concentrations of active ice nuclei in the stem were estimated from quantitative analyses. Stem INA was localized mainly in the bark while the xylem and pith had much lower INA. Bark INA was located mostly in the cell wall fraction (cell walls and intercellular structural components). Intracellular fractions had much less INA. Some cultivar differences were identified. The results corresponded closely with the intrinsic freezing behaviour (extracellular freezing) of the bark, icicle accumulation in the bark and initial ice nucleation in the stem under dry surface conditions. Stem INA was resistant to various antimicrobial treatments. These properties and specific localization imply that high INA in blueberry stems is of intrinsic origin and contributes to the spontaneous initiation of freezing in extracellular spaces of the bark by acting as a subfreezing temperature sensor. PMID:25082142

  4. The freezing and supercooling of garlic (Allium sativum L.)

    Energy Technology Data Exchange (ETDEWEB)

    James, Christian; Seignemartin, Violaine; James, Stephen J. [Food Refrigeration and Process Engineering Research Centre (FRPERC), University of Bristol, Churchill Building, Langford, Bristol BS40 5DU (United Kingdom)

    2009-03-15

    This work shows that peeled garlic cloves demonstrate significant supercooling during freezing under standard conditions and can be stored at temperatures well below their freezing point (-2.7 C) without freezing. The nucleation point or 'metastable limit temperature' (the point at which ice crystal nucleation is initiated) of peeled garlic cloves was found to be between -7.7 and -14.6 C. Peeled garlic cloves were stored under static air conditions at temperatures between -6 and -9 C for up to 69 h without freezing, and unpeeled whole garlic bulbs and cloves were stored for 1 week at -6 C without freezing. (author)

  5. [Studies of viability and vitality after freezing of the probiotic yeast Saccharomyces boulardii: physiological preconditioning effect].

    Science.gov (United States)

    Pardo, Silvina; Galvagno, Miguel Angel; Cerrutti, Patricia

    2009-06-30

    The aim of this study was to evaluate the vitality and viability of the probiotic yeast Saccharomyces boulardii after freezing/thawing and the physiological preconditioning effect on these properties. The results indicate that the specific growth rate (0.3/h(-1)) and biomass (2-3 x10(8)cells/ml) of S. boulardii obtained in flasks shaken at 28 degrees C and at 37 degrees C were similar. Batch cultures of the yeast in bioreactors using glucose or sugar-cane molasses as carbon sources, reached yields of 0.28 g biomass/g sugar consumed, after 10h incubation at 28 degrees C; the same results were obtained in fed batch fermentations. On the other hand, in batch cultures, the vitality of cells recovered during the exponential growth phase was greater than the vitality of cells from the stationary phase of growth. Vitality of cells from fed-batch fermentations was similar to that of stationary growing cells from batch fermentations. Survival to freezing at -20 degrees C and subsequent thawing of cells from batch cultures was 0.31% for cells in exponential phase of growth and 11.5% for cells in stationary phase. Pre-treatment of this yeast in media with water activity (a(w)) 0.98 increased the survival to freezing of S. boulardii cells stored at -20 degrees C for 2 months by 10 fold. Exposure of the yeast to media of reduced a(w) and/or freezing/thawing process negatively affected cell vitality. It was concluded that stress conditions studied herein decrease vitality of S. boulardii. Besides, the yeast strain studied presented good tolerance to bile salts even at low pH values. PMID:19631167

  6. Ultrastructure and energy-dispersive x-ray microanalysis of cartilage after rapid freezing, low temperature freeze drying, and embedding in Spurr's resin

    Energy Technology Data Exchange (ETDEWEB)

    Appleton, J.; Lyon, R.; Swindin, K.J.; Chesters, J.

    1985-10-01

    In order to undertake meaningful high-resolution x-ray microanalysis of tissues, methods should be used that minimize the introduction of artefacts produced by loss or translocation of ions. The most ideal method is rapid freezing but the subsequent sectioning of frozen tissues is technically difficult. An alternative method is to freeze dry the tissues at a low temperature, and then embed them in resin. This facilitates the rapid production of reproducible thin sections. With freeze-dried, embedded hypertrophic cartilage, the morphology was similar to that seen using aqueous fixatives even when no additional electron density is introduced by the use of osmium vapor. Energy-dispersive analysis of specific areas show that little or no loss or migration of ions occurs from structures such as mitochondria. Mitochondrial granules consisting of calcium and phosphorus precipitates were not observed except where the cells were damaged as a result of the freezing process. This may suggest that these granules only appear when tissue is damaged because of inadequate preservation.

  7. Assessment of three generations of mice derived by ICSI using freeze-dried sperm

    Science.gov (United States)

    Li, Ming-Wen; Willis, Brandon J.; Griffey, Stephen M.; Spearow, Jimmy L.; Lloyd, K.C. Kent

    2016-01-01

    Summary Although the derivation of mice by intracytoplasmic sperm injection (ICSI) using freeze-dried sperm has been demonstrated previously, a comprehensive analysis of their viability, health, and fertility has not. The purpose of the present study was to determine the extent to which ICSI using freeze-dried sperm stored at 4 °C for 1–2 months from mice on either an inbred (C57BL/6J) or hybrid (B6D2F1/J) genetic background results in genomic instability and/or phenotypic abnormality in mice and two generations of their progeny. Fertilization rates (number of 2-cells per injected oocytes) using ICSI of fresh and freeze-dried sperm were similar within and between mouse strains, although fewer freeze-dried sperm-derived embryos than fresh sperm-derived embryos developed to blastocysts in vitro (C57BL/6J and B6D2F1/J) and liveborn pups in vivo (B6D2F1/J only). Nevertheless, once born, mice derived by ICSI using freeze-dried sperm in both mouse strains were healthy and reproductively sound. No major differences in litter size, weaning rate, and sex ratio were noted in the two generations of progeny (F2 and F3) of ICSI-derived offspring using freeze-dried sperm compared with that in the natural mating (control) group. Further, there was no evidence that either ICSI or freeze drying induced genomic instability, as determined by microsatellite analysis of the derived mice and subsequent generations when compared with both parental genotypes, nor were there differences in the number or types of pathological changes in any of the three generations of progeny. We conclude that viable, healthy and genomically stable mice can be derived by ICSI using freeze-dried mouse sperm stored in the refrigerator for at least 2 months. Further, because freeze drying is a simpler and more economical technique compared with embryo and sperm cryopreservation, the results of this study justify additional research to continue to develop and enhance the technique for the preservation

  8. Ground freezing for containment of hazardous waste

    Energy Technology Data Exchange (ETDEWEB)

    Sayles, F.N.; Iskandar, I.K.

    1998-07-01

    The freezing of ground for the containment of subsurface hazardous waste is a promising method that is environmentally friendly and offers a safe alternative to other methods of waste retention in many cases. The frozen soil method offers two concepts for retaining waste. One concept is to freeze the entire waste area into a solid block of frozen soil thus locking the waste in situ. For small areas where the contaminated soil does not include vessels that would rupture from frost action, this concept may be simpler to install. A second concept, of course, is to create a frozen soil barrier to confine the waste within prescribed unfrozen soil boundaries; initial research in this area was funded by EPA, Cincinnati, OH, and the Army Corps of Engineers. The paper discusses advantages and limitations, a case study from Oak Ridge, TN, and a mesh generation program that simulates the cryogenic technology.

  9. Cold acclimation induced accumulation of phenolic compounds and freezing tolerance in Ammopiptanthus mongolicus

    Institute of Scientific and Technical Information of China (English)

    Liu Mei-qin; Chen Yi-yin; Lu Cun-fu; Zhang Hui; Yin Wei-lun

    2007-01-01

    Ammopiptanthus mongolicus, the only freezing tolerant evergreen broad-leaved shrub, local species of the Alashan desert,northwest sand area of China, can survive -30℃ or even lower temperature in winter. In the present study, the secondary products phenolics in A. mongolicus cotyledons were determined to study the effects ofphenolics on cold tolerance. Cytochemical localization of phenolics in cotyledon cells was observed by electron microscopy and the content of phenolic compounds was assayed by spectrophotometric measurement. The results showed that the freezing tolerance of A. mongolicus seedlings increased after acclimation at 2-6℃ for 14 days, which accompanied the increase of the content of phenolic compounds in cotyledons. Cytochemical observation showed that phenolic deposits were mainly localized in vacuoles and in close proximity to tonoplast, and also in the cytoplasm. The amount and the size of phenolics droplets increased obviously in cytoplasm and vacuoles after cold acclimation, predominantly aggregated along membranes of vacuoles and tonoplast. No phenolic deposits were found in cell walls. As hydrogen- or electron-donating agents, phenolics may protect plant cells against reactive oxygen species formed during chilling or freezing stress and improve the freezing tolerance of cold-acclimated A. mongolicus seedlings.

  10. Freezing enhancement around a horizontal tube using copper foil disks

    Science.gov (United States)

    Sugawara, M.; Komatsu, Y.; Takahashi, Y.; Beer, H.

    2011-12-01

    Freezing of water saturated in circumferentially arranged copper foils around a cooling tube is studied experimentally and numerically. The copper foils need not to be welded to the cooling tube but are merely placed around the tube so that the freezing system is easily arranged. Copper foils greatly enhance freezing compared with that of a bare tube, even with a small copper volume fraction in the freezing system. Numerical calculations by means of a continuum model predict well freezing enhancement. The effect of the copper foils is also considered numerically for the melting process in order to compare with freezing. It is seen that copper foils contribute more to the melting enhancement than to the increase of the freezing rate.

  11. Scaling-Up Eutectic Freeze Crystallization

    OpenAIRE

    Genceli, F.E.

    2008-01-01

    A novel crystallization technology, Eutectic Freeze Crystallization (EFC) has been investigated and further developed in this thesis work. EFC operates around the eutectic temperature and composition of aqueous solutions and can be used for recovery of (valuable) dissolved salts (and/or or acids) and water from a wide variety of aqueous process streams. Using EFC, processes producing large quantities of saline solutions could be carried out in an ecologically and economically attractive way. ...

  12. 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.

  13. 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.

  14. Atmospheric freeze drying assisted by power ultrasound

    Science.gov (United States)

    Santacatalina, J. V.; Cárcel, J. A.; Simal, S.; Garcia-Perez, J. V.; Mulet, A.

    2012-12-01

    Atmospheric freeze drying (AFD) is considered an alternative to vacuum freeze drying to keep the quality of fresh product. AFD allows continuous drying reducing fix and operating costs, but presents, as main disadvantage, a long drying time required. The application of power ultrasound (US) can accelerate AFD process. The main objective of the present study was to evaluate the application of power ultrasound to improve atmospheric freeze drying of carrot. For that purpose, AFD experiments were carried out with carrot cubes (10 mm side) at constant air velocity (2 ms-1), temperature (-10°C) and relative humidity (10%) with (20.5 kWm-3,USAFD) and without (AFD) ultrasonic application. A diffusion model was used in order to quantify the influence of US in drying kinetics. To evaluate the quality of dry products, rehydration capacity and textural properties were determined. The US application during AFD of carrot involved the increase of drying rate. The effective moisture diffusivity identified in USAFD was 73% higher than in AFD experiments. On the other hand, the rehydration capacity was higher in USAFD than in AFD and the hardness of dried samples did not show significant (p<0.05) differences. Therefore, US application during AFD significantly (p<0.05) sped-up the drying process preserving the quality properties of the dry product.

  15. The influence of freezing rates on bovine pericardium tissue Freeze-drying

    Directory of Open Access Journals (Sweden)

    Camila Figueiredo Borgognoni

    2009-12-01

    Full Text Available The bovine pericardium has been used as biomaterial in developing bioprostheses. Freeze-drying is a drying process that could be used for heart valve's preservation. The maintenance of the characteristics of the biomaterial is important for a good heart valve performance. This paper describes the initial step in the development of a bovine pericardium tissue freeze-drying to be used in heart valves. Freeze-drying involves three steps: freezing, primary drying and secondary drying. The freezing step influences the ice crystal size and, consequently, the primary and secondary drying stages. The aim of this work was to investigate the influence of freezing rates on the bovine pericardium tissue freeze-drying parameters. The glass transition temperature and the structural behaviour of the lyophilized tissues were determined as also primary and secondary drying time. The slow freezing with thermal treatment presented better results than the other freeze-drying protocols.O pericárdio bovino é um material utilizado na fabricação de biopróteses. A liofilização é um método de secagem que vem sendo estudado para a conservação de válvulas cardíacas. A preservação das características do biomaterial é de fundamental importância no bom funcionamento das válvulas. Este artigo é a primeira etapa do desenvolvimento do ciclo de liofilização do pericárdio bovino. Liofilização é o processo de secagem no qual a água é removida do material congelado por sublimação e desorção da água incongelável, sob pressão reduzida. O congelamento influencia o tamanho do cristal de gelo e, consequentemente, a secagem primária e secundária. O objetivo deste estudo foi verificar a influência das taxas de congelamento nos parâmetros de liofilização do pericárdio bovino. Determinou-se a temperatura de transição vítrea e o comportamento estrutural do pericárdio bovino liofilizado. Determinou-se o tempo da secagem primária e secundária. O

  16. Food freezing with simultaneous surface dehydration: approximate prediction of weight loss during freezing and storage

    Energy Technology Data Exchange (ETDEWEB)

    Campanone, Laura A.; Salvadori, Viviana O.; Mascheroni, Rodolfo H. [Centro de Investigacion Desarollo en Criotecnologia de Alimentos (CIDCA), Facultad de Ciencias Exactas, La Plata (Argentina); MODIAL, Facultad de Ingenieria, La Plata (Argentina)

    2005-03-01

    Weight loss of unpackaged foods during freezing and later storage is an important quality and economic issue. It is originated on surface ice sublimation due to differences in water activity between food surface and the refrigerating air. Weight loss rate is determined by refrigerating conditions and product characteristics. The modelling of this phenomenon has merited very little attention; at present there are no simplified methods to predict weight losses during the freezing and the storage of unpackaged foods. In previous studies we developed a detailed model for the simultaneous heat and mass transfer during food freezing and storage with ice sublimation. Based on the information of this numerical model, simplified analytical methods for the prediction of weight loss during the freezing and the storage of unpackaged frozen foods were developed. The methods account for product characteristics and storage conditions. The prediction equations are very simple and results of their use - simulating usual freezing and storage conditions for different products - give very good accuracy when tested against the previously cited numerical model and experimental data. (Author)

  17. Development of metal-carbon eutectic cells for application as high temperature reference points in nuclear reactor severe accident tests: Results on the Fe-C, Co-C, Ti-C and Ru-C alloys' melting/freezing transformation temperature under electromagnetic induction heating

    International Nuclear Information System (INIS)

    With the aim of reducing the high temperature measurement uncertainty of nuclear reactor severe accident experimental tests at the PLINIUS platform in Cadarache Research Centre, France, a variety of graphite cells containing a metal-carbon eutectic mix have been tested to assess the melting/freezing temperature reproducibility and their feasibility as calibration cells for thermometers. The eutectic cells have been thermally cycled in an induction furnace to assess the effect of heating/cooling rate, metal purity, graphite crucible design, and binary system constituents on the eutectic transformation temperature. A bi-chromatic pyrometer was used to perform temperature measurements in the graphite cell black cavity containing the metal-carbon eutectic mix. The eutectic points analyzed are all over 1100 C and cover an almost thousand degree span, i.e. from the Fe-Fe3C to the Ru-C eutectic. The induction heating permitted the attainment of heating and cooling rates of over 200 C/min under an inert atmosphere. The conducted tests allowed the determination of general trends and peculiarities of the solid. liquid transformation temperature under non-equilibrium and non-steady-state conditions of a variety of eutectic alloys (Fe-C, Co-C, Ti-C and Ru-C binary systems). (authors)

  18. Leavening ability and freeze tolerance of yeasts isolated from traditional corn and rye bread doughs.

    Science.gov (United States)

    Almeida, M J; Pais, C

    1996-12-01

    Strains of Saccharomyces cerevisiae and Torulaspora delbrueckii isolated from traditional bread doughs displayed dough-raising capacities similar to the ones found in baker's yeasts. During storage of frozen doughs, strains of T. delbrueckii (IGC 5321, IGC 5323, and IGC 4478) presented approximately the same leavening ability for 30 days. Cell viability was not significantly affected by freezing, but when the dough was submitted to a bulk fermentation before being stored at -20 degrees C, there was a decrease in the survival ratio which depended on the yeast strain. Furthermore, the leavening ability after 4 days of storage decreased as the prefermentation period of the dough before freezing increased, except for strains IGC 5321 and IGC 5323. These two strains retained their fermentative activity after 15 days of storage and 2.5 h of prefermentation, despite showing a reduction of viable cells under the same conditions. The intracellular trehalose content was higher than 20% (wt/wt) in four of the yeasts tested: the two commercial strains of baker's yeast (S. cerevisiae IGC 5325 and IGC 5326) and the two mentioned strains of T. delbrueckii (IGC 5321 and IGC 5323). However, the strains of S. cerevisiae were clearly more susceptible to freezing damages, indicating that other factors may contribute to the freeze tolerance of these yeasts. PMID:8953712

  19. Survival and Growth of Epidemically Successful and Nonsuccessful Salmonella enterica Clones after Freezing and Dehydration

    DEFF Research Database (Denmark)

    Müller, Karoline; Aabo, Søren; Birk, Tina;

    2012-01-01

    contribute. During epidemiological studies, a mathematical model was developed that allowed estimation of a factor (q) accounting for the relative ability of Salmonella serovars with different antimicrobial resistances to survive in the food chain and cause human disease. Based on this q-factor, 26...... Salmonella isolates were characterized as successful or nonsuccessful. We studied the survival and growth of stationary- and exponential-phase cells of these isolates after freezing for up to 336 days in minced meat. We also investigated survival and growth after dehydration at 10°C and 82% relative humidity...... (RH) and 25°C and 49% RH for 112 days. Stationary-phase cells were reduced by less than 1 log unit during 1 year of freezing, and growth was initiated with an average lag phase of 1.7 h. Survival was lower in exponentialphase cells, but lag phases tended to be shorter. High humidity and low...

  20. Stability evaluation of freeze-dried Lactobacillus paracasei subsp. tolerance and Lactobacillus delbrueckii subsp. bulgaricus in oral capsules.

    Science.gov (United States)

    Jalali, M; Abedi, D; Varshosaz, J; Najjarzadeh, M; Mirlohi, M; Tavakoli, N

    2012-01-01

    Freeze-drying is a common preservation technology in the pharmaceutical industry. Various studies have investigated the effect of different cryoprotectants on probiotics during freeze-drying. However, information on the effect of cryoprotectants on the stability of some Lactobacillus strains during freeze-drying seems scarce. Therefore, the aim of the present study was to establish production methods for preparation of oral capsule probiotics containing Lactobacillus paracasei subsp. tolerance and Lactobacillus delbrueckii subsp. Bulgaricus. It was also of interest to examine the effect of various formulations of cryoprotectant media containing skim milk, trehalose and sodium ascorbate on the survival rate of probiotic bacteria during freeze-drying at various storage temperatures. Without any cryoprotectant, few numbers of microorganisms survived. However, microorganisms tested maintained higher viability after freeze-drying in media containing at least one of the cryoprotectants. Use of skim milk in water resulted in an increased viability after lyophilization. Media with a combination of trehalose and skim milk maintained a higher percentage of live microorganisms, up to 82%. In general, bacteria retained a higher number of viable cells in capsules containing freeze-dried bacteria with sodium ascorbate after three months of storage. After this period, a marked decline was observed in all samples stored at 23°C compared to those stored at 4°C. The maximum survival rate (about 72-76%) was observed with media containing 6% skim milk, 8% trehalose and 4% sodium ascorbate.

  1. Improving freeze-tolerance of baker's yeast through seamless gene deletion of NTH1 and PUT1.

    Science.gov (United States)

    Dong, Jian; Chen, Didi; Wang, Guanglu; Zhang, Cuiying; Du, Liping; Liu, Shanshan; Zhao, Yu; Xiao, Dongguang

    2016-06-01

    Baker's yeast strains with freeze-tolerance are highly desirable to maintain high leavening ability after freezing. Enhanced intracellular concentration of trehalose and proline in yeast is linked with freeze-tolerance. In this study, we constructed baker's yeast with enhanced freeze-tolerance by simultaneous deletion of the neutral trehalase-encoded gene NTH1 and the proline oxidase-encoded gene PUT1. We first used the two-step integration-based seamless gene deletion method to separately delete NTH1 and PUT1 in haploid yeast. Subsequently, through two rounds of hybridization and sporulation-based allelic exchange and colony PCR-mediated tetrad analysis, we obtained strains with restored URA3 and deletion of NTH1 and/or PUT1. The resulting strain showed higher cell survival and dough-leavening ability after freezing compared to the wild-type strain due to enhanced accumulation of trehalose and/or proline. Moreover, mutant with simultaneous deletion of NTH1 and PUT1 exhibits the highest relative dough-leavening ability after freezing compared to mutants with single-gene deletion perhaps due to elevated levels of both trehalose and proline. These results verified that it is applicable to construct frozen dough baker's yeast using the method proposed in this paper. PMID:26965428

  2. Lipophilic components of the brown seaweed, Ascophyllum nodosum, enhance freezing tolerance in Arabidopsis thaliana.

    Science.gov (United States)

    Rayirath, Prasanth; Benkel, Bernhard; Mark Hodges, D; Allan-Wojtas, Paula; Mackinnon, Shawna; Critchley, Alan T; Prithiviraj, Balakrishnan

    2009-06-01

    Extracts of the brown seaweed Ascophyllum nodosum enhance plant tolerance against environmental stresses such as drought, salinity, and frost. However, the molecular mechanisms underlying this improved stress tolerance and the nature of the bioactive compounds present in the seaweed extracts that elicits stress tolerance remain largely unknown. We investigated the effect of A. nodosum extracts and its organic sub-fractions on freezing tolerance of Arabidopsis thaliana. Ascophyllum nodosum extracts and its lipophilic fraction significantly increased tolerance to freezing temperatures in in vitro and in vivo assays. Untreated plants exhibited severe chlorosis, tissue damage, and failed to recover from freezing treatments while the extract-treated plants recovered from freezing temperature of -7.5 degrees C in in vitro and -5.5 degrees C in in vivo assays. Electrolyte leakage measurements revealed that the LT(50) value was lowered by 3 degrees C while cell viability staining demonstrated a 30-40% reduction in area of damaged tissue in extract treated plants as compared to water controls. Moreover, histological observations of leaf sections revealed that extracts have a significant effect on maintaining membrane integrity during freezing stress. Treated plants exhibited 70% less chlorophyll damage during freezing recovery as compared to the controls, and this correlated with reduced expression of the chlorphyllase genes AtCHL1 and AtCHL2. Further, the A. nodosum extract treatment modulated the expression of the cold response genes, COR15A, RD29A, and CBF3, resulting in enhanced tolerance to freezing temperatures. More than 2.6-fold increase in expression of RD29A, 1.8-fold increase of CBF3 and two-fold increase in the transcript level of COR15A was observed in plants treated with lipophilic fraction of A. nodosum at -2 degrees C. Taken together, the results suggest that chemical components in A. nodosum extracts protect membrane integrity and affect the expression of

  3. Dynamical freeze-out in event-by-event hydrodynamics

    CERN Document Server

    Holopainen, Hannu

    2012-01-01

    In hydrodynamical modeling of the ultrarelativistic heavy-ion collisions the freeze-out is typically performed at a constant temperature or density. In this work we apply a dynamical freeze-out criterion, which compares the hydrodynamical expansion rate with the pion scattering rate. Recently many calculations have been done using event-by-event hydrodynamics where the initial density profile fluctuates from event to event. In these event-by-event calculations the expansion rate fluctuates strongly as well, and thus it is interesting to check how the dynamical freeze-out changes hadron distributions with respect to the constant temperature freeze-out. We present hadron spectra and elliptic flow calculated using (2+1)-dimensional ideal hydrodynamics, and show the differences between constant temperature and dynamical freeze-out criteria. We find that the differences caused by different freeze-out criteria are small in all studied cases.

  4. Well-plate freeze-drying

    DEFF Research Database (Denmark)

    Trnka, Hjalte; Rantanen, Jukka; Grohganz, Holger

    2015-01-01

    , cake collapse and reconstitution time. Results: Samples freeze-dried in well-plates had an acceptable visual cake appearance. Solid form analysis by high throughput X-ray powder diffraction indicated comparable polymorphic outcome independent of the container. The expected increase in moisture level...... due to increasing amount of amorphous matter in the samples was observed in both vials and well plates. Cake collapse was found to be representative in well plates and could be effectively quantified using image analysis. Reconstitution time was also found to be equal in all three platforms. Finally...

  5. Mechano-freezing of the ambient water

    CERN Document Server

    Zhang, Xi; Zou, Bo; Sun, Chang Q

    2013-01-01

    Raman spectroscopy examination of the 25 deg-C water freezing under compression revealed transition from 1.35 GPa to 0.86 GPa upon ice being formed at continued volume change. The transition is associated with a slight blue shift of the high-frequency phonon (omiga_H ~ 3120 cm-1) and creation of the low-frequency phonons (Omiga_L ~ 200 cm-1). In the liquid and in the solid phase, the increased pressure softens the Omiga_H and stiffens the Omida_L, which indicates the presence of the inter-electron-pair repulsion in both liquid and solid water.

  6. PREPARATION OF POROUS NANOCOMPOSITE SCAFFOLDS WITH HONEYCOMB MONOLITH STRUCTURE BY ONE PHASE SOLUTION FREEZE DRYING METHOD

    Institute of Scientific and Technical Information of China (English)

    Yang Xu; Duo Zhang; Zong-liang Wang; Zhan-tuan Gao; Pei-biao Zhang; Xue-si Chen

    2011-01-01

    Biodegradable porous nanocomposite scaffolds of poly(lactide-co-glycolide) (PLGA) and L-lactic acid (LAc) oligomer surface-grafted hydroxyapatite nanoparticles (op-HA) with a honeycomb monolith structure were fabricated with the single-phase solution freeze-drying method. The effects of different freezing temperatures on the properties of the scaffolds, such as microstructures, compressive strength, cell penetration and cell proliferation were studied. The highly porous and well interconnected scaffolds with a tunable pore structure were obtained. The effect of different freezing temperature (4℃, -20℃, -80℃ and -196℃) was investigated in relation to the scaffold morphology, the porosity varied from 91.2% to 83.0% and the average pore diameter varied from (167.2 ± 62.6) pm to (11.9 ± 4.2) μm while the σ10 increased significantly. The cell proliferation were decreased and associated with the above-mentioned properties. Uniform distribution of op-HA particles and homogeneous roughness of pore wall surfaces were found in the 4℃ frozen scaffold. The 4℃ frozen scaffold exhibited better cell penetration and increased cell proliferation because of its larger pore size, higher porosity and interconnection. The microstmctures described here provide a new approach for the design and fabrication of op-HA/PLGA based scaffold materials with potentially broad applicability for replacement of bone defects.

  7. Freeze-Thaw Durability of Air-Entrained Concrete

    OpenAIRE

    Huai-Shuai Shang; Ting-Hua Yi

    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. T...

  8. FEM Simulation on Artificial Freezing of Seepage Ground

    Institute of Scientific and Technical Information of China (English)

    ZHOU Xiao-min

    2005-01-01

    The coupling mechanism in freezing process of seepage ground was studied and a simplified coupling math model was proposed. The nonlinear and coupling problems of PDEs were well solved using the exponential function, error function and normal distribution function, and a series of FEM equations of coupled fields of temperature and seepage were deduced and put forward. With the example of shaft ground freezing, the formation of freezing wall in seepage ground was simulated.

  9. SOME STUDIES ABOUT CEREALS BEHAVIOR DURING FREEZE DRYING PROCESS

    OpenAIRE

    GABRIELA-VICTORIA MNERIE; D. MNERIE; D. ŢUCU

    2013-01-01

    The paper presents some special method and equipment and the principal advantages of freeze-dried food. The freeze drying is a good method of freeze-drying for make some experiments with many kind of cereals, for the improvement that in food production. It is necessary and is possible to study the corn oil extract, wheat flour, the maltodextrin from corn, modified cornstarch, spice extracts, soy sauce, hydrolyzed wheat gluten, partially hydrogenated soybean and cottonseed oil etc. That is ver...

  10. Identifying freezing of gait in Parkinson's disease during freezing provoking tasks using waist-mounted accelerometry

    NARCIS (Netherlands)

    Zach, H.; Janssen, A.M.; Snijders, A.H.; Delval, A.; Ferraye, M.U.; Auff, E.; Weerdesteyn, V.G.; Bloem, B.R.; Nonnekes, J.H.

    2015-01-01

    BACKGROUND: Freezing of gait (FOG) is a common and debilitating phenomenon in Parkinson's disease (PD). Wearable accelerometers might help to assess FOG in the research setting. Here, we evaluate whether accelerometry can detect FOG while executing rapid full turns and while walking with rapid short

  11. Experimental quantification of contact freezing in an electrodynamic balance

    Directory of Open Access Journals (Sweden)

    N. Hoffmann

    2013-04-01

    Full Text Available Heterogeneous nucleation of ice in a supercooled water droplet induced by an external contact with a dry aerosol particle has long been known to be more effective than freezing induced by the same nucleus immersed in the droplet. However, the experimental quantification of contact freezing is challenging. Here we report an experimental method allowing to determine the temperature dependent ice nucleation probability of size selected aerosol particles. The method uses supercooled charged water droplets suspended in a laminar flow of air containing aerosol particles as contact freezing nuclei. The rate of droplet–particle collisions is calculated numerically with account for Coulomb attraction, drag force and induced dipole interaction between charged droplet and aerosol particles. The calculation is verified by direct counting of aerosol particles collected by a levitated droplet. By repeating the experiment on individual droplets for a sufficient number of times, we are able to reproduce the statistical freezing behavior of a large ensemble of supercooled droplets and measure the average rate of freezing events. The freezing rate is equal to the product of the droplet–particle collision rate and the probability of freezing on a single contact, the latter being a function of temperature, size and composition of the contact ice nuclei. Based on these observations, we show that for the types of particles investigated so far, contact freezing is the dominating freezing mechanism on the time scale of our experiment.

  12. Experimental quantification of contact freezing in an electrodynamic balance

    Directory of Open Access Journals (Sweden)

    N. Hoffmann

    2013-09-01

    Full Text Available Heterogeneous nucleation of ice in a supercooled water droplet induced by external contact with a dry aerosol particle has long been known to be more effective than freezing induced by the same nucleus immersed in the droplet. However, the experimental quantification of contact freezing is challenging. Here we report an experimental method to determine the temperature-dependent ice nucleation probability of size-selected aerosol particles. The method is based on the suspension of supercooled charged water droplets in a laminar flow of air containing aerosol particles as contact freezing nuclei. The rate of droplet–particle collisions is calculated numerically with account for Coulomb attraction, drag force and induced dipole interaction between charged droplet and aerosol particles. The calculation is verified by direct counting of aerosol particles collected by a levitated droplet. By repeating the experiment on individual droplets for a sufficient number of times, we are able to reproduce the statistical freezing behavior of a large ensemble of supercooled droplets and measure the average rate of freezing events. The freezing rate is equal to the product of the droplet–particle collision rate and the probability of freezing on a single contact, the latter being a function of temperature, size and composition of the contact ice nuclei. Based on these observations, we show that for the types of particles investigated so far, contact freezing is the dominating freezing mechanism on the timescale of our experiment.

  13. A molecular dynamics study of freezing in a confined geometry

    Science.gov (United States)

    Ma, Wen-Jong; Banavar, Jayanth R.; Koplik, Joel

    1992-01-01

    The dynamics of freezing of a Lennard-Jones liquid in narrow channels bounded by molecular walls is studied by computer simulation. The time development of ordering is quantified and a novel freezing mechanism is observed. The liquid forms layers and subsequent in-plane ordering within a layer is accompanied by a sharpening of the layer in the transverse direction. The effects of channel size, the methods of quench, the liquid-wall interaction and the roughness of walls on the freezing mechanism are elucidated. Comparison with recent experiments on freezing in confined geometries is presented.

  14. Study on Freeze-drying Process of Dumpling Wrappers

    OpenAIRE

    Wanren Chen; Hua Li; Xingli Jiao; Xiang Gui

    2015-01-01

    The freeze-drying process of frozen dumpling wrappers is studied in this study. And the effects of drying time, drying temperature and the capacity of unit area in the freezing process on the drying rate and rehydration rate of freeze-drying dumpling wrappers are investigated. The result shows that, in the process of freeze-drying dumpling wrappers, the optimal condition is: drying time is 3 h, drying temperature is 45°C and the loadage of per unit area is 4.0 kg/m2.

  15. A“TIME-SPACE” RELATED DESIGNMETHOD OF FREEZING WALL

    Institute of Scientific and Technical Information of China (English)

    陈湘生

    1996-01-01

    Artificially ground freezing (AGF) is one of the main methods to establish temporarysupport for shaft sinking in unstable water bearing strata. Domde (1915) formula based on frozensoil strength has widely been used for designing freezing wall thickness. However, it can not en-sure the stability of freezing wall, nor guarantee the safety of shaft construction as frozen depth in-creases in unstable water bearing strata. F. A. Auld (1985, 1988)[1'2] presented a designmethod of freezing wall, which is on the basis of strength and stability, together with deformationof freezing wall. This paper, according to the practice in China, describes a "time -space" relat-ed design method for deep freezing wall. The method is based on "time-space" concept, whichincludes influence of excavation rate of advance, unsupported length of freezing wall and the sumpstate on inward deformation of freezing wall, and the allowable pipe deformation caused by in-ward deformation of freezing wall. Finally, successful application of this method to the large scalecoal mine-Jining No. 2 Mine[3] in Shandong Province of China is presented. It saved much invest-ment compared with F. A. Auld's design for the same mine.

  16. Study on Freeze-drying Process of Dumpling Wrappers

    Directory of Open Access Journals (Sweden)

    Wanren Chen

    2015-06-01

    Full Text Available The freeze-drying process of frozen dumpling wrappers is studied in this study. And the effects of drying time, drying temperature and the capacity of unit area in the freezing process on the drying rate and rehydration rate of freeze-drying dumpling wrappers are investigated. The result shows that, in the process of freeze-drying dumpling wrappers, the optimal condition is: drying time is 3 h, drying temperature is 45°C and the loadage of per unit area is 4.0 kg/m2.

  17. Parameter Sensitivity of the Microdroplet Vacuum Freezing Process

    Directory of Open Access Journals (Sweden)

    Zhijun Zhang

    2015-01-01

    Full Text Available The vacuum freezing process of microdroplets (1 mm. Pressure and droplet diameter have an effect on cooling and freezing stages, but initial temperature only affects the cooling stage. The thermal conductivity coefficient kl affected the cooling stage, whereas ki affected the freezing stage. Heat capacity Cl affected the cooling stage, but Ci has virtually no effect on all stages. The actual latent heat of freezing ΔH was also affected. Higher density corresponds to lower cooling rate in the cooling stage.

  18. Freezing of water droplets colliding with kaolinite particles

    DEFF Research Database (Denmark)

    Svensson, Erik Anders; Delval, Christophe Eric Ludovic; Freiherr von Und zu Hessberg, P J H;

    2009-01-01

    Contact freezing of single supercooled water droplets colliding with kaolinite dust particles has been investigated. The experiments were performed with droplets levitated in an electrodynamic balance at temperatures from 240 to 268 K. Under dry conditions freezing 5 was observed to occur below 249...... K, while a freezing threshold of 267 K was observed at high relative humidity. The effect of relative humidity is attributed to an influence on the contact freezing process for the kaolinite-water droplet system, and it is not related to the lifetime of the droplets in the electrodynamic balance...

  19. Automated assessment of Pavlovian conditioned freezing and shock reactivity in mice using the VideoFreeze system

    Directory of Open Access Journals (Sweden)

    Stephan G Anagnostaras

    2010-09-01

    Full Text Available The Pavlovian conditioned freezing paradigm has become a prominent mouse and rat model of learning and memory, as well as of pathological fear. Due to its efficiency, reproducibility, and well-defined neurobiology, the paradigm has become widely adopted in large-scale genetic and pharmacological screens. However, one major shortcoming of the use of freezing behavior has been that it has required the use of tedious hand scoring, or a variety of proprietary automated methods that are often poorly validated or difficult to obtain and implement. Here we report an extensive validation of the Video Freeze system in mice, a turn-key all-inclusive system for fear conditioning in small animals. Using digital video and near-infrared lighting, the system achieved outstanding performance in scoring both freezing and movement. Given the large-scale adoption of the conditioned freezing paradigm, we encourage similar validation of other automated systems for scoring freezing, or other behaviors.

  20. Influence of freezing on strata behaviour

    Energy Technology Data Exchange (ETDEWEB)

    Stoss, K.; Oellers, T.

    1985-10-10

    Freezing the ground has not only the desired beneficial effects but can also have unavoidable deleterious effects on strata properties and behaviour. The usual calculation for dimensioning the ice wall does provide some valuable indications about strata behaviour during sinking. But the effects from fissure and ice lentil formation caused by temperature are not recorded, nor can they be in practical terms. For certain beds it is advisable to complement the preliminary investigations carried out on core samples to establish strata properties and behaviour when frozen by investigations into the degree of frost endangerment (ice lentils). In critical beds regular convergence measurement in the shaft with continuous evaluation are the most important means of avoiding unpleasant surprises during sinking. A properly tested array of additional support measures has to be ready to keep strata movement around the shaft within acceptable limits even in unfavourable conditions. To sum up: Success in sinking a freezing shaft does not only depend on the care during preliminary examinations, planning and preparation but also to a large degree on the ability and experience of the shaft construction engineers and on the decisions which these will make at the site in each case on the basis of what they see and measure. (orig./MOS).

  1. Asset Freezing: Smart Sanction or Criminal Charge?

    Directory of Open Access Journals (Sweden)

    Wouter de Zanger

    2011-02-01

    Full Text Available In this article the question is asked whether asset freezing can be qualified as a criminal charge within the meaning of Article6 ECHR and if yes, what effects this qualification may have on the legislative framework on so called smart sanctions. Byanalysing Community and EU law and case law of the European Court of Human Rights, General Court of Instance andCourt of Justice of the European Communities the authors give an overview of the notion and possible qualification of assetfreezing as a criminal charge. The article further focusses on the consequenses of qualifying asset freezing as a criminal chargeunder ECHR and EC/EU law and concludes by answering the aforementioned question.This article is a rewrite of a research paper written under supervision of prof. dr. J.A.E. Vervaele and prof. dr. C.H. Brants(Willem Pompe Institute for Criminal Law and Criminology, Utrecht University School of Law, whom the authors wouldlike to thank for their useful comments and supervision.

  2. Immersion freezing of birch pollen washing water

    Directory of Open Access Journals (Sweden)

    S. Augustin

    2012-12-01

    Full Text Available In the present study, the immersion freezing behavior of birch pollen, i.e. its ice nucleating active (INA macromolecules, was investigated at the Leipzig Aerosol Cloud Interaction Simulator (LACIS. For that, washing water of two different birch pollen samples with different regional origin (Northern birch and Southern birch were used. The immersion freezing of droplets generated from the pollen washing water was already observed at temperatures higher than −20 °C, for both samples. Main differences between the Northern birch pollen and the Southern birch pollen were obvious in a temperature range, between −18 °C and −24 °C, where the ice fraction increased with decreasing temperature. There, the Northern birch pollen washing water featured two different slopes, with one being steeper and one being similar to the slope of the Southern birch pollen washing water. As we assume single INA macromolecules being the reason for the ice nucleation, we concluded that the Northern birch pollen are able to produce at least two different types of INA macromolecules. We were able to determine the heterogeneous nucleation rates for both INA macromolecule types and so could explain the ice nucleation behavior of both, the Southern and the Northern birch pollen washing water.

  3. Time-dependent freezing rate parcel model

    Directory of Open Access Journals (Sweden)

    G. Vali

    2014-11-01

    Full Text Available The Time-Dependent Freezing Rate (TDFR model here described represents the formation of ice particles by immersion freezing within an air parcel. The air parcel trajectory follows an adiabatic ascent and includes a period at time with the parcel remaining stationary at the top of its ascent. The description of the ice nucleating particles (INPs in the air parcel is taken from laboratory experiments with cloud and precipitation samples and is assumed to represent the INP content of the cloud droplets in the parcel. Time-dependence is included to account for variations in updraft velocity and for the continued formation of ice particles at isothermal conditions. The magnitudes of these factors are assessed on the basis of laboratory measurements. Results show that both factors give rise to factors of about 3 variations in ice concentration for a realistic range of the input parameters. Refinements of the parameters specifying time-dependence and INP concentrations are needed to make the results more specific to different atmospheric aerosol types. The simple model framework described in this paper can be adapted to more elaborate cloud models. The results here presented can help guide decisions on whether to include a time-dependent ice nucleation scheme or a simpler singular description in models.

  4. Production of freeze-dried yeast culture for the brewing of traditional sorghum beer, tchapalo.

    Science.gov (United States)

    N'Guessan, Florent K; Coulibaly, Hermann W; Alloue-Boraud, Mireille W A; Cot, Marlène; Djè, Koffi Marcellin

    2016-01-01

    Freeze-drying is a well-known dehydration method widely used to preserve microorganisms. In order to produce freeze-dried yeast starter culture for the brewing purpose of African sorghum beer, we tested protective agents (sucrose, glucose, glycerol) in combination with support materials (millet, maize, sorghum, and cassava flours) at 1:1 ratio (v/v). The yeast strains Saccharomyces cerevisiae F 12-7 and Candida tropicalis C 0-7 previously isolated from sorghum beer were used in a mixed culture at a ratio of 2:1 (C. tropicalis/S. cerevisiae). After the freeze-drying, the residual water contents were between 0.78 -2.27%, 0.55 -4.09%, and 0.40-2.61%, respectively, with sucrose, glucose and glycerol. The dried yeasts viabilities were between 4.0% and 10.6%. Among the protective agents used, sucrose was found to be the best protectant giving cell viabilities of 8.4-10.6%. Considering the support materials, millet flour was the best support after drying. When the freeze-dried yeast powders were stored at 4°C and room temperature (25-28°C) for up to 3 months, the survival rates were the highest with cassava flour as the support material.

  5. Production of freeze-dried yeast culture for the brewing of traditional sorghum beer, tchapalo.

    Science.gov (United States)

    N'Guessan, Florent K; Coulibaly, Hermann W; Alloue-Boraud, Mireille W A; Cot, Marlène; Djè, Koffi Marcellin

    2016-01-01

    Freeze-drying is a well-known dehydration method widely used to preserve microorganisms. In order to produce freeze-dried yeast starter culture for the brewing purpose of African sorghum beer, we tested protective agents (sucrose, glucose, glycerol) in combination with support materials (millet, maize, sorghum, and cassava flours) at 1:1 ratio (v/v). The yeast strains Saccharomyces cerevisiae F 12-7 and Candida tropicalis C 0-7 previously isolated from sorghum beer were used in a mixed culture at a ratio of 2:1 (C. tropicalis/S. cerevisiae). After the freeze-drying, the residual water contents were between 0.78 -2.27%, 0.55 -4.09%, and 0.40-2.61%, respectively, with sucrose, glucose and glycerol. The dried yeasts viabilities were between 4.0% and 10.6%. Among the protective agents used, sucrose was found to be the best protectant giving cell viabilities of 8.4-10.6%. Considering the support materials, millet flour was the best support after drying. When the freeze-dried yeast powders were stored at 4°C and room temperature (25-28°C) for up to 3 months, the survival rates were the highest with cassava flour as the support material. PMID:26788308

  6. Of plants and other pets: practical aspects of freeze-substitution and resin embedding.

    Science.gov (United States)

    Hess, M W

    2003-10-01

    Representative tissues from higher plants (e.g. developing pollen, somatic anther tissues from the monocotyledonous angiosperm Ledebouria) and mammalian cell cultures were successfully cryoimmobilized by means of high-pressure freezing. Various substitution and embedding protocols were then evaluated considering the preservation of ultrastructural details, membrane staining, immunolabelling properties, as well as reproducibility and ease of use. Two types of recipe proved to be highly suitable for most applications, regardless of type, developmental stage or physiological conditions of the cells: (i) the best choice for morphology is still osmium in acetone (optionally supplemented with uranyl acetate) followed by embedding in Epon and/or Araldite; (ii) feasible approaches for immunocytochemistry are freeze-substitution with ethanol containing uranyl acetate and formaldehyde, or with pure acetone (in the case of fixation-sensitive antigens), followed by embedding with LR-white acrylic resin; though being far from optimal, these combinations represent, in my opinion, an acceptable compromise between labelling intensity, section stability, structural preservation and health hazards. Notably, the patterns observed in Ledebouria were consistent with data obtained from a broad range of other specimens from all kingdoms (e.g. leaves and callus cultures from angiosperms, gymnosperm roots with their ectomycorrhizal fungi, mammalian cell cultures and eubacteria). Finally, a warning is given as to the extractive potentials of embedding resins (Spurr's mixture, LR-white, but also Epon) being sometimes the cause of unacceptable artefacts, both in plant and in mammalian cells prepared by cryoimmobilization and freeze-substitution.

  7. Criopreservação de medula óssea e células pluripotentes periféricas utilizando um congelador programável: experiência em 86 congelamentos Cryopreservation of bone marrow and peripheral blood stem cells using a controlled rate freezing system. Experience on 86 procedures

    Directory of Open Access Journals (Sweden)

    C.M. Massumoto

    1997-06-01

    Full Text Available A infusão de células hematopoéticas totipotentes criopreservadas permite a recuperação da hematopoese após quimioterapia mieloablativa. OBJETIVO. A formação de cristais de gelo durante o processo de congelamento é o fator principal que causa ruptura das estruturas celulares. A criopreservação dessas células a uma taxa constante preveniria os danos causados pelo congelamento brusco. MÉTODOS. Vinte e três pacientes com mediana de 25 anos (variação 3-57 tiveram a medula óssea e/ou células-tronco periféricas (CTP coletadas no período de março de 1993 a outubro de 1994, totalizando 86 congelamentos. Os pacientes apresentavam as seguintes neoplasias: linfoma não-Hodgkin (n=5, leucemia mielóide aguda (n=8, leucemia linfóide aguda (n=6, doença de Hodgkin (n=3 e mieloma múltiplo (n=1. O congelamento foi controlado por um computador, acoplado ao sistema, às seguintes temperaturas: -1°C/min até -45°C e depois a -10°C/min até -80°C. Após o congelamento, as células foram mantidas em freezer a -110°C até o momento da infusão. Para obtenção das CTP, empregou-se o fator de crescimento estimulante de granulócitos (G-CSF. RESULTADOS. Uma mediana de 3,16 x 10(8 céls./kg (variação 0,86-24,22 de CTP e 2,03 x 10(8 céls./kg (variação 0,19-12,21 de medula óssea foi congelada. A mediana para atingir granulócitos maior ou igual a 500/µL e plaquetas maior que 20.000/µL foi de 12 dias (variação 8-40 e 31 dias (variação 8-80, respectivamente. Todos os pacientes tiveram recuperação hematopoética após a infusão das células criopreservadas. CONCLUSÃO. A criopreservação em congelador programável permite o armazenamento de células hematopoéticas e, potencialmente, pode causar menor dano celular.The cryopreservation of hematopoietic stem cells can be used for rescuing the hematopoiesis after high dose chemotherapy. PURPOSE. The ice cristal formation during the freezing procedure is the key point that can be

  8. Theoretical and experimental studies of heat transfer with moving phase-change interface in freezing and thawing of porous potting soil

    Institute of Scientific and Technical Information of China (English)

    Michael K.H.LEUNG; Kit-ying CHAN

    2009-01-01

    Soil in a cold region is subject to frequent freezing and thawing cycles. Soil frozen for a prolonged period may cause adverse freeze damage to the plants due to cell dehydration or root cell rupture. It is important to understand the detailed heat transfer behaviors of the freezing and thawing processes to prevent freeze damage, and to devise proper mitigation measures for effective pot planting in cold regions. A theoretical model was developed to analyze the transient moving phase-change interface heat transfer in the freezing and thawing of porous potting soil. The theoretical derivation is based on the assumption that the soil freezes completely at a single temperature. Microscopic poromechanic effects on heat transfer behavior were ignored. The spatial domain of the problem was simplified to a 1D spherical coordinate system with variation in the radial direction. Green's function was applied to solve for the time-dependent body temperature. Experiments were conducted for validation of the theoretical model. Reasonable agreement between the theoretical predictions and experimental measurements was obtained. The theoretical model developed can be easily used to determine the sensitivity of various parameters in the freezing/thawing processes, e.g., thermal properties of soil, ambient temperature, and planting pot size.

  9. Specific anti-tumor immune responses of dendritic cells pulsed with recombinant human rhHSP70 and freeze-thaw cellular lysates derived from breast cancer%rhHSP70联合冻融抗原修饰树突状细胞诱导的抗乳腺癌作用*

    Institute of Scientific and Technical Information of China (English)

    李斌; 陈鹏; 郑建云

    2013-01-01

      目的:利用rhHSP70联合树突状细胞递呈肿瘤抗原的特性提高细胞毒T淋巴细胞(CTLs)对乳腺癌细胞的杀伤活性。方法:外周血单个核细胞体外经GM-CSF和IL-4诱导产生树突状细胞,负载冻融抗原肽的同时加入新型热休克蛋白(rhHSP70),不同分组分别诱导自体CTLs产生。ELISA测定CTLs杀伤活性和细胞因子的分泌。结果:冻融抗原肽致敏的DCs促进CTLs增殖,上调CTLs中CD3+和CD8+T细胞群及Th1型细胞因子的分泌;体外实验中具有对人乳腺癌细胞MCF-7的杀伤活性,在加入rhHSP70后效果更加明显,并能显著增强CTLs对肿瘤细胞的杀伤率。结论:rhHSP70联合肝癌冻融抗原修饰DCs,能够促进DCs的成熟,增强DCs刺激淋巴细胞增殖的能力,诱导的CTLs在体外对乳腺癌细胞能产生高效杀伤力。rhHSP70增强DCs抗肿瘤能力的机制可能与其促进DCs成熟有关。%Objective:This work aims to use the characteristics of dendritic cells (DCs) pulsed with recombinant human HSP70, which can present and process tumor antigens, to enhance the killing activity of cytotoxic t lymphocytes (CTLs) against breast neoplasms. Methods:Autologous DCs were isolated from peripheral blood mononuclear cells and then stimulated in vitro with granulocyte macrophage-colony stimulating factor and interleukin-4. The DCs were loaded with A549 tumor cell freeze-thaw lysate, and rhHSP70 was added as an immune adjuvant. The specific groups were subjected to tumor-specific cytotoxic assay, enzyme-linked immunosorbent assay, and fluores-cence-activated cell sorting. Results:DCs pulsed with A549 tumor cell lysate enhanced the growth expansion of CTLs, upregulated CD40 and CD80 populations in CTLs, and augmented Th1 cytokines. In addition, the cytotoxicity of specific CTLs against A549 was highly enhanced. The above indications became more obvious after the addition of rhHSP70. Conclusion:DCs pulsed with freeze-thaw cell

  10. 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.

  11. 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. PMID:26585223

  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. Understanding freeze stress in biological tissues: thermodynamics of interfacial water

    Science.gov (United States)

    A thermodynamic approach to distinguish forms of freeze energy that injure plants as the temperature decreases is developed. The pattern resulting from this analysis dictated the sequence of thermal requirements for water to exist as an independent state. Improvement of freezing tolerance in biolo...

  14. Understanding freeze stress in biological tissues: Thermodynamics of interfacial water

    Energy Technology Data Exchange (ETDEWEB)

    Olien, C. Robert [USDA-ARS (retired), Crop and Soil Sciences, Michigan State University, East Lansing, MI 48824-1325 (United States); Livingston, David P. [USDA and North Carolina State University, Crop Science, 840 Method Road, Unit 3, Raleigh, NC 27502 (United States)]. E-mail: dpl@unity.ncsu.edu

    2006-12-01

    A thermodynamic approach to distinguish forms of freeze energy that injure plants as the temperature decreases is developed. The pattern resulting from this analysis dictated the sequence of thermal requirements for water to exist as an independent state. Improvement of freezing tolerance in biological systems depends on identification of a specific form of stress, just as control of a disease depends on identification of the pathogen causing the disease. The forms of energy that stress hydrated systems as temperature decreases begin with disruption of biological function from chill injury that occurs above freezing. Initiation of non-equilibrium freezing with sufficient free energy to drive disruptive effects can occur in a supercooled system. As the temperature continues to decrease and freezing occurs in an equilibrium manner, adhesion at hydrated interfaces contributes to disruptive effects as protoplasts contract by freeze-dehydration. If protective systems are able to prevent injury from direct interactions with ice, passive effects of freeze-dehydration may cause injury at lower temperatures. The temperature range in which an injury occurs is an indicator of the form of energy causing stress. The form of energy is thus a primary guide for selection of a protective mechanism. An interatomic force model whose response to temperature change corresponds with the enthalpy pattern might help define freeze stress from a unique perspective.

  15. Physical Stability of Freeze-Dried Isomalt Diastereomer Mixtures

    DEFF Research Database (Denmark)

    Koskinen, Anna-Kaisa; Fraser-Miller, Sara J.; Bøtker, Johan P.;

    2016-01-01

    Isomalt is a sugar alcohol used as an excipient in commercially available solid oral dosage forms. The potential of isomalt as a novel freeze-drying excipient was studied in order to increase knowledge of the behavior of isomalt when it is freeze-dried....

  16. DOES GLUTATHIONE PLAY A ROLE IN FREEZING TOLERANCE OF PLANTS

    NARCIS (Netherlands)

    Stuiver, C.E.E.; De Kok, Luit J.; Kuiper, P.J.C.

    1992-01-01

    During low temperature hardening enhanced levels of glutathione (GSH) are generally observed in plant shoots and are often related to the development of freezing tolerance. The present communication shows that there is no direct relation between an increased GSH content and freezing tolerance of lea

  17. Cooling method with automated seasonal freeze protection

    Energy Technology Data Exchange (ETDEWEB)

    Cambell, Levi; Chu, Richard; David, Milnes; Ellsworth, Jr, Michael; Iyengar, Madhusudan; Simons, Robert; Singh, Prabjit; Zhang, Jing

    2016-05-31

    An automated multi-fluid cooling method is provided for cooling an electronic component(s). The method includes obtaining a coolant loop, and providing a coolant tank, multiple valves, and a controller. The coolant loop is at least partially exposed to outdoor ambient air temperature(s) during normal operation, and the coolant tank includes first and second reservoirs containing first and second fluids, respectively. The first fluid freezes at a lower temperature than the second, the second fluid has superior cooling properties compared with the first, and the two fluids are soluble. The multiple valves are controllable to selectively couple the first or second fluid into the coolant in the coolant loop, wherein the coolant includes at least the second fluid. The controller automatically controls the valves to vary first fluid concentration level in the coolant loop based on historical, current, or anticipated outdoor air ambient temperature(s) for a time of year.

  18. Cooling system with automated seasonal freeze protection

    Energy Technology Data Exchange (ETDEWEB)

    Campbell, Levi A.; Chu, Richard C.; David, Milnes P.; Ellsworth, Jr., Michael J.; Iyengar, Madhusudan K.; Simons, Robert E.; Singh, Prabjit; Zhang, Jing

    2016-05-24

    An automated multi-fluid cooling system and method are provided for cooling an electronic component(s). The cooling system includes a coolant loop, a coolant tank, multiple valves, and a controller. The coolant loop is at least partially exposed to outdoor ambient air temperature(s) during normal operation, and the coolant tank includes first and second reservoirs containing first and second fluids, respectively. The first fluid freezes at a lower temperature than the second, the second fluid has superior cooling properties compared with the first, and the two fluids are soluble. The multiple valves are controllable to selectively couple the first or second fluid into the coolant in the coolant loop, wherein the coolant includes at least the second fluid. The controller automatically controls the valves to vary first fluid concentration level in the coolant loop based on historical, current, or anticipated outdoor air ambient temperature(s) for a time of year.

  19. Hadron freeze-out and Unruh radiation

    International Nuclear Information System (INIS)

    In this paper, we consider hadron production in high energy collisions as an Unruh radiation phenomenon. This mechanism describes the production pattern of newly formed hadrons and is directly applicable at vanishing baryon chemical potential, μ ≃ 0. It had already been found to correctly yield the hadronization temperature, Th = √σ/2π ≃ 165 MeV in terms of the string tension σ. Here, we show that the Unruh mechanism also predicts hadronic freeze-out conditions, giving s/Th3 = 3π2/4 ≃ 7.4 in terms of the entropy density s and 〈E〉/〈N〉 = √2πσ ≃ 1.09 for the average energy per hadron. These predictions provide a theoretical basis for previous phenomenological results and are also in accord with recent lattice studies. (author)

  20. Hot water can freeze faster than cold?!?

    CERN Document Server

    Jeng, M

    2005-01-01

    We review the Mpemba effect, where intially hot water freezes faster than initially cold water. While the effect appears impossible at first sight, it has been seen in numerous experiments, was reported on by Aristotle, Francis Bacon, and Descartes, and has been well-known as folklore around the world. It has a rich and fascinating history, which culminates in the dramatic story of the secondary school student, Erasto Mpemba, who reintroduced the effect to the twentieth century scientific community. The phenomenon, while simple to describe, is deceptively complex, and illustrates numerous important issues about the scientific method: the role of skepticism in scientific inquiry, the influence of theory on experiment and observation, the need for precision in the statement of a scientific hypothesis, and the nature of falsifiability. We survey proposed theoretical mechanisms for the Mpemba effect, and the results of modern experiments on the phenomenon. Studies of the observation that hot water pipes are more ...

  1. Characterization of freeze-fractured epithelial plasma membranes on nanometer scale with ToF-SIMS.

    Science.gov (United States)

    Draude, Felix; Körsgen, Martin; Pelster, Andreas; Schwerdtle, Tanja; Müthing, Johannes; Arlinghaus, Heinrich F

    2015-03-01

    Time-of-flight secondary ion mass spectrometry (ToF-SIMS) was used to characterize the freeze-fracturing process of human epithelial PANC-1 and UROtsa cells. For this purpose, phosphatidylcholine, sphingomyelin, phosphatidylethanolamine, and phosphatidylserine standard samples were investigated to find specific signals with both high specificity and signal intensity. The results were used to investigate single cells of subconfluent cell layers prepared with a special silicon wafer sandwich preparation technique. This freeze-fracturing technique strips cell membranes off the cells, isolating them on opposing silicon wafer substrates. Criteria were found for defining regions with stripped off cell membranes and, on the opposing wafer, complementary regions with the remaining cells. Measured ethanolamine/choline and serine/choline ratios in these regions clearly showed that in the freeze-fracturing process, the lipid bilayer of the plasma membrane is split along its central zone. Accordingly, only the outer lipid monolayer is stripped off the cell, while the inner lipid monolayer remains attached to the cell on the opposing wafer, thus allowing detailed analysis of a single lipid monolayer. Furthermore, it could be shown that using different washing procedures did not influence the transmembrane lipid distribution. Under optimized preparation conditions, it became feasible to detect lipids with a lateral resolution of approximately 100 nm. The data indicate that ToF-SIMS would be a very useful technique to study with very high lateral resolution changes in lipid composition caused, for example, by lipid storage diseases or pharmaceuticals that interfere with the lipid metabolism.

  2. Pyrazine analogues are active components of wolf urine that induce avoidance and freezing behaviours in mice.

    Directory of Open Access Journals (Sweden)

    Kazumi Osada

    Full Text Available BACKGROUND: The common grey wolf (Canis lupus is found throughout the entire Northern hemisphere and preys on many kinds of mammals. The urine of the wolf contains a number of volatile constituents that can potentially be used for predator-prey chemosignalling. Although wolf urine is put to practical use to keep rabbits, rodents, deer and so on at bay, we are unaware of any prior behavioural studies or chemical analyses regarding the fear-inducing impact of wolf urine on laboratory mice. METHODOLOGY/PRINCIPAL FINDINGS: Three wolf urine samples harvested at different times were used in this study. All of them induced stereotypical fear-associated behaviors (i.e., avoidance and freezing in female mice. The levels of certain urinary volatiles varied widely among the samples. To identify the volatiles that provoked avoidance and freezing, behavioural, chemical, and immunohistochemical analyses were performed. One of the urine samples (sample C had higher levels of 2,6-dimethylpyrazine (DMP, trimethylpyrazine (TMP, and 3-ethyl-2,5-dimethyl pyrazine (EDMP compared with the other two urine samples (samples A and B. In addition, sample C induced avoidance and freezing behaviours more effectively than samples A and B. Moreover, only sample C led to pronounced expression of Fos-immunoreactive cells in the accessory olfactory bulb (AOB of female mice. Freezing behaviour and Fos immunoreactivity were markedly enhanced when the mice were confronted with a mixture of purified DMP, TMP, and EDMP vs. any one pyrazine alone. CONCLUSIONS/SIGNIFICANCE: The current results suggest that wolf urinary volatiles can engender aversive and fear-related responses in mice. Pyrazine analogues were identified as the predominant active components among these volatiles to induce avoidance and freezing behaviours via stimulation of the murine AOB.

  3. A novel and versatile flash-freezing approach for evaluating the health of Delta Smelt.

    Science.gov (United States)

    Teh, Swee J; Baxa, Dolores V; Hammock, Bruce G; Gandhi, Saikrithika A; Kurobe, Tomofumi

    2016-01-01

    A common approach used to assess environmental impacts in aquatic environments is to measure indicators of stress (biomarkers) and condition of fish within ecosystems. Particularly in estuarine ecosystems with multiple stressors, it is often desirable to quantify a suite of biological endpoints that (1) reflect fish condition at several levels of biological organization and time scales and (2) are sensitive to a range of environmental stressors. However, established methods of preservation and processing of fish for specific endpoints are often incompatible. Here, we developed a novel flash-freezing approach for assessing the health of a small, sensitive fish, the endangered Delta Smelt (Hypomesus transpacificus) after collections from the San Francisco Estuary (SFE). We assess whether flash-freezing the entire fish ensures effective preservation of multiple tissues for subsequent biomarker analyses by comparing measurements of fresh to frozen tissue. Tissues included brain, gill, and liver for enzyme activity, kidney and spleen for pathogens, and gills, liver, and gonads for histopathology and reproduction. Although flash-freezing in liquid nitrogen altered the length, weight, and condition factor of Delta Smelt, the percent changes were small (artefacts were observed in ovaries, but did not hinder the identification and interpretation of cell types and oocyte stages. Freezing did not alter bacterial isolation or the activities of ethoxyresorufin-0-deethylase (EROD) or acetylcholinesterase (AChE), but had a small, negative influence on sodium potassium adenosine triphosphatase (ATPase) activity. Thus, flash-freezing in the field is a versatile preservation method for Delta Smelt, allowing for multiple tissue collections and bioassays from an individual tiny fish exposed to a wide range of natural and anthropogenic stressors. Similar methodology may be applicable to other species for which a range of biological endpoints and histopathology data are needed. PMID

  4. PREPARATION AND CHARACTERIZATION OF FREEZE DRIED CRYSTALS OF IBUPROFEN

    Directory of Open Access Journals (Sweden)

    Dixit Mudit

    2011-12-01

    Full Text Available Ibuprofen, an anti-inflammatory drug, exhibits poor water solubility and flow properties.. Crystallization medium used for freeze dried crystals of Ibuprofen consisted of Isopropyl alcohol and water (50:50% respectively. Freeze dried crystals were characterized by differential scanning calorimetry, Infrared spectroscopy, X-ray diffractometry and scanning electron microscopy. Micromeritic and mechanical property and dissolution behavior studies were carried out. Process variables such as amount of bridging liquid, stirring time and duration of stirring were optimized. Dissolution profile of the freeze dried crystals was compared with pure sample and recrystallized sample. Freeze dried crystals exhibited decreased crystallinity and improved micromeritic properties. The dissolution of the freeze dried crystals was improved compared with pure sample.

  5. Versatile Aerogel Fabrication by Freezing and Subsequent Freeze-Drying of Colloidal Nanoparticle Solutions.

    Science.gov (United States)

    Freytag, Axel; Sánchez-Paradinas, Sara; Naskar, Suraj; Wendt, Natalja; Colombo, Massimo; Pugliese, Giammarino; Poppe, Jan; Demirci, Cansunur; Kretschmer, Imme; Bahnemann, Detlef W; Behrens, Peter; Bigall, Nadja C

    2016-01-18

    A versatile method to fabricate self-supported aerogels of nanoparticle (NP) building blocks is presented. This approach is based on freezing colloidal NPs and subsequent freeze drying. This means that the colloidal NPs are directly transferred into dry aerogel-like monolithic superstructures without previous lyogelation as would be the case for conventional aerogel and cryogel fabrication methods. The assembly process, based on a physical concept, is highly versatile: cryogelation is applicable for noble metal, metal oxide, and semiconductor NPs, and no impact of the surface chemistry or NP shape on the resulting morphology is observed. Under optimized conditions the shape and volume of the liquid equal those of the resulting aerogels. Also, we show that thin and homogeneous films of the material can be obtained. Furthermore, the physical properties of the aerogels are discussed.

  6. Versatile Aerogel Fabrication by Freezing and Subsequent Freeze-Drying of Colloidal Nanoparticle Solutions.

    Science.gov (United States)

    Freytag, Axel; Sánchez-Paradinas, Sara; Naskar, Suraj; Wendt, Natalja; Colombo, Massimo; Pugliese, Giammarino; Poppe, Jan; Demirci, Cansunur; Kretschmer, Imme; Bahnemann, Detlef W; Behrens, Peter; Bigall, Nadja C

    2016-01-18

    A versatile method to fabricate self-supported aerogels of nanoparticle (NP) building blocks is presented. This approach is based on freezing colloidal NPs and subsequent freeze drying. This means that the colloidal NPs are directly transferred into dry aerogel-like monolithic superstructures without previous lyogelation as would be the case for conventional aerogel and cryogel fabrication methods. The assembly process, based on a physical concept, is highly versatile: cryogelation is applicable for noble metal, metal oxide, and semiconductor NPs, and no impact of the surface chemistry or NP shape on the resulting morphology is observed. Under optimized conditions the shape and volume of the liquid equal those of the resulting aerogels. Also, we show that thin and homogeneous films of the material can be obtained. Furthermore, the physical properties of the aerogels are discussed. PMID:26638874

  7. Viability of ligaments after freezing: an experimental study in a rabbit model

    Energy Technology Data Exchange (ETDEWEB)

    Frank, C.; Edwards, P.; McDonald, D.; Bodie, D.; Sabiston, P.

    1988-01-01

    Our purpose in this study was to assess ligament fibroblast viability after freezing by quantifying the subsequent ability of fibroblasts to synthesize collagen in vitro. Both medial collateral ligament (MCL) complexes from 40 adolescent rabbits were studied. Collagen production was determined by in vitro incubation of ligaments in /sup 3/H-proline (a collagen precursor) and subsequent analysis of /sup 3/H-hydroxyproline (a marker of newly synthesized collagen). Autoradiographs determined the distributions of ligament cell activity. All right MCL complexes served as fresh controls, providing a baseline of collagen production. Each left MCL was assigned to an experimental group and was either incubated fresh (10 animals); killed by drying, multiple freeze thawing, or cycloheximide (six animals); or slowly frozen at -70 degrees C without cryoprotection (24 animals). Collagen production of rapidly thawed ligaments was studied by proline incubation at 1 day, 9 days, or 6 weeks after freezing and was compared with that of contralateral fresh controls. Results demonstrate that some cells in the substance of these rabbit ligaments retained the ability to synthesize collagen in vitro after being frozen for up to 6 weeks. Mean collagen production of frozen ligaments was decreased, but tests of mean and median values as well as ratios were statistically similar to fresh contralateral ligaments in all animals. This postfreezing ligament cell survival and collagen production after -70 degrees C storage may have implications for ligament transplantation.

  8. Freezing Precipitation and Freezing Events over Northern Eurasia and North America

    Science.gov (United States)

    Groisman, Pavel; Yin, Xungang; Bulygina, Olga; Partasenok, Irina; Zolina, Olga; Hanssen-Bauer, Inger

    2016-04-01

    With global climate change in the extratropics, the 0°C isotherm will not disappear and associated precipitation events will continue to occur. The near-0°C temperatures should generally move poleward and arrive at many locations earlier in spring or later in autumn. This could potentially affect the seasonal cycle of near-0°C precipitation. The overall warming, together with a larger influx of the water vapor in the winter atmosphere from the oceans (including ice-free portions of the Arctic Ocean) can also affect the amount of near-0°C precipitation. The issue of near 0°C precipitation is linked with several hazardous phenomena including heavy snowfall/rainfall transition around °C; strong blizzards; rain-on-snow events causing floods; freezing rain and freezing drizzle; and ice load on infrastructure. In our presentation using more than 1,500 long-term time series of synoptic observations for the past four decades, we present climatology and the empirical evidence about changes in occurrence, timing, and intensity of freezing rains and freezing drizzles over several countries of Northern Eurasia and North America. In the former Soviet Union, instrumental monitoring of ice load has been performed by ice accretion indicator that in addition to the type, intensity and duration of ice deposits reports also their weight and size. Estimates of climatology and changes in ice load based on this monitoring at 958 Russian stations will be also presented. The work was supported by the Ministry of Education and Science of the Russian Federation (grant 14.B25.31.0026) and NASA LCLUC Program (grant "How Environmental Change in Central Asian Highlands Impacts High Elevation Communities").

  9. Effect of freezing and thawing rates on sperm motility in Bocachico Prochilodus magdalenae (Pisces, Characiformes)

    OpenAIRE

    José G. Martínez; Sandra Pardo C.

    2015-01-01

    ABSTRACTObjective. To determine the freezing and thawing rates necessary to maintain sperm viability during cryopreservation of Bocachico semen. Materials and methods. Four interactional treatments were implemented between two freezing (rapid and slow) and two thawing (rapid and slow) curves, in a 2x2 factorial as follows: rapid freezing-rapid thawing, rapid freezing-slow thawing, slow freezing-rapid thawing, and slow freezing-slow thawing. After thawing by Sperm Class Analyzer (SCA) curvilin...

  10. Effects of industrial pre-freezing processing and freezing handling on glucosinolates and antioxidant attributes in broccoli florets.

    Science.gov (United States)

    Cai, Congxi; Miao, Huiying; Qian, Hongmei; Yao, Leishuan; Wang, Bingliang; Wang, Qiaomei

    2016-11-01

    The effects of industrial pre-freezing processing and freezing handling on the contents of glucosinolates and antioxidants (vitamin C, polyphenols, carotenoid and chlorophyll), as well as the antioxidant capacity in broccoli (Brassica oleracea L. var. italica) florets were investigated in the present study. Our results showed that the glucosinolate accumulations were significantly decreased after pre-freezing processing, whereas elevated levels of phenols, carotenoids, chlorophyll, and also antioxidant capacity were observed in frozen broccoli florets. The contents of vitamin C remained constant during above mentioned processing. In conclusion, the current industrial freezing processing method is a good practice for the preservation of main antioxidant nutrients in broccoli florets, although some improvements in pre-freezing processing, such as steam blanching and ice-water cooling, are needed to attenuate the decrease in glucosinolate content. PMID:27211670

  11. Preservation of tissue microstructure and functionality during freezing by modulation of cytoskeletal structure.

    Science.gov (United States)

    Park, Seungman; Seawright, Angela; Park, Sinwook; Craig Dutton, J; Grinnell, Frederick; Han, Bumsoo

    2015-05-01

    Cryopreservation is one of the key enabling technologies for tissue engineering and regenerative medicine, which can provide reliable long-term storage of engineered tissues (ETs) without losing their functionality. However, it is still extremely difficult to design and develop cryopreservation protocols guaranteeing the post-thaw tissue functionality. One of the major challenges in cryopreservation is associated with the difficulty of identifying effective and less toxic cryoprotective agents (CPAs) to guarantee the post-thaw tissue functionality. In this study, thus, a hypothesis was tested that the modulation of the cytoskeletal structure of cells embedded in the extracellular matrix (ECM) can mitigate the freezing-induced changes of the functionality and can reduce the amount of CPA necessary to preserve the functionality of ETs during cryopreservation. In order to test this hypothesis, we prepared dermal equivalents by seeding fibroblasts in type I collagen matrices resulting in three different cytoskeletal structures. These ETs were exposed to various freeze/thaw (F/T) conditions with and without CPAs. The freezing-induced cell-fluid-matrix interactions and subsequent functional properties of the ETs were assessed. The results showed that the cytoskeletal structure and the use of CPA were strongly correlated to the preservation of the post-thaw functional properties. As the cytoskeletal structure became stronger via stress fiber formation, the ET's functionality was preserved better. It also reduced the necessary CPA concentration to preserve the post-thaw functionality. However, if the extent of the freezing-induced cell-fluid-matrix interaction was too excessive, the cytoskeletal structure was completely destroyed and the beneficial effects became minimal.

  12. Versatile wedge-based system for the construction of unidirectional collagen scaffolds by directional freezing: practical and theoretical considerations

    NARCIS (Netherlands)

    Pot, M.W.; Faraj, K.A.; Adawy, A.; Enckevort, W.J.P. van; Moerkerk, H.T. van; Vlieg, E.; Daamen, W.F.; Kuppevelt, T.H. van

    2015-01-01

    Aligned unidirectional collagen scaffolds may aid regeneration of those tissues where alignment of cells and extracellular matrix is essential, as for instance in cartilage, nerve bundles, and skeletal muscle. Pores can be introduced by ice crystal formation followed by freeze-drying, the pore archi

  13. [Effect of freezing and cooking on the texture and electrophoretic pattern of the proteins of octopus arms (Octopus vulgaris)].

    Science.gov (United States)

    Reyes, Genara; Nirchio, Mauro; Bello, Rafael; Borderías, Javier

    2014-09-01

    Texture is the most valuable feature in cephalopods. Factors that mainly affect the texture of octopus are: freezing, scalding and cooking. The aim of this study was to assess the effect of freezing, scalding and length of cooking time on the texture and electrophoretic pattern of proteins of octopus arms. Octopuses were trapped near Margarita Island and carried with ice to the laboratory where they were packed and subjected to: a) freezing at -27 degrees C or at -20 degrees C b) scalding c) cooking for 25 min, 35 min or 45 min. Shear force was determined by Kramer cell on strips of octopus arms. SDS-PAGE was done according to the Laemmli method with 12% polyacrilamide gels. A sensory evaluation of the preference of texture was carried out using a hedonic scale of 7-points and a non-trained panel. Octopus texture was not affected by freezing temperature or scalding. Frozen octopus was softer after cooking than fresh. The longer the cooking time was, the softer the octopus was. Myosin heavy chain (MHC) was not significantly affected by scalding or cooking; however large aggregates heavier than MHC, new bands and loss of resolution of the bands appeared. Myosin and paramyosin bands were more affected by freezing prior to cooking.

  14. Disruption of the Arabidopsis Defense Regulator Genes SAG101, EDS1, and PAD4 Confers Enhanced Freezing Tolerance.

    Science.gov (United States)

    Chen, Qin-Fang; Xu, Le; Tan, Wei-Juan; Chen, Liang; Qi, Hua; Xie, Li-Juan; Chen, Mo-Xian; Liu, Bin-Yi; Yu, Lu-Jun; Yao, Nan; Zhang, Jian-Hua; Shu, Wensheng; Xiao, Shi

    2015-10-01

    In Arabidopsis, three lipase-like regulators, SAG101, EDS1, and PAD4, act downstream of resistance protein-associated defense signaling. Although the roles of SAG101, EDS1, and PAD4 in biotic stress have been extensively studied, little is known about their functions in plant responses to abiotic stresses. Here, we show that SAG101, EDS1, and PAD4 are involved in the regulation of freezing tolerance in Arabidopsis. With or without cold acclimation, the sag101, eds1, and pad4 single mutants, as well as their double mutants, exhibited similarly enhanced tolerance to freezing temperatures. Upon cold exposure, the sag101, eds1, and pad4 mutants showed increased transcript levels of C-REPEAT/DRE BINDING FACTORs and their regulons compared with the wild type. Moreover, freezing-induced cell death and accumulation of hydrogen peroxide were ameliorated in sag101, eds1, and pad4 mutants. The sag101, eds1, and pad4 mutants had much lower salicylic acid (SA) and diacylglycerol (DAG) contents than the wild type, and exogenous application of SA and DAG compromised the freezing tolerance of the mutants. Furthermore, SA suppressed the cold-induced expression of DGATs and DGKs in the wild-type leaves. These findings indicate that SAG101, EDS1, and PAD4 are involved in the freezing response in Arabidopsis, at least in part, by modulating the homeostasis of SA and DAG.

  15. Metabolic changes in Avena sativa crowns recovering from freezing.

    Directory of Open Access Journals (Sweden)

    Cynthia A Henson

    Full Text Available Extensive research has been conducted on cold acclimation and freezing tolerance of fall-sown cereal plants due to their economic importance; however, little has been reported on the biochemical changes occurring over time after the freezing conditions are replaced by conditions favorable for recovery and growth such as would occur during spring. In this study, GC-MS was used to detect metabolic changes in the overwintering crown tissue of oat (Avena sativa L. during a fourteen day time-course after freezing. Metabolomic analysis revealed increases in most amino acids, particularly proline, 5-oxoproline and arginine, which increased greatly in crowns that were frozen compared to controls and correlated very significantly with days after freezing. In contrast, sugar and sugar related metabolites were little changed by freezing, except sucrose and fructose which decreased dramatically. In frozen tissue all TCA cycle metabolites, especially citrate and malate, decreased in relation to unfrozen tissue. Alterations in some amino acid pools after freezing were similar to those observed in cold acclimation whereas most changes in sugar pools after freezing were not. These similarities and differences suggest that there are common as well as unique genetic mechanisms between these two environmental conditions that are crucial to the winter survival of plants.

  16. Metabolic changes in Avena sativa crowns recovering from freezing.

    Science.gov (United States)

    Henson, Cynthia A; Duke, Stanley H; Livingston, David P

    2014-01-01

    Extensive research has been conducted on cold acclimation and freezing tolerance of fall-sown cereal plants due to their economic importance; however, little has been reported on the biochemical changes occurring over time after the freezing conditions are replaced by conditions favorable for recovery and growth such as would occur during spring. In this study, GC-MS was used to detect metabolic changes in the overwintering crown tissue of oat (Avena sativa L.) during a fourteen day time-course after freezing. Metabolomic analysis revealed increases in most amino acids, particularly proline, 5-oxoproline and arginine, which increased greatly in crowns that were frozen compared to controls and correlated very significantly with days after freezing. In contrast, sugar and sugar related metabolites were little changed by freezing, except sucrose and fructose which decreased dramatically. In frozen tissue all TCA cycle metabolites, especially citrate and malate, decreased in relation to unfrozen tissue. Alterations in some amino acid pools after freezing were similar to those observed in cold acclimation whereas most changes in sugar pools after freezing were not. These similarities and differences suggest that there are common as well as unique genetic mechanisms between these two environmental conditions that are crucial to the winter survival of plants.

  17. 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.

  18. Freeze gelated porous membranes for periodontal tissue regeneration.

    Science.gov (United States)

    Qasim, Saad B; Delaine-Smith, Robin M; Fey, Tobias; Rawlinson, Andrew; Rehman, Ihtesham Ur

    2015-09-01

    Guided tissue regeneration (GTR) membranes have been used for the management of destructive forms of periodontal disease as a means of aiding regeneration of lost supporting tissues, including the alveolar bone, cementum, gingiva and periodontal ligaments (PDL). Currently available GTR membranes are either non-biodegradable, requiring a second surgery for removal, or biodegradable. The mechanical and biofunctional limitations of currently available membranes result in a limited and unpredictable treatment outcome in terms of periodontal tissue regeneration. In this study, porous membranes of chitosan (CH) were fabricated with or without hydroxyapatite (HA) using the simple technique of freeze gelation (FG) via two different solvents systems, acetic acid (ACa) or ascorbic acid (ASa). The aim was to prepare porous membranes to be used for GTR to improve periodontal regeneration. FG membranes were characterized for ultra-structural morphology, physiochemical properties, water uptake, degradation, mechanical properties, and biocompatibility with mature and progenitor osteogenic cells. Fourier transform infrared (FTIR) spectroscopy confirmed the presence of hydroxyapatite and its interaction with chitosan. μCT analysis showed membranes had 85-77% porosity. Mechanical properties and degradation rate were affected by solvent type and the presence of hydroxyapatite. Culture of human osteosarcoma cells (MG63) and human embryonic stem cell-derived mesenchymal progenitors (hES-MPs) showed that all membranes supported cell proliferation and long term matrix deposition was supported by HA incorporated membranes. These CH and HA composite membranes show their potential use for GTR applications in periodontal lesions and in addition FG membranes could be further tuned to achieve characteristics desirable of a GTR membrane for periodontal regeneration.

  19. Sucrose Diffusion in Decellularized Heart Valves for Freeze-Drying.

    Science.gov (United States)

    Wang, Shangping; Oldenhof, Harriëtte; Goecke, Tobias; Ramm, Robert; Harder, Michael; Haverich, Axel; Hilfiker, Andres; Wolkers, Willem Frederik

    2015-09-01

    Decellularized heart valves can be used as starter matrix implants for heart valve replacement therapies in terms of guided tissue regeneration. Decellularized matrices ideally need to be long-term storable to assure off-the-shelf availability. Freeze-drying is an attractive preservation method, allowing storage at room temperature in a dried state. However, the two inherent processing steps, freezing and drying, can cause severe damage to extracellular matrix (ECM) proteins and the overall tissue histoarchitecture and thus impair biomechanical characteristics of resulting matrices. Freeze-drying therefore requires a lyoprotective agent that stabilizes endogenous structural proteins during both substeps and that forms a protective glassy state at room temperature. To estimate incubation times needed to infiltrate decellularized heart valves with the lyoprotectant sucrose, temperature-dependent diffusion studies were done using Fourier transform infrared spectroscopy. Glycerol, a cryoprotective agent, was studied for comparison. Diffusion of both protectants was found to exhibit Arrhenius behavior. The activation energies of sucrose and glycerol diffusion were found to be 15.9 and 37.7 kJ·mol(-1), respectively. It was estimated that 4 h of incubation at 37°C is sufficient to infiltrate heart valves with sucrose before freeze-drying. Application of a 5% sucrose solution was shown to stabilize acellular valve scaffolds during freeze-drying. Such freeze-dried tissues, however, displayed pores, which were attributed to ice crystal damage, whereas vacuum-dried scaffolds in comparison revealed no pores after drying and rehydration. Exposure to a hygroscopic sucrose solution (80%) before freeze-drying was shown to be an effective method to diminish pore formation in freeze-dried ECMs: matrix structures closely resembled those of control samples that were not freeze-dried. Heart valve matrices were shown to be in a glassy state after drying, suggesting that they can

  20. Changes in freezing point of blood and milk during dehydration and rehydration in lactating cows

    DEFF Research Database (Denmark)

    Bjerg, M.; Rasmussen, M.D.; Nielsen, Mette Olaf

    2005-01-01

    We studied the influence of short-term changes in water intake in 4 lactating Holstein cows on diurnal fluctuation of packed cell volume (PCV), freezing point of blood (FP blood), freezing point of milk ( FP milk), and the relationship between changes in FP blood and FP milk. The experiment lasted...... the jugular vein hourly throughout the experiment, and at 0, 15, 30, 60, 90, 120, 150, 180, 210, and 240 min after initiated rehydration following the 8-h dehydration sequences. Intakes of free water and water in feed were recorded every hour. The PCV was negatively affected by water intake within the hour...... before sampling. Dehydration lowered FP blood steadily, whereas FP blood increased by 0.024°C within 30 min following a large water intake in the rehydration period. The FP blood was not significantly influenced by actual water intake, but was highly correlated with the available water pool at time...

  1. SOME STUDIES ABOUT CEREALS BEHAVIOR DURING FREEZE DRYING PROCESS

    Directory of Open Access Journals (Sweden)

    GABRIELA-VICTORIA MNERIE

    2013-07-01

    Full Text Available The paper presents some special method and equipment and the principal advantages of freeze-dried food. The freeze drying is a good method of freeze-drying for make some experiments with many kind of cereals, for the improvement that in food production. It is necessary and is possible to study the corn oil extract, wheat flour, the maltodextrin from corn, modified cornstarch, spice extracts, soy sauce, hydrolyzed wheat gluten, partially hydrogenated soybean and cottonseed oil etc. That is very porous, since it occupies the same volume as the original and so rehydrates rapidly. There is less loss of flavour and texture than with most other methods of drying.

  2. Prototype device for converting freezing energy into mechanical work

    Energy Technology Data Exchange (ETDEWEB)

    Habeebullah, B.; Zaki, G.M.; Akyurt, M. [Fakieh Center for Applied Research, Makkah Al-Mukarramah (Saudi Arabia)

    2003-01-01

    Certain properties of ice are reviewed. The phenomenon of freezing of water is discussed, with special emphasis on the blockage and bursting of water pipes. Several concepts are discussed that are related to achieving the freezing of water, the design of the container where freezing occurs and the conversion into mechanical energy. A conceptual design is presented for one type of ice energy converter. Certain design details for this machine are provided. Initial test results are furnished for this device, which was manufactured and tested. Conclusions are drawn and recommendations are made. (author)

  3. Freeze-thaw bond properties of new-old concrete

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Using composite cubic specimens of new-old concrete, the bond splitting tensile strength and failure mechanism for the interface of new-old concrete in saturating state were explored when exposed to freeze-thaw cycling. Specimens were experienced for 0,25,50,75,100 and 125 freezing cycles. The roughness and ad hesion agent including cement paste, cement mortar and cement paste with 10 percent of UEA expanding agent were also investigated. The test results indicate that the bond splitting tensile strength decreases with increased numbers of freezing-and-thawing cycles. The roughness and adhesion agent have different effects on the bond strength.

  4. Chloroplast Membrane Remodeling during Freezing Stress Is Accompanied by Cytoplasmic Acidification Activating SENSITIVE TO FREEZING21[OPEN

    Science.gov (United States)

    Barnes, Allison C.

    2016-01-01

    Low temperature is a seasonal abiotic stress that restricts native plant ranges and crop distributions. Two types of low-temperature stress can be distinguished: chilling and freezing. Much work has been done on the mechanisms by which chilling is sensed, but relatively little is known about how plants sense freezing. Recently, Arabidopsis (Arabidopsis thaliana) SENSITIVE TO FREEZING2 (SFR2) was identified as a protein that responds in a nontranscriptional manner to freezing. Here, we investigate the cellular conditions that allow SFR2 activation. Using a combination of isolated organelle, whole-tissue, and whole-plant assays, we provide evidence that SFR2 is activated by changes in cytosolic pH and Mg2+. Manipulation of pH and Mg2+ in cold-acclimated plants is shown to cause changes similar to those of freezing. We conclude that pH and Mg2+ are perceived as intracellular cues as part of the sensing mechanism for freezing conditions. This evidence provides a specific molecular mechanism to combat freezing. PMID:27233750

  5. Amplitude Manipulation Evokes Upper Limb Freezing during Handwriting in Patients with Parkinson’s Disease with Freezing of Gait

    Science.gov (United States)

    Heremans, Elke; Nackaerts, Evelien; Vervoort, Griet; Vercruysse, Sarah; Broeder, Sanne; Strouwen, Carolien; Swinnen, Stephan P.; Nieuwboer, Alice

    2015-01-01

    Background Recent studies show that besides freezing of gait (FOG), many people with Parkinson’s disease (PD) also suffer from freezing in the upper limbs (FOUL). Up to now, it is unclear which task constraints provoke and explain upper limb freezing. Objective To investigate whether upper limb freezing and other kinematic abnormalities during writing are provoked by (i) gradual changes in amplitude or by (ii) sustained amplitude generation in patients with and without freezing of gait. Methods Thirty-four patients with PD, including 17 with and 17 without FOG, performed a writing task on a touch-sensitive writing tablet requiring writing at constant small and large size as well as writing at gradually increasing and decreasing size. Patients of both groups were matched for disease severity, tested while ‘on’ medication and compared to healthy age-matched controls. Results Fifty upper limb freezing episodes were detected in 10 patients, including 8 with and 2 without FOG. The majority of the episodes occurred when participants had to write at small or gradually decreasing size. The occurrence of FOUL and the number of FOUL episodes per patient significantly correlated with the occurrence and severity of FOG. Patients with FOUL also showed a significantly smaller amplitude in the writing parts outside the freezing episodes. Conclusions Corroborating findings of gait research, the current study supports a core problem in amplitude control underlying FOUL, both in maintaining as well as in flexibly adapting the cycle size. PMID:26580556

  6. Adaptation to seasonality and the winter freeze

    Directory of Open Access Journals (Sweden)

    Jill Christine Preston

    2013-06-01

    Full Text Available Flowering plants initially diversified during the Mesozoic era at least 140 million years ago in regions of the world where temperate seasonal environments were not encountered. Since then several cooling events resulted in the contraction of warm and wet environments and the establishment of novel temperate zones in both hemispheres. In response, less than half of modern angiosperm families have members that evolved specific adaptations to cold seasonal climates, including cold acclimation, freezing tolerance, endodormancy, and vernalization responsiveness. Despite compelling evidence for multiple independent origins, the level of genetic constraint on the evolution of adaptations to seasonal cold is not well understood. However, the recent increase in molecular genetic studies examining the response of model and crop species to seasonal cold offers new insight into the evolutionary lability of these traits. This insight has major implications for our understanding of complex trait evolution, and the potential role of local adaptation in response to past and future climate change. In this review, we discuss the biochemical, morphological, and developmental basis of adaptations to seasonal cold, and synthesize recent literature on the genetic basis of these traits in a phylogenomic context. We find evidence for multiple genetic links between distinct physiological responses to cold, possibly reinforcing the coordinated expression of these traits. Furthermore, repeated recruitment of the same or similar ancestral pathways suggests that land plants might be somewhat pre-adapted to dealing with temperature stress, perhaps making inducible cold traits relatively easy to evolve.

  7. Drying a tuberculosis vaccine without freezing.

    Science.gov (United States)

    Wong, Yun-Ling; Sampson, Samantha; Germishuizen, Willem Andreas; Goonesekera, Sunali; Caponetti, Giovanni; Sadoff, Jerry; Bloom, Barry R; Edwards, David

    2007-02-20

    With the increasing incidence of tuberculosis and drug resistant disease in developing countries due to HIV/AIDS, there is a need for vaccines that are more effective than the present bacillus Calmette-Guérin (BCG) vaccine. We demonstrate that BCG vaccine can be dried without traditional freezing and maintained with remarkable refrigerated and room-temperature stability for months through spray drying. Studies with a model Mycobacterium (Mycobacterium smegmatis) revealed that by removing salts and cryoprotectant (e.g., glycerol) from bacterial suspensions, the significant osmotic pressures that are normally produced on bacterial membranes through droplet drying can be reduced sufficiently to minimize loss of viability on drying by up to 2 orders of magnitude. By placing the bacteria in a matrix of leucine, high-yield, free-flowing, "vial-fillable" powders of bacteria (including M. smegmatis and M. bovis BCG) can be produced. These powders show relatively minor losses of activity after maintenance at 4 degrees C and 25 degrees C up to and beyond 4 months. Comparisons with lyophilized material prepared both with the same formulation and with a commercial formulation reveal that the spray-dried BCG has better overall viability on drying.

  8. GPR utilization in artificial freezing engineering

    International Nuclear Information System (INIS)

    To utilize ground penetrating radar (GPR) in artificial freezing engineering (AFE), the electromagnetic parameters (EMP) of frozen soil were measured using a vector network analyser, which showed that the dielectric permittivity and electric conductivity change abruptly at the boundary between the frozen and the non-frozen soil. Then similarity criteria of GPR model experiments were deduced, and GPR laboratory model experiments and field explorations of AFE were carried out. It was found that for AFE, the GPR travel time and profile characters of anomalies in model experiments were similar to those in field explorations, while the amplitude of GPR signals in laboratory model experiments were much stronger than those in field explorations. Numerical simulations were also implemented to analyse the relationship between model experiments and field explorations, which further told us why we could easily find the targets by GPR in the laboratory but not in field explorations. The outputs showed that GPR could be used to detect the thickness of the frozen wall and to find unfrozen soil defects, even though the amplitude of the reflective signals were much weaker than those of laboratory experiments. The research findings have an important theoretical value for AFE and permafrost region engineering, and the deduced GPR similarity criteria could be widely used in other GPR model experiments. (paper)

  9. Biological activity and mechanical stability of sol-gel-based biofilters using the freeze-gelation technique for immobilization of Rhodococcus ruber.

    Science.gov (United States)

    Pannier, Angela; Mkandawire, Martin; Soltmann, Ulrich; Pompe, Wolfgang; Böttcher, Horst

    2012-02-01

    Biofilters with long lifetime and high storage stability are very important for bioremediation processes to ensure the readiness at the occurrence of sudden contaminations. By using the freeze-gelation technique, living cells can be immobilized within a mechanically and chemically stable ceramic-like matrix. Due to a freeze-drying step, the embedded microorganisms are converted into a preserved form. In that way, they can be stored under dry conditions, which comply better with storage, transport, and handling requirements. Thus, in contrast to other immobilization techniques, there is no need for storage in liquid or under humid atmosphere. The biological activity, mechanical strength, and the structure of the biologically active ceramic-like composites (biocers) produced by freeze gelation have been investigated by using the phenol-degrading bacteria Rhodococcus ruber as model organism. Samples of freeze-gelation biocers have been investigated after defined storage periods, demonstrating nearly unchanged mechanical strength of the immobilization matrix as well as good storage stability of the activity of the immobilized cells over several months of storage at 4 °C. Repeated-batch tests demonstrated further that the freeze-gelation biocers can be repeatedly used over a period of more than 12 months without losing its bioactivity. Thus, these results show that freeze-gelation biocers have high potential of being scaled up from laboratory test systems to applications in real environment because of their long bioactivity as well as mechanical stability.

  10. Effect of Boundary Conditions on Freezing in Porous Media

    Directory of Open Access Journals (Sweden)

    Rahul Basu

    2004-07-01

    Full Text Available This paper examines a model for coupled heat and mass transfer for freezing in a porous media with Dirichlet and convective boundary conditions. Variables include porosity, heat transfer coefficients, thermal and mass diffusivity, density, latent heat, and boundary temperatures. A simulation for the slab illustrates the appearance of undercooling. A stability criterion for the phase interface is linked with well-known metallurgical parameters like undercooling and freezing rate. A possible mechanism for freckling in ingots of niobium-rich superalloys is examined. It has been shown that heat and mass transfer balance at the interface can affect stability. The effect of boundary conditions on the velocity of freezing is computed for some cases, including the self-freezing process.

  11. Microwave Enhanced Freeze Drying of Solid Waste Project

    Data.gov (United States)

    National Aeronautics and Space Administration — Development of technology for Microwave Enhanced Freeze Drying of Solid Waste (MEFDSW) is proposed. The present state of the art for solid waste stabilization using...

  12. Universality of Tip Singularity Formation in Freezing Water Drops

    CERN Document Server

    Marin, Alvaro G; Brunet, Philipe; Colinet, Pierre; Snoeijer, Jacco H

    2014-01-01

    A drop of water deposited on a cold plate freezes into an ice drop with a pointy tip. While this phenomenon clearly finds its origin in the expansion of water upon freezing, a quantitative description of the tip singularity has remained elusive. Here we demonstrate how the geometry of the freezing front, determined by heat transfer considerations, is crucial for the tip formation. We perform systematic measurements of the angles of the conical tip, and reveal the dynamics of the solidification front in a Hele-Shaw geometry. It is found that the cone angle is independent of substrate temperature and wetting angle, suggesting a universal, self-similar mechanism that does not depend on the rate of solidification. We propose a model for the freezing front and derive resulting tip angles analytically, in good agreement with observations.

  13. REDOX ASPECTS REGARDING PRESERVATION OF VEGETAL FOOD PRODUCTS BY FREEZING

    Directory of Open Access Journals (Sweden)

    Cristinel Zanoaga

    2007-08-01

    Full Text Available Freezing, at least of the vegetable products, no matter of the anatomic part involved – root, stalk, leaf, flower, fruit or seed – induces a biochemical changes manifested, implicitly, by modification of the tissular rH. More precisely, the more distant the native rH of the product under consideration is from a certain value – apparently the optimum/compatible to the animal, consuming organism – (rH ~ 26, the deeper are the transformations it suffers as a result of freezing, namely: if the native rH takes values below 26, the product gets oxidized by freezing while, in the opposite case, it is reduced, as if the whole group of vegetal products would tend to attain – by means of freezing – a (redox homeostasis

  14. Microwave Enhanced Freeze Drying of Solid Waste Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The development of advanced methods for Microwave Enhanced Freeze Drying of Solid Waste (MEFDSW) is proposed. Methods for the recovery of relatively pure water as a...

  15. CURRENT TECHNOLOGY AND INNOVATIONS IN FREEZE SHAFT SINKING IN CHINA

    Institute of Scientific and Technical Information of China (English)

    张铭; 翁家杰; 夏正明

    1991-01-01

    This paper gives a brief review of the development of shaft sinking by artificial ground freezing since 1949 when new China was founded. Several shaft freezing schemes which have been successfully applied from the economic and safe viewpoints are presented. Current technology and some innovative techniques,especially the shah lining which have experienced major improvements over the last four decades,are briefly reviewed. The technique of the in-situ pour concrete incorporating ailica fume with higher early strength under low temperature curing conditions is described. The temperature field in shah freezing and its finite difference solution are given in this paper. A recently developed method combining freeze wall model test with back analysis technique based on numerical simulation is also described.

  16. An approximation for homogeneous freezing temperature of water droplets

    Science.gov (United States)

    O, K.-T.; Wood, R.

    2015-11-01

    In this work, based on the well-known formulae of classical nucleation theory (CNT), the temperature TNc = 1 at which the mean number of critical embryos inside a droplet is unity is derived and proposed as a new approximation for homogeneous freezing temperature of water droplets. Without consideration of time dependence and stochastic nature of the ice nucleation process, the approximation TNc = 1 is able to reproduce the dependence of homogeneous freezing temperature on drop size and water activity of aqueous drops observed in a wide range of experimental studies. We use the TNc = 1 approximation to argue that the distribution of homogeneous freezing temperatures observed in the experiments may largely be explained by the spread in the size distribution of droplets used in the particular experiment. It thus appears that this approximation is useful for predicting homogeneous freezing temperatures of water droplets in the atmosphere.

  17. Freezing Resistance of HPC with Nano-SiO2

    Institute of Scientific and Technical Information of China (English)

    WANG Baomin; WANG Lijiu; F C Lai

    2008-01-01

    The test of fast-freezing method is employed to research the freezing resistance of C60and C80 high-performance concrete with nanometer-SiO2,and XRD and SEM are employed to analyze the mechanism of its influence.The experimental results show that the freezing resistance of high-performance concrete has been effectively improved.The compressive strength of C60,C70 and C80 high-performance concrete added with nano-SiO2 can be improved by 8%,6% and 5% respectively,the weight loss can be reduced by 1%,0.6% and 0.3% respectively and the relative dynamic elastic modulus can be improved by 7%,4% and 6%respectively after the 300 times freeze-thaw cycle.

  18. PREPARATION AND CHARACTERIZATION OF FREEZE DRIED CRYSTALS OF IBUPROFEN

    OpenAIRE

    Dixit Mudit; Kulkarni Parthasarathi Keshavarao; Panner Selvam; Syed Mohsin Ali

    2011-01-01

    Ibuprofen, an anti-inflammatory drug, exhibits poor water solubility and flow properties.. Crystallization medium used for freeze dried crystals of Ibuprofen consisted of Isopropyl alcohol and water (50:50%) respectively. Freeze dried crystals were characterized by differential scanning calorimetry, Infrared spectroscopy, X-ray diffractometry and scanning electron microscopy. Micromeritic and mechanical property and dissolution behavior studies were carried out. Process variables such as amou...

  19. Freeze concentration of dairy products Phase 2. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Best, D.E.; Vasavada, K.C.

    1993-09-01

    An efficient, electrically driven freeze concentration system offers potential for substantially increasing electricity demand while providing the mature dairy industry with new products for domestic and export markets together with enhanced production efficiencies. Consumer tests indicate that dairy products manufactured from freeze-concentrated ingredients are either preferred or considered equivalent in quality to fresh milk-based products. Economic analyses indicate that this technology should be competitive with thermal evaporation processes on a commercial basis.

  20. Study on the Efficiency of the Vegetables Freezing Process

    Directory of Open Access Journals (Sweden)

    Simona Paulikienė

    2014-10-01

    Full Text Available Changes taking place during the storage of the products of vegetable origin are inevitable. Products can deteriorate and change their shelf appearance, nutritional value, texture, taste and aroma. Freezing is one of the most appropriate technological operations for the product storage. By freezing products of vegetable origin, cleaner production technology can be used to consume less energy and to preserve quality of the product as well as valuable nutritional/ biologically active substances.The study of vegetable freezing process was carried out by freezing red beets. Beetroot was frozen in a chamber with controlled climatic environment - the Feutron.A portion of moisture content always evaporates from products of vegetable origin at freezing. Air flow velocity has a great influence on freezing rate of vegetables. Intensity of moisture evaporation also increases up to the moment of freezing-up of the surface layers.Intensity of retardation of the evaporation process depends on a peel and on stomata on it. Moisture evaporation’s retardation rate of the red beet peel εevp.ap = 0,50 ± 0,02. When air velocity was 0,15 m/s - εevp varied from 0,47 to 0,53, whilst at the air flow velocity 0,65 m/s - εevp = 0,37 – 0,45.Purpose of the study - to analyse the freezing process of vegetables (by the red beetroot example under the influence of different environment factors.DOI: http://dx.doi.org/10.5755/j01.erem.69.3.6064

  1. Stabilized, Freeze-Dried PCR Mix for Detection of Mycobacteria

    Science.gov (United States)

    Klatser, Paul R.; Kuijper, Sjoukje; van Ingen, Cor W.; Kolk, Arend H. J.

    1998-01-01

    We report here the development of a freeze-drying procedure allowing stabilization at ambient temperature of preoptimized, premixed, and predispensed PCR mixes aimed at the detection of mycobacteria in clinical materials. The freeze-dried mixes retained activity at 4°C and at 20°C for 1 year and for 3 months at 37°C, as judged by their performance with 50 and 500 fg of purified Mycobacterium bovis BCG target DNA. PMID:9620427

  2. Winter railcar unloading: heat thawing vs FCA. [Freeze conditioning agents

    Energy Technology Data Exchange (ETDEWEB)

    Walsh, R.J.

    1983-11-01

    The use of freeze conditioning agents and thawing by radiant heat are compared as methods for freezing coal for unloading from railcars in severe weather. The costs of the methods are evaluated and it is shown that, although heating systems require capital expenditure, if a sufficient volume of coal is to be treated, the cost per ton compares very favourably with the use of chemical agents.

  3. STUDY OF HIGH INTENSITY, AIRBORNE ULTRASOUND IN ATMOSPHERIC FREEZE DRYING

    OpenAIRE

    Bantle, Michael

    2011-01-01

    Atmospheric or convective freeze drying (AFD) is a dehydration process mostly employed in the food industry. It is also a promising process for the freeze drying of high-end products in the biological and pharmaceutical industry. Proteolytic and lipolytic activity as well as changes in lipid classes and lipid oxidation for products from AFD were investigated and the general potential of the drying method is described.The drying rate in AFD is generally low. In order to accelerate the drying p...

  4. Freezing and Thawing Durability of Very High Strength Concrete

    Directory of Open Access Journals (Sweden)

    Sameer Hamoush

    2011-01-01

    Full Text Available Problem statement: The newly developed Very High Strength Concrete (VHSC, having compressive strengths of 29 ksi and flexural strengths of 6 ksi, represents a breakthrough in concrete technology. Study to further enhance the properties of this new concrete is continuing. Approach: The objective of this study is to investigate the effect of exposing Very High Strength Concrete (VHSC specimens to rapid freeze/thaw cycles. Twenty one specimens were tested according to the Standards of the American Society for Testing and Materials ASTM C215, ASTM C666 and ASTM C78. Results: One hundred freeze/thaw cycles were performed on the VHSC specimens. Change in specimen’s dimensions and material’s properties were recorded at zero, forty, seventy and one hundred cycles. Dimensions and properties considered were: dimension of cross section, length, weight, Dynamic Moduli, Poisson’s Ratio, durability factor and Modulus of Rupture. Conclusion/Recommendations: The test results indicated that VHSC is good freeze-thaw resistance (durability factor > 85% and can avoid freeze/thaw damage. Freeze- thaw cycling did not significantly affect VHSC specimens’ cross sectional dimensions, length, or Poisson’s Ratio. However, there was a decrease in the specimens’ weight with the increase in number of freeze/thaw cycles, but the decrease was very slim indicating little or no deterioration has occur. Moreover, the fine voids exist in VHSC greatly lower the freezing point of any trapped water, making the material less susceptible to Freeze- Thaw damage.

  5. Effect of Surface Energy on Freezing Temperature of Water.

    Science.gov (United States)

    Zhang, Yu; Anim-Danso, Emmanuel; Bekele, Selemon; Dhinojwala, Ali

    2016-07-13

    Previous studies have found that superhydrophobic surfaces are effective in delaying freezing of water droplets. However, the freezing process of water droplets on superhydrophobic surfaces depends on factors such as droplet size, surface area, roughness, and cooling rate. The role of surface energy, independent of any other parameters, in delaying freezing of water is not understood. Here, we have used infrared-visible sum frequency generation spectroscopy (SFG) to study the freezing of water next to solid substrates with water contact angles varying from 5° to 110°. We find that the freezing temperature of water decreases with increasing surface hydrophobicity only when the sample volume is small (∼10 μL). For a larger volume of water (∼300 μL), the freezing temperature is independent of surface energy. For water next to the surfaces with contact angle ≥54°, we observe a strong SFG peak associated with highly coordinated water. This research sheds new light on understanding the key factors in designing new anti-icing coatings. PMID:27314147

  6. Mechanisms of deterioration of nutrients. [improved quality of freeze-dried foods

    Science.gov (United States)

    Karel, M.; Flink, J. M.

    1978-01-01

    Methods for improving the quality of freeze-dried foods were investigated. Areas discussed include: (1) microstructure of freeze-dried systems, (2) structural changes in freeze-dried systems, (3) artificial food matrices, and (4) osmotic preconcentration to yield improved freeze-dried products.

  7. Ketahanan Hidup Sel Acetobacter xylinum pada Pengawetan secara Kering-Beku Menggunakan Medium Pembawa (Viability of A. xylinum Subjected to Freeze Drying Using Carrier Media

    Directory of Open Access Journals (Sweden)

    Noor Aini Habibah

    2009-03-01

    Full Text Available A research on the use of sucrose and lactose as carrier media to protect Acetobacter xylinum cell subjected to freeze drying has been done. The aim of the research was to know the number of the viable cells from dried culture and to know the concentration of the carrier medium that would give best result. The best result is sucrose at the concentration of 15% that produced 28.2 x 106 viable cells/ ml of rehidrated culture. The rehidrated culture used in the research was Schramm & Herstin medium. Key words : Acetobacter xylinum, freeze drying, carrier media

  8. Effects of freezing, freeze drying and convective drying on in vitro gastric digestion of apples.

    Science.gov (United States)

    Dalmau, Maria Esperanza; Bornhorst, Gail M; Eim, Valeria; Rosselló, Carmen; Simal, Susana

    2017-01-15

    The influence of processing (freezing at -196°C in liquid N2, FN sample; freeze-drying at -50°C and 30Pa, FD sample; and convective drying at 60°C and 2m/s, CD sample) on apple (var. Granny Smith) behavior during in vitro gastric digestion was investigated. Dried apples (FD and CD samples) were rehydrated prior to digestion. Changes in carbohydrate composition, moisture, soluble solids, acidity, total polyphenol content (TPC), and antioxidant activity (AA) of apple samples were measured at different times during digestion. Processing resulted in disruption of the cellular structure during digestion, as observed by scanning electron microscopy, light microscopy, and changes in carbohydrate composition. Moisture content increased (6-11% dmo), while soluble solids (55-78% dmo), acidity (44-72% dmo), total polyphenol content (30-61% dmo), and antioxidant activity (41-87%) decreased in all samples after digestion. Mathematical models (Weibull and exponential models) were used to better evaluate the influence of processing on apple behavior during gastric digestion. PMID:27542444

  9. Strength and stiffness variation of frozen soils according to confinement during freezing

    Institute of Scientific and Technical Information of China (English)

    SangYeob Kim; JongSub Lee

    2015-01-01

    When water between soil particles is frozen, the strength and stiffness behavior of soils significantly change. Thus, nu-merous experimental studies in the laboratory have been carried out to characterize the strength and stiffness of frozen soils. The goals of this study are to evaluate the strength characteristics of frozen soils, which underwent confinement in freezing and shearing stages, and to estimate the stiffness variation by shear wave velocity during shear phase. The specimens are prepared in a brass cell by mixing sand and silt with 10% degree of saturation at a relative density of 60%. The applied normal stresses as confining stresses are 5, 10, 25 and 50 kPa. When the temperature of the specimens is lowered up to−5 °C, direct shear tests are carried out. Furthermore, shear waves are continuously measured through bender elements during shearing stage for the investigation of stiffness change. Test results show that shear strength and stiffness are significantly affected by the confining stress in freezing and shearing phases. This study suggests that the strength and stiffness of frozen soils may be dependent on the confining stresses applied during freezing and shearing.

  10. 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.

  11. Antioxidant, antiproliferative and antimicrobial activity of freeze-dried raspberry

    Directory of Open Access Journals (Sweden)

    Vulić Jelena J.

    2014-01-01

    Full Text Available The main chemical composition, i.e. the total content of bioactive compounds (phenolics 2209.86 ± 70.32 mg GAE/100g FDR, flavonoids 831.87 ± 12.61 mg R/100g FDR and anthocyanins 144.55 ± 0.39 mg CGE/100g FDR, in freeze-dried raspberry (FDR was evaluated spectrophotometrically. Vitamin C content was determined by HPLC analysis (88.81 ± 4.38 mg vit C/100g FDR. Antioxidant activities of FDR extract were evaluated spectrophotometrically on stable 2,2-diphenyl-1-picrylhydrazyl (DPPH free radicals and by electron spin resonance spectroscopy (ESR method on hydroxyl radicals (•OH. EC50 values were evaluated. EC50 DPPH• was 0.127 ± 0.013 mg/ml, while EC50 •OH was 1.366 ± 0.026 mg/ml. Antiproliferative activity of the FDR extract was evaluated in vitro in three human cell lines by colorimetric sulphorhodamine B (SRB assay. The most pronounced effects were obtained in the breast adenocarcinoma cell line (MCF7. EC50 value was 395.07 ± 96.38 μg/ml. Antimicrobial activity was determined by disk diffusion method. The FDR extract produced a clear inhibition zone (without visible colonies only toward Staphylococcus aureus. The minimal inhibitory (MIC and minimal bactericidal (MBC concentrations of FDR extract were evaluated. The values MIC were in the range of 4.7 - 100 mg/ml, and of MBC in the range of 6.3 - > 100 mg/ml.[ Projekat Ministarstva nauke Republike Srbije, br. TR 31044

  12. Preparation and Characterization ofβ-TCP/CS Scaffolds by Freeze-extraction and Freeze-gelation

    Institute of Scientific and Technical Information of China (English)

    HAO Ruiran; WANG Deping; YAO Aihua; HUANG Wenhai

    2011-01-01

    The freeze-extraction and freeze-gelation methods were used to prepare highly porous β-TCP/CS scaffolds with different β-TCP/CS ratio. In these methods, the suspending mixture of β-TCP and chitosan was frozen, the frozen mixture was immersed in a non-solvent(0.05 mol/L NaOH/ehanol aqueous) bath to allow the exchange between solvent(acetic-acid aqueous) and non-solvent at a temperature lower than the freezing point of the acetic-acid. Then, the β-TCP/CS scaffolds were formed and dried at room temperature. Scanning electron microscopy (SEM), X-ray diffraction (XRD), gas chromatography (GC) and omnipotence material testing machine were employed to characterize theβ-TCP/CS scaffolds. The results of GC show that the freeze extraction of the β-TCP/chitosan scaffolds was completed when the extraction time is above 24 h. The SEM results show that the β-TCP/CS scaffolds are composed of interconnected pore network. The porosity of the β-TCP/CS scaffolds decrease with the increase of the content of the β-TCP. The β-TCP/CS scaffolds have a highest compressive strength when the chitosan/β-TCP ratio is 30:70. The present work displays that the β-TCP/CS composite scaffolds with appropriate mechanical properties and high porosity can be successfully prepared by the freeze-extraction and freeze-gelation methods.

  13. Antifreeze proteins govern the precipitation of trehalose in a freezing-avoiding insect at low temperature

    OpenAIRE

    WEN, Xin; Wang, Sen; Duman, John G.; Arifin, Josh Fnu; Juwita, Vonny; Goddard, William A.; Rios, Alejandra; Liu, Fan; Kim, Soo-Kyung; Abrol, Ravinder; DeVries, Arthur L.; Henling, Lawrence M.

    2016-01-01

    The remarkable adaptive strategies of insects to extreme environments are linked to the biochemical compounds in their body fluids. Trehalose, a versatile sugar molecule, can accumulate to high levels in freeze-tolerant and freeze-avoiding insects, functioning as a cryoprotectant and a supercooling agent. Antifreeze proteins (AFPs), known to protect organisms from freezing by lowering the freezing temperature and deferring the growth of ice, are present at high levels in some freeze-avoiding ...

  14. Repeated vitrification/warming of human sperm gives better results than repeated slow programmable freezing

    Institute of Scientific and Technical Information of China (English)

    Teraporn Vutyavanich; Worashorn Lattiwongsakorn; Waraporn Piromlertamorn; Sudarat Samchimchom

    2012-01-01

    In this study,we compared the effects of repeated freezing/thawing of human sperm by our in-house method of rapid freezing with slow programmable freezing.Sperm samples from 11 normozoospermic subjects were processed through density gradients and divided into three aliquots:non-frozen,rapid freezing and slow programmable freezing.Sperm in the rapid freezing group had better motility and viability than those in the slow freezing group (P<O.01) after the first,second and third cycles of freezing/thawing,but there was no difference in morphology.In the second experiment,rapid freezing was repeated three times in 20 subjects.The samples from each thawing cycle were evaluated for DNA fragmentation using the alkaline comet assay.DNA fragmentation began to increase considerably after the second cycle of freezing/thawing,but to a level that was not clinically important.In the third experiment,rapid freezing was done repeatedly in 10 subjects,until no motile sperm were observed after thawing.The median number of repeated freezing/thawing that yielded no motile sperm was seven (range:5-8,mean:6.8).In conclusion,we demonstrated that repeated freezing/thawing of processed semen using our rapid freezing method gave better results than standard slow programmable freezing.This method can help maximize the usage of precious cryopreserved sperm samples in assisted reproduction technology.

  15. Comparison of cell components in low sugar adaptation and freeze-tolerance baker's yeast with common yeast%低糖适应性耐冻酵母菌与普通酵母菌细胞构成成分的比较

    Institute of Scientific and Technical Information of China (English)

    宋振玉; 李楠; 张姝; 王雷; 居勇

    2009-01-01

    采用气相、液相色谱法和分光光度比色法,对筛选出的低糖适应性耐冻面包酵母BY-03和FY-03与普通高糖面包酵母(PT)细胞构成成分,即胞内海藻糖含量,脂肪酸组成,麦角固醇,磷脂和氨基酸组成进行了分析与比较,进而了解其对酵母耐冻性的影响.结果表明,FY-03海藻糖含量是PT含量的1.40倍,BY-03的海藻糖含量与PT含量基本持平.从3种酵母中皆检出18种氨基酸,其中有助于提高酵母耐冻性的3种带电荷氨基酸即精氨酸,脯氨酸和谷氨酸的含量,BY-03和FY-03均高于PT.从3种酵母中检出了8种饱和与不饱和的脂肪酸,其中BY-03、FY-03的不饱和脂肪酸比例分别为79.82%和78.62%,明显高于PT 64.96%,其中棕榈油酸C16∶ 1和油酸C18∶ 1的含量相差较大.麦角固醇的含量BY-03和FY-03分别为4.99mg/g和4.77mg/g显著高于PT的2.56mg/g,而三者的磷脂含量没有明显差别.%The cell components, including trehalose, fatty acid component, ergosterol, phospholipid and amino acid in the BY-03 and FY-03 were qualitatively and quantitatively determined and compared with the common baker's yeast (PT) by GC, HPLC and UV-spectrophotometer in order to find out the effects of these index on freeze-tolerance of yeast.The results suggest that the content of the trehalose in FY-03 was 1.4 times more than that in PT, while the one in BY-03 was similar to PT.Meanwhile, eighteen kinds of known amino acid were detected from three kinds of yeast.The content of the arginine, proline and glutamate, which were helpful for enhancing the ability of freeze-tolerance, in BY-03 and FY-03 were more than those in PT.Eight kinds of known saturated and unsaturated fatty acid components were detected in three kinds of yeasts. The unsaturated fatty acid in BY-03 and FY-03 were 79.82% and 78.62%, 4.99 mg/g and 4.77 mg/g, but 2.57 mg/g in PT, while the phospholipid contents were similar in all samples.

  16. 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.

  17. Preparation of Chitosan Nanocompositeswith a Macroporous Structure by Unidirectional Freezing and Subsequent Freeze-Drying

    Directory of Open Access Journals (Sweden)

    Inmaculada Aranaz

    2014-11-01

    Full Text Available Chitosan is the N-deacetylated derivative of chitin, a naturally abundant mucopolysaccharide that consists of 2-acetamido-2-deoxy-β-d-glucose through a β (1→4 linkage and is found in nature as the supporting material of crustaceans, insects, etc. Chitosan has been strongly recommended as a suitable functional material because of its excellent biocompatibility, biodegradability, non-toxicity, and adsorption properties. Boosting all these excellent properties to obtain unprecedented performances requires the core competences of materials chemists to design and develop novel processing strategies that ultimately allow tailoring the structure and/or the composition of the resulting chitosan-based materials. For instance, the preparation of macroporous materials is challenging in catalysis, biocatalysis and biomedicine, because the resulting materials will offer a desirable combination of high internal reactive surface area and straightforward molecular transport through broad “highways” leading to such a surface. Moreover, chitosan-based composites made of two or more distinct components will produce structural or functional properties not present in materials composed of one single component. Our group has been working lately on cryogenic processes based on the unidirectional freezing of water slurries and/or hydrogels, the subsequent freeze-drying of which produce macroporous materials with a well-patterned structure. We have applied this process to different gels and colloidal suspensions of inorganic, organic, and hybrid materials. In this review, we will describe the application of the process to chitosan solutions and gels typically containing a second component (e.g., metal and ceramic nanoparticles, or carbon nanotubes for the formation of chitosan nanocomposites with a macroporous structure. We will also discuss the role played by this tailored composition and structure in the ultimate performance of these materials.

  18. Burkholderia phytofirmans PsJN reduces damages to freezing temperature in Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Fan eSU

    2015-10-01

    Full Text Available Several plant growth-promoting rhizobacteria (PGPR are known to improve plant tolerance to multiple stresses, including low temperatures. However, mechanisms underlying this protection are still poorly understood. The aim of this study was to evaluate the role of the endophytic PGPR, Burkholderia phytofirmans strain PsJN (Bp PsJN, on Arabidopsis thaliana cold tolerance using photosynthesis parameters as physiological markers.Under standard conditions, our results indicated that Bp PsJN inoculation led to growth promotion of Arabidopsis plants without significant modification on photosynthesis parameters and chloroplast organization. However, bacterial colonization induced a cell wall strengthening in the mesophyllImpact of inoculation modes (either on seeds or by soil irrigation and their effects overnight at 0, -1 or -3°C, were investigated by following photosystem II (PSII activity and gas exchanges. Following low temperatures stress, a decrease of photosynthesis parameters was observed. In addition, during three consecutive nights or days at -1°C, PSII activity was monitored. Pigment contents, RuBisCO protein abundance, expression of several genes including RbcS, RbcL, CBF1, CBF2, CBF3, ICE1, COR15a, and COR78 were evaluated at the end of exposure. To assess the impact of the bacteria on cell ultrastructure under low temperatures, microscopic observations were achieved. Results indicated that freezing treatment induced significant changes in PSII activity as early as the first cold day, whereas the same impact on PSII activity was observed only during the third cold night. The significant effects conferred by PsJN were differential accumulation of pigments, and reduced expression of RbcL and COR78. Microscopical observations showed an alteration/disorganization in A. thaliana leaf mesophyll cells independently of the freezing treatments. The presence of bacteria during the three successive nights or days did not significantly improved A

  19. Freeze-In Dark Matter with Displaced Signatures at Colliders

    CERN Document Server

    Co, Raymond T; Hall, Lawrence J; Pappadopulo, Duccio

    2015-01-01

    Dark matter, $X$, may be generated by new physics at the TeV scale during an early matter-dominated (MD) era that ends at temperature $T_R \\ll {\\rm TeV}$. Compared to the conventional radiation-dominated (RD) results, yields from both Freeze-Out and Freeze-In processes are greatly suppressed by dilution from entropy production, making Freeze-Out less plausible while allowing successful Freeze-In with a much larger coupling strength. Freeze-In is typically dominated by the decay of a particle $B$ of the thermal bath, $B \\rightarrow X$. For a large fraction of the relevant cosmological parameter space, the decay rate required to produce the observed dark matter abundance leads to displaced signals at LHC and future colliders, for any $m_X$ in the range ${\\rm keV} < m_X < m_B$ and for values of $m_B$ accessible to these colliders. This result applies whether the early MD era arises after conventional inflation, when $T_R$ is the usual reheat temperature, or is a generic MD era with an alternative origin. I...

  20. Enhanced enzymatic conversion with freeze pretreatment of rice straw

    Energy Technology Data Exchange (ETDEWEB)

    Chang, Ken-Lin; Thitikorn-amorn, Jitladda; Ou, Bay-Ming; Chen, Shan-He; Huang, Po-Jung [Institute of Biological Chemistry and Genomics Research Center Academia Sinica, Nankang, Taipei 115 (China); Hsieh, Jung-Feng [Department of Food Science, Fu Jen Catholic University, Xin Zhuang, Taipei 242 (China); Ratanakhanokchai, Khanok [School of Bioresources and Technology, King Mongkut' s University of Technology Thonburi, Bangkok 10150 (Thailand); Chen, Shui-Tein [Institute of Biological Chemistry and Genomics Research Center Academia Sinica, Nankang, Taipei 115 (China); Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 106 (China)

    2011-01-15

    Production of bioethanol by the conversion of lignocellulosic waste has attracted much interest in recent years, because of its low cost and great potential availability. The pretreatment process is important for increasing the enzymatic digestibility of lignocellulosic materials. Enzymatic conversion with freeze pretreatment of rice straw was evaluated in this study. The freeze pretreatment was found to significantly increase the enzyme digestibility of rice straw from 48% to 84%. According to the results, enzymatic hydrolysis of unpretreated rice straw with 150 U cellulase and 100 U xylanase for 48 h yielded 226.77 g kg{sup -1} and 93.84 g kg{sup -1} substrate-reducing sugars respectively. However, the reducing sugar yields from freeze pretreatment under the same conditions were 417.27 g kg{sup -1} and 138.77 g kg{sup -1} substrate, respectively. In addition, hydrolyzates analysis showed that the highest glucose yield obtained during the enzymatic hydrolysis step in the present study was 371.91 g kg{sup -1} of dry rice straw, following pretreatment. Therefore, the enhanced enzymatic conversion with freeze pretreatment of rice straw was observed in this study. This indicated that freeze pretreatment was highly effective for enzymatic hydrolysis and low environmental impact. (author)

  1. 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......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...... and cod and rainbow trout seem potential candidates for freeze-chilled meal elements. (C) 2009 Society of Chemical Industry...

  2. Freezing in porous media: Phase behavior, dynamics and transport phenomena

    Energy Technology Data Exchange (ETDEWEB)

    Wettlaufer, John S. [Yale Univ., New Haven, CT (United States)

    2012-12-21

    This research was focused on developing the underlying framework for the mechanisms that control the nature of the solidification of a broad range of porous media. To encompass the scope of porous media under consideration we considered material ranging from a dilute colloidal suspension to a highly packed saturated host matrix with a known geometry. The basic physical processes that occur when the interstitial liquid phase solidifies revealed a host of surprises with a broad range of implications from geophysics to materials science and engineering. We now understand that ostensibly microscopic films of unfrozen liquid control both the equilibrium and transport properties of a highly packed saturated host matrix as well as a rather dilute colloidal suspension. However, our description of the effective medium behavior in these settings is rather different and this sets the stage for the future research based on our past results. Once the liquid phase of a saturated relatively densely packed material is frozen, there is a rich dynamical behavior of particles for example due to the directed motion driven by thermomolecular pressure gradients or the confined Brownian motion of the particles. In quite striking contrast, when one freezes a dilute suspension the behavior can be rather more like that of a binary alloy with the particles playing the role of a ``solute''. We probed such systems quantitatively by (i) using X ray photon correlation spectroscopy (XPCS) and Small Angle X-ray Scattering (SAXS) at the Advanced Photon Source at Argonne (ii) studying the Argonne cell in the laboratory using optical microscopy and imagery (because it is not directly visible while in the vacuum can). (3) analyzed the general transport phenomena within the framework of both irreversible thermodynamics and alloy solidification and (4) applied the results to the study of the redistribution of solid particles in a frozen interstitial material. This research has gone a long way

  3. Group 1 LEA proteins contribute to the desiccation and freeze tolerance of Artemia franciscana embryos during diapause.

    Science.gov (United States)

    Toxopeus, Jantina; Warner, Alden H; MacRae, Thomas H

    2014-11-01

    Water loss either by desiccation or freezing causes multiple forms of cellular damage. The encysted embryos (cysts) of the crustacean Artemia franciscana have several molecular mechanisms to enable anhydrobiosis-life without water-during diapause. To better understand how cysts survive reduced hydration, group 1 late embryogenesis abundant (LEA) proteins, hydrophilic unstructured proteins that accumulate in the stress-tolerant cysts of A. franciscana, were knocked down using RNA interference (RNAi). Embryos lacking group 1 LEA proteins showed significantly lower survival than control embryos after desiccation and freezing, or freezing alone, demonstrating a role for group 1 LEA proteins in A. franciscana tolerance of low water conditions. In contrast, regardless of group 1 LEA protein presence, cysts responded similarly to hydrogen peroxide (H2O2) exposure, indicating little to no function for these proteins in diapause termination. This is the first in vivo study of group 1 LEA proteins in an animal and it contributes to the fundamental understanding of these proteins. Knowing how LEA proteins protect A. franciscana cysts from desiccation and freezing may have applied significance in aquaculture, where Artemia is an important feed source, and in the cryopreservation of cells for therapeutic applications. PMID:24846336

  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 activity of lactic acid bacteria is related to the glycolytic pathway. To investigate the phenomenon of this stress-related viability improvement in L. bulgaricus, the activities and corresponding genes of key enzymes in glycolysis during 2% NaCl stress were studied. NaCl stress significantly enhanced (p activities of glycolytic 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. PMID:26691481

  5. Physiological and biochemical responses of Yarrowia lipolytica to dehydration induced by air-drying and freezing.

    Directory of Open Access Journals (Sweden)

    Caroline Pénicaud

    Full Text Available Organisms that can withstand anhydrobiosis possess the unique ability to temporarily and reversibly suspend their metabolism for the periods when they live in a dehydrated state. However, the mechanisms underlying the cell's ability to tolerate dehydration are far from being fully understood. The objective of this study was to highlight, for the first time, the cellular damage to Yarrowia lipolytica as a result of dehydration induced by drying/rehydration and freezing/thawing. Cellular response was evaluated through cell cultivability determined by plate counts, esterase activity and membrane integrity assessed by flow cytometry, and the biochemical composition of cells as determined by FT-IR spectroscopy. The effects of the harvesting time (in the log or stationary phase and of the addition of a protective molecule, trehalose, were investigated. All freshly harvested cells exhibited esterase activity and no alteration of membrane integrity. Cells freshly harvested in the stationary phase presented spectral contributions suggesting lower nucleic acid content and thicker cell walls, as well as longer lipid chains than cells harvested in the log phase. Moreover, it was found that drying/rehydration induced cell plasma membrane permeabilization, loss of esterase activity with concomitant protein denaturation, wall damage and oxidation of nucleic acids. Plasma membrane permeabilization and loss of esterase activity could be reduced by harvesting in the stationary phase and/or with trehalose addition. Protein denaturation and wall damage could be reduced by harvesting in the stationary phase. In addition, it was shown that measurements of loss of membrane integrity and preservation of esterase activity were suitable indicators of loss and preservation of cultivability, respectively. Conversely, no clear effect of freezing/thawing could be observed, probably because of the favorable operating conditions applied. These results give insights into Y

  6. Surviving freezing in plant tissues by oomycetous snow molds.

    Science.gov (United States)

    Murakami, Ryo; Yajima, Yuka; Kida, Ken-ichi; Tokura, Katsuyuki; Tojo, Motoaki; Hoshino, Tamotsu

    2015-04-01

    Oomyceteous snow molds, Pythium species, were reported to be less tolerant to chilling and freezing temperatures than other snow mold taxa. However, Pythium species are often found to be pathogenic on mosses in Polar Regions. We investigated the frost resistance of Pythium species from Temperate (Hokkaido, Japan) and Subantarctic Regions. Free mycelia and hyphal swellings, structures for survival, of Pythium iwayamai and Pythium paddicum lost viability within freeze-thaw 3 cycles; however, mycelia in host plants survived the treatment. It was reported that fungi in permafrost are characterized both by the presence of natural cryoprotectants in these ecotopes and by the ability to utilize their inherent mechanisms of protection. It is conceivable that plant substrates or derivatives thereof are natural cryoprotectants, enabling them to provide advantageous conditions to microorganisms under freezing conditions. Our results are the first to experimentally support this hypothesis.

  7. Theory of freezing of alkali halides and binary alloys

    International Nuclear Information System (INIS)

    Using the basic equations of classical statistical mechanics relating the singlet densities rho1 and rho2 of a binary system to the three partial direct correlation functions csub(ij), a theory of freezing is developed. Though the theory is set up for arbitrary concentration, we focus on the freezing of the alkali halides. In particular, we show that periodic solutions of the equations for rho1 and rho2 can coexist with homogeneous solutions. The difference in free energy between periodic and homogeneous phases is built up in terms of (i) the volume difference and (ii) the Fourier components of rho1, rho2 and csub(ij). To lowest order, it is stressed that the freezing transition is determined by the charge-charge structure factor at the principal peak and by the compressibility. (author)

  8. Freezing point depression in model Lennard-Jones solutions

    Science.gov (United States)

    Koschke, Konstantin; Jörg Limbach, Hans; Kremer, Kurt; Donadio, Davide

    2015-09-01

    Crystallisation of liquid solutions is of uttermost importance in a wide variety of processes in materials, atmospheric and food science. Depending on the type and concentration of solutes the freezing point shifts, thus allowing control on the thermodynamics of complex fluids. Here we investigate the basic principles of solute-induced freezing point depression by computing the melting temperature of a Lennard-Jones fluid with low concentrations of solutes, by means of equilibrium molecular dynamics simulations. The effect of solvophilic and weakly solvophobic solutes at low concentrations is analysed, scanning systematically the size and the concentration. We identify the range of parameters that produce deviations from the linear dependence of the freezing point on the molal concentration of solutes, expected for ideal solutions. Our simulations allow us also to link the shifts in coexistence temperature to the microscopic structure of the solutions.

  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 damages......, however, in most part unknown. In this work samples of concrete at different water to cement ratios and air bubble contents subjected to freeze/thaw cycles with the lowest temperature at about -80 oC are investigated. By adopting a novel technique a scanning calorimeter is used to obtain data from which...... 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 bubble structure was investigated using...

  10. Encapsulation and sustained release from biodegradable microcapsules made by emulsification/freeze drying and spray/freeze drying.

    Science.gov (United States)

    Yin, Weisi; Yates, M Z

    2009-08-01

    Hollow biodegradable poly(DL-lactide) (PLA) particles with porous shell walls were prepared by freeze drying small droplets of PLA solution formed by emulsification or spraying. The hollow freeze-dried particles were dispersed in water, and the resulting aqueous suspensions were exposed to plasticizing solvents, either dichloromethane or compressed carbon dioxide. The plasticizing solvent causes the pores in the shell wall to close, forming microcapsules surrounding an aqueous core. A water soluble drug, procaine hydrochloride, was successfully encapsulated in the microcapsule core. The encapsulation efficiency is affected by the hollow particle morphology, amount of solvent used, solvent exposure time, surfactant, and method of dispersing the freeze-dried particles in water. The encapsulation process is explained in terms of interfacial free energy of the hollow particles and mobility of the plasticized polymer. Controlled release of procaine hydrochloride from the microcapsules into phosphate buffer solution was observed. The microcapsules had a small burst release, with the remainder of encapsulated drug slowly released over 9 days. The novel hollow PLA particles produced by emulsification/freeze drying and spray/freeze drying can potentially be used as vehicles for controlled release. PMID:19423128

  11. Simulation of in situ freezing damage of the photosynthetic apparatus by freezing in vitro of thylakoids suspended in complex media.

    Science.gov (United States)

    Grafflage, S; Krause, G H

    1986-05-01

    Chloroplast thylakoid membranes were isolated from leaves of unhardened and cold-acclimated spinach (Spinacia oleracea L.). For freezethaw treatment, the membranes were suspended in complex media composed to simulate the solute concentrations in the chloroplast stroma in the unhardened and hardened states of the leaves. In particular, high concentrations of amino acids were applied for simulating the hardened state. After frost treatment, photosynthetic activities and chlorophyll fluorescence parameters of the thylakoids were tested to determine the degree of freezing damage. The results revealed a pattern of freezing injury similar to that observed upon frost treatment of thylakoids in situ. A major manifestation of damage was the inhibition of photosynthetic electron transport. Uncoupling of photophosphorylation, which is the dominating effect of freezing of thylakoids suspended in binary solutions (e.g., containing one sugar and one inorganic salt), was also visible but less pronounced in the complex media. Thylakoids obtained from cold-acclimated leaves did not exhibit an increased frost tolerance in vitro, as compared with thylakoids from unhardened plants. The results, furthermore, indicated a strong protective effect of free amino acids at the concentrations and composition found in chloroplasts of hardened leaves. The presence of inorganic salts in the complex media slightly stabilized rather than damaged the membranes during freezing. It is concluded that inactivation of thylakoids in situ may be understood as the destabilizing action of the combined solutes surrounding the thylakoids, occurring when solute concentration is raised due to freezing of water. PMID:24233737

  12. Encapsulation and subsequent freeze-drying of Lactobacillus reuteri CRL 1324 for its potential inclusion in vaginal probiotic formulations.

    Science.gov (United States)

    Juárez Tomás, María Silvina; De Gregorio, Priscilla Romina; Leccese Terraf, María Cecilia; Nader-Macías, María Elena Fátima

    2015-11-15

    Probiotic formulations must include a high number of viable and active microorganisms. In this work, the survival of human vaginal Lactobacillus reuteri CRL 1324 during encapsulation, lyophilization and storage, and the activity of encapsulated and/or freeze-dried bacterial cells were evaluated. Extrusion-ionic gelation technique was applied to encapsulate L. reuteri CRL 1324, using xanthan and gellan. Encapsulated and free bacterial cells were freeze-dried with or without lactose and skim milk as lyoprotectors. The different systems obtained were stored at room temperature and at 4°C for 150days. The following determinations were performed: L. reuteri CRL 1324 viability, microorganism released from capsules, survival in a medium simulating the vaginal fluid and maintenance of beneficial properties (growth inhibition of opportunistic pathogenic Streptococcus agalactiae NH 17 and biofilm formation). L. reuteri CRL 1324 encapsulation was efficient, allowing the recovery of a high number of entrapped lactobacilli. The survival of encapsulated L. reuteri during lyophilization and storage was significantly higher in the presence of lyoprotectors. At the end of storage, the highest numbers of viable cells were obtained in free or encapsulated cells freeze-dried with lyoprotectors, stored at 4°C. Encapsulated and/or lyophilized L. reuteri cells maintained their viability in simulated vaginal fluid as well as the ability to inhibit S. agalactiae NH 17 growth and to form biofilm. Encapsulated and freeze-dried L. reuteri CRL 1324 can be included in a suitable pharmaceutical form for vaginal application to prevent or treat urogenital infections in women. PMID:26299342

  13. Morphology and Morphogenesis of Sindbis Virus as Seen with Freeze-Etching Techniques

    Science.gov (United States)

    Brown, Dennis T.; Waite, Marilynn R. F.; Pfefferkorn, Elmer R.

    1972-01-01

    Freeze-etch electron microscope studies of the morphogenesis and morphology of Sindbis virus confirmed results obtained by other workers employing thin-sectioning techniques. The 68-nm virion was found to have a nucleocapsid 36 nm in diameter surrounded by a double-layered, unit membrane. The membranous envelope is acquired as the capsid buds through the plasma membrane of the infected cell. The freeze-etch technique also provided the following new information. (i) At any one time, budding occurs in patches rather than evenly over the cell surface. (ii) The nucleocapsid is composed of capsomers 7 nm in diameter. (iii) The capsid interacts strongly with the membrane, both prior to budding and after maturation. (iv) The 7- to 10-nm particles characteristic of the internal faces of plasma membranes, which presumably represent host membrane proteins, are present in early stages of budding but disappear as morphogenesis progresses. (v) Fusion of the cell membrane at the base of the budding virion is a two-step process; the inner leaflet fuses into a sphere before the outer one. (vi) The outer surface of the viral envelope is covered with 4-nm subunits with a center-to-center spacing of 6 nm. Images PMID:4672393

  14. Social Egg Freezing: Developing Countries Are Not Exempt.

    Science.gov (United States)

    Allahbadia, Gautam N

    2016-08-01

    Non-medical egg freezing has only been available for about the last 5 years, as new vitrification techniques have made the success rates for actual conception more reliable than the earlier method of slow freezing. The improved outcomes of new technologies of vitrification and intra-cytoplasmic sperm injection (ICSI) have led to the marketing of egg freezing for non-medical reasons, whereby women are offered the possibility of preserving their eggs until such time as they wish to have a child. For many women today, it is not cancer but the simple passage of time that robs them of their chance of motherhood. Social, educational, emotional and financial pressures often lead them to delay trying to start a family until their late thirties, by which time the chance of success is very low. Women at age 40 face a 40 % chance of miscarriage if they can get pregnant at all, and by the age of 45, the risk of miscarriage is 75 %. Donor eggs are not an option for many because of supply constraints and ethical and cultural concerns. Freezing a woman's eggs at age 30 literally "freezes in time" her fertility potential and gives her the chance of a healthy pregnancy at a time of her choosing. Despite the initial reactions of disapproval, more and more fertility clinics are now offering oocyte cryopreservation to healthy women in order to extend their reproductive options. This procedure is now becoming popular even in developing economies, and egg freezing in major Indian Metros is now routine. PMID:27382212

  15. Dynamics and Sizes of the Fireball at Freeze-out

    CERN Document Server

    Tomasik, Boris; Heinz, Ulrich W; Tomasik, Boris; Wiedemann, Urs Achim; Heinz, Ulrich

    2000-01-01

    Analyzing the m_t-spectrum and two-particle correlations of negative hadrons from 158 AGeV/c Pb+Pb collisions at slightly forward rapidities we find a (thermal) freeze-out temperature of about 100 MeV and transverse flow with approximately 0.55c. The M_t-dependence of the correlation radii prefers a box-like transverse density profile over a Gaussian. From an analysis of the pion phase-space density we find pion chemical potential of approximately 60 MeV at thermal freeze-out.

  16. Large tundra methane burst during onset of freezing

    DEFF Research Database (Denmark)

    Mastepanov, Mikhail; Sigsgaard, Charlotte; Dlugokencky, Edward J.;

    2008-01-01

    Terrestrial wetland emissions are the largest single source of the greenhouse gas methane1. Northern high-latitude wetlands contribute significantly to the overall methane emissions from wetlands, but the relative source distribution between tropical and high-latitude wetlands remains uncertain2...... after the growing season but then increase significantly during the freeze-in period. The integral of emissions during the freeze-in period is approximately equal to the amount of methane emitted during the entire summer season. Three-dimensional atmospheric chemistry and transport model simulations...

  17. Field application of freezing technology for social infrastructures

    Institute of Scientific and Technical Information of China (English)

    EunChul Shin; YuRa Gong; CheolGyu Jeong

    2015-01-01

    There are numerous methods to prevent seepage flow and ground improvement methodologies such as cement grouting, sheet piling, and the membrane method. In this paper, we present case histories of freezing technology applications in the construction of a deep tunnel sewerage system, undersea highway tunnel, and liquefied natural gas tank. Heaving pressure measurements for various soil types around a liquefied natural tank are compared with existing data. In this paper, we present temperature variations at the bottom and side of a liquefied natural gas tank. Our findings show that ground-freezing technology is very effective in controlling ground water infiltration into underground structures as well providing soil reinforcement in the soft ground.

  18. Freeze-out conditions from fluctuations of conserved charges

    Energy Technology Data Exchange (ETDEWEB)

    Ratti, C. [Dipartimento di Fisica, Universitá di Torino e INFN, Sezione di Torino (Italy); Physics Department, University of Houston, Houston, TX 77204 (United States); Borsanyi, S. [Department of Physics, Wuppertal University, Gaussstr. 20, D-42119 Wuppertal (Germany); Fodor, Z. [Department of Physics, Wuppertal University, Gaussstr. 20, D-42119 Wuppertal (Germany); Inst. for Theoretical Physics, Eötvös University, Pázmány P. sétány 1/A, H-1117 Budapest (Hungary); Jülich Supercomputing Centre, Forschungszentrum Jülich, D-52425 Jülich (Germany); Katz, S.D. [Inst. for Theoretical Physics, Eötvös University, Pázmány P. sétány 1/A, H-1117 Budapest (Hungary); MTA-ELTE “Lendület” Lattice Gauge Theory Research Group, Pázmány P. sétány 1/A, H-1117 Budapest (Hungary); Krieg, S.; Szabo, K.K. [Department of Physics, Wuppertal University, Gaussstr. 20, D-42119 Wuppertal (Germany); Jülich Supercomputing Centre, Forschungszentrum Jülich, D-52425 Jülich (Germany)

    2014-11-15

    The latest results on fluctuations of electric charge and baryon number, simulated on the lattice by the Wuppertal–Budapest Collaboration, are compared to the moments of multiplicity distribution of the corresponding conserved charges, measured in heavy ion collision experiments by the STAR Collaboration. The purpose of this study is to extract the chemical freeze-out parameters (temperature and chemical potential) as a function of the collision energy, from first principles. Consistency between the freeze-out parameters obtained through the two different conserved charges used in the analysis is discussed.

  19. On freeze-out problem in relativistic hydrodynamics

    CERN Document Server

    Ivanov, Yu B

    2008-01-01

    A finite unbound system which is equilibrium in one reference frame is in general nonequilibrium in another frame. This is a consequence of the relative character of the time synchronization in the relativistic physics. This puzzle was a prime motivation of the Cooper--Frye approach to the freeze-out in relativistic hydrodynamics. Solution of the puzzle reveals that the Cooper--Frye recipe is far not a unique phenomenological method that meets requirements of energy-momentum conservation. Alternative freeze-out recipes are considered and discussed.

  20. Empirical formula for the refractive index of freezing brine

    DEFF Research Database (Denmark)

    Frisvad, Jeppe Revall

    2009-01-01

    The refractive index of freezing brine is important for example in order to estimate oceanic scattering as sea ice develops. Previously, no simple continuous expression was available for estimating the refractive index of brine at subzero temperatures. I show that extrapolation of the empirical...... formula for the refractive index of seawater by Quan and Fry [Appl. Opt. 34(18), 3477-3480 (1995)] provides a good fit to the refractive index of freezing brine for temperatures above -24 degrees celsius and salinities below 180 parts per thousand....

  1. Versatile wedge-based system for the construction of unidirectional collagen scaffolds by directional freezing: practical and theoretical considerations.

    Science.gov (United States)

    Pot, Michiel W; Faraj, Kaeuis A; Adawy, Alaa; van Enckevort, Willem J P; van Moerkerk, Herman T B; Vlieg, Elias; Daamen, Willeke F; van Kuppevelt, Toin H

    2015-04-29

    Aligned unidirectional collagen scaffolds may aid regeneration of those tissues where alignment of cells and extracellular matrix is essential, as for instance in cartilage, nerve bundles, and skeletal muscle. Pores can be introduced by ice crystal formation followed by freeze-drying, the pore architecture reflecting the ice crystal morphology. In this study we developed a wedge-based system allowing the production of a wide range of collagen scaffolds with unidirectional pores by directional freezing. Insoluble type I collagen suspensions were frozen using a custom-made wedge system, facilitating the formation of a horizontal as well as a vertical temperature gradient and providing a controlled solidification area for ice dendrites. The system permitted the growth of aligned unidirectional ice crystals over a large distance (>2.5 cm), an insulator prolonging the freezing process and facilitating the construction of crack-free scaffolds. Unidirectional collagen scaffolds with tunable pore sizes and pore morphologies were constructed by varying freezing rates and suspension media. The versatility of the system was indicated by the construction of unidirectional scaffolds from albumin, poly(vinyl alcohol) (a synthetic polymer), and collagen-polymer blends producing hybrid scaffolds. Macroscopic observations, temperature measurements, and scanning electron microscopy indicated that directed horizontal ice dendrite formation, vertical ice crystal nucleation, and evolutionary selection were the basis of the aligned unidirectional ice crystal growth and, hence, the aligned unidirectional pore structure. In conclusion, a simple, highly adjustable freezing system has been developed allowing the construction of large (hybrid) bioscaffolds with tunable unidirectional pore architecture. PMID:25822583

  2. Cryopreservation of human failed-matured oocytes followed by in vitro maturation: vitrification is superior to the slow freezing method

    Directory of Open Access Journals (Sweden)

    Zhang ZhiGuo

    2011-12-01

    Full Text Available Abstract Background Oocyte cryopreservation is an important method used in a number of human fertility circumstances. Here, we compared the survival, in vitro maturation, fertilization, and early embryonic development rates of frozen-thawed human immature oocytes using two different cryopreservation methods. Methods A total of 454 failed-matured oocytes [germinal vesicle (GV and metaphase I (MI stages] were collected from 135 patients (mean age 33.84 +/- 5.0 y who underwent intracytoplasmic sperm injection (ICSI cycles between February 2009 and December 2009 and randomly divided into a slow freezing group [1.5 mol/L-1, 2-propanediol (PROH + 0.2 mol/l sucrose] and vitrification group [20% PROH + 20% ethylene glycol (EG + 0.5 mol/l sucrose]. Results The vitrification protocol yielded a better survival rate than the slow freezing protocol at each maturation stage assessed. Regardless of the maturation stage (GV + MI, the slow freezing protocol had a significantly lower survival rate than the vitrification protocol (p in vitro maturation (21.2 vs. 54.0%, respectively; p 0.05. For the GV-matured oocytes, no fertilized eggs were obtained in the slow-freezing group, while a 19.0% (4/21 fertilization rate was observed in the vitrification group. For the MI-matured oocytes, fertilization rates for the slow freezing and vitrified groups were 36% and 61.1%, respectively, but no significant difference was found between the two groups (PIn the Methods section in the MS, all procedures were compliant with ethical guidelines, i.e. approved by the Ethical Committee of our university and Informed Consent signed by each patient. > 0.05. In the GV vitrification group, no embryo formed; however, in the MI slow freezing group, 12 oocytes were fertilized, but only two achieved cleavage and were subsequently blocked at the 2-cell stage. In the MI vitrification group, a total of 22 embryos were obtained, five of which developed to the blastocyst stage. Conclusions

  3. Capital stock management during a recession that freezes credit markets

    NARCIS (Netherlands)

    Caulkins, J.P.; Feichtinger, G.; Grass, D.; Hartl, R.F.; Kort, Peter; Seidl, A.

    2015-01-01

    This paper considers the problem of how to price a conspicuous product while maintaining liquidity during a recession which both reduces demand and freezes credit markets. Reducing price would help maintain cash flow, but low prices can erode brand image and, hence, long-term sales. The paper extend

  4. How Circulation of Water Affects Freezing in Ponds

    Science.gov (United States)

    Moreau, Theresa; Lamontagne, Robert; Letzring, Daniel

    2007-01-01

    One means of preventing the top of a pond from freezing involves running a circulating pump near the bottom to agitate the surface and expose it to air throughout the winter months. This phenomenon is similar to that of the flowing of streams in subzero temperatures and to the running of taps to prevent pipe bursts in winter. All of these cases…

  5. Freeze desalination of seawater using LNG cold energy.

    Science.gov (United States)

    Chang, Jian; Zuo, Jian; Lu, Kang-Jia; Chung, Tai-Shung

    2016-10-01

    With the aid of cold energy from regasification of liquefied natural gas (LNG), freeze desalination (FD) is an emerging technology for seawater desalination because of its low energy characteristics and insensitivities to fouling problems. This work aims to investigate the major operating parameters of FD such as coolant temperature, freezing duration, supercooling, seeding, agitation, crystallizer material and subsequent washing procedure on ice production and water quality. It was found that the optimal freezing duration per batch was 1 h for an iron crystallizer and 1.5 h for a glass crystallizer. The optimal coolant temperature should be around -8 °C. The optimal amount of washing water to clean the raw ice was about 50 wt% of the raw ice. Over 50 wt% of the feed could be recovered as raw ice within 1 h, which means an overall ice recovery rate of higher than 25% (of the original seawater), considering the consumption of washing water. Both artificial and real seawater were tested under the optimized conditions. The total dissolved solid in the product ice was around 300 ppm, which met the World Health Organization (WHO) potable water salinity standard of 500 ppm. Therefore, the process parameters optimized in this study can be directly used for the freeze desalination of seawater.

  6. Growing bubbles and freezing drops: depletion effects and tip singularities

    NARCIS (Netherlands)

    Enríquez Paz y Puente, Oscar Raúl

    2015-01-01

    In this thesis, the author investigates the growth of gas bubbles in a supersaturated solution and the freezing of water drops when placed on a cold plate. Supersaturated solutions are common in nature and industry; perhaps the best know examples are carbonated drinks, such as beer or soda. These ar

  7. Cash Freeze for E-Rate Hits Schools

    Science.gov (United States)

    Borja, Rhea R.

    2004-01-01

    A freeze on $3.28 billion in requests for aid under the federal E-rate program has left hundreds of school districts scrambling to pay for their technology needs--and, in some cases, crippled classroom instruction. The E-rate program, which has disbursed more than $8 billion since its inception in 1997, helps link public and private schools as…

  8. Anatomy of a Tuition Freeze: The Case of Ontario

    Science.gov (United States)

    Rexe, Deanna

    2015-01-01

    Using two conceptual frameworks from political science--Kingdon's (2003) multiple streams model and the advocacy coalition framework (Sabatier & Jenkins-Smith, 1993)--this case study examines the detailed history of a major tuition policy change in Ontario in 2004: a tuition freeze. The paper explores the social, political, and economic…

  9. Freeze-drying of filamentous fungi and yeasts

    NARCIS (Netherlands)

    Tan, C.S.

    2011-01-01

    The aim of this thesis was to optimize the freeze-drying protocol for fungi in general and for those genera that do not survive this preservation method, in particular. To this end, the influence of the cooling rate, the lyoprotectant and the drying process itself was examined. Since most fungi belo

  10. Obstacle avoidance to elicit freezing of gait during treadmill walking.

    NARCIS (Netherlands)

    Snijders, A.H.; Weerdesteijn, V.G.M.; Hagen, Y.J.; Duysens, J.E.J.; Giladi, N.; Bloem, B.R.

    2010-01-01

    Freezing of gait (FOG) is a common and disabling feature of Parkinson's disease (PD). Detailed pathophysiological studies are hampered by the fact that FOG episodes are difficult to elicit in a gait laboratory. We evaluated whether the need to avoid sudden obstacles on a treadmill can provoke FOG. W

  11. 40 CFR 52.1135 - Regulation for parking freeze.

    Science.gov (United States)

    2010-07-01

    ... from acts of God, strikes, litigation, or other matters beyond the control of the owner shall be... of any such existing facility in the freeze area unless and until he has obtained from the Governor... conform to the requirements of 40 CFR 51.4(b); and the agency rules or procedures may provide that if...

  12. IMPACT OF FREEZING OF FCAS: THE CASE OF PAKISTAN

    Directory of Open Access Journals (Sweden)

    Hafiz Abdur Rashi

    2011-09-01

    Full Text Available Foreign investment and foreign exchange reserves have ample importance for developing countries. So, there is a needed to encourage the foreign and domestic investors whose confidence was suffer by the unexpected decision of freezing of FCAs. The purpose of this study was to identify the areas that were affected after the decision of freezing of FCAs. Moreover, the impact of freezing decision on economy of Pakistan also indicated. More sophisticated impact on banking sector, balance of payment, foreign exchange reserves, foreign debt, and foreign investment. Study found the negative impact of freezing decision of FCAs on foreign banking and positive impact on domestic banking but Pakistan banking sector was disconnected from the international banking; insatiability in balance of payment was increased; the foreign investment and exports was reduced.Resultantly, foreign exchange reserves were reduced and foreign exchange rates was increased. Therefore, it is suggested to increase the confidence of foreign investors in order to increase the foreign investment and foreign exchange reserves. Discussion of conclusions and recommendations were also provided.

  13. A heat equation for freezing processes with phase change

    DEFF Research Database (Denmark)

    Backi, Christoph Josef; Bendtsen, Jan Dimon; Leth, John-Josef;

    2016-01-01

    In this work, the stability properties as well as possible applications of a partial differential equation (PDE) with state-dependent parameters are investigated. Among other things, the PDE describes freezing of foodstuff, and is closely related to the (potential) Burgers’ equation. We show that...

  14. Freeze desalination of seawater using LNG cold energy.

    Science.gov (United States)

    Chang, Jian; Zuo, Jian; Lu, Kang-Jia; Chung, Tai-Shung

    2016-10-01

    With the aid of cold energy from regasification of liquefied natural gas (LNG), freeze desalination (FD) is an emerging technology for seawater desalination because of its low energy characteristics and insensitivities to fouling problems. This work aims to investigate the major operating parameters of FD such as coolant temperature, freezing duration, supercooling, seeding, agitation, crystallizer material and subsequent washing procedure on ice production and water quality. It was found that the optimal freezing duration per batch was 1 h for an iron crystallizer and 1.5 h for a glass crystallizer. The optimal coolant temperature should be around -8 °C. The optimal amount of washing water to clean the raw ice was about 50 wt% of the raw ice. Over 50 wt% of the feed could be recovered as raw ice within 1 h, which means an overall ice recovery rate of higher than 25% (of the original seawater), considering the consumption of washing water. Both artificial and real seawater were tested under the optimized conditions. The total dissolved solid in the product ice was around 300 ppm, which met the World Health Organization (WHO) potable water salinity standard of 500 ppm. Therefore, the process parameters optimized in this study can be directly used for the freeze desalination of seawater. PMID:27371931

  15. 7 CFR 58.638 - Freezing the mix.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Freezing the mix. 58.638 Section 58.638 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) REGULATIONS AND STANDARDS UNDER...

  16. Deformation of freezing water droplets on a cold copper surface

    Institute of Scientific and Technical Information of China (English)

    WANG Jieteng; LIU Zhongliang; GOU Yujun; ZHANG Xinhua; CHENG Shuiyuan

    2006-01-01

    Freezing processes of water and peanut oil droplets on a cold surface are investigated in this paper. We observed during our experiments that the base surface of a water droplet that is in direct contact with the cold surface keeps its original shape, but the other part of the droplet shows an obvious growth along the direction normal to the base surface. One small protrusion appears on the top of the water droplet at the end of the freezing process. The experimental observations also show that no obvious shape change happens during the freezing of peanut oil droplets. It is postulated that the effects of surface tension and volume dilatation resulted from liquid-to-solid phase change cause the shape change and protrusions formation. Based on this postulation, a physical and mathematical model is developed. The results of the model of a water droplet's freezing process correspond with our experimental observations. The observed phenomenon that frost-growth speed on the protrusion is higher than that on the other part of the water droplet is also analyzed.

  17. Freeze desalination of seawater using LNG cold energy

    KAUST Repository

    Chang, Jian

    2016-06-23

    With the aid of cold energy from regasification of liquefied natural gas (LNG), freeze desalination (FD) is an emerging technology for seawater desalination because of its low energy characteristics and insensitivities to fouling problems. This work aims to investigate the major operating parameters of FD such as coolant temperature, freezing duration, supercooling, seeding, agitation, crystallizer material and subsequent washing procedure on ice production and water quality. It was found that the optimal freezing duration per batch was 1 h for an iron crystallizer and 1.5 h for a glass crystallizer. The optimal coolant temperature should be around −8 °C. The optimal amount of washing water to clean the raw ice was about 50 wt% of the raw ice. Over 50 wt% of the feed could be recovered as raw ice within 1 h, which means an overall ice recovery rate of higher than 25% (of the original seawater), considering the consumption of washing water. Both artificial and real seawater were tested under the optimized conditions. The total dissolved solid in the product ice was around 300 ppm, which met the World Health Organization (WHO) potable water salinity standard of 500 ppm. Therefore, the process parameters optimized in this study can be directly used for the freeze desalination of seawater.

  18. Freezing Point Depressions of Phase Change CO2 Solvents

    DEFF Research Database (Denmark)

    Arshad, Muhammad Waseem; Fosbøl, Philip Loldrup; von Solms, Nicolas;

    2013-01-01

    Freezing point depressions (FPD) in phase change solvents containing 2-(diethylamino)ethanol (DEEA) and 3-(methylamino)propylamine (MAPA) were measured using a modified Beckmann apparatus. The measurements were performed for the binary aqueous DEEA and MAPA solutions, respectively......, in the concentration ranges of (0 to 55) mass percent and (0 to 32.5) mass percent of amine. For the ternary aqueous DEEA–MAPA solutions, freezing points were measured for 5:1, 3:1, 1:1, 1:3, and 1:5 molar ratios of DEEA/MAPA. The FPD method was extended for easy and accurate measurement of freezing points in the CO2...... loaded systems. It is based on saturation of the solution by CO2 and then dilution by using a batch of the original unloaded solution in order to get the solutions with different CO2 loadings. Freezing point measurements were then carried out for (12, 20, 30, and 33) mass percent DEEA solutions and (10...

  19. Measurement of Freezing Point Depression of Selected Food Solutions

    Science.gov (United States)

    Murata, Satoshi; Tanaka, Fumihiko; Matsuoka, Takahisa

    Freezing point depression of selected food solutions were measured at various concentrations in order to reveal the characteristics of solid-liquid phase equilibrium. The measurement were carried out on a hand made apparatus that was calibrated before the measurement by some of selected reagents (acid and sugar) with known thermal properties. The results revealed that the freezing point depression of selected food solutions deviated from the behavior of the ideal solution with increasing solute concentration, so the water activity for non-ideal solution were introduced to the freezing point depression equation. Further, assuming that the heat of fusion was a equation of temperature, thus the following new equation was led, ln {(1-Xs)/(l-Xs+α·Xs + β·Xs2)} = A(1/To - 1/Tf) - Bln(To/Tf) The goodness of fit of the equation showed the best results. Futhermore, by using the parameters a formula of freezing ratio and the relative water activities, which showed deviation from the ideal solution, were derived.

  20. Freeze Drying of Fruits and Vegetables: A Laboratory Experiment.

    Science.gov (United States)

    Noble, Richard D.

    1979-01-01

    Describes a laboratory experiment for freeze-drying fruits and vegetables which aims to expose college students to the principles of drying and simultaneous heat and mass transfer. The experimental apparatus, procedure of the experiment, and data analysis are also included. (HM)

  1. Review. Freeze-casting: Fabrication of highly porous and hierarchical ceramic supports for energy applications

    Energy Technology Data Exchange (ETDEWEB)

    Gaudillere, C.; Serra, J. M.

    2016-05-01

    The manufacture of structured ceramic porous support knows an important boom since more than a decade with the development of new shaping techniques. Among the most promising ones, the freeze-casting also called Ice-Tem plating allows the fabrication of ceramic parts exhibiting high porosity (>50%) and vertically aligned and hierarchically organized pores. Such structures were firstly conceived for biomedical applications like bone substitute and tissue engineering, but the distinctive features of freeze-cast structures have attracted the attention of diverse scientific fields, especially in high temperature ceramic-based energy production systems. Indeed, technologies like (a) Solid Oxide Fuel Cell (SOFC) and Electrolyser Cell (SOEC), (b) gas separation (O{sub 2}, H{sub 2}) by asymmetric supported membranes based on mixed ionic and electronic conductors (MIEC) or hydrogen-permeable metals, and (c) Catalytic Membrane Reactor (CMR) systems present a porous component in their physical structure. This latest, presenting a tortuous pathway for gas access and as a consequence, a high transport limitation, is known to be a limiting component for the operation at high flow streams that would enable to reach industrial target. (Author)

  2. Fast high-pressure freezing of protein crystals in their mother liquor

    International Nuclear Information System (INIS)

    Protein crystals were vitrified using high-pressure freezing in their mother liquor at 210 MPa and 77 K without cryoprotectants or oil coating. The method was successfully applied to photosystem II, which is representative of a membrane protein with a large unit cell and weak crystal contacts. High-pressure freezing (HPF) is a method which allows sample vitrification without cryoprotectants. In the present work, protein crystals were cooled to cryogenic temperatures at a pressure of 210 MPa. In contrast to other HPF methods published to date in the field of cryocrystallography, this protocol involves rapid sample cooling using a standard HPF device. The fast cooling rates allow HPF of protein crystals directly in their mother liquor without the need for cryoprotectants or external reagents. HPF was first attempted with hen egg-white lysozyme and cubic insulin crystals, yielding good to excellent diffraction quality. Non-cryoprotected crystals of the membrane protein photosystem II have been successfully cryocooled for the first time. This indicates that the presented HPF method is well suited to the vitrification of challenging systems with large unit cells and weak crystal contacts

  3. Freeze-Out Parameters in Heavy-Ion Collisions at AGS, SPS, RHIC, and LHC Energies

    International Nuclear Information System (INIS)

    We review the chemical and kinetic freeze-out conditions in high energy heavy-ion collisions for AGS, SPS, RHIC, and LHC energies. Chemical freeze-out parameters are obtained using produced particle yields in central collisions while the corresponding kinetic freeze-out parameters are obtained using transverse momentum distributions of produced particles. For chemical freeze-out, different freeze-out scenarios are discussed such as single and double/flavor dependent freeze-out surfaces. Kinetic freeze-out parameters are obtained by doing hydrodynamic inspired blast wave fit to the transverse momentum distributions. The beam energy and centrality dependence of transverse energy per charged particle multiplicity are studied to address the constant energy per particle freeze-out criteria in heavy-ion collisions

  4. Freezing point of milk in a herd of high yielding dairy cows

    Directory of Open Access Journals (Sweden)

    Slavica Golc Teger

    2005-04-01

    Full Text Available Factors affecting the freezing point of milk in a herd of 200 Black and White cows with the average milk yield of 8 386 kg in the lactation and 8 328 kg in the standard lactation were examinated. Over the period of one year (2002 and based upon 1 773 individual monthly collected milk samples with the average contents of 3.91% fat, 3.26% protein, 4.54% lactose, 33.4 mg/100 ml urea and 331000 somatic cells per ml in milk were determined. The average freezing point of milk (n = 1 680 was estimated to be –0.527 ºC, with a range from -0.562 ºC to -0.423 ºC. In 210 (12.5% samples was higher than -0.515 ºC. The lowest freezing point (-0.532 ºC was found in the samples collected in the first month after calving and highest (-0.522 ºC in the samples of 12th month of lactation. The differences between the freezing point of milk after the first and the second calving (-0.530 ºC; P < 0.05 and those after the fifth calving (-0.523 ºC; P < 0.05 were also significant. The samples collected in month from January to April (-0.538 ºC to -0.532 ºC were significantly lower in comparison to samples collected in May and June (-0.517 ºC and -0.519 ºC. The following statistically significant correlation coefficients between cows' properties, milk composition and the freezing point of milk were found: month of lactation r = 0.233 (P < 0.001; lactation number r = 0.196 (P < 0.001; age of cows (years r = 0.231 (P < 0.001; month of the year r = 0.0253 (P < 0.001; milk yield per milking day r = -0.106 (P < 0.001; fat corrected milk content (FCM per milking day r = -0.234 (P < 0.001; lactose % r = -0.530 (P < 0.001; fat % r = -0.351 (P < 0.001; protein % r = 0.058 (P < 0.05; urea mg/100 mL r = 0.091 (P < 0.001 and somatic cell count r = 0.154 (P < 0.001. The sum of effects (month of the year, lactation lenght and fat, protein and lactose content of milk was found to account for about 70% variability of the total depression of milk freezing point (R2 = 0.698.

  5. Freezing-Thawing Characteristics of Botanical Tissues and Influence of Water Morphology

    Institute of Scientific and Technical Information of China (English)

    WU Hai-Ling; MA Yuan; PENG Xiao-Feng

    2004-01-01

    @@ A series of visualization experiments were conducted to investigate the transport phenomena and interface behaviour during the freezing-thawing process of typical botanical tissues. Attention was paid to the growth of ice crystals and the advance of the phase-change interface. A comparison was made to identify the freezing/thawing behaviour for different tissues under various freezing conditions. Based on the experimental observation, analyses were conducted to explore the influence of water morphology on the freezing/thawing characteristics.

  6. Effects of freezing on soil temperature, frost propagation and moisture redistribution in peat: laboratory investigations

    OpenAIRE

    R. M. Nagare; R. A. Schincariol; W. L. Quinton; Hayashi, M

    2011-01-01

    The effects of freezing on soil temperature and water movement were monitored in four peat Mesocosms subjected to bidirectional freezing. Temperature gradients were applied by bringing the Mesocosm tops in contact with sub-zero air temperature while maintaining a continuously frozen layer at the bottom (proxy permafrost). Soil water movement towards the freezing front (from warmer to colder regions) was inferred from soil freezing curves and from the total water content of frozen core samples...

  7. Technology Advances and Mechanistic Modelling in Freeze-drying and Dehydration of Food

    OpenAIRE

    Wanren Chen; Xiang Gui; Hua Li

    2015-01-01

    Aim of study is to introduce some advanced freeze-drying technology and mechanistic modelling in freeze-drying and dehydration of food, freeze-drying is based on the dehydration by sublimation of a frozen product, due to very low temperature, all the deterioration activity and microbiological activity are stopped and provide better quality to the final product. Meanwhile the main problems of the freeze-dried food were proposed and its prospect and outlook was also analyzed, expecting to obtai...

  8. Effect of freezing and thawing rates on denaturation of proteins in aqueous solutions.

    Science.gov (United States)

    Cao, Enhong; Chen, Yahuei; Cui, Zhanfeng; Foster, Peter R

    2003-06-20

    The freeze denaturation of model proteins, LDH, ADH, and catalase, was investigated in absence of cryoprotectants using a microcryostage under well-controlled freezing and thawing rates. Most of the experimental data were obtained from a study using a dilute solution with an enzyme concentration of 0.025 g/l. The dependence of activity recovery of proteins on the freezing and thawing rates showed a reciprocal and independent effect, that is, slow freezing (at a freezing rate about 1 degrees C/min) and fast thawing (at a thawing rate >10 degrees C/min) produced higher activity recovery, whereas fast freezing with slow thawing resulted in more severe damage to proteins. With minimizing the freezing concentration and pH change of buffer solution by using a potassium phosphate buffer, this phenomenon could be ascribed to surface-induced denaturation during freezing and thawing process. Upon the fast freezing (e.g., when the freezing rate >20 degrees C/min), small ice crystals and a relatively large surface area of ice-liquid interface are formed, which increases the exposure of protein molecules to the ice-liquid interface and hence increases the damage to the proteins. During thawing, additional damage to proteins is caused by recrystallization process. Recrystallization exerts additional interfacial tension or shear on the entrapped proteins and hence causes additional damage to the latter. When buffer solutes participated during freezing, the activity recovery of proteins after freezing and thawing decreased due to the change of buffer solution pH during freezing. However, the patterns of the dependence on freezing and thawing rates of activity recovery did not change except for that at extreme low freezing rates (solutions could be reduced by changing the buffer type and composition and by optimizing the freezing-thawing protocol.

  9. Cellulose and Their Characteristic Ice Nucleation Activity- Freezing on a Chip

    Science.gov (United States)

    Häusler, Thomas; Felgitsch, Laura; Grothe, Hinrich

    2016-04-01

    The influence of clouds on the Earth's climate system is well known (IPCC, 2013). Cloud microphysics determines for example cloud lifetime and precipitation properties. Clouds are cooling the climate system by reflecting incoming solar radiation and warm its surface by trapping outgoing infrared radiation (Baker and Peter, 2008). In all these processes, aerosol particles play a crucial role by acting as cloud condensation nuclei (CCN) for liquid droplets and as an ice nucleation particle (INP) for the formation of ice particles. Freezing processes at higher temperatures than -38°C occur heterogeneously (Pruppacher and Klett 1997). Therefore aerosol particles act like a catalyst, which reduces the energy barrier for nucleation. The nucleation mechanisms, especially the theory of functional sites are not entirely understood. It remains unclear which class of compound nucleates ice. Here we present a unique technique to perform drop- freezing experiments in a more efficient way. A self-made freezing- chip will be presented. Measurements done to proof the efficiency of our setup as well as advantages compared with other setups will be discussed. Furthermore we present a proxy for biological INPs, microcrystalline cellulose. Cellulose is the main component of herbal cell walls (about 50 wt%). It is a polysaccharide consisting of a linear chain of several hundred to many thousands of β(1→4) linked D-glucose units. Cellulose can contribute to the diverse spectrum of ice nucleation particles. We present results of the nucleation activity measurements of MCCs as well as the influence of concentration, preparation or chemical modification.

  10. Impact of the industrial freezing process on selected vegetables -Part II. Colour and bioactive compounds

    NARCIS (Netherlands)

    Mazzeo, Teresa; Paciulli, Maria; Chiavaro, Emma; Visconti, Attilio; Fogliano, Vincenzo; Ganino, Tommaso; Pellegrini, Nicoletta

    2015-01-01

    In the present study, the impact of the different steps (i.e. blanching, freezing, storage following the industrial freezing process and the final cooking prior to consumption) of the industrial freezing process was evaluated on colour, chlorophylls, lutein, polyphenols and ascorbic acid content

  11. The impact of freeze-drying on microstructure and rehydration properties of carrot

    NARCIS (Netherlands)

    Voda, A.; Homan, N.; Witek, M.; Duijster, A.; Dalen, van G.; Sman, van der R.G.M.; Nijsse, J.; Vliet, van L.J.; As, van H.; Duynhoven, van J.P.M.

    2012-01-01

    The impact of freeze-drying, blanching and freezing rate pre-treatments on the microstructure and on the rehydration properties of winter carrots were studied by µCT, SEM, MRI and NMR techniques. The freezing rate determines the size of ice crystals being formed that leave pores upon drying. Their a

  12. Effects of freezing on soil temperature, frost propagation and moisture redistribution in peat: laboratory investigations

    Directory of Open Access Journals (Sweden)

    R. M. Nagare

    2011-05-01

    Full Text Available The effects of freezing on soil temperature and water movement were monitored in four peat Mesocosms subjected to bidirectional freezing. Temperature gradients were applied by bringing the Mesocosm tops in contact with sub-zero air temperature while maintaining a continuously frozen layer at the bottom (proxy permafrost. Soil water movement towards the freezing front (from warmer to colder regions was inferred from soil freezing curves and from the total water content of frozen core samples collected at the end of freezing cycle. This study illustrates how differences in initial water content influence the hydrologic functions of active layer in permafrost terrains covered with thick peat during soil freezing. A substantial amount of water, enough to raise the upper surface of frozen saturated soil within 15 cm of the soil surface at the end of freezing period, appeared to have moved upwards during freezing. Effects of temperature on soil matric potential, at least in the initial freezing period, appear to drive such movement as seen from analysis of soil freezing curves. Vapour movement from warmer to colder regions also appears to contribute in moisture movement. Frost propagation is controlled by latent heat for a long time during freezing. A simple conceptual model describing freezing of an organic active layer initially resembling a variable moisture landscape is proposed based upon the results of this study. The results of this study will help in understanding, and ultimately forecasting, the hydrologic response of wetland-dominated terrain underlain by discontinuous permafrost.

  13. 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…

  14. Vapor Pressure Plus: An Experiment for Studying Phase Equilibria in Water, with Observation of Supercooling, Spontaneous Freezing, and the Triple Point

    Science.gov (United States)

    Tellinghuisen, Joel

    2010-01-01

    Liquid-vapor, solid-vapor, and solid-liquid-vapor equilibria are studied for the pure substance water, using modern equipment that includes specially fabricated glass cells. Samples are evaporatively frozen initially, during which they typically supercool to -5 to -10 [degrees]C before spontaneously freezing. Vacuum pumping lowers the temperature…

  15. A novel collagen film with micro-rough surface structure for corneal epithelial repair fabricated by freeze drying technique

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Yang [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China); Guangdong Province Key Laboratory of Biomedical Engineering, South China University of Technology, Guangzhou 510006 (China); Ren, Li, E-mail: psliren@scut.edu.cn [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China); Guangdong Province Key Laboratory of Biomedical Engineering, South China University of Technology, Guangzhou 510006 (China); Wang, Yingjun, E-mail: imwangyj@163.com [School of Materials Science and Engineering, South China University of Technology, Guangzhou 510641 (China); National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou 510006 (China); Guangdong Province Key Laboratory of Biomedical Engineering, South China University of Technology, Guangzhou 510006 (China)

    2014-05-01

    Highlights: • Collagen film with micro-rough surface is fabricated by freeze drying technique. • The film has suitable water uptake capability and toughness performance. • The film has good optical performance. • Human corneal epithelial cells studies confirmed the biocompatibility of the film. - Abstract: Corneal epithelial defect is a common disease and keratoplasty is a common treatment method. A collagen film with micro-rough surface was fabricated through a simple freeze drying technique in this study. Compared with the air-dried collagen film (AD-Col), this freeze-dried collagen film (FD-Col) has a more suitable water uptake capability (about 85.5%) and toughness performance. Both of the two films have good optical properties and the luminousness of them is higher than 80%. Besides, the adhesion and proliferation rate of human corneal epithelial cells on the micro-rough surface of FD-Col film is higher than that on the smooth surface of AD-Col film. The results indicate that this FD-Col film may have potential applications for corneal epithelial repair.

  16. Effects of pressure-shift freezing and pressure-assisted thawing on sea bass (Dicentrarchus labrax) quality.

    Science.gov (United States)

    Tironi, V; LeBail, A; de Lamballerie, M

    2007-09-01

    Effects of pressure-shift freezing and/or pressure-assisted thawing on the quality of sea bass muscle were evaluated and compared with conventional (air-blast) frozen and thawed samples. Microstructural analysis showed a marked decrease of muscle cell damage for pressure-assisted frozen samples. According to differential scanning calorimetry (DSC), protein extractability, and SDS-PAGE results, high-pressure treatment (200 MPa) produced a partial denaturation with aggregation and insolubilization of the myosin, as well as alterations of the sarcoplasmic proteins. Only small differences between high-pressure processes (freezing or/and thawing) were registered. High-pressure-treated systems led to a decrease of water holding capacity but differences between high-pressure and conventional methods disappeared after cooking. Muscle color showed important alterations due to high-pressure treatments (increasing L* and b*).

  17. Distribution of coniferin in freeze-fixed stem of Ginkgo biloba L. by cryo-TOF-SIMS/SEM

    Science.gov (United States)

    Aoki, Dan; Hanaya, Yuto; Akita, Takuya; Matsushita, Yasuyuki; Yoshida, Masato; Kuroda, Katsushi; Yagami, Sachie; Takama, Ruka; Fukushima, Kazuhiko

    2016-08-01

    To clarify the role of coniferin in planta, semi-quantitative cellular distribution of coniferin in quick-frozen Ginkgo biloba L. (ginkgo) was visualized by cryo time-of-flight secondary ion mass spectrometry and scanning electron microscopy (cryo-TOF-SIMS/SEM) analysis. The amount and rough distribution of coniferin were confirmed through quantitative chromatography measurement using serial tangential sections of the freeze-fixed ginkgo stem. The lignification stage of the sample was estimated using microscopic observations. Coniferin distribution visualized at the transverse and radial surfaces of freeze-fixed ginkgo stem suggested that coniferin is stored in the vacuoles, and showed good agreement with the assimilation timing of coniferin to lignin in differentiating xylem. Consequently, it is suggested that coniferin is stored in the tracheid cells of differentiating xylem and is a lignin precursor.

  18. Distribution of coniferin in freeze-fixed stem of Ginkgo biloba L. by cryo-TOF-SIMS/SEM

    Science.gov (United States)

    Aoki, Dan; Hanaya, Yuto; Akita, Takuya; Matsushita, Yasuyuki; Yoshida, Masato; Kuroda, Katsushi; Yagami, Sachie; Takama, Ruka; Fukushima, Kazuhiko

    2016-01-01

    To clarify the role of coniferin in planta, semi-quantitative cellular distribution of coniferin in quick-frozen Ginkgo biloba L. (ginkgo) was visualized by cryo time-of-flight secondary ion mass spectrometry and scanning electron microscopy (cryo-TOF-SIMS/SEM) analysis. The amount and rough distribution of coniferin were confirmed through quantitative chromatography measurement using serial tangential sections of the freeze-fixed ginkgo stem. The lignification stage of the sample was estimated using microscopic observations. Coniferin distribution visualized at the transverse and radial surfaces of freeze-fixed ginkgo stem suggested that coniferin is stored in the vacuoles, and showed good agreement with the assimilation timing of coniferin to lignin in differentiating xylem. Consequently, it is suggested that coniferin is stored in the tracheid cells of differentiating xylem and is a lignin precursor. PMID:27510918

  19. Expression of a bacterial ice nucleation gene in a yeast Saccharomyces cerevisiae and its possible application in food freezing processes.

    Science.gov (United States)

    Hwang, W Z; Coetzer, C; Tumer, N E; Lee, T C

    2001-10-01

    A 3.6 kb ice nucleation gene (ina) isolated from Erwinia herbicola was placed under control of the galactose-inducible promoter (GAL1) and introduced into Saccharomyces cerevisiae. Yeast transformants showed increased ice nucleation activity over untransformed controls. The freezing temperature of a small volume of water droplets containing yeast cells was increased from approximately -13 degrees C in the untransformed controls to -6 degrees C in ina-expressing (Ina(+)) transformants. Lower temperature growth of Ina(+) yeast at temperatures below 25 degrees C was required for the expression of ice nucleation activity. Shift of temperature to 5-20 degrees C could induce the ice nucleation activity of Ina(+) yeast when grown at 25 degrees C, and maximum ice nucleation activity was achieved after induction at 5 degrees C for approximately 12 h. The effects of Ina(+) yeast on freezing and texturization of several food materials was also demonstrated. PMID:11600004

  20. Summer Freezing Resistance: A Critical Filter for Plant Community Assemblies in Mediterranean High Mountains.

    Science.gov (United States)

    Pescador, David S; Sierra-Almeida, Ángela; Torres, Pablo J; Escudero, Adrián

    2016-01-01

    Assessing freezing community response and whether freezing resistance is related to other functional traits is essential for understanding alpine community assemblages, particularly in Mediterranean environments where plants are exposed to freezing temperatures and summer droughts. Thus, we characterized the leaf freezing resistance of 42 plant species in 38 plots at Sierra de Guadarrama (Spain) by measuring their ice nucleation temperature, freezing point (FP), and low-temperature damage (LT50), as well as determining their freezing resistance mechanisms (i.e., tolerance or avoidance). The community response to freezing was estimated for each plot as community weighted means (CWMs) and functional diversity (FD), and we assessed their relative importance with altitude. We established the relationships between freezing resistance, growth forms, and four key plant functional traits (i.e., plant height, specific leaf area, leaf dry matter content (LDMC), and seed mass). There was a wide range of freezing resistance responses and more than in other alpine habitats. At the community level, the CWMs of FP and LT50 responded negatively to altitude, whereas the FD of both traits increased with altitude. The proportion of freezing-tolerant species also increased with altitude. The ranges of FP and LT50 varied among growth forms, and only leaf dry matter content was negatively correlated with freezing-resistance traits. Summer freezing events represent important abiotic filters for assemblies of Mediterranean high mountain communities, as suggested by the CWMs. However, a concomitant summer drought constraint may also explain the high freezing resistance of species that thrive in these areas and the lower FD of freezing resistance traits at lower altitudes. Leaves with high dry matter contents may maintain turgor at lower water potential and enhance drought tolerance in parallel to freezing resistance. This adaptation to drought seems to be a general prerequisite for plants

  1. Isolation and characterization of a freeze-tolerant diploid derivative of an industrial baker's yeast strain and its use in frozen doughs.

    Science.gov (United States)

    Teunissen, Aloys; Dumortier, Françoise; Gorwa, Marie-Françoise; Bauer, Jürgen; Tanghe, An; Loïez, Annie; Smet, Peter; Van Dijck, Patrick; Thevelein, Johan M

    2002-10-01

    The routine production and storage of frozen doughs are still problematic. Although commercial baker's yeast is highly resistant to environmental stress conditions, it rapidly loses stress resistance during dough preparation due to the initiation of fermentation. As a result, the yeast loses gassing power significantly during storage of frozen doughs. We obtained freeze-tolerant mutants of polyploid industrial strains following screening for survival in doughs prepared with UV-mutagenized yeast and subjected to 200 freeze-thaw cycles. Two strains in the S47 background with a normal growth rate and the best freeze tolerance under laboratory conditions were selected for production in a 20-liter pilot fermentor. Before frozen storage, the AT25 mutant produced on the 20-liter pilot scale had a 10% higher gassing power capacity than the S47 strain, while the opposite was observed for cells produced under laboratory conditions. AT25 also retained more freeze tolerance during the initiation of fermentation in liquid cultures and more gassing power during storage of frozen doughs. Other industrially important properties (yield, growth rate, nitrogen assimilation, and phosphorus content) were very similar. AT25 had only half of the DNA content of S47, and its cell size was much smaller. Several diploid segregants of S47 had freeze tolerances similar to that of AT25 but inferior performance for other properties, while an AT25-derived tetraploid, TAT25, showed only slightly improved freeze tolerance compared to S47. When AT25 was cultured in a 20,000-liter fermentor under industrial conditions, it retained its superior performance and thus appears to be promising for use in frozen dough production. Our results also show that a diploid strain can perform at least as well as a tetraploid strain for commercial baker's yeast production and usage. PMID:12324320

  2. Technology Advances and Mechanistic Modelling in Freeze-drying and Dehydration of Food

    Directory of Open Access Journals (Sweden)

    Wanren Chen

    2015-08-01

    Full Text Available Aim of study is to introduce some advanced freeze-drying technology and mechanistic modelling in freeze-drying and dehydration of food, freeze-drying is based on the dehydration by sublimation of a frozen product, due to very low temperature, all the deterioration activity and microbiological activity are stopped and provide better quality to the final product. Meanwhile the main problems of the freeze-dried food were proposed and its prospect and outlook was also analyzed, expecting to obtain technical and theoretical support for the production of freeze-drying food.

  3. Experimental investigations of freezing soils at ground conditions of Astana, Kazakhstan

    Institute of Scientific and Technical Information of China (English)

    Zhussupbekov Askar; Shakhmov Zhanbolat

    2015-01-01

    Kazakhstan regions is seasonal climatic with transient freezing of soil ground during the winter. Roadbed integrity is important to resist the sustained load transmitted by traffic on the road surface. Freezing of soil ground could significantly influence roadbed integrity in the seasonal freezing climate of Kazakhstan. The proper determination magnitude of frost heave and heaving pressure by the influence of freezing temperatures during the winter season are necessary for design and construction of highways. Thus, experimental tests were conducted on specimens obtained from Astana (Kazakhstan) to determine the freezing pressure and magnitude of frost heaving.

  4. Multiscale model of a freeze-thaw process for tree sap exudation.

    Science.gov (United States)

    Graf, Isabell; Ceseri, Maurizio; Stockie, John M

    2015-10-01

    Sap transport in trees has long fascinated scientists, and a vast literature exists on experimental and modelling studies of trees during the growing season when large negative stem pressures are generated by transpiration from leaves. Much less attention has been paid to winter months when trees are largely dormant but nonetheless continue to exhibit interesting flow behaviour. A prime example is sap exudation, which refers to the peculiar ability of sugar maple (Acer saccharum) and related species to generate positive stem pressure while in a leafless state. Experiments demonstrate that ambient temperatures must oscillate about the freezing point before significantly heightened stem pressures are observed, but the precise causes of exudation remain unresolved. The prevailing hypothesis attributes exudation to a physical process combining freeze-thaw and osmosis, which has some support from experimental studies but remains a subject of active debate. We address this knowledge gap by developing the first mathematical model for exudation, while also introducing several essential modifications to this hypothesis. We derive a multiscale model consisting of a nonlinear system of differential equations governing phase change and transport within wood cells, coupled to a suitably homogenized equation for temperature on the macroscale. Numerical simulations yield stem pressures that are consistent with experiments and provide convincing evidence that a purely physical mechanism is capable of capturing exudation. PMID:26400199

  5. Experimental Study on the Characteristics of CO2 Hydrate Formation in Porous Media below Freezing Point

    Institute of Scientific and Technical Information of China (English)

    Zhang Xuemin; Li Jinping; Wu Qingbai; Wang Chunlong; Nan Junhu

    2015-01-01

    Porous medium has an obvious effect on the formation of carbon dioxide hydrate. In order to study the character-istics of CO2 hydrate formation in porous medium below the freezing point, the experiment of CO2 hydrate formation was conducted in a high-pressure 1.8-L cell in the presence of porous media with a particle size of 380μm, 500μm and 700μm, respectively. The test results showed that the porous medium had an important inlfuence on the process of CO2 hydrate for-mation below the freezing point. Compared with porous media with a particle size of 500μm and 700μm, respectively, the average hydrate formation rate and gas storage capacity of carbon dioxide hydrate in the porous medium with a particle size of 380μm attained 0.016 14 mol/h and 65.094 L/L, respectively. The results also indicated that, within a certain range of particle sizes, the smaller the particle size of porous medium was, the larger the average hydrate formation rate and the gas storage capacity of CO2 hydrate during the process of hydrate formation would be.

  6. Freeze-drying of HI-6-loaded recombinant human serum albumin nanoparticles for improved storage stability.

    Science.gov (United States)

    Dadparvar, Miriam; Wagner, Sylvia; Wien, Sascha; Worek, Franz; von Briesen, Hagen; Kreuter, Jörg

    2014-10-01

    Severe intoxications with organophosphates require the immediate administration of atropine in combination with acetyl cholinesterase (AChE) reactivators such as HI-6. Although this therapy regimen enables the treatment of peripheral symptoms, the blood-brain barrier (BBB) restricts the access of the hydrophilic antidotes to the central nervous system which could lead to a fatal respiratory arrest. Therefore, HI-6-loaded albumin nanoparticles were previously developed to enhance the transport across this barrier and were able to reactivate organophosphate-(OP)-inhibited AChE in an in vitro BBB model. Since HI-6 is known to be moisture-sensitive, the feasibility of freeze-drying of the HI-6-loaded nanoparticles was investigated in the present study using different cryo- and lyoprotectants at different concentrations. Trehalose and sucrose (3%, w/v)-containing formulations were superior to mannitol concerning the physicochemical parameters of the nanoparticles whereas trehalose-containing samples were subject of a prolonged storage stability study at temperatures between -20°C and +40°C for predetermined time intervals. Shelf-life computations of the freeze-dried HI-6 nanoparticle formulations revealed a shelf-life time of 18 months when stored at -20°C. The formulations' efficacy was proven in vitro by reactivation of OP-inhibited AChE after transport over a porcine brain capillary endothelial cell layer model.

  7. Effects of Low Temperature on Freezing Injury of Various Winter Wheat Cultivars at Different Sowing Time

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    Freezing injury is one of the major disasters for the production of winter wheat in the North China Plain, which leads to a significant decrease of wheat yield. This study was conducted to investigate the impact of subfreezing temperature on freezing injury of various winter wheat cultivars at different sowing time. Three wheat cultivars, including Zhengmai 9023, Wanmai 48 and Wanmai 50, were sowed on 25 September and 5 October, respectively. Plant anatomy was applied to investi- gate the impact of subfreezing temperature on cells of three wheat cultivars, results showed that severe plasmolysis occurred in wheat sowed earlier suddenly encoun- tered subfreezing temperature without cold acclimation, compared with wheat sowed at proper sowing time. The degree of plasmolysis of different cultivars and tissues of wheat had significant differences and showed positive correlation with subfreezing temperature. Wanmai 50 had the highest cold resistance compared with Zhengmai 9023 and Wanmai 48, and there was no significant difference between Zhengmai 9023 and Wanmai 48. This study concluded that wheat cold resistance may be im- proved by adopting proper cultivars and sowing dates.

  8. Ribosome binding sites visualized on freeze-fractured membranes of the rough endoplasmic reticulum.

    Science.gov (United States)

    Giddings, T H; Staehelin, L A

    1980-04-01

    Freeze-fracture micrographs of cells of the green alga Micrasterias denticulata stabilized by ultrarapid freezing reveal imprints of polysomes on the rough endoplasmic reticulum membranes. The imprints appear as broad, spiral ridges on the P faces and as corresponding wide grooves on the E faces of the membranes. Distinct 110-A particles with a spacing of 270 +/- 45 A are associated with the P-face ridges. Where imprints of individual ribosomes can be discerned, it is seen that there is a 1:1 relationship between the ribosomes and the 110-A particles, and that the 110-A particles are located in a peripheral position with respect to the polysome spirals. We propose that the 110-A particles could be structural equivalents of ribosome-binding sites, consisting of a molecule each of ribophorins I and II and a nascent polypeptide chain. These observations suggest that the spiral form of polysomes could result from the forces generated by the extrusion of the growing polypeptide chains to one side of the polysome. PMID:7364870

  9. Calculation of Effective Freezing Time in Lung Cancer Cryosurgery Based on Godunov Simulation

    Directory of Open Access Journals (Sweden)

    Т.G. Kotova

    2016-03-01

    Full Text Available There have been presented the results of lung cancer cryosurgery simulation using numerical solutions of enthalpy equation according to Godunov method. For the cryodestruction improvement purposes we successfully calculated the effective freezing time taking into account the evolution of an ice ball covering the tumor area. Geometrical transformation parameters of an ice ball have been measured by calculating the temperature distribution and the interface position in biological tissue. Mathematical cryosurgical procedures are described by heat transfer equations in solid and liquid phases. Numerical results for one-dimensional case were verified by comparing with exact solutions. In twodimensional modeling an effective cryotherapy time, which corresponds to freezing time of all tumor parts, was calculated as the area of forming ice balls covering all tumor region. The findings enable to set the effective time of a cryosurgical procedure in lung cancer. The knowledge of temperature distribution and interface position in biological tissue offers an opportunity to a cryosurgeon to finish the procedure within a certain time period to minimize the healthy tissue damage and destroy tumor cells to the maximum. Simulation application enables to schedule cryotherapy in lung cancer more effectively and to a good quality.

  10. Chemical Freeze-Out and Higher Order Multiplicity Moments

    CERN Document Server

    Tawfik, A

    2013-01-01

    We calculate the non-normalized moments of the particle multiplicity within the framework of the hadron resonance gas (HRG) model. At finite chemical potential $\\mu$, a non-monotonic behavior is observed in the thermal evolution of third order moment (skewness $S$) and the higher order ones as well. Among others, this observation likely reflects dynamical fluctuations and strong correlations. The signatures of non-monotonicity in the normalized fourth order moment (kurtosis $\\kappa$) and its products get very clear. Based on these findings, we introduce a novel condition characterizing the universal freeze-out curve. The chemical freeze-out parameters $T$ and $\\mu$ are described by vanishing $\\kappa\\, \\sigma^2$ or equivalently $m_4=3\\,\\chi^2$, where $\\sigma$, $\\chi$ and $m_4$ are the standard deviation, susceptibility and fourth order moment, respectively. The fact that the HRG model is not able to release information about criticality related to the confinement and chiral dynamics should not veil the observa...

  11. Characterization of freeze dried egg melange long stored after irradiation

    Science.gov (United States)

    Bakalivanov, Stefan; Tsvetkova, Eli; Bakalivanova, Todorka; Tsvetkov, Tsvetan; Kaloyanov, Nikolay; Grigorova, Stoyanka; Alexieva, Vanja

    2008-01-01

    During the 4-year period of storage at room temperature of the freeze-dried (control group) and the freeze-dried and gamma-irradiated (2.0 and 3.5 kGy) whole hen's egg mélange, no significant changes were found into the sensory and functional characteristics till the 28th month. The change in the number of SH groups was not unidirectional up to the 28th month and then it started to decrease in all investigated samples. During the entire period of investigation the amount of malondialdehyde in all three groups of egg mélange was considerably below the allowed limit for foodstuffs. The most significant fractions of the protein spectra showed a general tendency of decrease during the storage.

  12. Encapsulation of black carrot juice using spray and freeze drying.

    Science.gov (United States)

    Murali, S; Kar, Abhijit; Mohapatra, Debabandya; Kalia, Pritam

    2015-12-01

    Black carrot juice extracted using pectinase enzyme was encapsulated in three different carrier materials (maltodextrin 20DE, gum arabic and tapioca starch) using spray drying at four inlet temperatures (150, 175, 200 and 225 ℃) and freeze drying at a constant temperature of - 53 ℃ and vacuum of 0.22-0.11 mbar with the constant feed mixture. The products were analyzed for total anthocyanin content, antioxidant activity, water solubility index, encapsulation efficiency and total colour change. For both the drying methods followed in this study, maltodextrin 20DE as the carrier material has proven to be better in retaining maximum anthocyanin and antioxidant activity compared to gum arabic and tapioca starch. The best spray dried product, was obtained at 150 ℃. The most acceptable was the freeze dried product with maximum anthocyanin content, antioxidant activity, water solubility index, encapsulation efficiency and colour change. PMID:25367889

  13. Study on Semen Freezing Preservation of German Shepherd Dogs

    Institute of Scientific and Technical Information of China (English)

    Huang He; Tian Ya-guang; Zheng Peng; Zhang Gui-xue

    2012-01-01

    In order to prolong semen preservation and improve pregnancy rate, semen freezing was studied with German shepherd dogs for experimental animals. The semen of four dogs was collected 40 times in four dilution frozen into two formulations, according to the sperm motility to compare the advantages and disadvantages. The result indicated that the sperm motility of the pellet frozen semen in dilute 2 was significantly higher than that in dilution 1, 3, and 4 (P0.01). The sperm motility of dogs semen with fried smoked method was notablely higher than that of frozen semen of program method (P0.01). The dilution which contained yolk-Tris mainly was the best; the pellet semen frozen with fried smoked method was superior to tuble semen frozen with program freezing method; sperm motility of pellet semen was higher than that of tuble semen in the same dilution. The conception rate and litter size of the natural matting were higher than AI.

  14. INFLUENCE OF CYCLIC FREEZING AND THAWING UPON SPRUCE WOOD PROPERTIES

    Directory of Open Access Journals (Sweden)

    Maria Bernadett SZMUTKU

    2012-03-01

    Full Text Available The paper presents the results concerning the change of some physical and mechanical properties of spruce wood (Picea abies L., after repeated freezing and thawing, respectively after being subjected to temperature variation from positive valuea (+100C to negative values (-250C. Through this approach, the authors aimed at simulating normal temperature variations that occur in winter between night and daytime and which fresh cut timber has to stand if being stored in an open yard for one week before being kilndried. By comparing these results with the ones obtained after continuous freezing at -250C for one week, it was established that the temperature variation and the repeated changing phase of water inside wood increase dimensional instability and reduce the mechanical strengths much more than the simple exposure to theconstant negative temperature.

  15. Preventing freezing of condensate inside tubes of air cooled condenser

    Energy Technology Data Exchange (ETDEWEB)

    Joo, Jeong A; Hwang, In Hwan; Lee, Dong Hwan [Chonbuk Nat' l Univ., Jeonju (Korea, Republic of); Cho, Young Il [Drexel Univ., Philadelphia (United States)

    2012-08-15

    An air cooled condenser is a device that is used for converting steam into condensate by using ambient air. The air cooled condenser is prone to suffer from a serious explosion when the condensate inside the tubes of a heat exchanger is frozen; in particular, tubes can break during winter. This is primarily due to the structural problem of the tube outlet of an existing conventional air cooled condenser system, which causes the backflow of residual steam and noncondensable gases. To solve the backflow problem in such condensers, such a system was simulated and a new system was designed and evaluated in this study. The experimental results using the simulated condenser showed the occurrence of freezing because of the backflow inside the tube. On the other hand, no backflow and freezing occurred in the advanced new condenser, and efficient heat exchange occurred.

  16. Variable Conductance Heat Pipe Performance after Extended Periods of Freezing

    Science.gov (United States)

    Ellis, Michael C.; Anderson, William G.

    2009-03-01

    Radiators operating in lunar or Martian environments must be designed to reject the maximum heat load at the maximum sink temperature, while maintaining acceptable temperatures at lower powers or sink temperatures. Variable Conductance Heat Pipe (VCHP) radiators can passively adjust to these changing conditions. Due to the presence of non-condensable gas (NCG) within each VCHP, the active condensing section adjusts with changes in either thermal load or sink temperature. In a Constant Conductance Heat Pipe (CCHP) without NCG, it is possible for all of the water to freeze in the condenser, by either sublimation or vaporization. With a dry evaporator, startup is difficult or impossible. Several previous studies have shown that adding NCG suppresses evaporator dryout when the condenser is frozen. These tests have been for relatively short durations, with relatively short condensers. This paper describes freeze/thaw experiments involving a VCHP with similar dimensions to the current reactor and cavity cooling radiator heat pipe designs.

  17. [Effect of freezing on cord blood serum proteins].

    Science.gov (United States)

    Nardid, E O; Rozanova, E D; Tsymbal, L V; Zinchenko, A V; Nardid, O A; Grishchenko, V I

    2009-01-01

    The effect of freezing regimes and storage temperatures on protein conformation and the spectrum of cord blood serum has been investigated. Changes in the parameters of ESR spectra of spin probes in cord blood serum after slow freezing and subsequent thawing were established, indicating protein conformational changes characterized by loosening. This fact is confirmed by an earlier process, the first stage of albumin heat denaturation, as indicated by calorimetric data. It was shown that slow cooling results in the aggregation of serum protein in which serum albumin and immunoglobulins play an important role. It was concluded that, for retaining the properties, of cord blood serum proteins, it is preferable to perform cooling at a rate not lower than 100 degrees C/min and a storage temperature of -80 degrees C and lower. PMID:19894629

  18. Heterogeneous ice nucleation: bridging stochastic and singular freezing behavior

    Directory of Open Access Journals (Sweden)

    D. Niedermeier

    2011-01-01

    Full Text Available Heterogeneous ice nucleation, a primary pathway for ice formation in the atmosphere, has been described alternately as being stochastic, in direct analogy with homogeneous nucleation, or singular, with ice nuclei initiating freezing at deterministic temperatures. We present an idealized model that bridges these stochastic and singular descriptions of heterogeneous ice nucleation. This "soccer ball" model treats statistically similar particles as being covered with surface sites (patches of finite area characterized by different nucleation barriers, but with each surface site following the stochastic nature of ice embryo formation. The model provides a phenomenological explanation for seemingly contradictory experimental results obtained in our research groups. We suggest that ice nucleation is fundamentally a stochastic process but that for realistic atmospheric particle populations this process can be masked by the heterogeneity of surface properties. Full evaluation of the model will require experiments with well characterized ice nucleating particles and the ability to vary both temperature and waiting time for freezing.

  19. CFD modelling of condensers for freeze-drying processes

    Indian Academy of Sciences (India)

    Miriam Petitti; Antonello A Barresi; Daniele L Marchisio

    2013-12-01

    The aim of the present research is the development of a computational tool for investigating condensation processes and equipment with particular attention to freeze-dryers. These condensers in fact are usually operated at very low pressures, making it difficult to experimentally acquire quantitative knowledge of all the variables involved. Mathematical modelling and CFD (Computational Fluid Dynamics) simulations are used here to achieve a better comprehension of the flow dynamics and of the process of ice condensation and deposition in the condenser, in order to evaluate condenser efficiency and gain deeper insights of the process to be used for the improvement of its design. Both a complete laboratory-scale freeze-drying apparatus and an industrial-scale condenser have been investigated in this work, modelling the process of water vapour deposition. Different operating conditions have been considered and the influence exerted by the inert gas as well as other parameters has been investigated.

  20. Optimization Of Freeze-Dried Starter For Yogurt By Full Factorial Experimental Design

    Directory of Open Access Journals (Sweden)

    Chen He

    2015-12-01

    Full Text Available With the rapidly development of fermented milk product, it is significant for enhancing the performance of starter culture. This paper not only investigated the influence of anti-freeze factors and freeze-drying protective agents on viable count, freeze-drying survival rate and yield of Lactobacillus bulgaricus (LB and Streptococcus thermophilus (ST, but also optimized the bacteria proportion of freeze-dried starter culture for yogurt by full factorial experimental design. The results showed as following: the freeze-drying protective agents or anti-freeze factors could enhanced survival rate of LB and ST; the freeze-dried LB and ST powders containing both of anti-freeze factors and freeze-drying protective agents had higher viable count and freeze-drying survival rate that were 84.7% and 79.7% respectively; In terms of fermentation performance, the best group of freeze-dried starter for yogurt was the compound of LB3 and ST2.

  1. Importance of freeze-thaw events in low temperature ecotoxicology of cold tolerant enchytraeids.

    Science.gov (United States)

    Silva, Ana L Patrício; Enggrob, Kirsten; Slotsbo, Stine; Amorim, Mónica J B; Holmstrup, Martin

    2014-08-19

    Due to global warming it is predicted that freeze-thaw cycles will increase in Arctic and cold temperate regions. The effects of this variation becomes of particular ecological importance to freeze-tolerant species when it is combined with chemical pollutants. We compared the effect of control temperature (2 °C), daily freeze-thaw cycles (2 to -4 °C) and constant freezing (-2 °C) temperatures on the cold-tolerance of oligochaete worms (Enchytraeus albidus) and tested how survival was influenced by pre-exposure to 4-nonylphenol (4-NP), a common nonionic detergent found in sewage sludge amended soils. Results showed that combined effect of 4-NP and daily freeze-thaw cycles can cause higher mortality to worms as compared with sustained freezing or control temperature. Exposure to 4-NP caused a substantial depletion of glycogen reserves which is catabolized during freezing to produce cryoprotective concentrations of free glucose. Further, exposure to freeze-thaw cycles resulted in higher concentrations of 4-NP in worm tissues as compared to constant freezing or control temperature (2 °C). Thus, worms exposed to combined effect of freeze-thaw cycles and 4-NP suffer higher consequences, with the toxic effect of the chemical potentiating the deleterious effects of freezing and thawing.

  2. Petroleum refinery secondary effluent polishing using freezing processes--toxicity and organic contaminant removal.

    Science.gov (United States)

    Gao, W; Smith, D W; Habib, M

    2008-06-01

    A petroleum refinery secondary effluent was treated using two freezing techniques--spray freezing and unidirectional downward freezing (UDF). The freezing processes were effective to remove toxicity and total organic carbon (TOC)- and chemical oxygen demand (COD)-causing materials in the effluent. Agitation of the liquid during UDF significantly improved the impurity separation efficiency; 85 to 96% removal of TOC and COD was achieved without any pretreatment and freezing only 70% of the feed water. The treatment efficiency of the spray freezing was at the same level as that of UDF without mixing. The spray ice with longer storage time released more contaminants with early meltwater. The initial contaminant concentration of the feed water and the freezing temperatures (-10 degrees C and -25 degrees C) had no significant influence on the treatment efficiency. A small fluctuation in effluent TOC concentration caused a dramatic change in effluent toxicity (Microtox). The effective concentration (EC20) (Microtox) was effective in detecting effluent toxicity. PMID:18686927

  3. Freeze-drying and quality evaluation of protein drugs.

    Science.gov (United States)

    Inazu, K; Shima, K

    1992-01-01

    Gamma interferon from genetic recombination (IFN) has been found to have an optimal pH at about 7. An increase in IFN concentration may cause a decrease in solution clarity. A proper selection of isotonizing agent, as well as the addition of sugars, is effective in improving the clarity. The amount of IFN adsorbed on filter membranes varies with the membrane materials: cellulose acetate adsorbs much IFN, 2-fluorovinylidene is the next, followed by polysulfon, and polycarbonate adsorbs it least of all materials tested. Stainless steel adsorbs little IFN, and the level can be lowered even more by electropolishing. Silicone coating can decrease the amount adsorbed to about 1 microgram per vial of 10 ml. The effect of pressure given to the IFN solution during filtration is negligible. Transfer of IFN solution through pipings of conventional shape may result in partial deactivation by bubbling. At around pH 7, a lower pH of IFN solution causes a higher moisture level of the freeze-dried product. Moisture levels up to 3% have no effect on IFN stability. Upon reconstitution of freeze-dried IFN by vigorous shaking with distilled water, filtration of the solution may become difficult because polymers might have been formed during vigorous shaking. The addition of L-cysteine, maltose, and human serum albumin, has been found to be as effective in preventing such unfavorable reactions. Fatty acids in human serum albumin, which is effective in stabilizing IFN, has been found to participate in preventing denaturation of human serum albumin upon freezing and freeze-drying; however, the denaturation prevention mechanisms are not clear yet. PMID:1592181

  4. Satellite freeze forecast system. Operating/troubleshooting manual

    Science.gov (United States)

    Martsolf, J. D. (Principal Investigator)

    1983-01-01

    Examples of operational procedures are given to assist users of the satellites freeze forecasting system (SFFS) in logging in on to the computer, executing the programs in the menu, logging off the computer, and setting up the automatic system. Directions are also given for displaying, acquiring, and listing satellite maps; for communicating via terminal and monitor displays; and for what to do when the SFFS doesn't work. Administrative procedures are included.

  5. Texture of onions before and after freeze-drying.

    OpenAIRE

    Guiné, Raquel; Barroca, Maria João

    2010-01-01

    Dehydrated onion is widely used as flavor additive in the manufacture of processed foods and many other convenience foods. However, when drying shelf-stable vegetables it is absolutely essential to preserve their desired quality attributes. In the present work fresh and freeze-dried onions were analysed in terms of Texture Profile Analysis (TPA) to evaluate in what extent this operation influences the texture attributes of onions. From the ana...

  6. Universal pion freeze-out phase-space density

    CERN Document Server

    Ferenc, D; Tomasik, Boris; Wiedemann, Urs Achim; Cramer, J G

    1999-01-01

    Results on the pion phase-space density at freeze-out in sulphur- nucleus. Pb-Pb and pi -p collisions at the CERN SPS are presented. All heavy-ion reactions are consistent with the thermal Bose-Einstein distribution f=[exp(E/T)-1]/sup -1/ at T~120 MeV, modified for radial expansion. pi -p data are also consistent with f, but at T~180 MeV and without radial flow. (18 refs).

  7. Freeze-Etch of Emulsified Cake Batters During Baking

    OpenAIRE

    Cloke, J. D.; Gordon, J; Davis, E.A.

    1982-01-01

    Cryofixation, freeze-etch techniques were used to study the structure of cake batters made from a lean cake formulation before heating and after heating to temperatures up to l00-l02°C. Batters were prepared without added emulsifiers and with saturated and unsaturated monoglycerides replacing 5 and l 0% of the oil. Unsaturated monoglyceri des were more effective than saturated monoglycerides in dispersing oil droplets through the batter. Saturated monoglycerides formed liquid crystals during ...

  8. Cold, no sweat: Eutectic freezing beats evaporation hands down

    OpenAIRE

    Hartmann, L

    2001-01-01

    Researchers at the Laboratory for Process Equipment at Delft University of Technology have developed an energy-friendly method to separate saline solutions into clean water and pure salt crystals. The method works by crystallizing the water and salt simultaneously though independently from each other. One of the many useful applications of what is known as eutectic freeze crystallisation is the extraction of valuable salts from waste water from the potato processing industry. The technique ca...

  9. Immersion freezing of ice nucleation active protein complexes

    OpenAIRE

    Hartmann, S.; Augustin, S.; T. Clauss; Wex, H.; T. Šantl-Temkiv; Voigtländer, J.; D. Niedermeier; Stratmann, F.

    2013-01-01

    Utilising the Leipzig Aerosol Cloud Interaction Simulator (LACIS), the immersion freezing behaviour of droplet ensembles containing monodisperse particles, generated from a Snomax™ solution/suspension, was investigated. Thereto ice fractions were measured in the temperature range between −5 °C to −38 °C. Snomax™ is an industrial product applied for artificial snow production and contains Pseudomonas syringae} bacteria which have long been used as model organism for atmospheric relevant ice nu...

  10. An elastoplastic model for the THM analysis of freezing soils

    OpenAIRE

    Satoshi,Nishimura; Gens Solé, Antonio; Olivella Pastallé, Sebastià; Jardine, Richard

    2009-01-01

    By employing a combination of ice pressure, liquid pressure and total stress as state variables, a new thermoplastic constitutive model has been developed that encompasses frozen and unfrozen behaviour within a unified effective-stress-based framework. It has been incorporated into a fully coupled thermo-hydro-mechanical (THM) finite element formulation that considers freezing and thawing in water-saturated soils and applied to a large-scale pipeline frost heave test. Peer Reviewed

  11. Comparison of thawing and freezing dark energy parametrizations

    Science.gov (United States)

    Pantazis, G.; Nesseris, S.; Perivolaropoulos, L.

    2016-05-01

    Dark energy equation of state w (z ) parametrizations with two parameters and given monotonicity are generically either convex or concave functions. This makes them suitable for fitting either freezing or thawing quintessence models but not both simultaneously. Fitting a data set based on a freezing model with an unsuitable (concave when increasing) w (z ) parametrization [like Chevallier-Polarski-Linder (CPL)] can lead to significant misleading features like crossing of the phantom divide line, incorrect w (z =0 ), incorrect slope, etc., that are not present in the underlying cosmological model. To demonstrate this fact we generate scattered cosmological data at both the level of w (z ) and the luminosity distance DL(z ) based on either thawing or freezing quintessence models and fit them using parametrizations of convex and of concave type. We then compare statistically significant features of the best fit w (z ) with actual features of the underlying model. We thus verify that the use of unsuitable parametrizations can lead to misleading conclusions. In order to avoid these problems it is important to either use both convex and concave parametrizations and select the one with the best χ2 or use principal component analysis thus splitting the redshift range into independent bins. In the latter case, however, significant information about the slope of w (z ) at high redshifts is lost. Finally, we propose a new family of parametrizations w (z )=w0+wa(z/1 +z )n which generalizes the CPL and interpolates between thawing and freezing parametrizations as the parameter n increases to values larger than 1.

  12. Promising freeze protection alternatives in solar domestic hot water systems

    Energy Technology Data Exchange (ETDEWEB)

    Bradley, D.E.

    1997-12-31

    Since the gains associated with solar thermal energy technologies are comparatively small in relation to the required capital investment, it is vital to maximize conversion efficiency. While providing the necessary function of freeze protection, the heat exchanger commonly included in solar domestic water heating systems represents a system inefficiency. This thesis explores two alternate methods of providing freeze protection without resorting to a heat exchanger. Commonly, collectors are made of rigid copper tubes separated by copper or aluminum fins. Cracking damage can occur when water is allowed to freeze and expand inside the non compliant tubes. The possibility of making collectors out of an elastic material was investigated and shown to be effective. Since unlike copper, elastomers typically have low thermal conductivities, the standard collector performance prediction equations do not apply. Modified thermal performance prediction equations were developed which can be used for both low and high thermal conductivity materials to provide accurate predictions within a limited range of plate geometries. An elastomeric collector plate was then designed and shown to have comparable performance to a copper plate collector whose aperture area is approximately 33% smaller. Another options for providing freeze protection to an SDHW system is to turn it off during the winter. Choosing a three-season operating period means two things. First, the system will have different optimums such as slope and collector area. Second, the wintertime solar energy incident on the collector is unavailable for meeting a heating load. However, the system`s heat exchanger becomes unnecessary and removing it increases the amount of energy that arrives at the storage tank during those periods in which the system is operating.

  13. Chemical freeze-out and higher order multiplicity moments

    Energy Technology Data Exchange (ETDEWEB)

    Tawfik, A., E-mail: a.tawfik@eng.mti.edu.eg

    2014-02-15

    We calculate the non-normalized moments of the particle multiplicity within the framework of the hadron resonance gas (HRG) model. At finite chemical potential μ, a non-monotonic behavior is observed in the thermal evolution of third order moment (skewness S) and the higher order ones as well. The signatures of non-monotonicity in the normalized fourth order moment (kurtosis κ) and its products get very clear. Among others, this observation likely reflects dynamical fluctuations and strong correlations. Based on these findings, we introduce the physics of the universal freeze-out curve. It is found that the chemical freeze-out parameters T and μ are described by vanishing κσ{sup 2} or equivalently m{sub 4}=3χ{sup 2}, where σ, χ and m{sub 4} are the standard deviation, susceptibility and fourth order moment, respectively. The fact that the HRG model is not able to release information about criticality related to the confinement and chiral dynamics should not veil the observations related to the chemical freeze-out. Recent lattice QCD studies strongly advocate the main conclusion of the present paper.

  14. Tuning towards dynamic freezing using a two-rate protocol

    Science.gov (United States)

    Kar, Satyaki; Mukherjee, Bhaskar; Sengupta, K.

    2016-08-01

    We study periodically driven closed quantum systems where two parameters of the system Hamiltonian are varied periodically in time with frequencies ω1 and ω2=r ω1 . We show that such drives may be used to tune towards dynamics-induced freezing where the wave function of the state of the system after a drive cycle at time T =2 π /ω1 has almost perfect overlap with the initial state. We locate regions in the (ω1,r ) plane where the freezing is near exact for a class of integrable models and a specific nonintegrable model. The integrable models that we study encompass Ising and XY models in d =1 , Kitaev model in d =2 , and Dirac fermions in graphene and atop a topological insulator surface, whereas the nonintegrable model studied involves the experimentally realized one-dimensional tilted Bose-Hubbard model in an optical lattice. In addition, we compute the relevant correlation functions of such driven systems and describe their characteristics in the region of the (ω1,r ) plane where the freezing is near exact. We supplement our numerical analysis with semianalytic results for integrable driven systems within adiabatic-impulse approximation and discuss experiments which may test our theory.

  15. Preparation and Characterization of Directionally Freeze-cast Copper Foams

    Directory of Open Access Journals (Sweden)

    Aurelia I. Cuba Ramos

    2012-08-01

    Full Text Available Because of their excellent thermal and electric conductivities, copper foams are ideally suited for applications such as heat exchangers, catalyst supports and EMI-shields. Here, we demonstrate the preparation of copper with ~80% aligned, elongated, interconnected pores via directional freeze casting, a well established processing technique for porous ceramics. First, an aqueous slurry of 40−80 nm cupric oxide powders was directionally solidified, resulting in a preform consisting of elongated, aligned dendrites of pure ice separated by interdendritic ice walls with high oxide powder content. Oxide rather than metallic nanometric particles are used, as the latter would oxidize rapidly and uncontrollably when suspended in the aqueous solution used during directional casting. The preforms were then freeze-dried to sublimate the ice and sintered in a hydrogen-bearing atmosphere to reduce the copper oxide to metallic copper particles and densify these copper particles. Microstructural analysis of the copper foams shows that three types of porosities are present: (i aligned, elongated pores replicating the ice dendrites created during the freeze-casting process; (ii micro-porosity in the partially sintered copper walls separating the elongated pores; and (iii cracks in these copper walls, probably created because of shrinkage associated with the reduction of the oxide powders.

  16. On the transition between heterogeneous and homogeneous freezing

    Directory of Open Access Journals (Sweden)

    K. Gierens

    2002-12-01

    Full Text Available Box model simulations of an uplifting and adiabatically cooling cloud of aerosol have been performed in order to study the transition between cirrus formation dominated by homogeneous nucleation of ice to that dominated by heterogeneous nucleation. The aerosol was assumed to consist of an internal mixture of sulfuric acid solution droplets with inclusions of soot. The simulations show that the transition from heterogeneous to homogeneous nucleation occurs over a narrow range of soot concentration. Thus it seems to be possible to fix critical concentrations of heterogeneous ice nuclei which must be exceeded if heterogeneous freezing dominates cirrus formation. A formula has been derived that allows to compute the critical concentrations of heterogeneous ice nuclei as a function of temperature, updraft speed, ambient pressure, and supersaturation at which heterogeneous freezing occurs. Generally, homogeneous nucleation dominates in regions with updrafts stronger than 20 cmps, with the exception of heavily polluted areas which could be common in the northern hemisphere due to air traffic, where updrafts of the order 1 mps may be necessary to render heterogeneous nucleation unimportant. According to the present results it cannot be excluded that heterogeneous nucleation plays a more important role for cirrus formation in the northern midlatitudes than anywhere else. A possible consequence of these results is that air pollution may lead to a higher coverage of cirrus clouds, but then these clouds will be optically thinner than clouds formed by homogeneous freezing, with the exception of regions where condensation trails are frequent.

  17. Antifreeze proteins enable plants to survive in freezing conditions

    Indian Academy of Sciences (India)

    Ravi Gupta; Renu Deswal

    2014-12-01

    Overwintering plants secrete antifreeze proteins (AFPs) to provide freezing tolerance. These proteins bind to and inhibit the growth of ice crystals that are formed in the apoplast during subzero temperatures. Antifreeze activity has been detected in more than 60 plants and AFPs have been purified from 15 of these, including gymnosperms, dicots and monocots. Biochemical characterization of plant antifreeze activity, as determined by the high ice recrystallization inhibition (IRI) activities and low thermal hysteresis (TH) of AFPs, showed that their main function is inhibition of ice crystal growth rather than the lowering of freezing temperatures. However, recent studies showed that antifreeze activity with higher TH also exists in plants. Calcium and hormones like ethylene and jasmonic acid have been shown to regulate plant antifreeze activity. Recent studies have shown that plant AFPs bind to both prism planes and basal planes of ice crystals by means of two flat ice binding sites. Plant AFPs have been postulated to evolve from the OsLRR-PSR gene nearly 36 million years ago. In this review, we present the current scenario of plant AFP research in order to understand the possible potential of plant AFPs in generation of freezing-tolerant crops.

  18. A Comparison of Thawing and Freezing Dark Energy Parametrizations

    CERN Document Server

    Pantazis, G; Perivolaropoulos, L

    2016-01-01

    Dark energy equation of state $w(z)$ parametrizations with two parameters and given monotonicity are generically either convex or concave functions. This makes them suitable for fitting either freezing or thawing quintessence models but not both simultaneously. Fitting a dataset based on a freezing model with an unsuitable (concave when increasing) $w(z)$ parametrization (like CPL) can lead to significant misleading features like crossing of the phantom divide line, incorrect $w(z=0)$, incorrect slope \\etc that are not present in the underlying cosmological model. To demonstrate this fact we generate scattered cosmological data both at the level of $w(z)$ and the luminosity distance $D_L(z)$ based on either thawing or freezing quintessence models and fit them using parametrizations of convex and of concave type. We then compare statistically significant features of the best fit $w(z)$ with actual features of the underlying model. We thus verify that the use of unsuitable parametrizations can lead to misleadin...

  19. Recent changes in the frequency of freezing precipitation in North America and Northern Eurasia

    Science.gov (United States)

    Groisman, Pavel Ya; Bulygina, Olga N.; Yin, Xungang; Vose, Russell S.; Gulev, Sergey K.; Hanssen-Bauer, Inger; Førland, Eirik

    2016-04-01

    Freezing rain and freezing drizzle events represent a critical feature of many regions of the world. Even at low intensities, these events often result in natural hazards that cause damage to housing, communication lines, and other man-made infrastructure. These events usually occur near the 0 °C isotherm. In a changing climate, this isotherm will not disappear, but its position in space and time will likely change as will the geography of freezing precipitation. A larger influx of water vapor into the continents from the oceans may also increase the amount and frequency of freezing precipitation events. This paper assesses our current understanding of recent changes in freezing precipitation for the United States, Canada, Norway, and Russia. The research is part of a larger GEWEX Cross-Cut Project addressing ‘cold/shoulder season precipitation near 0 °C’. Using an archive of 874 long-term time series (40 years of data) of synoptic observations for these four countries, we document the climatology of daily freezing rain and freezing drizzle occurrences as well as trends therein. The regions with the highest frequency of freezing rains (from 3 to 8 days per year) reside in the northeastern quadrant of the conterminous United States and adjacent areas of southeastern Canada south of 50 °N and over the south and southwest parts of the Great East European Plain. The frequency of freezing drizzle exceeds the frequency of freezing rain occurrence in all areas. During the past decade, the frequency of freezing rain events somewhat decreased over the southeastern US. In North America north of the Arctic Circle, it increased by about 1 day yr-1. Over Norway, freezing rain occurrences increased substantially, especially in the Norwegian Arctic. In European Russia and western Siberia, the frequency of freezing rain somewhat increased (except the southernmost steppe regions and the Arctic regions) while freezing drizzle frequency decreased over entire Russia.

  20. Monitoring Winter Wheat Freeze Injury Using Multi-Temporal MODIS Data

    Institute of Scientific and Technical Information of China (English)

    FENG Mei-chen; YANG Wu-de; CAO Liang-liang; DING Guang-wei

    2009-01-01

    Freeze injury is an usual disaster for winter wheat in Shanxi Province,China,and monitoring freeze injury is of important economic significance.The aim of this article is to monitor and analyze the winter wheat freeze injury using remote sensmg data,to monitor the occurrence and spatial distribution of winter wheat freeze in time.as well as the severity of the damage.The winter wheat freeze injury was monitored using multi-temporal moderate-resolution imaging spectroradiometer (MODIS) data,combined with ground meteorological data and field survey data,the change of normalized difference vegetation index (NDVI) before and after freeze injury was analyzed,as well as the effect of winter wheat growth recovery rate on yield.The results showed that the NDVI of winter wheat decreased dramatically after the suffering from freeze injury,which was the prominent feature for the winter wheat freeze injury monitoring.The degrees of winter wheat freeze injury were different in the three regions,of which,Yuncheng was the worst severity and the largest freeze injury area,the severity of freeze injury correlates with the breeding stage of the winter wheat.The yield of winter wheat showed positive correlation with its growth recovery rate (r=0.659**) which can be utilized to monitor the severity of winter wheat freeze injury as well as its impact on yield.It can effectively monitor the occurrence and severity of winter wheat freeze injury using horizontal and vertical profile distribution and growth recovery rate,and provide a basis for monitoring the winter wheat freeze injury in Shanxi Province.

  1. Freeze-In dark matter with displaced signatures at colliders

    Energy Technology Data Exchange (ETDEWEB)

    Co, Raymond T.; D’Eramo, Francesco; Hall, Lawrence J.; Pappadopulo, Duccio [Berkeley Center for Theoretical Physics, Department of Physics,and Theoretical Physics Group, Lawrence Berkeley National Laboratory,University of California,Berkeley, CA 94720 (United States)

    2015-12-11

    Dark matter, X, may be generated by new physics at the TeV scale during an early matter-dominated (MD) era that ends at temperature T{sub R}≪ TeV. Compared to the conventional radiation-dominated (RD) results, yields from both Freeze-Out and Freeze-In processes are greatly suppressed by dilution from entropy production, making Freeze-Out less plausible while allowing successful Freeze-In with a much larger coupling strength. Freeze-In is typically dominated by the decay of a particle B of the thermal bath, B→X. For a large fraction of the relevant cosmological parameter space, the decay rate required to produce the observed dark matter abundance leads to displaced signals at LHC and future colliders, for any m{sub X} in the range keV

  2. Studies of heterogeneous freezing by three different desert dust samples

    Directory of Open Access Journals (Sweden)

    P. J. Connolly

    2009-01-01

    Full Text Available We present results of experiments at the aerosol interactions and dynamics in the atmosphere (AIDA chamber facility looking at the freezing of water by three different types of mineral particles at temperatures between −12°C and −33°C. The three different dusts are Asia Dust-1 (AD1, Sahara Dust-2 (SD2 and Arizona test Dust (ATD. The dust samples used had particle concentrations of sizes that were log-normally distributed with mode diameters between 0.3 and 0.5 μm and standard deviations, σg, of 1.6–1.9. The results from the freezing experiments are consistent with the singular hypothesis of ice nucleation. The dusts showed different nucleation abilities, with ATD showing a rather sharp increase in ice-active surface site density at temperatures less than −24°C. AD1 was the next most efficient freezing nuclei and showed a more gradual increase in activity than the ATD sample. SD2 was the least active freezing nuclei.

    We used data taken with particle counting probes to derive the ice-active surface site density forming on the dust as a function of temperature for each of the three samples and polynomial curves are fitted to this data. The curve fits are then used independently within a bin microphysical model to simulate the ice formation rates from the experiments in order to test the validity of parameterising the data with smooth curves. Good agreement is found between the measurements and the model for AD1 and SD2; however, the curve for ATD does not yield results that agree well with the observations. The reason for this is that more experiments between −20 and −24°C are needed to quantify the rather sharp increase in ice-active surface site density on ATD in this temperature regime. The curves presented can be used as parameterisations in atmospheric cloud models where cooling rates of approximately 1°C min−1 or more are present to predict the concentration of ice crystals forming by

  3. Studies of heterogeneous freezing by three different desert dust samples

    Directory of Open Access Journals (Sweden)

    P. J. Connolly

    2009-04-01

    Full Text Available We present results of experiments at the aerosol interactions and dynamics in the atmosphere (AIDA chamber facility looking at the freezing of water by three different types of mineral particles at temperatures between −12°C and −33°C. The three different dusts are Asia Dust-1 (AD1, Sahara Dust-2 (SD2 and Arizona test Dust (ATD. The dust samples used had particle concentrations of sizes that were log-normally distributed with mode diameters between 0.3 and 0.5 μm and standard deviations, σg, of 1.6–1.9. The results from the freezing experiments are consistent with the singular hypothesis of ice nucleation. The dusts showed different nucleation abilities, with ATD showing a rather sharp increase in ice-active surface site density at temperatures less than −24°C. AD1 was the next most efficient freezing nuclei and showed a more gradual increase in activity than the ATD sample. SD2 was the least active freezing nuclei.

    We used data taken with particle counting probes to derive the ice-active surface site density forming on the dust as a function of temperature for each of the three samples and polynomial curves are fitted to this data. The curve fits are then used independently within a bin microphysical model to simulate the ice formation rates from the experiments in order to test the validity of parameterising the data with smooth curves. Good agreement is found between the measurements and the model for AD1 and SD2; however, the curve for ATD does not yield results that agree well with the observations. The reason for this is that more experiments between −20 and −24°C are needed to quantify the rather sharp increase in ice-active surface site density on ATD in this temperature regime. The curves presented can be used as parameterisations in atmospheric cloud models where cooling rates of approximately 1°C min−1 or more are present to predict the concentration of ice crystals forming by the

  4. 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

    During its lifetime, a KBS-3 repository will be subject to various ambient temperatures. Backfilled tunnels, shafts and investigation bore holes closest to ground level will experience periods of temperature below 0 deg C. From a safety assessment perspective, it is therefore essential to understand the behavior of compacted bentonite below 0 deg C. A theoretical framework for predicting the pressure response in compacted water saturated bentonite due to temperature changes has been developed based on thermodynamics and a single pore-type. This model predicts an approximately linear temperature dependence of swelling pressure P{sub s}(w,DELTAT) = P{sub s}(w,0 deg C) + DELTAs(w)DELTAT/nu{sub clay}(w) where DELTAT denotes a temperature difference from 0 deg C, DELTAs(w) is the difference in partial molar entropy between clay water and bulk water, nu{sub clay} (w) is the partial molar volume of the clay water and w denotes the water/solid mass ratio of the clay. As bulk water changes phase at 0 deg C, DELTAs(w) has a different value dependent on whether DELTAT is negative or positive. Above 0 deg C DELTAs(w) is a small value for all relevant densities which means that the pressure response due to temperature changes is small. A further consequence of this fact is that DELTAs(w) is a large positive number below 0 deg C when the external water phase is transformed to ice. Consequently, the model predicts a large drop of swelling pressure with temperature below 0 deg C, in the order of 1.2 MPa/deg C. Specifically, the swelling pressure is zero at a certain (negative) temperature T{sub C}. T{sub C} also quantifies the freezing point of the bentonite sample under consideration, as ice formation in the bentonite does not occur until swelling pressure is lost. A large set of laboratory tests have been performed where fully water saturated samples of bentonites have been exposed to temperatures in the range -10 deg C to +25 deg C. The swelling pressure response has been

  5. Flash-and-freeze: coordinating optogenetic stimulation with rapid freezing to visualize membrane dynamics at synapses with millisecond resolution

    Directory of Open Access Journals (Sweden)

    Shigeki Watanabe

    2016-08-01

    Full Text Available Electron microscopy depicts subcellular structures at synapses exquisitely but only captures static images. To visualize membrane dynamics, we have developed a novel technique, called flash-and-freeze, which induces neuronal activity with a flash of light and captures the membrane dynamics by rapid freezing. For characterizing membrane movements during synaptic transmission, a light-sensitive cation channel, channelrhodopsin, is heterologously expressed in mouse hippocampal neurons or in Caenorhabditis elegans motor neurons. A brief pulse of blue light activates channelrhodopsin and induces an action potential, leading to synaptic transmission. Following the light stimulation, neurons are frozen at different time intervals ranging from 10 ms to 20 s. Electron micrographs are then acquired from each time point to visualize the morphological changes. Using this approach, we have characterized a novel form of endocytosis, ultrafast endocytosis, which rapidly removes excess membrane added to the surface during neurotransmission. The flash-and-freeze approach can be adapted to study other cellular phenomena that can be induced by light-sensitive genetic or pharmacological tools.

  6. Flash-and-Freeze: Coordinating Optogenetic Stimulation with Rapid Freezing to Visualize Membrane Dynamics at Synapses with Millisecond Resolution

    Science.gov (United States)

    Watanabe, Shigeki

    2016-01-01

    Electron microscopy depicts subcellular structures at synapses exquisitely but only captures static images. To visualize membrane dynamics, we have developed a novel technique, called flash-and-freeze, which induces neuronal activity with a flash of light and captures the membrane dynamics by rapid freezing. For characterizing membrane movements during synaptic transmission, a light-sensitive cation channel, channelrhodopsin, is heterologously expressed in mouse hippocampal neurons or in Caenorhabditis elegans motor neurons. A brief pulse of blue light activates channelrhodopsin and induces an action potential, leading to synaptic transmission. Following the light stimulation, neurons are frozen at different time intervals ranging from 10 ms to 20 s. Electron micrographs are then acquired from each time point to visualize the morphological changes. Using this approach, we have characterized a novel form of endocytosis, ultrafast endocytosis, which rapidly removes excess membrane added to the surface during neurotransmission. The flash-and-freeze approach can be adapted to study other cellular phenomena that can be induced by light-sensitive genetic or pharmacological tools. PMID:27594835

  7. Immersion freezing of ice nucleation active protein complexes

    Science.gov (United States)

    Hartmann, S.; Augustin, S.; Clauss, T.; Wex, H.; Šantl-Temkiv, T.; Voigtländer, J.; Niedermeier, D.; Stratmann, F.

    2013-06-01

    Utilising the Leipzig Aerosol Cloud Interaction Simulator (LACIS), the immersion freezing behaviour of droplet ensembles containing monodisperse particles, generated from a Snomax™ solution/suspension, was investigated. Thereto ice fractions were measured in the temperature range between -5 °C to -38 °C. Snomax™ is an industrial product applied for artificial snow production and contains Pseudomonas syringae} bacteria which have long been used as model organism for atmospheric relevant ice nucleation active (INA) bacteria. The ice nucleation activity of such bacteria is controlled by INA protein complexes in their outer membrane. In our experiments, ice fractions increased steeply in the temperature range from about -6 °C to about -10 °C and then levelled off at ice fractions smaller than one. The plateau implies that not all examined droplets contained an INA protein complex. Assuming the INA protein complexes to be Poisson distributed over the investigated droplet populations, we developed the CHESS model (stoCHastic modEl of similar and poiSSon distributed ice nuclei) which allows for the calculation of ice fractions as function of temperature and time for a given nucleation rate. Matching calculated and measured ice fractions, we determined and parameterised the nucleation rate of INA protein complexes exhibiting class III ice nucleation behaviour. Utilising the CHESS model, together with the determined nucleation rate, we compared predictions from the model to experimental data from the literature and found good agreement. We found that (a) the heterogeneous ice nucleation rate expression quantifying the ice nucleation behaviour of the INA protein complex is capable of describing the ice nucleation behaviour observed in various experiments for both, Snomax™ and P. syringae bacteria, (b) the ice nucleation rate, and its temperature dependence, seem to be very similar regardless of whether the INA protein complexes inducing ice nucleation are attached

  8. Immersion freezing of ice nucleation active protein complexes

    Directory of Open Access Journals (Sweden)

    S. Hartmann

    2013-06-01

    Full Text Available Utilising the Leipzig Aerosol Cloud Interaction Simulator (LACIS, the immersion freezing behaviour of droplet ensembles containing monodisperse particles, generated from a Snomax™ solution/suspension, was investigated. Thereto ice fractions were measured in the temperature range between −5 °C to −38 °C. Snomax™ is an industrial product applied for artificial snow production and contains Pseudomonas syringae} bacteria which have long been used as model organism for atmospheric relevant ice nucleation active (INA bacteria. The ice nucleation activity of such bacteria is controlled by INA protein complexes in their outer membrane. In our experiments, ice fractions increased steeply in the temperature range from about −6 °C to about −10 °C and then levelled off at ice fractions smaller than one. The plateau implies that not all examined droplets contained an INA protein complex. Assuming the INA protein complexes to be Poisson distributed over the investigated droplet populations, we developed the CHESS model (stoCHastic modEl of similar and poiSSon distributed ice nuclei which allows for the calculation of ice fractions as function of temperature and time for a given nucleation rate. Matching calculated and measured ice fractions, we determined and parameterised the nucleation rate of INA protein complexes exhibiting class III ice nucleation behaviour. Utilising the CHESS model, together with the determined nucleation rate, we compared predictions from the model to experimental data from the literature and found good agreement. We found that (a the heterogeneous ice nucleation rate expression quantifying the ice nucleation behaviour of the INA protein complex is capable of describing the ice nucleation behaviour observed in various experiments for both, Snomax™ and P. syringae bacteria, (b the ice nucleation rate, and its temperature dependence, seem to be very similar regardless of whether the INA protein complexes inducing ice

  9. 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 sod1 mutant could be made more resistant by treatment with the superoxide anion scavenger MnCl2, or by freezing in the absence of oxygen, or by the generation of a rho0 petite. Increased expression of SOD2 conferred freeze-thaw tolerance on the sod1 mutant indicating the ability of the mitochondrial...

  10. Influence of uniconazole on freezing stress resistance of cold acclimating (S. commersonii) and of non acclimating (S. tuberosum) potato species

    Energy Technology Data Exchange (ETDEWEB)

    Saavedra, J.A.; Palta, J.P.; Stang, E.J. (Univ. of Wisconsin, Madison (United States))

    1991-05-01

    Uniconazole (UCZ) is an effective growth suppressant of the group of triazoles. Triazoles have been used as fungicides (inhibiting sterol biosynthesis) or plant growth regulators (inhibiting gibberellin biosynthesis). Some reports suggest that triazoles can help to alleviate some environmental stresses, including SO{sub 2}, drought, and low and high temperatures. Using soil drench or foliar spray procedure the authors investigated the influence of UCZ 95-500 mg L{sup {minus}1} on freezing tolerance of leaf tissue. Freezing tolerance was evaluated 1-10 days following UCZ treatment by using electrolyte leakage method. They also measured internode length, chlorophyll content, and the size of palisade layer on treated leaves. The highest UCZ rate (500 mg L{sup {minus}1}) did not cause injury to the plants. Chlorophyll content and length of palisade cells increased by 30 and 14%, respectively, in the treated leaves. The internode length was suppressed in 22% by UCZ. Thus UCZ was effective in dwarfing the plants. However, no dramatic effects were found on the freezing stress resistance in both potato species.

  11. Effects of sugar alcohol and proteins on the survival of Lactobacillus bulgaricus LB6 during freeze drying

    Directory of Open Access Journals (Sweden)

    He Chen

    2015-06-01

    Full Text Available Background. Lactobacillus bulgaricus LB6 is a bacterium which was selected in the commercial yoghurt with high angiotensin converting enzyme (ACE inhibitory activity. Preparation of concentrated starter cultures via freeze drying is of practical importance to dairy and food industries. Material and methods. We optimized the optimal sugar alcohol and proteins for Lactobacillus bulgaricus LB6 during the process of freeze drying using a Plackett-Burman design. In our initial tests survival rate and the number of viable cells were associated with the type of lyoprotectant used and so our optimization protocol focused on increasing survival rate. Substances that had previously had a protective effect during freeze drying were investigated, for example: mannitol, sorbitol, xylitol, meso-erythritol, lactitol, whey protein isolate 90, bovine serum albumin, and whey protein concentrate 80 and soy protein isolate 70. Results. We found that the optimum sugar alcohol and proteins for survival of Lactobacillus bulgaricus LB6 were whey protein concentrate (p = 0.0040 for survival rate, xylitol (p = 0.0067 for survival rate and sorbitol (p = 0.0073 for survival rate, they showed positive effect (whey protein concentrate and sorbitol or negative effect (xylitol. Discussion. The effectiveness of three chosen sugar alcohols and protein implied that they could be used as lyoprotectant for Lactobacillus bulgaricus LB6 in the further research, the optimal composition of sugar alcohol and protein for the lyoprotectant use must be established.

  12. Evaluation of three methods for preservation of Azotobacter: freeze-drying, cryopreservation, and immobilization in dry polymers

    Directory of Open Access Journals (Sweden)

    Daniel Fernando Rojas Tapias

    2013-04-01

    Full Text Available Because the use of bacteria for biotechnological processes requires maintaining their viability and geneticstability, preserving them becomes essential. Here, we evaluated three preservation methods for A.chroococcum C26 and A. vinelandii C27; preservation methods: cryopreservation and immobilization in drypolymers for 60 days, and freeze-drying for 30. We evaluated their efficiency by counting viable cells andmeasuring nitrogen fixation activity. Additionally, we assessed the effect of three protective agents forfreeze-drying, three for cryopreservation, and four polymers. Freeze-drying proved the best technique tomaintain viability and activity, followed by immobilization and cryopreservation. Bacterial nitrogen fixingability remained unchanged using the freeze-drying method, and bacterial survival exceeded 80%; S/BSAwas the best protective agent. Immobilization maintained bacterial survival over 80%, but nitrogen fixationwas decreased by 20%. Lastly, cryopreservation resulted in a dramatic loss of viability for C26 (BSRapprox. 70%, whereas C27 was well preserved. Nitrogen fixation for both strains decreased regardless ofthe cryoprotective agent used (P < 0.05. In conclusion, the success of Azotobacter preservation methodsdepend on the technique, the protective agent, and the strain used. Our results also indicated that freezedryingusing S/BSA is the best technique to preserve bacteria of this genus.

  13. Micro-PIXE investigations of apoplastic iron in freeze-dried root cross-sections of soil grown barley

    International Nuclear Information System (INIS)

    Freeze-dried cryo-sections of barley roots (Hordeum vulgare L. cv. Alexis) were used to investigate the possible role of the root apoplast as an iron-storage pool for plants; this possibility has been a matter of controversy. Micro-PIXE analyzes in pixel mode were complemented by the STIM technique. Data were analyzed using the new Heidelberg software package BIOPIXE, which provides true elemental maps of inhomogeneous samples such as freeze-dried cross-sections of roots. The maps clearly show a high heterogeneity of the iron distribution in roots between adjacent cell layers. Accumulations of iron were observed in the cell walls of the outermost cell layers of the roots and at the endodermis. Based on the correlation between iron and soil related elements like titanium, aluminum and silicon, most of the iron located at the root surface could be attributed to soil contamination. It could also be shown that these soil contaminations lead to an overestimation of the apoplastic iron concentrations determined by methods commonly used in the botanical field. Besides this, low accumulations of iron were observed in the cell walls of the outmost cell layers of the roots. This may indicate that the root apoplast may have a minor function in iron nutrition

  14. AC Electric Field Enhances Cryopreservation Efficiency of Sprague-Dawley Rat Liver During a Slow Freezing Procedure.

    Science.gov (United States)

    Ma, Ya H; Qin, Guo F; Li, Jing; Ding, Gui R; Xu, Sheng L; Zhou, Yan; Guo, Guo Z

    2016-02-01

    Slow freezing coupled with an AC electric field (ACEF) has been demonstrated to miniaturize the ice crystals of a 0.9% (w/v) NaCl solution in a prior study. The aim of this study was to assess the effect of ACEF on Sprague-Dawley (SD) rat liver in vitro during the slow cooling procedure. SD rat liver exposed to an oscillating electric field was frozen in a programmed freezer initially down to -30°C at a cooling rate of -1°C/min and continuing down to -80°C at a cooling rate of -5°C/min. The cryovials were finally transferred into liquid nitrogen for 7 days. The frequency range was 0-20 MHz, and peak field strength was 1,000 V/m. For the sham and electric-exposed groups, the freezing solution consisted of 0%, 2.5%, 5.0%, 7.5%, or 10% (v/v) dimethyl sulfoxide (DMSO) Dulbecco's modified Eagles' medium culture solution, and fresh tissue was selected as the control group. The changes in cell survival rate, adenosine triphosphate (ATP) content, and morphology of fresh and frozen-thawed liver tissue were examined. Compared with the sham group with 5.0% DMSO, the result showed that slow freezing coupled with 2.45 MHz or 5 MHz ACEF significantly increased the relative survival rate by 43.27% and 26.31% (P membrane and some organelles (e.g., mitochondria) in the electric-exposed group appeared to be more intact according to the transmission electron microscopy images. The underlying mechanism might be that the ACEF affects the formation and growth of the ice crystallization, and thus inhibits cryoinjury. These results show that ACEF would provide an efficient method for cryopreservation banking with a low concentration of CPA during the slow freezing process. PMID:26828902

  15. Impact of vacuum-induced surface freezing on inter- and intra-vial heterogeneity.

    Science.gov (United States)

    Oddone, Irene; Van Bockstal, Pieter-Jan; De Beer, Thomas; Pisano, Roberto

    2016-06-01

    This paper aimed to study the impact of freezing on both within-batch (inter-vial) and within-product (intra-vial) heterogeneity. This analysis has been carried out using two freezing protocols, the conventional shelf-ramped method and the Vacuum Induced Surface Freezing, and placebo formulations containing both crystallizing (mannitol) and amorphous (lactose and sucrose) excipients. The freezing conditions (i.e., the temperature of freezing, the temperature and time of the equilibration phase, and the filling volume) were found to have a dramatic impact on both the within-batch and the within-product homogeneity. Overall, we observed that the control of freezing can effectively minimize the variability in product characteristics, and moisture content, within the same batch. In addition to more uniform production, the control of freezing was found to be fundamental to achieve a more uniform product than that produced by the shelf-ramped freezing method. The influence of the freezing protocol on the crystallization process of mannitol was also investigated, showing that the temperature of freezing plays a key role in the formation of the mannitol polymorphs. PMID:27063591

  16. Microaspiration for high-pressure freezing: a new method for ultrastructural preservation of fragile and sparse tissues for TEM and electron tomography

    Energy Technology Data Exchange (ETDEWEB)

    Auer, Manfred; Triffo, W.J.; Palsdottir, H.; McDonald, K.L.; Inman, J.L.; Bissell, M.J.; Raphael, R.M.; Auer, M.; Lee, J.K.

    2008-02-13

    High-pressure freezing is the preferred method to prepare thick biological specimens for ultrastructural studies. However, the advantages obtained by this method often prove unattainable for samples that are difficult to handle during the freezing and substitution protocols. Delicate and sparse samples are difficult to manipulate and maintain intact throughout the sequence of freezing, infiltration, embedding, and final orientation for sectioning and subsequent TEM imaging. An established approach to surmount these difficulties is the use of cellulose microdialysis tubing to transport the sample. With an inner diameter of 200 micrometers, the tubing protects small and fragile samples within the thickness constraints of high-pressure freezing, and the tube ends can be sealed to avoid loss of sample. Importantly, the transparency of the tubing allows optical study of the specimen at different steps in the process. Here, we describe the use of a micromanipulator and microinjection apparatus to handle and position delicate specimens within the tubing. We report two biologically significant examples that benefit from this approach, 3D cultures of mammary epithelial cells and cochlear outer hair cells. We illustrate the potential for correlative light and electron microscopy as well as electron tomography.

  17. Impact of the freeze-drying process on product appearance, residual moisture content, viability, and batch uniformity of freeze-dried bacterial cultures safeguarded at culture collections.

    Science.gov (United States)

    Peiren, Jindrich; Hellemans, Ann; De Vos, Paul

    2016-07-01

    In this study, causes of collapsed bacterial cultures in glass ampoules observed after freeze-drying were investigated as well as the influence of collapse on residual moisture content (RMC) and viability. Also, the effect of heat radiation and post freeze-drying treatments on the RMC was studied. Cake morphologies of 21 bacterial strains obtained after freeze-drying with one standard protocol could be classified visually into four major types: no collapse, porous, partial collapse, and collapse. The more pronounced the collapse, the higher residual moisture content of the freeze-dried product, ranging from 1.53 % for non-collapsed products to 3.62 % for collapsed products. The most important cause of collapse was the mass of the inserted cotton plug in the ampoule. Default cotton plugs with a mass between 21 and 30 mg inside the ampoule did not affect the viability of freeze-dried Aliivibrio fischeri LMG 4414(T) compared to ampoules without cotton plugs. Cotton plugs with a mass higher than 65 mg inside the ampoule induced a full collapsed product with rubbery look (melt-back) and decreasing viability during storage. Heat radiation effects in the freeze-drying chamber and post freeze-drying treatments such as exposure time to air after freeze-drying and manifold drying time prior to heat sealing of ampoules influenced the RMC of freeze-dried products. To produce uniform batches of freeze-dried bacterial strains with intact cake structures and highest viabilities, inserted cotton plugs should not exceed 21 mg per ampoule. Furthermore, heat radiation effects should be calculated in the design of the primary drying phase and manifold drying time before heat sealing should be determined as a function of exposure time to air.

  18. Freeze-drying behaviour of pasteurized whole egg

    International Nuclear Information System (INIS)

    Because it provides full nutritional and certain desirable functional attributes, egg products are widely used as ingredients in many food products. Dried egg is especially valuable for being stable, easily mixable and having a long shelf life. It is necessary to know the effects of drying conditions onto the moisture removal behaviour and the functional properties of the powder product, to serve the egg powder as an alternative. An experimental study was conducted to achieve an understanding of the freeze-drying behaviour of pasteurized whole egg having 24% dry solids. In order to determine the moisture removal behaviour; the percent moisture loss (w/w), the average moisture content and the drying rates were obtained, the drying curves were developed and total drying times were determined, also the movement of the dry-wet boundary between the frozen layer and the dry porous layer formed by sublimation of ice crystals were investigated during a complete process. The physical properties of pasteurized whole egg such as; colour, water activity (aw), the morphological structure (through SEM analysis) and functional properties (foam stability and dissolubility) were determined. The net colour change (ΔE) was about 22, independent of layer thickness. The water activity decreased to 0.22 at the end of drying. The SEM images of freeze-dried and slightly milled egg powder samples at magnification levels of 500 and 1000 showed the porous structure caused by sublimation of ice crystals generated within the egg structure during air blast freezing. The dissolubility and foaming capacity of powder egg were observed to be lower compared to those of pasteurized liquid egg. (author)

  19. Dynamics of a model colloidal suspension from dilute to freezing

    Science.gov (United States)

    Hannam, S. D. W.; Daivis, P. J.; Bryant, G.

    2016-07-01

    Molecular dynamics simulation was used to study a model colloidal suspension at a range of packing fractions from the dilute limit up to the freezing point. This study builds on previous work by the authors which modeled the colloidal particles with a hard core surrounded by a Weeks-Chandler-Anderson potential with modified interaction parameters, and included an explicit solvent. In this work, we study dynamical properties of the model by first calculating the velocity autocorrelation function, the self-diffusion coefficient, and the mutual diffusion coefficient. We also perform detailed calculations of the colloidal particle intermediate scattering function to study the change in dynamics leading up to the freezing point, and to determine whether the current model can be used to interpret light scattering experiments. We then perform a multiexponential analysis on the intermediate scattering function results and find that the data are fitted well by the sum of two exponentials, which is in line with previous analysis of experimental colloidal suspensions. The amplitudes and decay coefficients of the two modes are determined over a large range of wave vectors at packing fractions leading up to the freezing point. We found that the maximum wave vector at which macroscopic diffusive behavior was observed decreased as the packing fraction increased, and a simple extrapolation shows the maximum wave vector going to zero at the melting point. Lastly, the ratio of the two decay coefficients is compared to the scaling law proposed by Segrè and Pusey [Phys. Rev. Lett. 77, 771 (1996), 10.1103/PhysRevLett.77.771]. It was found that the ratio was not constant, but instead was wave vector dependent.

  20. On the transition between heterogeneous and homogeneous freezing

    Directory of Open Access Journals (Sweden)

    K. Gierens

    2003-01-01

    Full Text Available Box model simulations of an uplifting and adiabatically cooling cloud of aerosol have been performed in order to study the transition between cirrus formation dominated by homogeneous nucleation of ice to that dominated by heterogeneous nucleation. The aerosol was assumed to consist of an internal mixture of sulfuric acid solution droplets with inclusions of soot. The parametrisation of De Mott et al. (1997 was used to simulate the heterogeneous nucleation of ice in such droplets with soot inclusions. The simulations show that the transition from heterogeneous to homogeneous nucleation occurs over a narrow range of soot concentration. Thus it seems to be possible to fix critical concentrations of heterogeneous ice nuclei which must be exceeded if heterogeneous freezing dominates cirrus formation. A formula has been derived that allows to compute the critical concentrations of heterogeneous ice nuclei as a function of temperature, updraft speed, ambient pressure, and supersaturation at which heterogeneous freezing occurs. Generally, homogeneous nucleation dominates in regions with updrafts stronger than 20 cm s -1, with the exception of heavily polluted areas which could be common in the northern hemisphere due to air traffic, where updrafts of the order 1 m s-1 may be necessary to render heterogeneous nucleation unimportant. According to the present results it cannot be excluded that heterogeneous nucleation plays a more important role for cirrus formation in the northern midlatitudes than anywhere else. A possible consequence of these results is that air pollution may lead to a higher coverage of cirrus clouds, but then these clouds will be optically thinner than clouds formed by homogeneous freezing, with the exception of regions where condensation trails are frequent.

  1. Demonstrating Functional Equivalence of Pilot and Production Scale Freeze-Drying of BCG

    Science.gov (United States)

    ten Have, R.; Reubsaet, K.; van Herpen, P.; Kersten, G.; Amorij, J.-P.

    2016-01-01

    Process analytical technology (PAT)-tools were used to monitor freeze-drying of Bacille Calmette-Guérin (BCG) at pilot and production scale. Among the evaluated PAT-tools, there is the novel use of the vacuum valve open/close frequency for determining the endpoint of primary drying at production scale. The duration of primary drying, the BCG survival rate, and the residual moisture content (RMC) were evaluated using two different freeze-drying protocols and were found to be independent of the freeze-dryer scale evidencing functional equivalence. The absence of an effect of the freeze-dryer scale on the process underlines the feasibility of the pilot scale freeze-dryer for further BCG freeze-drying process optimization which may be carried out using a medium without BCG. PMID:26981867

  2. Diurnal freeze/thaw cycles of the ground surface on the Tibetan Plateau

    Institute of Scientific and Technical Information of China (English)

    YANG MeiXue; YAO TanDong; GOU XiaoHua; HIROSE Nozomu; FUJII Hide Yuki; HAO LiSheng; D.F.LEVIA

    2007-01-01

    The exchange of energy and water between the lithosphere and atmosphere mainly takes place at the ground surface. Therefore, freeze/thaw condition at the ground surface is an important factor in examining the interactions between the land surface and atmosphere. Based on the observation data obtained by CEOP/CAMP-Tibet, the diurnal freeze/thaw cycles of the ground surface near Naqu, central Tibetan Plateau was preliminarily analyzed. The results show that the surface layer was completely frozen for approximately one month. However, the time that the ground surface experienced diurnal freeze/thaw cycles was about 6 months. The high frequency of freeze/thaw cycles at the ground surface significantly influences water and energy exchanges between ground and atmosphere over half a year. The interaction processes between the ground and atmosphere under different soil conditions (such as complete thaw, complete freeze and diurnal freeze/thaw cycles) are issues worthy of further examination.

  3. Demonstrating Functional Equivalence of Pilot and Production Scale Freeze-Drying of BCG.

    Science.gov (United States)

    Ten Have, R; Reubsaet, K; van Herpen, P; Kersten, G; Amorij, J-P

    2016-01-01

    Process analytical technology (PAT)-tools were used to monitor freeze-drying of Bacille Calmette-Guérin (BCG) at pilot and production scale. Among the evaluated PAT-tools, there is the novel use of the vacuum valve open/close frequency for determining the endpoint of primary drying at production scale. The duration of primary drying, the BCG survival rate, and the residual moisture content (RMC) were evaluated using two different freeze-drying protocols and were found to be independent of the freeze-dryer scale evidencing functional equivalence. The absence of an effect of the freeze-dryer scale on the process underlines the feasibility of the pilot scale freeze-dryer for further BCG freeze-drying process optimization which may be carried out using a medium without BCG. PMID:26981867

  4. Fast-freezing with liquid nitrogen preserves bulk dissolved organic matter concentrations, but not its composition

    Science.gov (United States)

    Thieme, Lisa; Graeber, Daniel; Kaupenjohann, Martin; Siemens, Jan

    2016-08-01

    Freezing can affect concentrations and spectroscopic properties of dissolved organic matter (DOM) in water samples. Nevertheless, water samples are regularly frozen for sample preservation. In this study we tested the effect of different freezing methods (standard freezing at -18 °C and fast-freezing with liquid nitrogen) on DOM concentrations measured as organic carbon (DOC) concentrations and on spectroscopic properties of DOM from different terrestrial ecosystems (forest and grassland). Fresh and differently frozen throughfall, stemflow, litter leachate and soil solution samples were analyzed for DOC concentrations, UV-vis absorption and fluorescence excitation-emission matrices combined with parallel factor analysis (PARAFAC). Fast-freezing with liquid nitrogen prevented a significant decrease of DOC concentrations observed after freezing at -18 °C. Nonetheless, the share of PARAFAC components 1 (EXmax nitrogen for preservation of bulk DOC concentrations of samples from terrestrial sources, whereas immediate measuring is preferable to preserve spectroscopic properties of DOM.

  5. Damage Evaluation on Freeze-Thawing Process of Food by Using NMR

    Science.gov (United States)

    Andou, Hiroko; Fukuoka, Mika; Miyawaki, Osato; Suzuki, Toru

    Freeze-thawing process gives significant damages for food structure. Several new techniques have been attempted for quantitative evaluation of the damages. In this study, using NMR (nuclear magnetic resonance) with a stimulated echo method, restricted diffusion phenomena of water molecules was measured for damaged food (onion and tuna) tissues that were subjected to the repeat of freeze-thawing, Through experiments, water permeability of tissue membrane was calculated. The water permeability of fresh tissues for onion showed clearly restricted diffusion, but after freeze-thawing, it disappeared. On the other hand, the water permeability of fresh tuna tissue was small significantly, even though it was a little higher after freeze-thawing. After all, the damage level after freeze-thawing showed a significant difference between onion and tuna. These results support the view that plant tissue is very sensitive to freeze-thawing and that the water permeability of plant is much lower than that of animal.

  6. Numerical Simulation on Freezing Process of Saturated Granlar Soil

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    The relation between ice pressure and load as a criterion of segregated ice initiation is introduced into the rigid ice model to simulate frost heave in saturated and granular soil. The calculated results show that unfrozen water content, thermal conductivity and hydraulic conductivity change greatly in frozen fringe. In numerical simulations, the influence of load, hydraulic conductivity and property of soil containing water on the process of soil freezing are analyzed, and the simulation curves such as cumulative heave,the change of depth of frozen and the distributions of water content are similar to the observations reported elsewhere.

  7. Freezing of capillary waves at the glass transition

    International Nuclear Information System (INIS)

    The freezing of capillary waves on glycerol surfaces is studied by in situ x-ray reflectivity measurements. A wide temperature range around the calorimetric glass transition temperature at Tg≅186 K is investigated. For T>250 K the obtained surface roughness as a function of the temperature differs significantly from the value predicted by the classical capillary waves theory. Below the temperature T≅250 K the magnitude of the roughness remains constant. Furthermore, a large hysteresis, i.e., a large difference of the roughnesses measured during cooling and heating of the sample, is observed. These findings are discussed in terms of viscosity effects

  8. Numerical modeling of seasonally freezing ground and permafrost

    Science.gov (United States)

    Nicolsky, Dmitry J.

    2007-12-01

    This thesis represents a collection of papers on numerical modeling of permafrost and seasonally freezing ground dynamics. An important problem in numerical modeling of temperature dynamics in permafrost and seasonally freezing ground is related to parametrization of already existing models. In this thesis, a variation data assimilation technique is presented to find soil properties by minimizing the discrepancy between in-situ measured temperatures and those computed by the models. The iterative minimization starts from an initial approximation of the soil properties that are found by solving a sequence of simple subproblems. In order to compute the discrepancy, the temperature dynamics is simulated by a new implementation of the finite element method applied to the heat equation with phase change. Despite simplifications in soil physics, the presented technique was successfully applied to recover soil properties, such as thermal conductivity, soil porosity, and the unfrozen water content, at several sites in Alaska. The recovered properties are used in discussion on soil freezing/thawing and permafrost dynamics in other parts of this thesis. Another part of this thesis concerns development of a numerical thermo-mechanical model of seasonal soil freezing on the lateral scale of several meters. The presented model explains observed differential frost heave occurring in non-sorted circle ecosystems north of the Brooks Range in the Alaskan tundra. The model takes into account conservation principles for energy, linear momentum and mass of three constituents: liquid water, ice and solid particles. The conservation principles are reduced to a computationally convenient system of coupled equations for temperature, liquid water pressure, porosity, and the velocity of soil particles in a three-dimensional domain with cylindrical symmetry. Despite a simplified rheology, the model simulates the ground surface motion, temperature, and water dynamics in soil and explains

  9. Cost shifting and the freezing of corporate pension plans

    OpenAIRE

    Joshua Rauh; Irina Stefanescu; Stephen Zeldes

    2013-01-01

    Many U.S. corporations have frozen defined benefit (DB) pension plans, replacing new DB promises with contributions to defined contribution (DC) plans. We estimate expected DB accruals from the age-service and salary distributions of a large sample of U.S. corporate pension plans with more than 1,000 employees. Comparing the counterfactual DB accruals to the actual increase in 401(k) and other DC contributions for firms that freeze, we find only partial compensation to employees for the lost ...

  10. Deformed liquid marbles: Freezing drop oscillations with powders

    KAUST Repository

    Marston, Jeremy

    2012-09-01

    In this work we show that when a liquid drop impacts onto a fine-grained hydrophobic powder, the final form of the drop can be very different from the spherical form with which it impacts. In all cases, the drop rebounds due to the hydrophobic nature of the powder. However, we find that above a critical impact speed, the drop undergoes a permanent deformation to a highly non-spherical shape with a near-complete coverage of powder, which then freezes the drop oscillations during rebound. © 2012 Elsevier B.V.

  11. Heterogeneous freezing of water droplets containing kaolinite particles

    Directory of Open Access Journals (Sweden)

    B. J. Murray

    2011-05-01

    Full Text Available Clouds composed of both ice particles and supercooled liquid water droplets exist at temperatures above ~236 K. These mixed phase clouds, which strongly impact climate, are very sensitive to the presence of solid particles that can catalyse freezing. In this paper we describe experiments to determine the conditions at which the clay mineral kaolinite nucleates ice when immersed within water droplets. These are the first immersion mode experiments in which the ice nucleating ability of kaolinite has been determined as a function of clay surface area, cooling rate and also at constant temperatures. Water droplets containing a known amount of clay mineral were supported on a hydrophobic surface and cooled at rates of between 0.8 and 10 K min−1 or held at constant sub-zero temperatures. The time and temperature at which individual 10–50 μm diameter droplets froze were determined by optical microscopy. For a cooling rate of 10 K min−1, the median nucleation temperature of 10–40 μm diameter droplets increased from close to the homogeneous nucleation limit (236 K to 240.8 ± 0.6 K as the concentration of kaolinite in the droplets was increased from 0.005 wt% to 1 wt%. This data shows that the probability of freezing scales with surface area of the kaolinite inclusions. We also show that at a constant temperature the number of liquid droplets decreases exponentially as they freeze over time. The constant cooling rate experiments are consistent with the stochastic, singular and modified singular descriptions of heterogeneous nucleation; however, freezing during cooling and at constant temperature can be reconciled best with the stochastic approach. We report temperature dependent nucleation rate coefficients (nucleation events per unit time per unit area for kaolinite and present a general parameterisation for immersion nucleation which may be suitable for cloud modelling once nucleation by other important ice nucleating

  12. Calorimetric analysis of cryopreservation and freeze-drying formulations.

    Science.gov (United States)

    Sun, Wendell Q

    2015-01-01

    Differential scanning calorimetry (DSC) is a commonly used thermal analysis technique in cryopreservation and freeze-drying research. It has been used to investigate crystallization, eutectic formation, glass transition, devitrification, recrystallization, melting, polymorphism, molecular relaxation, phase separation, water transport, thermochemistry, and kinetics of complex reactions (e.g., protein denaturation). Such information can be used for the optimization of protective formulations and process protocols. This chapter gives an introduction to beginners who are less familiar with this technique. It covers the instrument and its basic principles, followed by a discussion of the methods as well as examples of specific applications.

  13. Static critical fluctuations on the freeze-out surface

    CERN Document Server

    Jiang, Lijia; Song, Huichao

    2016-01-01

    In this proceeding, we summarize the main results of our recent paper, which introduces a freeze-out scheme to the dynamical models near the QCD critical point. Within such framework of static critical fluctuations, the Beam Energy Scan (BES) data of $C_4$ and $\\kappa \\sigma^2$ for net protons within different $p_T$ ranges can be roughly described. Besides, the momentum acceptance dependence of higher cumulants at lower collision energies can also be qualitatively described. However, $C_2$ and $C_3$ are always over-predicted due to the positive static critical fluctuations.

  14. THE BEHAVIOR OF DIFFERENT FIBERS AT BREAD DOUGH FREEZING

    Directory of Open Access Journals (Sweden)

    Jelena Filipović

    2008-11-01

    Full Text Available Three different types of commercial fibers (inulin originated from artichoke with long and short molecule chains and Fibrex - originated from sugar beet were incorporated into the dough formula as flour supplements at the level of 5 %. The influence of fiber characteristics on yeast dough (proving time and stability and bread quality (volume and crumb quality during 60 days freezing is presented. Data show that the addition of fibers in frozen yeast dough is positively contributing to preserving the quality of the final product and their influence depends on the characteristics of fibers.

  15. Prognostic value of a pre-freeze hypo-osmotic swelling test on the post-thaw quality of dog semen.

    Science.gov (United States)

    Karger, S; Geiser, B; Grau, M; Burfeind, O; Heuwieser, W; Arlt, S P

    2016-03-01

    Throughout cryopreservation, sperm are exposed to major osmotic challenges. Only intact membranes of sperm cells are able to regulate these volumetric changes, which can be determined by the hypo-osmotic swelling test (HOS test). Correlations between the HOS test and conventional semen variables are inconsistent. Therefore, the objectives of this study were (1) to examine relationships between HOS test results and standard semen variables before freezing and after thawing and (2) to evaluate the prognostic value of the HOS assessments on post-thaw quality of dog semen. Semen of 35 dogs was collected and analyzed before freezing and after thawing following a 7-day freeze-thaw interval. Conventional semen variables such as sperm cell motility, membrane integrity morphology were evaluated and the HOS test was conducted with results from this test being recorded. In fresh semen the HOS test was positively correlated with progressive motility of sperm cells: r=0.52, sperm cell membrane integrity: r=0.50 and normal sperm cell morphology: r=0.46 (Pdog semen using the HOS test. PMID:26837622

  16. The influence of carboxymethylcellulose, xanthan and guar-gum addition in bread dough before freezing on metabolism and viability of Saccharomyces cerevisiae

    Directory of Open Access Journals (Sweden)

    Kocić-Tanackov Sunčica D.

    2009-01-01

    Full Text Available Doughs were prepared with different concentrations of carboxymethylcellulose, xanthan, and guar-gum (0.1, 0.2 and 0.3% in doughs, freezed at -20°C and analyzed after 0, 7, 15 and 30 days. Pure Saccharomyces cerevisiae culture was isolated from dough and was cultivated under optimal conditions during 24 hrs to determine the following parameters: specific growth rate, fermentative activities and cytochromes contents in intact cells with the aim of determining the respiration intensity. During freezing of dough for 30 days, the percentage of living cells from dough surface was 53.11% and from the middle 54.95%. Carboxymethylcellulose in concentration of 0.3 and 0.5% increased number of survived cells on the surface to 70.64, and 70.28% and in the middle to 74.79, and 76.54%, respectively. Guar-gum increased number of survived cells only in concentration of 0.1% on the surface to 70.17% and in the middle of the dough to 75.26%. The mean specific growth rate decreased by approximately 10% during 30 days of storage at -20°C. Content of cytochromes in intact cells decreased in all samples during freezing.

  17. Determination of End Point of Primary Drying in Freeze-Drying Process Control

    OpenAIRE

    Patel, Sajal M.; Doen, Takayuki; Pikal, Michael J.

    2010-01-01

    Freeze-drying is a relatively expensive process requiring long processing time, and hence one of the key objectives during freeze-drying process development is to minimize the primary drying time, which is the longest of the three steps in freeze-drying. However, increasing the shelf temperature into secondary drying before all of the ice is removed from the product will likely cause collapse or eutectic melt. Thus, from product quality as well as process economics standpoint, it is very crit...

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

    OpenAIRE

    Wang, Xiaoxiao; Shen, Xiangdong; Wang, Hailong; Gao, Chu; Zhang, Tong

    2016-01-01

    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 f...

  19. Exogenous silicon leads to increased antioxidant capacity in freezing-stressed pistachio leaves

    OpenAIRE

    Habibi, Ghader

    2015-01-01

      Freezing stress limits photosynthesis and growth of plants. This may be attributed to the enhancement of freezing-associated oxidative damage. In this study, we followed precisely changes in the extent of lipid peroxidation and oxidative damage in leaves of pistachio (Pistacia vera ‘Ahmadaghaii’) plants exposed to foliar-applied silicon (Si) under freezing stress. The foliar-applied Si decreased significantly damaging effects of cold on relative water content (RWC), accompanied by an increa...

  20. Freezing behavior of cement pastes saturated with NaCl solution

    OpenAIRE

    Zeng, Qiang; FEN CHONG, Teddy; Li, Kefei

    2014-01-01

    This study investigates the freezing behaviors of two cement pastes saturated with water and salt (NaCl) solutions of different concentrations. Special experimental set-up was designed to measure the freezing strains of cylindrical specimens in undrained condition. Using the interfacial curvature properties involved in mercury intrusion under pressure and ice penetration under freezing, the pore ice saturation degree is evaluated through mercury intrusion porosimetry (MIP) data. Experimental ...

  1. Experimental analysis and modeling of ultrasound assisted freezing of potato spheres.

    Science.gov (United States)

    Kiani, Hossein; Zhang, Zhihang; Sun, Da-Wen

    2015-09-01

    In recent years, innovative methods such as ultrasound assisted freezing have been developed in order to improve the freezing process. During freezing of foods, accurate prediction of the temperature distribution, phase ratios, and process time is very important. In the present study, ultrasound assisted immersion freezing process (in 1:1 ethylene glycol-water solution at 253.15K) of potato spheres (0.02 m diameter) was evaluated using experimental, numerical and analytical approaches. Ultrasound (25 kHz, 890 W m(-2)) was irradiated for different duty cycles (DCs=0-100%). A finite volume based enthalpy method was used in the numerical model, based on which temperature and liquid fraction profiles were simulated by a program developed using OpenFOAM® CFD software. An analytical technique was also employed to calculate freezing times. The results showed that ultrasound irradiation could decrease the characteristic freezing time of potatoes. Since ultrasound irradiation increased the heat transfer coefficient but simultaneously generated heat at the surface of the samples, an optimum DC was needed for the shortest freezing time which occurred in the range of 30-70% DC. DCs higher than 70% increased the freezing time. DCs lower than 30% did not provide significant effects on the freezing time compared to the control sample. The numerical model predicted the characteristic freezing time in accordance with the experimental results. In addition, analytical calculation of characteristic freezing time exhibited qualitative agreement with the experimental results. As the numerical simulations provided profiles of temperature and water fraction within potatoes frozen with or without ultrasound, the models can be used to study and control different operation situations, and to improve the understanding of the freezing process. PMID:25776740

  2. Practical limitations of ITS-90 from the mercury triple point to the silver freeze point

    International Nuclear Information System (INIS)

    The NPL published a forward to the ITS-90 text as follows:- 'The purpose of the ITS is to define procedures by which certain specified practical thermometers of the required quality can be calibrated in such a way that the values of temperature obtained from them can be precise and reproducible, while at the same time closely approximating the corresponding thermodynamic values.' [1]. The paper investigates the properties of thirty four lots of 6N pure metal used to make cells conforming to ITS-90 from mercury through silver over a period of twenty years. Three hundred individual cells are analysed by the impurities listed and supplied with each lot, melt and freeze curve slopes are also summarised for each lot and depressions calculated. These are then compared to the slopes and depressions suggested in the Supplementary Information for the ITS-90 and in CCT/2000-13 'Optimal Realizations'. Results are summarised, tabulated and discussed. Three lots of the thirty four were found to produce cells outside 6N expectations; however the remaining thirty one lots no matter how well or badly the accompanying certification was presented produced cells that conformed to 6N expectations as suggested in Supplementary Information to ITS-90 and CCT/2000-13

  3. Solid Material Characterization of Freeze-Dried Gabexate Mesilate Containing D-Mannitol by Terahertz Spectroscopy

    Science.gov (United States)

    Otsuka, Makoto; Fukura, Naomi; Abe, Hiroyuki

    2013-02-01

    The purpose of the present study is to characterize polymorphic forms and intermolecular interactions of freeze-dried pharmaceuticals containing additives by terahertz (THz) spectroscopy as a, process analytical technology tool in the pharmaceutical industry. Freeze-dried gabexate mesilate/D-mannitol products containing 17-75 mol% gabexate mesilate were obtained using a conventional freeze-dryer. Freeze-dried products and physical mixtures of gabexate mesilate and mannitol with various drug contents were characterized by X-ray powder diffraction (XRD) analysis, differential scanning calorimetry (DSC) and THz. The XRD and DSC results indicated that freeze-dried mannitol was obtained as a mixture of β and δ forms of mannitol from a plain solution, but the freeze-dried product of the gabexate mesilate/mannitol mixture consisted of crystalline gabexate mesilate and the pure δ form of mannitol. Similar to the results of XRD and DSC, THz before the freeze-drying of gabexate mesilate was almost the same as that after. In contrast, the THz of mannitol before freeze-drying had specific peaks due to the β form, but that after had peaks due to δ and β forms. To clarify the polymorphic forms of the freeze-dried products, the THz were analyzed by least squares regression. The calibration models used to predict the amounts of gabexate mesilate and mannitol had sufficient accuracy and linearity, respectively. Two decomposed THz in FGMs had specific peaks due to the δ form of mannitol or gabexate mesilate.

  4. Fast-freezing with liquid nitrogen preserves bulk dissolved organic matter concentrations, but not its composition

    DEFF Research Database (Denmark)

    Thieme, Lisa; Graeber, Daniel; Kaupenjohann, Martin;

    2016-01-01

    Freezing can affect concentrations and spectroscopic properties of dissolved organic matter (DOM) in water samples. Nevertheless, water samples are regularly frozen for sample preservation. In this study we tested the effect of different freezing methods (standard freezing at −18 °C and fast......-freezing with liquid nitrogen) on DOM concentrations measured as organic carbon (DOC) concentrations and on spectroscopic properties of DOM from different terrestrial ecosystems (forest and grassland). Fresh and differently frozen throughfall, stemflow, litter leachate and soil solution samples were analyzed...... spectroscopic properties of DOM....

  5. Effect of Anti-freezing Admixtures on Alkali-silica Reaction in Mortars

    Institute of Scientific and Technical Information of China (English)

    LIU Junzhe; LI Yushun; LV Lihua

    2005-01-01

    The influence of anti-freezing admixture on the alkali aggregate reaction in mortar was analyzed with accelerated methods. It is confirmed that the addition of sodium salt ingredients of anti-freezing admixture accelerates the alkali silica reaction to some extent, whereas calcium salt ingredient of anti-freezing admixture reduces the expansion of alkali silica reaction caused by high alkali cement. It is found that the addition of the fly ash considerably suppresses the expansion of alkali silica reaction induced by the anti-freezing admixtures.

  6. 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.

  7. An improved approach for measuring immersion freezing in large droplets over a wide temperature range.

    Science.gov (United States)

    Tobo, Yutaka

    2016-01-01

    Immersion freezing (ice nucleation by particles immersed in supercooled water) is a key process for forming ice in mixed-phase clouds. Immersion freezing experiments with particles in microliter-sized (millimeter-sized) water droplets are often applied to detecting very small numbers of ice nucleating particles (INPs). However, the application of such large droplets remains confined to the detection of INPs active at temperatures much higher than the homogeneous freezing limit, because of artifacts related to freezing of water droplets without added INPs at temperatures of -25 °C or higher on a supporting substrate. Here I report a method for measuring immersion freezing in super-microliter-sized droplets over a wide temperature range. To reduce possible artifacts, droplets are pipetted onto a thin layer of Vaseline and cooled in a clean booth. In the Cryogenic Refrigerator Applied to Freezing Test (CRAFT) system, freezing of pure (Milli-Q) water droplets are limited at temperatures above -30 °C. An intercomparison of various techniques for immersion freezing experiments with reference particles (Snomax and illite NX) demonstrates that despite the use of relatively large droplets, the CRAFT setup allows for evaluating the immersion freezing activity of the particles over almost the entire temperature range (about -30 °C to 0 °C) relevant for mixed-phase cloud formation. PMID:27596247

  8. The Adsorption and Desorption of Pb(2+) and Cd(2+) in Freeze-Thaw Treated Soils.

    Science.gov (United States)

    Li, Linhui; Ma, Jincai; Xu, Meng; Li, Xu; Tao, Jiahui; Wang, Guanzhu; Yu, Jitong; Guo, Ping

    2016-01-01

    Adsorption and desorption are important processes that influence the potential toxicity and bioavailability of heavy metals in soils. However, information regarding adsorption and desorption behavior of heavy metals in soils subjected to freeze-thaw cycles is poorly understood. In the current study, the effect of freeze-thaw cycles with different freezing temperature (-15, -25, -35°C) on soil properties was investigated. Then the adsorption and desorption behavior of Pb(2+) and Cd(2+) in freeze-thaw treated soils was studied. The adsorption amounts of Pb(2+) and Cd(2+) in freeze-thaw treated soils were smaller than those in unfrozen soils (p adsorption amounts of Pb(2+) and Cd(2+) in soils treated with lower freezing temperatures were higher than those in soils treated with higher freezing temperatures. Desorption percentages of Pb(2+) and Cd(2+) in unfrozen soils were smaller than those in freeze-thaw treated soils (p adsorption and desorption behavior of typical heavy metals in freeze-thaw treated soils located in seasonal frozen soils zone in northeast China.

  9. HSP90 expression correlation with the freezing resistance of bull sperm.

    Science.gov (United States)

    Wang, Peng; Wang, Yan-Feng; Wang, Hong; Wang, Chun-Wei; Zan, Lin-Sen; Hu, Jian-Hong; Li, Qing-Wang; Jia, Yong-Hong; Ma, Guo-Ji

    2014-05-01

    To date, there has been little improvement in cryopreservation of bull sperm due to lack of understanding of the freezing mechanisms. Therefore, this study set out to investigate expression levels of fertility-associated proteins in bull sperm, and in particular the relationship between the 90 kDa heat-shock protein (HSP90) and the sperm characteristics after freezing-thawing. Semen was collected from eight Holstein bulls by artificial vagina. Characteristics of these fresh semen, including sperm motility, morphology, viability and concentration, were evaluated. Sperm quality was also assessed after freezing-thawing. Eight ejaculates were divided into two groups based on freezing resistance and sperm motility. Sperm proteins were extracted and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis and western blotting were performed. SDS-PAGE results showed that there was substantial diversity in 90 kDa proteins in the frozen-thawed sperm and HSP90 was confirmed as one of the 90 kDa proteins by western blot. This study indicated that HSP90 expression correlated positively with sperm quality. The amount of expressed 90 kDa proteins in the high freezing resistance (HFR) group was significantly higher than that in the low freezing resistance (LFR) group (P HSP90 could probably lead to the higher motility and freezing resistance of sperm found after freezing-thawing. Therefore, we concluded that level of HSP90 expression could be used to predict reliably and simply the freezing resistance of bull sperm. PMID:23506739

  10. Behavior of High Water-cement Ratio Concrete under Biaxial Compression after Freeze-thaw Cycles

    Institute of Scientific and Technical Information of China (English)

    SHANG Huaishuai; SONG Yupu; OU Jinping

    2008-01-01

    The high water-cement ratio concrete specimens under biaxial compression that completed in a triaxial testing machine were experimentally studied.Strength and deformations of plain concrete specimens after 0,25,50 cycles of freeze-thaw.Influences of freeze-thaw cycles and stress ratio on the peak stress and deformation of this point were analyzed aecording to the experimental results.Based on the test data,the failure criterion expressed in terms of principal stress after difierent cycles of freeze-thaw,and the failure criterion with consideration of the influence of freeze-thaw cycle and sffess ratio were proposed respectively.

  11. Effect of Freeze-thaw Cycles on Bond Strength between Steel Bars and Concrete

    Institute of Scientific and Technical Information of China (English)

    JI Xiaodong; SONG Yupu; LIU Yuan

    2008-01-01

    The effect of freezing and thawing cycles on mechanical properties of concrete(compressive,splitting tensile strength)was experimentally investigated.According to the pullout test data of three kinds of deformed steel bars,the bond stress-slip curves after freezing and thawing were obtained.The empirical equations of peak bond strength were proposed that the damage accounted for effects of freezing and thawing cycle.Meanwhile,the mechanism of bond deterioration between steel bars and concrete after freezing and thawing cycles was discussed.All these conclusions will be useful to the durability design and reliability calculation of RC structures in cold region.

  12. Advances in studies on concrete durability and countermeasures against freezing-thawing effects

    Institute of Scientific and Technical Information of China (English)

    WuJian Yan; FuJun Niu; XianJun Zhang; Jing Luo; GuoAn Yin

    2014-01-01

    This-paper-is-a-meta-analysis-of-recent-domestic-and-foreign-research-on-freezing-thawing-effects-on-concrete-durability.-The-main-theories-on-the-mechanisms-of-freeze-thaw-damage-to-concrete-are-introduced:-the-hydrostatic-pressure-theory,-the-osmotic-pressure-theory,-the-critical-water-saturation-degree-theory,-the-dual-mechanism-theory,-and-the-mi-cro-ice-crystal-lens-model-theory.-The-influence-laws-of-freezing-thawing-on-the-mechanical-properties-of-concrete-are-summarized,-and-countermeasures-to-improve-concrete-durability-in-freezing-thawing-circumstances-are-presented.-This-work-provides-valuable-references-for-future-engineering-constructions-in-cold-regions.

  13. An improved approach for measuring immersion freezing in large droplets over a wide temperature range

    Science.gov (United States)

    Tobo, Yutaka

    2016-09-01

    Immersion freezing (ice nucleation by particles immersed in supercooled water) is a key process for forming ice in mixed-phase clouds. Immersion freezing experiments with particles in microliter-sized (millimeter-sized) water droplets are often applied to detecting very small numbers of ice nucleating particles (INPs). However, the application of such large droplets remains confined to the detection of INPs active at temperatures much higher than the homogeneous freezing limit, because of artifacts related to freezing of water droplets without added INPs at temperatures of ‑25 °C or higher on a supporting substrate. Here I report a method for measuring immersion freezing in super-microliter-sized droplets over a wide temperature range. To reduce possible artifacts, droplets are pipetted onto a thin layer of Vaseline and cooled in a clean booth. In the Cryogenic Refrigerator Applied to Freezing Test (CRAFT) system, freezing of pure (Milli-Q) water droplets are limited at temperatures above ‑30 °C. An intercomparison of various techniques for immersion freezing experiments with reference particles (Snomax and illite NX) demonstrates that despite the use of relatively large droplets, the CRAFT setup allows for evaluating the immersion freezing activity of the particles over almost the entire temperature range (about ‑30 °C to 0 °C) relevant for mixed-phase cloud formation.

  14. 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.

  15. Degradation of ATP and glycogen in cod ( Gadus morhua ) muscle during freezing

    DEFF Research Database (Denmark)

    Cappeln, Gertrud; Jessen, Flemming

    2001-01-01

    Changes in ATP, IMP, lactate and glycogen contents in the muscle of cod were followed during freezing at temperatures of -20C and -45C. ATP degradation was accompanied by a corresponding increase in IMP content. Simultaneous measurement of temperature showed that at both freezing rates......, the greatest decrease in ATP content was observed when the temperature reached -0.8C. Glycolysis occurred during freezing of cod as indicated by an increase in lactate content. The changes found in all measured metabolites were more pronounced when freezing was performed at a slow rate compared to a fast rate...

  16. Cold tolerance and freeze-induced glucose accumulation in three terrestrial slugs

    DEFF Research Database (Denmark)

    Slotsbo, Stine; Hansen, Lars Monrad; Jordaens, Kurt;

    2012-01-01

    Cold tolerance and metabolic responses to freezing of three slug species common in Scandinavia (Arion ater, Arion rufus and Arion lusitanicus) are reported. Autumn collected slugs were cold acclimated in the laboratory and subjected to freezing conditions simulating likely winter temperatures in....... Glucose increased from about 6 to 22 µg/mg dry tissue upon freezing in A. rufus, but less so in A. ater and A. lusitanicus. Glucose may thus act as a cryoprotectant in these slugs, although the concentrations are not as high as reported for other freeze tolerant invertebrates....

  17. Pulsed electric field in combination with vacuum impregnation with trehalose improves the freezing tolerance of spinach leaves

    OpenAIRE

    Phoon, Pui Yeu; Gómez Galindo, Federico; Vicente, A.A.; Dejmek, Petr

    2008-01-01

    Pulsed electric fields in combination with vacuum infusion have been utilized to impregnate cells with trehalose, aiming at substantially improving the freezing tolerance of spinach leaves. Spinach samples were first treated with ten trains of bi-polar, rectangular electric field pulses with a nominal electric field strength of 580 V/cm and immediately immersed in a 40% (w/w) solution of trehalose under vacuum for 20 min. The samples were kept in the trehalose solution for 2.5 h at a...

  18. Scanning Electron Microscopic Study of Rockfish Preserved at Either Ambient Temperature or by Isothermal Freeze-Fixation

    OpenAIRE

    Lampila, L. E.; Mohr, V.; Reid, D. S.

    1985-01-01

    Fresh rockfish (Bocaccio, Sebastus paucispinis) fillets were blast frozen and stored at either -5 °C or -20 °C for 60 days . At defined sampling intervals, speci mens were removed and chemically fixed at either ambient temperature or isothermally, at the respective storage temperatures (-5 °C or -20°C) . The results indicated that isothermally freeze - fixed specimens showed a degree of crushing and/or distortion of the muscle fiber, the deposition of presumed ice crystals in the cell membran...

  19. 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.

  20. Freeze-drying of emulsified systems: A review.

    Science.gov (United States)

    Morais, Andreza Rochelle do Vale; Alencar, Éverton do Nascimento; Xavier Júnior, Francisco Humberto; de Oliveira, Christian Melo; Marcelino, Henrique Rodrigues; Barratt, Gillian; Fessi, Hatem; do Egito, Eryvaldo Sócrates Tabosa; Elaissari, Abdelhamid

    2016-04-30

    Colloidal systems such as emulsions, microemulsions and nanoemulsions are able to transport active molecules, enhance their solubility and stability and minimize their side effects. However, since they are dispersions with an aqueous continuous phase they have some disadvantages such as the risk of microbiological contamination, degradation by hydrolysis, physico-chemical instability and loss of pharmacological activity of the drug. Freeze drying, in which the water is removed from the preparation by sublimation under vacuum, has been suggested as a means to resolve these problems. Lyophilized products are very stable and are easy to transport and store. However, there is very little information in the literature about the application of this technique to emulsified systems. The aim of this review is to evaluate the lyophilization process as a tool for increasing the shelf life of emulsified systems such as emulsions, microemulsions and nanoemulsions. In addition, the mechanism of cryoprotection and the techniques that can be used to characterize the freeze-dried systems are discussed. PMID:26943974

  1. Dynamics of freeze-thaw embolism in Smilax rotundifolia (Smilacaceae).

    Science.gov (United States)

    Cobb, Alexander R; Choat, Brendan; Holbrook, N Michele

    2007-04-01

    Freeze-thaw cycles pose a major physiological challenge for all temperate perennial plants, but monocotyledonous vines face a still greater risk because their few large vessels are especially susceptible to embolism and are not replaced by secondary growth. The genus Smilax is particularly remarkable because it is widespread in the tropics but includes species that survive the hard frosts of New England winters. Smilax rotundifolia was monitored for a year for evidence of stem xylem freeze-thaw cavitation and refilling. Embolism of metaxylem was complete by late November and was completely reversed by late April, when root pressures rose as high as 100 kPa. Protoxylem remained full of sap throughout the year in cryogenic scanning electron micrographs. Three methods were used to quantify embolism: percent loss conductivity (PLC), gravimetric air fraction (GAF: mass of water in stem xylem relative to capacity), and cryogenic scanning electron microscopy (cryo-SEM). The three methods corroborated one another well and gave quantitatively similar results. Osmolality of xylem sap extracted from exuding stems was 64 mol/kg (±7.0, N = 8), consistent with the root pressures observed. Strong root pressure can account for Smilax's survival in temperate regions with severe frosts, where few monocots with persistent aboveground organs are found. PMID:21636432

  2. Flux-freezing breakdown in high-conductivity magnetohydrodynamic turbulence.

    Science.gov (United States)

    Eyink, Gregory; Vishniac, Ethan; Lalescu, Cristian; Aluie, Hussein; Kanov, Kalin; Bürger, Kai; Burns, Randal; Meneveau, Charles; Szalay, Alexander

    2013-05-23

    The idea of 'frozen-in' magnetic field lines for ideal plasmas is useful to explain diverse astrophysical phenomena, for example the shedding of excess angular momentum from protostars by twisting of field lines frozen into the interstellar medium. Frozen-in field lines, however, preclude the rapid changes in magnetic topology observed at high conductivities, as in solar flares. Microphysical plasma processes are a proposed explanation of the observed high rates, but it is an open question whether such processes can rapidly reconnect astrophysical flux structures much greater in extent than several thousand ion gyroradii. An alternative explanation is that turbulent Richardson advection brings field lines implosively together from distances far apart to separations of the order of gyroradii. Here we report an analysis of a simulation of magnetohydrodynamic turbulence at high conductivity that exhibits Richardson dispersion. This effect of advection in rough velocity fields, which appear non-differentiable in space, leads to line motions that are completely indeterministic or 'spontaneously stochastic', as predicted in analytical studies. The turbulent breakdown of standard flux freezing at scales greater than the ion gyroradius can explain fast reconnection of very large-scale flux structures, both observed (solar flares and coronal mass ejections) and predicted (the inner heliosheath, accretion disks, γ-ray bursts and so on). For laminar plasma flows with smooth velocity fields or for low turbulence intensity, stochastic flux freezing reduces to the usual frozen-in condition.

  3. Freeze-out and coagulation in pre-protostellar collapse

    CERN Document Server

    Flower, D R

    2005-01-01

    We study the changes in physical and chemical conditions during the collapse of a pre--protostellar core, starting from initial conditions appropriate to a dense molecular cloud and proceeding to the ``completely depleted'' limit. We follow the evolution of the ionization degree and the ionic composition as functions of time and density. The processes of freeze--out on to the dust grains and coagulation of the grains were treated simultaneously with the chemical evolution of the medium.When proceeding at close to its maximum rate, coagulation has important consequences for the degree of ionization and the ionic composition of the medium, but its effect on the freeze--out of the neutral species is modest.An innovation of our study is to calculate the grain charge distribution which is significant because H+ ions recombine on the surfaces of negatively charged grains. We also consider the observational result that N-containing species, such as NH3 and N2H+,remain in the gas phase at densities for which CO and o...

  4. Influence of the Ultrasonic Power Applied on Freeze Drying Kinetics

    Science.gov (United States)

    Brines, C.; Mulet, A.; García-Pérez, J. V.; Riera, E.; Cárcel, J. A.

    The atmospheric freeze drying (AFD) constitutes an interesting alternative to vacuum freeze drying providing products with similar quality at lowest cost. However, the long process time needed represent an important drawback. In this sense, the application of high intensity ultrasound can enhance heat and mass transfer and intensify the operation. In hot air drying operation, the ultrasonic effects are dependent on the process variables such as air velocity, internal sample structure or ultrasonic power applied. However, in AFD processes, the internal structure of material or the air velocity has not significant influence on the magnitude of ultrasonic effects. The aim of this work was to determine the influence on drying kinetics of the ultrasonic power applied during the AFD of apple. For that purpose, AFD experiments (-10 °C, 2 m/s and 15% relative humidity) of apple slabs (cv. Granny Smith, 30 x 30 x 10 mm) were carried out with ultrasound application (21 kHz) at different power levels (0, 10.3, 20.5 and 30.8 kW/m3). The drying kinetics was obtained from the initial moisture content and the weight evolution of samples during drying. Experimental results showed a significant (peffective diffusion coefficient identified was 4.8 times higher when ultrasound was applied at the lowest power tested (10.3 kW/m3) that illustrated the high intensification potential of ultrasound application in the AFD.

  5. Chiral Condensate and Mott-Anderson Freeze-Out

    International Nuclear Information System (INIS)

    We present the idea of a Mott-Anderson freeze-out that suggests a key role of the localization of the hadron wave functions when traversing the hadronization transition. The extension of hadron wave functions in dense matter is governed by the behavior of the chiral quark condensate such that its melting at finite temperatures and chemical potentials entails an increase of the size of hadrons and thus their geometrical strong interaction cross sections. It is demonstrated within a schematic resonance gas model, that a kinetic freeze-out condition reveals a correlation with the reduction of the chiral condensate in the phase diagram up to 50% of its vacuum value. Generalizing the description of the chiral condensate by taking into account a full hadron resonance gas such correlation gets distorted. We discuss, that this may be due to our approximations in calculating the chiral condensate which disregard both, in-medium effects on hadron masses and hadron-hadron interactions. The latter, in particular due to quark exchange reactions, could lead to a delocalization of the hadron wave functions in accordance with the picture of a Mott-Anderson transition. (author)

  6. Ice encapsulation protects rather than disturbs the freezing lichen.

    Science.gov (United States)

    Bjerke, J W

    2009-03-01

    Arctic and alpine terricolous lichens are adapted to harsh environments and are tolerant to extremely low temperatures when metabolically inactive. However, there are reports indicating that freezing can be lethal to metabolically active lichens. With a projected warmer and more unstable climate, winter precipitation at high latitudes will fall more frequently as rain, causing snowmelt and encapsulating terricolous lichens in ice or exposing them to large temperature fluctuations. Lichens are a major winter food source for reindeer in most parts of the circumpolar region. A laboratory experiment tested how three hydrated reindeer forage lichen species covered by snow, encapsulated in ice, or uncovered responded to storage at freezing temperatures and subsequent warming. Photosynthetic performance (maximal fluorescence of dark-adapted samples and net photosynthetic rates) was significantly lower in lichens not insulated by snow or ice, whereas there were few differences between the snow and ice treatments. It is suggested that snow and ice provide sufficiently moist environments to improve extracellular and reduce intracellular ice nucleation activity. Ice encapsulation, which is often lethal to vascular plants, did not have any negative effects on the studied lichens. The results indicate that complete snow and ice melt followed by refreezing can be detrimental to terricolous lichen ecosystems. Reduced lichen biomass will have a negative effect both on reindeer winter survival and the indigenous peoples who herd reindeer. PMID:19228329

  7. A New Mechanism of Canopy Effect in Unsaturated Freezing Soils

    Directory of Open Access Journals (Sweden)

    Teng Jidong

    2016-01-01

    Full Text Available Canopy effect refers to the phenomenon where moisture accumulates underneath an impervious cover. Field observation reveals that canopy effect can take place in relatively dry soils where the groundwater table is deep and can lead to full saturation of the soil immediately underneath the impervious cover. On the other hand, numerical analysis based on existing theories of heat and mass transfer in unsaturated soils can only reproduce a minor amount of moisture accumulation due to an impervious cover, particularly when the groundwater table is relatively deep. In attempt to explain the observed canopy effect in field, this paper proposes a new mechanism of moisture accumulation in unsaturated freezing soils: vapour transfer in such a soil is accelerated by the process of vapour-ice desublimation. A new approach for modelling moisture and heat movements is proposed, in which the phase change of evaporation, condensation and de-sublimation of vapor flow are taken into account. The computed results show that the proposed model can indeed reproduce the unusual moisture accumulation observed in relatively dry soils. The results also demonstrate that soil freezing fed by vapour transfer can result in a water content close to full saturation. Since vapour transfer is seldom considered in geotechnical design, the canopy effect deserves more attention during construction and earth works in cold and arid regions.

  8. A new portable device for automatic controlled-gradient cryopreservation of blood mononuclear cells

    DEFF Research Database (Denmark)

    Hviid, L; Albeck, G; Hansen, B;

    1993-01-01

    Protection of the functional integrity of mononuclear cells stored in liquid N2 requires careful control of the freezing procedure. Consequently, optimal quality of cryopreserved cells is usually assured by freezing according to a specified time-temperature gradient generated by computer-controll......Protection of the functional integrity of mononuclear cells stored in liquid N2 requires careful control of the freezing procedure. Consequently, optimal quality of cryopreserved cells is usually assured by freezing according to a specified time-temperature gradient generated by computer......-controlled freezing devices. While such equipment offers large capacity and secures maximum survival and functional integrity of the lymphocytes upon thawing, it is quite costly and strictly stationary. We have previously developed and tested an alternative, manual device for controlled-gradient lymphocyte freezing...

  9. Measurements of the salt-removal of NaCl, KCl and MgCl using a carbon electrode prepared with freezing thawing method in capacitive deionization

    Science.gov (United States)

    Endarko, Sari, Intan Permata; Fatimah, Iim

    2016-04-01

    Carbon electrodes prepared with freezing thawing method for desalination purpose has been synthesized and characterized. The carbon electrodes were prepared with an activated carbon (700 - 1400 m2/g) and polyvinyl alcohol (PVA) as a binder using freezing thawing method with 3 and 4 cycles (1 cycle is 12 hours for freezing and 12 hours for thawing). Electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were used to analyze their electrochemical properties. The main study was to measure the salt-removal of 180 µS/cm NaCl, MgCl and KCl using a capacitive deionization (CDI) unit cell with five pairs of carbon electrodes. The applied potential of 2.0 V and a flow rate of 25 mL/min were used to desalination tests, the result showed that the salt-removal percentage of KCl solution has greater than NaCl and MgCl. The highest value for the salt-removal of NaCl, KCl and MgCl can be achieved for the freezing thawing method with 4 cycles. The salt-removal percentage of KCl was achieved at 64.10% whilst resulted in 54.30 and 54.47 % for NaCl and MgCl, respectively.

  10. Assessment of freshness and freeze-thawing of sea bream fillets (Sparus aurata) by a cytosolic enzyme: Lactate dehydrogenase.

    Science.gov (United States)

    Diop, Mamadou; Watier, Denis; Masson, Pierre-Yves; Diouf, Amadou; Amara, Rachid; Grard, Thierry; Lencel, Philippe

    2016-11-01

    The evaluation of freshness and freeze-thawing of fish fillets was carried out by assessment of autolysis of cells using a cytosolic enzyme lactate dehydrogenase. Autolysis plays an important role in spoilage of fish and postmortem changes in fish tissue are due to the breakdown of the cellular structures and release of cytoplasmic contents. The outflow of a cytosolic enzyme, lactate dehydrogenase, was studied in sea bream fillets and the Sparus aurata fibroblasts (SAF-1) cell-line during an 8day storage period at +4°C. A significant increase of lactate dehydrogenase release was observed, especially after 5days of storage. The ratio between the free and the total lactate dehydrogenase activity is a promising predictive marker to measure the quality of fresh fish fillets. The effect of freeze-thawing on cytosolic lactate dehydrogenase and lysosomal α-d-glucosidase activities was also tested. Despite the protecting effect of the tissue compared to the cell-line, a loss of lactate dehydrogenase activity, but not of α-d-glucosidase, was observed. In conclusion, lactate dehydrogenase may be used as a marker to both assess freshness of fish and distinguish between fresh and frozen-thawed fish fillets. PMID:27211667

  11. Freeze-dried eudragit-hyaluronan multicompartment liposomes to improve the intestinal bioavailability of curcumin.

    Science.gov (United States)

    Catalan-Latorre, Ana; Ravaghi, Maryam; Manca, Maria Letizia; Caddeo, Carla; Marongiu, Francesca; Ennas, Guido; Escribano-Ferrer, Elvira; Peris, José Esteban; Diez-Sales, Octavio; Fadda, Anna Maria; Manconi, Maria

    2016-10-01

    This work aimed at finding an innovative vesicle-type formulation able to improve the bioavailability of curcumin upon oral administration. To this purpose, phospholipid, Eudragit® S100 and hyaluronan sodium salt were combined to obtain eudragit-hyaluronan immobilized vesicles using an easy and environmentally-friendly method. For the first time, the two polymers were combined in a system intended for oral delivery, to enhance curcumin stability when facing the harsh environment of the gastrointestinal tract. Four different formulations were prepared, keeping constant the amount of the phospholipid and varying the eudragit-hyaluronan ratio. The freeze-drying of the samples, performed to increase their stability, led to a reduction of vesicle size and a good homogeneity of the systems, after simple rehydration with water. X-ray diffraction study demonstrated that after the freeze-drying process, curcumin remained successfully incorporated within the vesicles. All the vesicles displayed similar features: size ranging from 220 to 287nm, spherical or oval shape, multilamellar or large unilamellar morphology with a peculiar multicompartment organization involving 1-4 smaller vesicles inside. In vitro studies demonstrated the ability of the combined polymers to protect the vesicles from the harsh conditions of the gastro-intestinal tract (i.e., ionic strength and pH variation), which was confirmed in vivo by the greater deposition of curcumin in the intestinal region, as compared to the free drug in dispersion. This enhanced accumulation of curcumin provided by the eudragit-hyaluronan immobilized vesicles, together with an increase in Caco-2 cell viability exposed to hydrogen peroxide, indicated that vesicles can ensure a local protection against oxidative stress and an increase in its intestinal absorption. PMID:27349806

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

    Energy Technology Data Exchange (ETDEWEB)

    Iijima, Mika, E-mail: m-iijima@nagasaki-u.ac.jp [Faculty of Education, Nagasaki University, 1-14, Bunkyo-machi, Nagasaki 852-8521 (Japan); Hatakeyama, Tatsuko [Lignocell Research, 73-8 Yatsumata, Fukui 910-3558 (Japan); Hatakeyama, Hyoe [Department of Environment and Biotechnology, Faculty of Engineering, Fukui University of Technology, 3-6-1, Gakuen, Fukui 910-8505 (Japan)

    2012-03-20

    Research highlights: Black-Right-Pointing-Pointer Locust bean gum forms hydrogels by freezing and thawing. Black-Right-Pointing-Pointer Syneresis was observed when freezing and thawing cycle (n) increased. Black-Right-Pointing-Pointer Dynamic Young's modulus increased with increasing n. Black-Right-Pointing-Pointer Non-freezing water content restrained by hydrogels decreased with increasing n. Black-Right-Pointing-Pointer 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 Prime ) 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.

  13. A spatiotemporal analysis of gait freezing and the impact of pedunculopontine nucleus stimulation

    Science.gov (United States)

    Cole, Michael H.; Graepel, Cara L.; Hyam, Jonathan A.; Jenkinson, Ned; Brittain, John-Stuart; Coyne, Terry J.; Silburn, Peter A.; Aziz, Tipu Z.; Kerr, Graham; Brown, Peter

    2012-01-01

    Gait freezing is an episodic arrest of locomotion due to an inability to take normal steps. Pedunculopontine nucleus stimulation is an emerging therapy proposed to improve gait freezing, even where refractory to medication. However, the efficacy and precise effects of pedunculopontine nucleus stimulation on Parkinsonian gait disturbance are not established. The clinical application of this new therapy is controversial and it is unknown if bilateral stimulation is more effective than unilateral. Here, in a double-blinded study using objective spatiotemporal gait analysis, we assessed the impact of unilateral and bilateral pedunculopontine nucleus stimulation on triggered episodes of gait freezing and on background deficits of unconstrained gait in Parkinson’s disease. Under experimental conditions, while OFF medication, Parkinsonian patients with severe gait freezing implanted with pedunculopontine nucleus stimulators below the pontomesencephalic junction were assessed during three conditions; off stimulation, unilateral stimulation and bilateral stimulation. Results were compared to Parkinsonian patients without gait freezing matched for disease severity and healthy controls. Pedunculopontine nucleus stimulation improved objective measures of gait freezing, with bilateral stimulation more effective than unilateral. During unconstrained walking, Parkinsonian patients who experience gait freezing had reduced step length and increased step length variability compared to patients without gait freezing; however, these deficits were unchanged by pedunculopontine nucleus stimulation. Chronic pedunculopontine nucleus stimulation improved Freezing of Gait Questionnaire scores, reflecting a reduction of the freezing encountered in patients’ usual environments and medication states. This study provides objective, double-blinded evidence that in a specific subgroup of Parkinsonian patients, stimulation of a caudal pedunculopontine nucleus region selectively improves gait

  14. Arginine and proline applied as food additives stimulate high freeze tolerance in larvae of Drosophila melanogaster.

    Science.gov (United States)

    Koštál, Vladimír; Korbelová, Jaroslava; Poupardin, Rodolphe; Moos, Martin; Šimek, Petr

    2016-08-01

    The fruit fly Drosophila melanogaster is an insect of tropical origin. Its larval stage is evolutionarily adapted for rapid growth and development under warm conditions and shows high sensitivity to cold. In this study, we further developed an optimal acclimation and freezing protocol that significantly improves larval freeze tolerance (an ability to survive at -5°C when most of the freezable fraction of water is converted to ice). Using the optimal protocol, freeze survival to adult stage increased from 0.7% to 12.6% in the larvae fed standard diet (agar, sugar, yeast, cornmeal). Next, we fed the larvae diets augmented with 31 different amino compounds, administered in different concentrations, and observed their effects on larval metabolomic composition, viability, rate of development and freeze tolerance. While some diet additives were toxic, others showed positive effects on freeze tolerance. Statistical correlation revealed tight association between high freeze tolerance and high levels of amino compounds involved in arginine and proline metabolism. Proline- and arginine-augmented diets showed the highest potential, improving freeze survival to 42.1% and 50.6%, respectively. Two plausible mechanisms by which high concentrations of proline and arginine might stimulate high freeze tolerance are discussed: (i) proline, probably in combination with trehalose, could reduce partial unfolding of proteins and prevent membrane fusions in the larvae exposed to thermal stress (prior to freezing) or during freeze dehydration; (ii) both arginine and proline are exceptional among amino compounds in their ability to form supramolecular aggregates which probably bind partially unfolded proteins and inhibit their aggregation under increasing freeze dehydration. PMID:27489218

  15. Freeze/thaw-induced embolism: probability of critical bubble formation depends on speed of ice formation

    Directory of Open Access Journals (Sweden)

    Sanna eSevanto

    2012-06-01

    Full Text Available Bubble formation in the conduits of woody plants sets a challenge for uninterrupted water transportation from the soil up to the canopy. Freezing and thawing of stems has been shown to increase the number of air-filled (embolized conduits, especially in trees with large conduit diameters. Despite numerous experimental studies, the mechanisms leading to bubble formation during freezing have not been addressed theoretically. We used classical nucleation theory and fluid mechanics to show which mechanisms are most likely to be responsible for bubble formation during freezing and what parameters determine the likelihood of the process. Our results confirm the common assumption that bubble formation during freezing is most likely due to gas segregation by ice. If xylem conduit walls are not permeable to the salts expelled by ice during the freezing process, osmotic pressures high enough for air seeding could be created. The build-up rate of segregated solutes in front of the ice-water interface depends equally on conduit diameter and freezing velocity. Therefore, bubble formation probability depends on these variables. The dependence of bubble formation probability on freezing velocity means that the experimental results obtained for cavitation threshold conduit diameters during freeze/thaw cycles depend on the experimental setup; namely sample size and cooling rate. The velocity dependence also suggests that to avoid bubble formation during freezing trees should have narrow conduits where freezing is likely to be fast (e.g. branches or outermost layer of the xylem. Avoidance of bubble formation during freezing could thus be one piece of the explanation why xylem conduit size of temperate and boreal zone trees varies quite systematically.

  16. Freezing of gait in Parkinson's disease is improved by treatment with weak electromagnetic fields.

    Science.gov (United States)

    Sandyk, R

    1996-03-01

    Freezing, a symptom characterized by difficulty in the initiation and smooth pursuit of repetitive movements, is a unique and well known clinical feature of Parkinson's disease (PD). It usually occurs in patients with long duration and advanced stage of the disease and is a major cause of disability often resulting in falling. In PD patients freezing manifests most commonly as a sudden attack of immobility usually experienced during walking, attempts to turn while walking, or while approaching a destination. Less commonly it is expressed as arrest of speech or handwriting. The pathophysiology of Parkinsonian freezing, which is considered a distinct clinical feature independent of akinesia, is poorly understood and is believed to involve abnormalities in dopamine and norepinephrine neurotransmission in critical motor control areas including the frontal lobe, basal ganglia, locus coeruleus and spinal cord. In general, freezing is resistant to pharmacological therapy although in some patients reduction or increase in levodopa dose may improve this symptom. Three medicated PD patients exhibiting disabling episodes of freezing of gait are presented in whom brief, extracerebral applications of pulsed electromagnetic fields (EMFs) in the picotesla range improved freezing. Two patients had freezing both during "on" and "off" periods while the third patient experienced random episodes of freezing throughout the course of the day. The effect of each EMFs treatment lasted several days after which time freezing gradually reappeared, initially in association with "off" periods. These findings suggest that the neurochemical mechanisms underlying the development of freezing are sensitive to the effects of EMFs, which are believed to improve freezing primarily through the facilitation of serotonin (5-HT) neurotransmission at both junctional (synaptic) and nonjunctional neuronal target sites. PMID:8727687

  17. Effect of freezing and thawing rates on sperm motility in Bocachico Prochilodus magdalenae (Pisces, Characiformes

    Directory of Open Access Journals (Sweden)

    José G. Martínez

    2013-03-01

    Full Text Available Objective. To determine the freezing and thawing rates necessary to maintain sperm viability during cryopreservation of Bocachico semen. Materials and methods. Four interactional treatments were implemented between two freezing (rapid and slow and two thawing (rapid and slow curves, in a 2x2 factorial as follows: rapid freezing-rapid thawing, rapid freezing-slow thawing, slow freezing-rapid thawing, and slow freezing-slow thawing. After thawing by Sperm Class Analyzer (SCA curvilinear velocity (VCL and straight-line (VSL (μm sec-1 were analyzed; total, rapid, medium, and slow motility, were compared among treatments. Results. The rapid freezing-slow thawing treatment was lethal for all variables of velocity and motility, causing a significant (p<0.01 post-thaw inmotility of 100%. The slow freezing-rapid thawing interaction had a significantly higher effect than the other treatments (p<0.05, particularly on variables such as rapid motility (10.1 ± 1.1%, medium motility (30.16 ± 4.1%, and curvilinear velocity (51.5 ± 4.75 μm sec.-1 also decreased the percentage of sperm with slow motility (41.7 ± 4.45%. Independently of the applied thawing rate, the freezing rate generated the main significant effect on total motility. Conclusions. It is possible to conclude that the interaction effect between freezing and thawing rates is nil (except for slow motility during cryopreservation process. However, the independent effects of these factors (main effects on remaining motility variables are positively significant and decisive to the maintenance of these features, especially the freeze factor (when it is slow. This becomes the first successful report of sperm cryopreservation from Bocachico Prochilodus magdalenae in the world and may be used in conservation programs for this endangered species.

  18. Stress-induced activation of the AMP-activated protein kinase in the freeze-tolerant frog Rana sylvatica.

    Science.gov (United States)

    Rider, Mark H; Hussain, Nusrat; Horman, Sandrine; Dilworth, Stephen M; Storey, Kenneth B

    2006-12-01

    Survival in the frozen state depends on biochemical adaptations that deal with multiple stresses on cells including long-term ischaemia and tissue dehydration. We investigated whether the AMP-activated protein kinase (AMPK) could play a regulatory role in the metabolic re-sculpting that occurs during freezing. AMPK activity and the phosphorylation state of translation factors were measured in liver and skeletal muscle of wood frogs (Rana sylvatica) subjected to anoxia, dehydration, freezing, and thawing after freezing. AMPK activity was increased 2-fold in livers of frozen frogs compared with the controls whereas in skeletal muscle, AMPK activity increased 2.5-, 4.5- and 3-fold in dehydrated, frozen and frozen/thawed animals, respectively. Immunoblotting with phospho-specific antibodies revealed an increase in the phosphorylation state of eukaryotic elongation factor-2 at the inactivating Thr56 site in livers from frozen frogs and in skeletal muscles of anoxic frogs. No change in phosphorylation state of eukaryotic initiation factor-2alpha at the inactivating Ser51 site was seen in the tissues under any of the stress conditions. Surprisingly, ribosomal protein S6 phosphorylation was increased 2-fold in livers from frozen frogs and 10-fold in skeletal muscle from frozen/thawed animals. However, no change in translation capacity was detected in cell-free translation assays with skeletal muscle extracts under any of the experimental conditions. The changes in phosphorylation state of translation factors are discussed in relation to the control of protein synthesis and stress-induced AMPK activation. PMID:16973146

  19. 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

  20. PECULIARITIES OF WATER FREEZING IN CRYOPROTECTIVE MEDIUM IMPLEMENTED IN A MATRIX OF HYDROPHOBIC SILICA BULL SPERM

    Directory of Open Access Journals (Sweden)

    V. V. Turov

    2014-06-01

    Full Text Available The study of the process of melting water in lactose-glycerol-yolk kriomedium containing gametes bull, incorporated in the hydrophobic silica powder, which are adsorbed on the surface of fixed amounts of nonpolar hydrocarbon – n-decane was the aim of the work. The possibility of water polyassociates structuring with a solid surface of interfacial water and solubility of trifluoroethanoic acid in it have been studied. Thereat survival of the germ cell after contact with the surface was not analyzed. State of water in initial cryoprotective glycerol-lactose-yolk medium and hydrophobic nanosilica TS-100 containing n-decane additive adsorbed on its surface incorporated in a matrix was studied using low-temperature 1H-NMR spectroscopy method. It is shown that the solid matrix induces formation of 6–7 water molecules per each dean molecule at the interface, which do not take part in formation of hydrogen bonds, and a sharp radius decrease (from 100 to 20 nm of ice crystals formed in cell suspension at its freezing. The results could give rise to safety improving of their cells at their cryopreservation and low temperature storage conditions by incorporating into a powder composite environment.