Sample records for bk060042 cell freeze

  1. Improvement In Rabbit Corneal Cell Suspension Viability After Freezing With Gingko Biloba Extrakt

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    Murad Aktan


    Full Text Available We investigated whether the addition of Gingko Biloba extract (EGb 761 to rabbit corneal epithelial medium before cell freezing improved cell viability after freezing then thawing. After removal of corneas, they were treated with enzymes and the corneal epithelium was prepared as a single cell suspension in freezing media with or without EGb 761. After freezing for two weeks then thawing, a higher cell viability was found in the cornea cell suspensions which had been frozen pretreated with EGb 761 in the media. The improvement with corneal cell viability with EGb 761 pretreatment is postulated to be based on the antioxidant capacity of the plant extract.

  2. Freeze drying of red blood cells: the use of directional freezing and a new radio frequency lyophilization device. (United States)

    Arav, Amir; Natan, Dity


    Red blood cell (RBC) units are administered routinely into patients expressing a wide range of acute and chronic conditions (e.g., anemia, traumatic bleeding, chronic diseases, and surgery). The modern blood banking system has been designed to answer this need and assure a continuous, high quality blood supply to patients. However, RBCs units can be stored under hypothermic conditions for only up to 42 days, which leads to periodic shortages. Cryopreservation can solve these shortages, but current freezing methods employ high glycerol concentrations, which need to be removed and the cells washed prior to transfusion, resulting in a long (more than 1 hour) and cumbersome washing step. Thus, frozen RBCs have limited use in acute and trauma situations. In addition, transportation of frozen samples is complicated and costly. Freeze drying (lyophilization) of RBCs has been suggested as a solution for these problems, since it will allow for a low weight sample to be stored at room temperature, but reaching this goal is not a simple task. We studied the effect of different solutions (IMT2 and IMT3) containing trehalose and antioxidants or trehalose and human serum albumin, respectively, on freezing/thawing and freeze drying of RBCs. In addition, we evaluated the effect of cells concentrations and cooling rates on the post thaw and post rehydration recoveries of the RBCs. Finally, we developed a new radio frequency (RF) lyophilization device for a more rapid and homogeneous sublimation process of the frozen RBCs samples. Recovery and free Hb were measured as well as oxygen association/dissociation and cell's deformability. We found that IMT3 (0.3 M trehalose and 10% HSA) solution that was directionally frozen at a rapid interface velocity of 1 mm/sec (resulting in a cooling rate of 150°C/min) yielded the best results (better than IMT2 solution and slow interface velocity). Freeze thawing gave 100% survival, while freeze drying followed by rehydration with 20% dextran-40k

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

    International Nuclear Information System (INIS)

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


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

  4. Cryopreservation of putative pre-pubertal bovine spermatogonial stem cells by slow freezing. (United States)

    Kim, Ki-Jung; Lee, Yong-An; Kim, Bang-Jin; Kim, Yong-Hee; Kim, Byung-Gak; Kang, Hyun-Gu; Jung, Sang-Eun; Choi, Sun-Ho; Schmidt, Jonathan A; Ryu, Buom-Yong


    Development of techniques for the preservation of mammalian spermatogonial stem cells (SSCs) is a critical step in commercial application of SSC based technologies, including species preservation, amplification of agriculturally valuable germ lines, and human fertility preservations. The objective of this study was to develop an efficient cryopreservation protocol for preservation of bovine SSCs using a slow freezing technique. To maximize the efficiency of SSC cryopreservation, the effects of various methods (tissue vs. cell freezing) and cryoprotective agents (trehalose, sucrose, and polyethylene glycol [PEG]) were tested. Following thawing, cells were enriched for undifferentiated spermatogonia by differential plating and evaluated for recovery rate, proliferation capacity, and apoptosis. Additionally, putative stem cell activity was assessed using SSC xenotransplantation. The recovery rate, and proliferation capacity of undifferentiated spermatogonia were significantly greater for germ cells frozen using tissue freezing methods compared to cell freezing methods. Cryopreservation in the presence of 200 mM trehalose resulted in significantly greater recovery rate, proliferation capacity, and apoptosis of germ cells compared to control. Furthermore, cryopreservation using the tissue freezing method in the presence of 200 mM trehalose resulted in the production of colonies of donor-derived germ cells after xenotransplantation into recipient mouse testes, indicating putative stem cell function. Collectively, these data indicate that cryopreservation using tissue freezing methods in the presence of 200 mM trehalose is an efficient cryopreservation protocol for bovine SSCs. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Preliminary study on the freeze-drying of human bone marrow-derived mesenchymal stem cells* (United States)

    Zhang, Shao-zhi; Qian, Huan; Wang, Zhen; Fan, Ju-li; Zhou, Qian; Chen, Guang-ming; Li, Rui; Fu, Shan; Sun, Jie


    Long-term preservation and easy transportation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) will facilitate their application in medical treatment and bioengineering. A pilot study on the freeze-drying of hBM-MSCs was carried out. hBM-MSCs were loaded with trehalose. The glass transition temperature of the freeze-drying suspension was measured to provide information for the cooling and primary drying experiment. After freeze-drying, various rehydration processes were tested. The highest recovery rate of hBM-MSCs was (69.33±13.08)%. Possible methods to improve freeze-drying outcomes are discussed. In conclusion, the present study has laid a foundation for the freeze-drying hBM-MSCs. PMID:21043058

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

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    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.

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

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    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%.

  8. Ultrastructural and nuclear antigen preservation after high-pressure freezing/freeze-substitution and low-temperature LR White embedding of HeLa cells

    Czech Academy of Sciences Publication Activity Database

    Strádalová, Vendula; Kyselá, Katarína; Hozák, Pavel

    Roč. 130, - ( 2008 ), s. 1047-1052 ISSN 0948-6143 R&D Projects: GA AV ČR KAN200520704; GA MŠk LC545; GA MŠk 2B06063; GA ČR(CZ) GD204/05/H023 Institutional research plan: CEZ:AV0Z50390512; CEZ:AV0Z50520514 Keywords : Cryofixation * Freeze substitution * High-presszre freezing Subject RIV: EA - Cell Biology Impact factor: 2.320, year: 2008

  9. Proline: A Novel Cryoprotectant for the Freeze Preservation of Cultured Cells of Zea mays L (United States)

    Withers, Lyndsey A.; King, Patrick J.


    Proline is an effective cryoprotectant for the storage of cultured cells of Zea mays L. in liquid N2. Increased freeze tolerance can be achieved by pregrowth for 3 to 4 days in medium containing proline. Cells cryoprotected with proline have an increased recovery potential when compared with cells cryoprotected with dimethylsulfoxide and glycerol. They also show a reduced postthaw viability loss and greater tolerance of a range of postthaw culture conditions. It is suggested that the mechanism of action of proline may be similar to that in its putative role of conferring protection against natural stresses. It may be protecting the cell against solution effects caused by dehydration during freezing. These findings are discussed in relation to other freeze tolerance enhancing treatments. PMID:16661033

  10. The contribution of apoptosis and necrosis in freezing injury of sea urchin embryonic cells. (United States)

    Boroda, Andrey V; Kipryushina, Yulia O; Yakovlev, Konstantin V; Odintsova, Nelly A


    Sea urchins have recently been reported to be a promising tool for investigations of oxidative stress, UV light perturbations and senescence. However, few available data describe the pathway of cell death that occurs in sea urchin embryonic cells after cryopreservation. Our study is focused on the morphological and functional alterations that occur in cells of these animals during the induction of different cell death pathways in response to cold injury. To estimate the effect of cryopreservation on sea urchin cell cultures and identify the involved cell death pathways, we analyzed cell viability (via trypan blue exclusion test, MTT assay and DAPI staining), caspase activity (via flow cytometry and spectrophotometry), the level of apoptosis (via annexin V-FITC staining), and cell ultrastructure alterations (via transmission electron microscopy). Using general caspase detection, we found that the level of caspase activity was low in unfrozen control cells, whereas the number of apoptotic cells with activated caspases rose after freezing-thawing depending on cryoprotectants used, also as the number of dead cells and cells in a late apoptosis. The data using annexin V-binding assay revealed a very high apoptosis level in all tested samples, even in unfrozen cells (about 66%). Thus, annexin V assay appears to be unsuitable for sea urchin embryonic cells. Typical necrotic cells with damaged mitochondria were not detected after freezing in sea urchin cell cultures. Our results assume that physical cell disruption but not freezing-induced apoptosis or necrosis is the predominant reason of cell death in sea urchin cultures after freezing-thawing with any cryoprotectant combination. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Cell growth and resistance of Lactococcus lactis subsp. lactis TOMSC161 following freezing, drying and freeze-dried storage are differentially affected by fermentation conditions. (United States)

    Velly, H; Fonseca, F; Passot, S; Delacroix-Buchet, A; Bouix, M


    To investigate the effects of fermentation parameters on the cell growth and on the resistance to each step of the freeze-drying process of Lactococcus lactis subsp. lactis TOMSC161, a natural cheese isolate, using a response surface methodology. Cells were cultivated at different temperatures (22, 30 and 38°C) and pH (5·6, 6·2 and 6·8) and were harvested at different growth phases (0, 3 and 6 h of stationary phase). Cultivability and acidification activity losses of Lc. lactis were quantified after freezing, drying, 1 and 3 months of storage at 4 and 25°C. Lactococcus lactis was not damaged by freezing but was sensitive to drying and to ambient temperature storage. Moreover, the fermentation temperature and the harvesting time influenced the drying resistance of Lc. lactis. Lactococcus lactis cells grown in a whey-based medium at 32°C, pH 6·2 and harvested at late stationary phase exhibited both an optimal growth and the highest resistance to freeze-drying and storage. A better insight on the individual and interaction effects of fermentation parameters made it possible the freeze-drying and storage preservation of a sensitive strain of technological interest. Evidence on the particularly damaging effect of the drying step and the high-temperature storage is presented. © 2014 The Society for Applied Microbiology.

  12. Freezing tolerance of sea urchin embryonic cells: Differentiation commitment and cytoskeletal disturbances in culture. (United States)

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


    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%). Copyright © 2015 Elsevier Inc. All rights reserved.

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

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    Thirumala, Sreedhar [Department of Mechanical Engineering, Louisiana State University, Baton Rouge, LA (United States); Forman, Julianne M [Department of Biological and Agricultural Engineering, Louisiana State University and Agricultural Center, Baton Rouge, LA (United States); Monroe, W Todd [Department of Biological and Agricultural Engineering, Louisiana State University and Agricultural Center, Baton Rouge, LA (United States); Devireddy, Ram V [Department of Mechanical Engineering, Louisiana State University, Baton Rouge, LA (United States)


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

  14. The effect of isochoric freezing on mammalian cells in an extracellular phosphate buffered solution. (United States)

    Preciado, J; Rubinsky, B


    Isochoric (constant volume) freezing has been recently suggested as a new method for cell and organ preservation. As a first step in studying the effect of isochoric freezing on mammalian cells, Madin-Darby canine kidney epithelial cells (MDCK), were frozen in an isochoric system, in a simple extracellular phosphate buffered solution to -10 °C (96.5 MPa), - 15 °C (162 MPa) and -20 °C (205 MPa) for 60 and 120 min. Cell membrane integrity and cell metabolism were studied with a Live/Dead cell vitality assay and flow cytometry. We found that cell survival decreases with an increase in pressure (lower temperatures) and time of exposure. For example, 60% of cells survived 60 min at - 10 °C and only 18% survived 120 min at this temperature. Negligible survival was measured at - 20 °C. This study may serve as the baseline towards further research on techniques to optimize the effects of isochoric freezing on living biological matter. Copyright © 2018. Published by Elsevier Inc.

  15. Enhanced resolution of membranes in cultured cells by cryoimmobilization and freeze-substitution. (United States)

    Wild, P; Schraner, E M; Adler, H; Humbel, B M


    Investigations of cellular processes demand immediate arresting of the process at any given time and excellent retention of cellular material and excellent visibility of membranes. To achieve this goal we used cryofixation to arrest cellular processes instantly and tested diverse freeze-substitution protocols. Madin-Darby kidney cells and Vero cells were grown on carbon-coated sapphire disks. For cryofixation the sapphire disks covered with a cell monolayer were injected with the aid of a guillotine into liquid propane or ethane or a mixture of both cooled by liquid nitrogen. Freezing of the cryogen was prevented by using a partially insulated cylinder and by vigorous stirring that results in a substantial decrement of the freezing point of the cryogen. Cell monolayers can be cryofixed successfully using the guillotine in a safety hood at ambient temperature and humidity or at 37 degrees C and 45% humidity. The freezing unit can also be placed in a laminar flow for working under biohazard conditions. For visualizing cell membranes at high contrast and high resolution, cells were substituted in the presence of various concentrations of glutaraldehyde and osmium tetroxide and the temperature was raised to diverse final temperatures. Substitution for 4 hours at -90 degrees C in anhydrous acetone containing 0.25% anhydrous glutaraldehyde and 0.5% osmium tetroxide followed by a temperature rise of 5 degrees C/hour to 0 degrees C and final incubation for 1 hour at 0 degrees C resulted in high contrast and excellent visibility of subcellular components at the level of the membrane bilayer. The high spatial and temporal resolution makes this methodology an excellent tool for studying cell membrane-bound processes, such as virus-cell interactions. Copyright 2001 Wiley-Liss, Inc.

  16. GMP cryopreservation of large volumes of cells for regenerative medicine: active control of the freezing process. (United States)

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


    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

  17. Effect of Glycerol, as Cryoprotectant in the Encapsulation and Freeze Drying of Microspheres Containing Probiotic Cells

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    Oana Lelia Pop


    Full Text Available It is reported that probiotics provide several health benefits as they help in maintaining a good balance and composition of intestinal flora, and increase the resistance against invasion of pathogens. Ensuring adequate dosages of probiotics at the time of consumption is a challenge, because several factors during processing and storage affect the viability of probiotic organisms. Major emphasis has been given to protect the microorganisms with the help of encapsulation technique, by addition of different protectants. In this study, probiotic cells (Bifidobacterium lactis 300B were entrapped in alginate/pullulan microspheres. In the encapsulation formula glycerol was used as cryoprotectant in the freeze drying process for long time storage. It was observed that the survival of Bifidobacterium lactis 300B when encapsulated without cryoprotectant was higher than the formula with glycerol in the fresh obtained microspheres. The addition of glycerol was in order to reduce the deep freezing and freeze drying damages. In the chosen formulations, glycerol did not proved protection for the entrapped probiotic cells in the freeze drying process, for which the use of glycerol as cryoprotectant for alginate/pullulan Bifidobacterium lactis 300B entrapment is not recommended.

  18. [Optimization of lyophilization procedures for freeze-drying of human red blood cells]. (United States)

    Chen, Lin-feng; Liu, Jing-han; Wang, De-qing; Ouyang, Xi-lin; Zhuang, Yuan; Che, Ji; Yu, Yang; Li, Hui


    To investigate the different parameters of the lyophilization procedures that affect the recovery of the rehydrated red blood cells (RBCs). Human RBCs loaded in tubes were cooled with 4 different modes and subjected to water bath at 25 degrees celsius;. The morphological changes of the RBCs were observed to assess the degree of vitrification, and the specimens were placed in the freeze-dryer with the temperature set up at 40, -50, -60, -70 and -80 degrees celsius;. The rates of temperature rise of the main and secondary drying in the lyophilization procedures were compared, and the water residue in the specimens was determined. The protectant did not show ice crystal in the course of freezing and thawing. No significant difference was found in the recovery rate of the rehydrated RBCs freeze-dried at the minimum temperature of -70 degrees celsius; and -80 degrees celsius; (P > 0.05). The E procedure resulted in the maximum recovery of the RBCs (83.14% ± 9.55%) and Hb (85.33% ± 11.42%), showing significant differences from the other groups(P < 0.01 or 0.05). The recovery of the RBCs showed a positive correlation to the water residue in the samples. Fast cooling in liquid nitrogen and shelf precooling at -70 degrees celsius; with a moderate rate of temperature rise in lyophylization and a start dry temperature close to the shelf equilibrium temperature produce optimal freeze-drying result of human RBCs.

  19. Cryo-scanning electron microscopic study on freezing behavior of xylem ray parenchyma cells in hardwood species (United States)

    Fujikawa; Kuroda


    Differential thermal analysis (DTA) has indicated that xylem ray parenchyma cells (XRPCs) of hardwood species adapt to freezing of apoplastic water either by deep supercooling or by extracellular freezing, depending upon the species. DTA studies indicated that moderately cold hardy hardwood species exhibiting deep supercooling in the XRPCs were limited in latitudinal distribution within the -40 degrees C isotherm, while very hardy hardwood species exhibiting extracellular freezing could distribute in colder areas beyond the -40 degrees C isotherm. Predictions based on the results of DTA, however, indicate that XRPCs exhibiting extracellular freezing may appear not only in very hardy woody species native to cold areas beyond the -40 degrees C isotherm but also in less hardy hardwood species native to tropical and subtropical zones as well as in a small number of moderately hardy hardwood species native to warm temperate zones. Cryo-scanning electron microscopic (cryo-SEM) studies on the freezing behavior of XRPCs have revealed some errors in DTA. These errors are originated mainly due to the overlap between exotherms produced by freezing of water in apoplastic spaces (high temperature exotherms, HTEs) and exotherms produced by freezing of intracellular water of XRPCs by breakdown of deep supercooling (low temperature exotherms, LTEs), as well as to the shortage of LTEs produced by intracellular freezing of XRPCs. In addition, DTA results are significantly affected by cooling rates employed. Further, cryo-SEM observations, which revealed the true freezing behavior of XRPCs, changed the previous knowledge of freezing behavior of XRPCs that had been obtained by freeze-substitution and transmission electron microscopic studies. Cryo-SEM results, in association with results obtained from DTA that were reconfirmed or changed by observation using a cryo-SEM, revealed a clear tendency of the freezing behavior of XRPCs in hardwood species to change with changes in the

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

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    Mazur, P.


    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

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    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)


    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. A modified differential scanning calorimetry for determination of cell volumetric change during the freezing process. (United States)

    Luo, Dawei; Han, Xu; He, Liqun; Cui, Xiangdong; Cheng, Shuxia; Lu, Caicheng; Liu, Jianghan; Gao, Dayong


    A modified analytical and experimental method using differential scanning calorimeter (DSC) was developed to determine the cell volume change during the freezing process. Two cell types were used in the study: human platelets and erythrocytes (red blood cells). Isotonic cell suspensions with different cytocrits were prepared and used in the DSC experiments. Low cooling rates were used to avoid intracellular ice formation. Cell suspensions were cooled from room temperature to -40 degrees C. Latent heat release from the freezing of cell suspensions was shown to be a linear function of cytocrit. From slope and intercept of the linear function, cell volume change was determined based on a developed theoretical model. From experimental data and theoretical analyses, it was revealed that (a) the final volume of a human platelet at -40 degrees C was 33.7% of its isotonic volume, and 15.2% of the original (at isotonic condition) intracellular water remained unfrozen inside platelets, and (b) the final volume of human erythrocyte at -40 degrees C was 50.0% of its isotonic volume, and 30.3% of the original intracellular water was kept inside cells as residual unfrozen water.

  3. DEA1, a circadian- and cold-regulated tomato gene, protects yeast cells from freezing death. (United States)

    Weyman, Philip D; Pan, Zhiqiang; Feng, Qin; Gilchrist, David G; Bostock, Richard M


    Cold and freezing damage to plants can be mitigated by inducible factors during an acclimation period. DEA1 is a circadian-regulated tomato (Solanum lycopersicum) gene with sequence similarity to EARLI1, an Arabidopsis thaliana gene that confers cold protection. To investigate whether DEA1 was responsive to environmental variables such as cold, cold-treated tomatoes were analyzed for DEA1 expression. DEA1 transcript accumulated in response to cold, and the rapidity of the cold-induced transcript accumulation was regulated by the circadian rhythm. To test whether DEA1 could protect cells from freezing damage, we transformed the yeast, Pichia pastoris, with an inducible DEA1 construct. Yeast cells transformed with the gene survived freezing at a significantly higher rate than control strains and a strain expressing the LacZ gene. Transgenic tomato plants over-expressing or knocking down DEA1 transcript levels did not have an altered phenotype with respect to cold- or pathogen-susceptibility relative to control plants.

  4. Cryo-scanning electron microscopic study on freezing behaviors of tissue cells in dormant buds of larch (Larix kaempferi). (United States)

    Endoh, Keita; Kasuga, Jun; Arakawa, Keita; Ito, Toshiaki; Fujikawa, Seizo


    The freezing behavior of dormant buds in larch, especially at the cellular level, was examined by a Cryo-SEM. The dormant buds exhibited typical extraorgan freezing. Extracellular ice crystals accumulated only in basal areas of scales and beneath crown tissues, areas in which only these living cells had thick walls unlike other tissue cells. By slow cooling (5 degrees C/day) of dormant buds to -50 degrees C, all living cells in bud tissues exhibited distinct shrinkage without intracellular ice formation detectable by Cryo-SEM. However, the recrystallization experiment of these slowly cooled tissue cells, which was done by further freezing of slowly cooled buds with LN and then rewarming to -20 degrees C, confirmed that some of the cells in the leaf primordia, shoot primordia and apical meristem, areas in which cells had thin walls and in which no extracellular ice accumulated, lost freezable water with slow cooling to -30 degrees C, indicating ability of these cells to adapt by extracellular freezing, whereas other cells in these tissues retained freezable water with slow cooling even to -50 degrees C, indicating adaptation of these cells by deep supercooling. On the other hand, all cells in crown tissues and in basal areas of scales, areas in which cells had thick walls and in which large masses of ice accumulated, had the ability to adapt by extracellular freezing. It is thought that the presence of two types of cells exhibiting different freezing adaptation abilities within a bud tissue is quite unique and may reflect sophisticated freezing adaptation mechanisms in dormant buds.

  5. A Vivens Ex Vivo Study on the Synergistic Effect of Electrolysis and Freezing on the Cell Nucleus. (United States)

    Lugnani, Franco; Zanconati, Fabrizio; Marcuzzo, Thomas; Bottin, Cristina; Mikus, Paul; Guenther, Enric; Klein, Nina; Rubinsky, Liel; Stehling, Michael K; Rubinsky, Boris


    Freezing-cryosurgery, and electrolysis-electrochemical therapy (EChT), are two important minimally invasive surgery tissue ablation technologies. Despite major advantages they also have some disadvantages. Cryosurgery cannot induce cell death at high subzero freezing temperatures and requires multiple freeze thaw cycles, while EChT requires high concentrations of electrolytic products-which makes it a lengthy procedure. Based on the observation that freezing increases the concentration of solutes (including products of electrolysis) in the frozen region and permeabilizes the cell membrane to these products, this study examines the hypothesis that there could be a synergistic effect between freezing and electrolysis in their use together for tissue ablation. Using an animal model we refer to as vivens ex vivo, which may be of value in reducing the use of animals for experiments, combined with a Hematoxylin stain of the nucleus, we show that there are clinically relevant protocols in which the cell nucleus appears intact when electrolysis and freezing are used separately but is affected by certain combinations of electrolysis and freezing.

  6. Transport phenomena during freezing of adipose tissue derived adult stem cells. (United States)

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


    In the present study a well-established differential scanning calorimeter (DSC) technique is used to measure the water transport phenomena during freezing of stromal vascular fraction (SVF) and adipose tissue derived adult stem (ADAS) cells at different passages (Passages 0 and 2). Volumetric shrinkage during freezing of adipose derived cells was obtained at a cooling rate of 20 degrees C/min in the presence of extracellular ice and two different, commonly used, cryoprotective agents, CPAs (10% DMSO and 10% Glycerol). The adipose derived cells were modeled as spheres of 50 microm diameter with an osmotically inactive volume (Vb) of 0.6Vo, where Vo is the isotonic cell volume. By fitting a model of water transport to the experimentally obtained volumetric shrinkage data, the "best-fit" membrane permeability parameters (reference membrane permeability to water, Lpg or Lpg[cpa] and the activation energy, ELp or ELp[cpa]) were determined. The "best-fit" membrane permeability parameters for adipose derived cells in the absence and presence of CPAs ranged from: Lpg=23.1-111.5x10(-15) m3/Ns (0.135-0.652 microm/min-atm) and ELp=43.1-168.8 kJ/mol (9.7-40.4 kcal/mol). Numerical simulations of water transport were then performed under a variety of cooling rates (5-100 degrees C/min) using the experimentally determined membrane permeability parameters. And finally, the simulation results were analyzed to predict the optimal rates of freezing adipose derived cells in the presence and absence of CPAs. Copyright (c) 2005 Wiley Periodicals, Inc.

  7. Accumulation of Poly(3-hydroxybutyrate Helps Bacterial Cells to Survive Freezing.

    Directory of Open Access Journals (Sweden)

    Stanislav Obruca

    Full Text Available Accumulation of polyhydroxybutyrate (PHB seems to be a common metabolic strategy adopted by many bacteria to cope with cold environments. This work aimed at evaluating and understanding the cryoprotective effect of PHB. At first a monomer of PHB, 3-hydroxybutyrate, was identified as a potent cryoprotectant capable of protecting model enzyme (lipase, yeast (Saccharomyces cerevisiae and bacterial cells (Cupriavidus necator against the adverse effects of freezing-thawing cycles. Further, the viability of the frozen-thawed PHB accumulating strain of C. necator was compared to that of the PHB non-accumulating mutant. The presence of PHB granules in cells was revealed to be a significant advantage during freezing. This might be attributed to the higher intracellular level of 3-hydroxybutyrate in PHB accumulating cells (due to the action of parallel PHB synthesis and degradation, the so-called PHB cycle, but the cryoprotective effect of PHB granules seems to be more complex. Since intracellular PHB granules retain highly flexible properties even at extremely low temperatures (observed by cryo-SEM, it can be expected that PHB granules protect cells against injury from extracellular ice. Finally, thermal analysis indicates that PHB-containing cells exhibit a higher rate of transmembrane water transport, which protects cells against the formation of intracellular ice which usually has fatal consequences.

  8. Accumulation of Poly(3-hydroxybutyrate) Helps Bacterial Cells to Survive Freezing (United States)

    Krzyzanek, Vladislav; Mravec, Filip; Hrubanova, Kamila; Samek, Ota; Kucera, Dan; Benesova, Pavla; Marova, Ivana


    Accumulation of polyhydroxybutyrate (PHB) seems to be a common metabolic strategy adopted by many bacteria to cope with cold environments. This work aimed at evaluating and understanding the cryoprotective effect of PHB. At first a monomer of PHB, 3-hydroxybutyrate, was identified as a potent cryoprotectant capable of protecting model enzyme (lipase), yeast (Saccharomyces cerevisiae) and bacterial cells (Cupriavidus necator) against the adverse effects of freezing-thawing cycles. Further, the viability of the frozen–thawed PHB accumulating strain of C. necator was compared to that of the PHB non-accumulating mutant. The presence of PHB granules in cells was revealed to be a significant advantage during freezing. This might be attributed to the higher intracellular level of 3-hydroxybutyrate in PHB accumulating cells (due to the action of parallel PHB synthesis and degradation, the so-called PHB cycle), but the cryoprotective effect of PHB granules seems to be more complex. Since intracellular PHB granules retain highly flexible properties even at extremely low temperatures (observed by cryo-SEM), it can be expected that PHB granules protect cells against injury from extracellular ice. Finally, thermal analysis indicates that PHB-containing cells exhibit a higher rate of transmembrane water transport, which protects cells against the formation of intracellular ice which usually has fatal consequences. PMID:27315285

  9. Two-step freezing of hybridoma cells in 96-well microculture plates. (United States)

    Pĕknicová, J; Kristofová, H


    Stabile hybridoma cells, colonies of hybridoma cells 14 days after fusion of immune spleen and myeloma cells, myeloma cells and fibroblasts cultured in 96-well microculture plates were frozen by the method of two-step freezing. The culture medium was aspirated, and 50 microliter of the medium containing a cryoprotectant (5% dimethyl sulphoxide) was added for 10 min at room temperature. The plates were put into microtene bags, placed at -25 degrees C in a freezer for 30 min and then stored at -100 degrees C in liquid nitrogen vapour. Plates with cells were thawed rapidly in a 50 degree C water bath. After thawing the hybrid cells were viable and continued to produce the specific antibody.

  10. Optimization of the freezing process for hematopoietic progenitor cells: effect of precooling, initial dimethyl sulfoxide concentration, freezing program, and storage in vapor-phase or liquid nitrogen on in vitro white blood cell quality. (United States)

    Dijkstra-Tiekstra, Margriet J; Setroikromo, Airies C; Kraan, Marcha; Gkoumassi, Effimia; de Wildt-Eggen, Janny


    Adding dimethyl sulfoxide (DMSO) to hematopoietic progenitor cells (HPCs) causes an exothermic reaction, potentially affecting their viability. The freezing method might also influence this. The aim was to investigate the effect of 1) precooling of DMSO and plasma (D/P) and white blood cell (WBC)-enriched product, 2) DMSO concentration of D/P, 3) freezing program, and 4) storage method on WBC quality. WBC-enriched product without CD34+ cells was used instead of HPCs. This was divided into six or eight portions. D/P (20 or 50%; precooled or room temperature [RT]) was added to the WBC-enriched product (precooled or RT), resulting in 10% DMSO, while monitoring temperature. The product was frozen using controlled-rate freezing ("fast-rate" or "slow-rate") and placed in vapor-phase or liquid nitrogen. After thawing, WBC recovery and viability were determined. Temperature increased most for precooled D/P to precooled WBC-enriched product, without influence of 20 or 50% D/P, but remained for all variations below 30°C. WBC recovery for both freezing programs was more than 95%. Recovery of WBC viability was higher for slow-rate freezing compared to fast-rate freezing (74% vs. 61%; p Effect of precooling D/P or WBC-enriched product and of storage in vapor-phase or liquid nitrogen was marginal. Based on these results, precooling is not necessary. Fifty percent D/P is preferred over 20% D/P. Slow-rate freezing is preferred over fast-rate freezing. For safety reasons storage in vapor-phase nitrogen is preferred over storage in liquid nitrogen. Additional testing using real HPCs might be necessary. © 2014 AABB.

  11. Transient warming events occurring after freezing impairs umbilical cord-derived mesenchymal stromal cells functionality. (United States)

    Chabot, Dominique; Tremblay, Tony; Paré, Isabelle; Bazin, Renée; Loubaki, Lionel


    Mesenchymal stromal cells (MSCs) have shown promising results for the treatment of refractory acute graft-versus-host disease. While safety of MSC infusion has been demonstrated, the use of cryopreserved MSCs in clinical trials has raised concerns regarding the retention of their functional activity. This has led to the recommendation by experts in the field to use freshly harvested MSCs, even though this approach is much less practical from a logistic point of view. In the present study, we revisited the impact of cryopreservation on MSC functionality and addressed the possibility that warming events on frozen cells rather than cryopreservation per se could impact MSC functionality. Following controlled-rate freezing to -130°C, umbilical cord-derived MSCs were left at room temperature (RT) for 2-10 min or on dry ice for 10 min, before being transferred into liquid nitrogen (LqN 2 ). MSCs of each group were subsequently tested (viability, functionality and cellular damage) and compared with their freshly harvested counterparts. We demonstrated that freshly harvested MSCs as well as cryopreserved MSCs that were left on dry ice following step-down freezing have comparable viability, functionality and integrity. In contrast, cryopreserved MSCs that were left at RT before being transferred into LqN 2 were functionally impaired and showed cellular damage upon thawing even though they exhibited high viability. Warming events after freezing and not cryopreservation per se significantly impair MSC functionality, indicating that cryopreserved MSCs can be an advantageous alternative to freshly harvested cells for therapeutic purposes. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  12. Cell surface damage and morphological changes in Oenococcus oeni after freeze-drying and incubation in synthetic wine. (United States)

    Bravo-Ferrada, Bárbara Mercedes; Gonçalves, Sónia; Semorile, Liliana; Santos, Nuno C; Brizuela, Natalia; Elizabeth Tymczyszyn, E; Hollmann, Axel


    The aim of the present study was to evaluate the effects of freeze-drying in the presence of trehalose as a cryoprotectant, followed by incubation in synthetic wine, on surface damage, viability and l-malic acid consumption of the oenological strain Oenococcus oeni UNQOe 73.2. After freeze-drying, no significant differences were observed in the number of viable cells (for both acclimated and non-acclimated cultures) respect to the fresh culture. In contrast, loss of viability was observed after wine incubation for 24 h, being acclimated freeze-dried cells the best conditions for this. After the preservation process, small changes in cell morphology were observed by Atomic Force Microscopy (AFM). The Zeta potential and AFM showed that 24 h of wine incubation was enough to induce several cell surface modifications. Plate count data allowed us to establish that surface damage is an important factor for loss of viability, regardless of the acclimation treatment. Although the number of surviving O. oeni cells decreased dramatically after incubation in synthetic wine for 15 days, the consumption of l-malic acid was higher than 70%, with freeze-dried cells showing a better performance than fresh cultures. These results demonstrate that O. oeni freeze-dried cultures could be applied to direct wine inoculation, to conduct malolactic fermentation, maintaining its technological properties and reducing the time and costs of the winemaking process. Copyright © 2018. Published by Elsevier Inc.

  13. 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:


    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.

  14. Extracellular freezing in leaves of freezing-sensitive species. (United States)

    Ashworth, Edward N; Pearce, Roger S


    Low-temperature scanning-electron microscopy was used to study the freezing of leaves of five species that have no resistance to freezing: bean (Phaseolus vulgaris L.), tobacco (Nicotiana tabacum L.), tomato (Lycopersicon esculentum L.), cucumber (Cucumis sativus L.), and corn (Zea mays L.). In the leaves of the four dicotyledonous species, ice was extracellular and the cells of all tissues were collapsed. In contrast, in maize leaves ice was extracellular in the mesophyll, and these cells were collapsed, but the epidermal and bundle-sheath cells apparently retained their original shapes and volume. It is concluded that the leaves of the freezing-sensitive dicotyledonous species tested were killed by cellular dehydration induced by extracellular freezing, and not by intracellular freezing. Freezing injury in maize leaves apparently resulted from a combination of freezing-induced cellular dehydration of some cells and intracellular ice formation in epidermal and bundle-sheath cells.

  15. Cryopreservation of chicken primordial germ cells by vitrification and slow freezing: A comparative study. (United States)

    Tonus, C; Connan, D; Waroux, O; Vandenhove, B; Wayet, J; Gillet, L; Desmecht, D; Antoine, N; Ectors, F J; Grobet, L


    In the present study, we compare a classical slow freezing (SLF) method and an aseptic vitrification (Vitrif) technique to cryopreserve a stable primordial germ cell (PGCs) line issued from the Ardennaise chicken breed. Viability immediately after warming was close to 80% and did not differ between the two cryopreservation methods. Proliferation tended to be slower for both cryopreservation methods compared with controls, but the difference was significant only for Vitrif. No difference was found between the two methods after flow cytometry analysis of stage-specific embryonic antigen-1 expression and reverse transcription-polymerase chain reaction on several factors related to PGC phenotype. After 1 week in culture, all cryopreserved cells reached controls' main morphologic and expanding (viability/proliferation) features. However, SLF generated more unwanted cells clusters than Vitrif. After injection of the PGCs into recipient embryos, vitrified PGCs reported a clear, yet not significant, tendency to colonize the gonad at a higher rate than slow frozen PGCs. SLF in cryovials remains simple, inexpensive, and less technically demanding than Vitrif. Nevertheless, the intrinsic advantages of our aseptic Vitrif method and the present study suggest that this should be considered as safer than classical SLF for cryopreserving chicken PGCs. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. Small molecule ice recrystallization inhibitors enable freezing of human red blood cells with reduced glycerol concentrations. (United States)

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


    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.

  17. 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: [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China)


    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.

  18. Microbial stratification structure within cathodic biofilm of the microbial fuel cell using the freezing microtome method. (United States)

    Li, Xiao; Lu, Yaobin; Luo, Haiping; Liu, Guangli; Zhang, Renduo


    The aim of this study was to investigate the microbial stratification structure within cathodic biofilm of the microbial fuel cell (MFC) using the freezing microtome method. Experiments were conducted in a single-chamber air-cathode MFC with 0.8g/L maltodextrin as substrate for ∼30d operation. The maximum power density was 945±10mW/m 2 in the MFC. Maltodextrin resulted in the relative abundance of Candidatus Saccharibacteria of 37.0% in the anodic biofilm. Different bacterial communities were identified in different layers within the cathodic biofilm. The relative abundance of Enterococcus was 3.7%, 10.5%, and 1.6% in the top (100-150μm), middle (50-100μm), and bottom (0-50μm) layers, respectively. Higher bacterial viability was observed within the top and bottom layers of the cathodic biofilm. Understanding the stratification of bacterial community in cathodic biofilm should be important to control the cathodic biofilm in the MFC. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

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


    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. Subzero water permeability parameters and optimal freezing rates for sperm cells of the southern platyfish, Xiphophorus maculatus. (United States)

    Pinisetty, D; Huang, C; Dong, Q; Tiersch, T R; Devireddy, R V


    This study reports the subzero water transport characteristics (and empirically determined optimal rates for freezing) of sperm cells of live-bearing fishes of the genus Xiphophorus, specifically those of the southern platyfish Xiphophorus maculatus. These fishes are valuable models for biomedical research and are commercially raised as ornamental fish for use in aquariums. Water transport during freezing of X. maculatus sperm cell suspensions was obtained using a shape-independent differential scanning calorimeter technique in the presence of extracellular ice at a cooling rate of 20 degrees C/min in three different media: (1) Hanks' balanced salt solution (HBSS) without cryoprotective agents (CPAs); (2) HBSS with 14% (v/v) glycerol, and (3) HBSS with 10% (v/v) dimethyl sulfoxide (DMSO). The sperm cell was modeled as a cylinder with a length of 52.35 microm and a diameter of 0.66 microm with an osmotically inactive cell volume (Vb) of 0.6 V0, where V0 is the isotonic or initial cell volume. This translates to a surface area, SA to initial water volume, WV ratio of 15.15 microm(-1). By fitting a model of water transport to the experimentally determined volumetric shrinkage data, the best fit membrane permeability parameters (reference membrane permeability to water at 0 degrees C, Lpg or Lpg [cpa] and the activation energy, E(Lp) or E(Lp) [cpa]) were found to range from: Lpg or Lpg [cpa] = 0.0053-0.0093 microm/minatm; E(Lp) or E(Lp) [cpa] = 9.79-29.00 kcal/mol. By incorporating these membrane permeability parameters in a recently developed generic optimal cooling rate equation (optimal cooling rate, [Formula: see text] where the units of B(opt) are degrees C/min, E(Lp) or E(Lp) [cpa] are kcal/mol, L(pg) or L(pg) [cpa] are microm/minatm and SA/WV are microm(-1)), we determined the optimal rates of freezing X. maculatus sperm cells to be 28 degrees C/min (in HBSS), 47 degrees C/min (in HBSS+14% glycerol) and 36 degrees C/min (in HBSS+10% DMSO). Preliminary empirical

  1. Serum-free freezing media support high cell quality and excellent ELISPOT assay performance across a wide variety of different assay protocols. (United States)

    Filbert, Helene; Attig, Sebastian; Bidmon, Nicole; Renard, Bernhard Y; Janetzki, Sylvia; Sahin, Ugur; Welters, Marij J P; Ottensmeier, Christian; van der Burg, Sjoerd H; Gouttefangeas, Cécile; Britten, Cedrik M


    Robust and sensitive ELISPOT protocols are commonly applied concomitant with the development of new immunotherapeutics. Despite the knowledge that individual serum batches differ in their composition and may change properties over time, serum is still commonly used in immunologic assays. Commercially available serum batches are expensive, limited in quantity and need to be pretested for suitability in immunologic assays, which is a laborious process. The aim of this study was to test whether serum-free freezing media can lead to high cell viability and favorable performance across multiple ELISPOT assay protocols. Thirty-one laboratories from ten countries participated in a proficiency panel organized by the Cancer Immunotherapy Immunoguiding Program to test the influence of different freezing media on cell quality and immunologic function. Each center received peripheral blood mononuclear cells which were frozen in three different media. The participants were asked to quantify antigen-specific CD8+ T-cell responses against model antigens using their locally established IFN-gamma ELISPOT protocols. Self-made and commercially available serum-free freezing media led to higher cell viability and similar cell recovery after thawing and resting compared to freezing media supplemented with human serum. Furthermore, the test performance as determined by (1) background spot production, (2) replicate variation, (3) frequency of detected antigen-specific spots and (4) response detection rate was similar for serum and serum-free conditions. We conclude that defined and accessible serum-free freezing media should be recommended for freezing cells stored for subsequent ELISPOT analysis.

  2. Freeze-thaw lysates of Plasmodium falciparum-infected red blood cells induce differentiation of functionally competent regulatory T cells from memory T cells. (United States)

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


    In addition to naturally occurring regulatory T (nTreg) cells derived from the thymus, functionally competent Treg cells can be induced in vitro from peripheral blood lymphocytes in response to TCR stimulation with cytokine costimulation. Using these artificial stimulation conditions, both naïve as well as memory CD4(+) T cells can be converted into induced Treg (iTreg) cells, but the cellular origin of such iTreg cells in vivo or in response to more physiologic stimulation with pathogen-derived antigens is less clear. Here, we demonstrate that a freeze/thaw lysate of Plasmodium falciparum schizont extract (PfSE) can induce functionally competent Treg cells from peripheral lymphocytes in a time- and dose-dependent manner without the addition of exogenous costimulatory factors. The PfSE-mediated induction of Treg cells required the presence of nTreg cells in the starting culture. Further experiments mixing either memory or naïve T cells with antigen presenting cells and CFSE-labeled Treg cells identified CD4(+) CD45RO(+) CD25(-) memory T cells rather than Treg cells as the primary source of PfSE-induced Treg cells. Taken together, these data suggest that in the presence of nTreg cells, PfSE induces memory T cells to convert into iTreg cells that subsequently expand alongside PfSE-induced effector T cells. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. 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


    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.

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

    DEFF Research Database (Denmark)

    Prentø, P


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

  5. Evaluation of sericin as a fetal bovine serum-replacing cryoprotectant during freezing of human mesenchymal stromal cells and human osteoblast-like cells. (United States)

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


    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 or reduce the presence of dimethyl sulfoxide (DMSO) in the medium. Sericin, a sticky protein derived from the silkworm cocoon, was investigated as a substitute for FBS and DMSO in the freezing medium. Cell viability (24 hours after thawing, both hMSC and SAOS-2) and colony-forming ability (2 weeks after thawing, only for hMSCs) were both determined. The FBS-free medium with 1% sericin in 10% DMSO was found to be a suitable freezing medium for primary hMSCs, in contrast to immortalized human osteoblasts. Surprisingly, the storage of hMSCs in a cultivation medium with only 10% DMSO also provided satisfactory results. Any drop in DMSO concentration led to significantly worse survival of cells, with little improvement in hMSC survival in the presence of sericin. Thus, sericin may substitute for FBS in the freezing medium for primary hMSCs, but cannot substitute for DMSO.

  6. Comparison of methods of high-pressure freezing and automated freeze-substitution of suspension cells combined with LR White embedding

    Czech Academy of Sciences Publication Activity Database

    Sobol, Margaryta; Philimonenko, Vlada; Hozák, Pavel


    Roč. 134, č. 6 (2010), s. 631-641 ISSN 0948-6143 R&D Projects: GA AV ČR KAN200520704; GA MŠk 2B06063; GA MŠk LC545 Institutional research plan: CEZ:AV0Z50520514 Keywords : cryofixation * automated freeze-substitution * LR White Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.727, year: 2010

  7. Generalized structural theory of freezing

    International Nuclear Information System (INIS)

    Yussouff, M.


    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)

  8. Effect of different freezing rates during cryopreservation of rat mesenchymal stem cells using combinations of hydroxyethyl starch and dimethylsulfoxide (United States)


    Background Mesenchymal stem cells (MSCs) are increasingly used as therapeutic agents as well as research tools in regenerative medicine. Development of technologies which allow storing and banking of MSC with minimal loss of cell viability, differentiation capacity, and function is required for clinical and research applications. Cryopreservation is the most effective way to preserve cells long term, but it involves potentially cytotoxic compounds and processing steps. Here, we investigate the effect of decreasing dimethyl sulfoxide (DMSO) concentrations in cryosolution by substituting with hydroxyethyl starch (HES) of different molecular weights using different freezing rates. Post-thaw viability, phenotype and osteogenic differentiation capacity of MSCs were analysed. Results The study confirms that, for rat MSC, cryopreservation effects need to be assessed some time after, rather than immediately after thawing. MSCs cryopreserved with HES maintain their characteristic cell surface marker expression as well as the osteogenic, adipogenic and chondrogenic differentiation potential. HES alone does not provide sufficient cryoprotection for rat MSCs, but provides good cryoprotection in combination with DMSO, permitting the DMSO content to be reduced to 5%. There are indications that such a combination would seem useful not just for the clinical disadvantages of DMSO but also based on a tendency for reduced osteogenic differentiation capacity of rat MSC cryopreserved with high DMSO concentration. HES molecular weight appears to play only a minor role in its capacity to act as a cryopreservation solution for MSC. The use of a ‘straight freeze’ protocol is no less effective in maintaining post-thaw viability of MSC compared to controlled rate freezing methods. Conclusion A 5% DMSO / 5% HES solution cryopreservation solution using a ‘straight freeze’ approach can be recommended for rat MSC. PMID:22889198

  9. Preservation of flavor in freeze dried green beans (United States)

    Huber, C. S.; Heidelbaugh, N. D.; Davis, D.


    Before freeze drying, green beans are heated to point at which their cell structure is altered. Beans freeze dried with altered cell structure have improved rehydration properties and retain color, flavor, and texture.

  10. Quantification of Intracellular Ice Formation and Recrystallization During Freeze-Thaw Cycles and Their Relationship with the Viability of Pig Iliac Endothelium Cells. (United States)

    Liu, Xiaoli; Zhao, Gang; Shu, Zhiquan; Niu, Dan; Zhang, Zhiguo; Zhou, Ping; Cao, Yunxia; Gao, Dayong


    Quantitative evaluation of the inherent correlation between cell cryoinjuries and intracellular ice formation (IIF) together with recrystallization (IIR) is of primary importance for both optimization of biopreservation and cryotherapy. The objective of this study is to thoroughly explore the roles of IIF on cell viability by using pig iliac endothelium cells (PIECs) as model cells during freezing and thawing. The experimental results indicated that both the probabilities of IIF (PIF) and IIR (PIR) increased along with the increase of cooling rates (p < 0.05) during the freeze-thaw cycles at cooling rates of 40, 60, 80, 100, and 150°C/min and the same warming rates of 100°C/min in phosphate-buffered saline-based solutions with or without 1 M DMSO. Viability evaluation with Hoechst 33342/propidium iodide double staining showed that most of the cells were killed (viability <20%) by the abovementioned freeze-thaw cycles, which indicated that the cooling rates investigated were all too rapid since large amounts of IIF and IIR were introduced. Another interesting phenomenon is that the presence of a low concentration of DMSO (1 M) tends to improve cell viability while increasing the PIF and PIR during freezing/thawing, contrary to the common belief that larger PIF corresponds to greater cryoinjury. This may be attributed to the intrinsic protection effect of DMSO by reduction of solution injury or other potential injuries. These findings may be of potential application value for both cryopreservation and cryosurgery by providing helpful additions to the existing studies on investigation of cryoinjuries of PIECs.

  11. 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


    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.

  12. A patient with squamous cell carcinoma developing as a secondary cancer after radiation therapy and carbon dioxide snow-freezing therapy

    International Nuclear Information System (INIS)

    Yoshida, Akihiro; Miki, Hirotoshi; Yokoyama, Toshiya; Katori, Nobutada


    We report on a 59-year-old male who developed skin cancer many years after receiving radiation therapy and carbon dioxide snow-freezing therapy. About 50 years ago, the patient underwent radiation therapy and carbon dioxide snow-freezing therapy for nevus of the Ota of the left buccal region, the orbital region, and the forehead (fractional dose and total dose unknown). About 6 months prior to evaluation at this hospital, the patient noted tumor growth on the skin in the areas previously treated, and protruding lesions were observed on the left buccal region, the orbital region, and the forehead. Histopathological examination showed squamous cell carcinoma. At surgery, protruding lesions and scar tissue were completely excised, and split-thickness skin graft was used in the areas of skin loss. After 2 years of follow-up, no local recurrence or metastasis has been observed. (author)

  13. Effect of chemical fixatives on accurate preservation of Escherichia coli and Bacillus subtilis structure in cells prepared by freeze-substitution

    International Nuclear Information System (INIS)

    Graham, L.L.; Beveridge, T.J.


    Five chemical fixatives were evaluated for their ability to accurately preserve bacterial ultrastructure during freeze-substitution of select Escherichia coli and Bacillus subtilis strains. Radioisotopes were specifically incorporated into the peptidoglycan, lipopolysaccharide, and nucleic acids of E. coli SFK11 and W7 and into the peptidoglycan and RNA of B. subtilis 168 and W23. The ease of extraction of radiolabels, as assessed by liquid scintillation counting during all stages of processing for freeze-substitution, was used as an indicator of cell structural integrity and retention of cellular chemical composition. Subsequent visual examination by electron microscopy was used to confirm ultrastructural conformation. The fixatives used were: 2% (wt/vol) osmium tetroxide and 2% (wt/vol) uranyl acetate; 2% (vol/vol) glutaraldehyde and 2% (wt/vol) uranyl acetate; 2% (vol/vol) acrolein and 2% (wt/vol) uranyl acetate; 2% (wt/vol) gallic acid; and 2% (wt/vol) uranyl acetate. All fixatives were prepared in a substitution solvent of anhydrous acetone. Extraction of cellular constituents depended on the chemical fixative used. A combination of 2% osmium tetroxide-2% uranyl acetate or 2% gallic acid alone resulted in optimum fixation as ascertained by least extraction of radiolabels. In both gram-positive and gram-negative organisms, high levels of radiolabel were detected in the processing fluids in which 2% acrolein-2% uranyl acetate, 2% glutaraldehyde-2% uranyl acetate, or 2% uranyl acetate alone were used as fixatives. Ultrastructural variations were observed in cells freeze-substituted in the presence of different chemical fixatives. We recommend the use of osmium tetroxide and uranyl acetate in acetone for routine freeze-substitution of eubacteria, while gallic acid is recommended for use when microanalytical processing necessitates the omission of osmium

  14. Scanning x-ray microscopy of living and freeze-dried blood cells in two vanadium-rich ascidian species, Phallusia mammillata and Ascidia sydneiensis samea. (United States)

    Ueki, Tatsuya; Takemoto, Kuniko; Fayard, Barbara; Salomé, Murielle; Yamamoto, Akitsugu; Kihara, Hiroshi; Susini, Jean; Scippa, Silvia; Uyama, Taro; Michibata, Hitoshi


    Some ascidians (sea squirts) accumulate the transitional metal vanadium in their blood cells at concentrations of up to 350 mM, about 10(7) times its concentration found in seawater. There are approximately 10 different types of blood cell in ascidians. The identity of the true vanadium-containing blood cell (vanadocyte) is controversial and little is known about the subcellular distribution of vanadium. A scanning x-ray microscope installed at the ID21 beamline of the European Synchroton Radiation Facility to visualize vanadium in ascidian blood cells. Without fixation, freezing or staining realized the visualization of vanadium localized in living signet ring cells and vacuolated amoebocytes of two vanadium-rich ascidian species, Phallusia mammillata and Ascidia sydneiensis samea. A combination of transmission and fluorescence images of signet ring cells suggested that in both species the vacuoles contain vanadium.

  15. Applicable technical method for lower temperature freeze ...

    African Journals Online (AJOL)

    Cryo-fixation and freeze substitution followed by microscopy are commonly used sample preparation methods for visualizing the morphology of intracellular organelles. Freeze substitution is an especially important preparative step because it enables the preservation of intracellular structures in cryo-fixed cells close to the ...

  16. A comparison study on the behavior of human endometrial stem cell-derived osteoblast cells on PLGA/HA nanocomposite scaffolds fabricated by electrospinning and freeze-drying methods. (United States)

    Namini, Mojdeh Salehi; Bayat, Neda; Tajerian, Roxana; Ebrahimi-Barough, Somayeh; Azami, Mahmoud; Irani, Shiva; Jangjoo, Saranaz; Shirian, Sadegh; Ai, Jafar


    An engineered tissue structure is an artificial scaffold combined with cells and signaling factors. Among various polymers, the polylactide-co-glycolide/hydroxyapatite (PLGA/HA) has attracted much attention due to their optimal properties. The aim of this study was to study the behavior of human endometrial stem cell (hEnSC)-derived osteoblast cells cultured on PLGA/HA nanocomposite scaffolds. hEnSCs were isolated and exposed to osteogenic media for 21 days. Differentiated cells were cultured on PLGA/HA synthetic scaffolds. The PLGA/HA-based nanocomposite scaffolds were fabricated using either electrospinning or freeze-drying methods. Behavior of the cells was evaluated a week after seeding hEnSC-derived osteoblast-like cells on these scaffolds. Osteogenesis was investigated in terms of alkaline phosphatase activity, gene expression, immunocytochemistry (ICC), proliferation, and scanning electron microscopy (SEM). Moreover, scaffold properties, such as pore size and morphology of the cells, onto the scaffolds were evaluated using SEM. Furthermore, biocompatibility of these scaffolds was confirmed by 3-(4,5-dimethylthiazoyl-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The matrix mineralization was proved by alizarin red staining, and the osteogenic media-treated cultures positively expressed osteocalcin and osteopontin markers. Moreover, qRT-PCR results confirmed the positive gene expression of osteopontin and osteonectin in the differentiated osteoblast-like cells. The results of behavior assessment of the cultured cells on electrospinning and freeze-dried scaffolds showed that the behavior of the cultured cells on the freeze-dried PLGA/HA scaffolds was significantly better than the electrospinning PLGA/HA scaffolds. It has been shown that the freeze-dried PLGA/HA nanocomposite scaffolds can appropriately support the attachment and proliferation of the differentiated osteoblast cells and are a suitable candidate for bone tissue engineering.

  17. Freezing for Love

    DEFF Research Database (Denmark)

    Carroll, Katherine; Kroløkke, Charlotte


    their eggs. Our analysis draws on semi-structured interviews with 16 women from the Midwest and East Coast regions of the USA who froze their eggs. Rather than freezing to balance career choices and ‘have it all’, the women in this cohort were largely ‘freezing for love’ and in the hope of having their ‘own...

  18. Freeze drying method

    International Nuclear Information System (INIS)

    Coppa, N.V.; Stewart, P.; Renzi, E.


    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

  19. Tris-egg yolk-glycerol (TEY) extender developed for freezing dog semen is a good option to cryopreserve bovine epididymal sperm cells. (United States)

    Lopes, G; Soares, L; Ferreira, P; Rocha, A


    Cryopreservation of epididymal spermatozoa is often performed after shipping the excised testis-epididymis complexes, under refrigeration, to a specialized laboratory. However, epididymal spermatozoa can be collected immediately after excision of the epididymis and sent extended and refrigerated to a laboratory for cryopreservation. In this experiment, we evaluated the effect of both methods of cold storage bovine epididymal spermatozoa as well as of two different extenders on spermatozoa characteristics after freeze-thawing. For that, spermatozoa collected from the caudae epididymis of 19 bulls were extended and cryopreserved in either AndroMed(®) or a Tris-egg yolk (TEY)-based extender. Cryopreservation of sperm cells was performed immediately after castration (Group A, n = 9) or after cold storage for 24 h diluted in the two extenders and (Group B, n = 9) and also after cold storage for 24 h within the whole epididymis (Group C, n = 10). Sperm subjective progressive motility (light microscopy), plasma membrane integrity (hypoosmotic swelling test) and sperm viability (eosin-nigrosin) were evaluated. In vitro fertilization and culture (IVF) was performed to assess the blastocyst rate. No differences (p > 0.05) were observed on post-thaw sperm parameters between samples from Group A, B and C. TEY extended samples presented a higher (p sperm, than those extended in AndroMed(®) . Blastocyst rate after IVF differed only (p epididymal sperm can be shipped chilled overnight either within the epididymal tail or after dilution without deleterious effect on post-thaw sperm quality. TEY extender was more suitable for cold storage and freezing bovine epididymal sperm, than the commercial extender AndroMed®. © 2014 Blackwell Verlag GmbH.

  20. No Correlates for Somatic Motility in Freeze-Fractured Hair-Cell Membranes of Lizards and Birds (United States)

    Köppl, C.; Forge, A.; Manley, G. A.


    It is not known whether active processes in mammals and non-mammals are due to the same underlying mechanism. To address this, we studied the size and density of particles in hair-cell membranes in mammals, in a lizard, the Tokay gecko, and in a bird, the barn owl. We surmised that if the prominent particles described in mammalian outer-hair-cell membranes are responsible for cochlear motility, a similar occurrence in non-mammalian hair cells would argue for similar mechanisms. Particle densities differed, however, substantially from those of mammals, suggesting that non-mammals have no membrane-based motility.

  1. Polymerization with freezing

    International Nuclear Information System (INIS)

    Ben-Naim, E; Krapivsky, P L


    Irreversible aggregation processes involving reactive and frozen clusters are investigated using the rate equation approach. In aggregation events, two clusters join irreversibly to form a larger cluster; additionally, reactive clusters may spontaneously freeze. Frozen clusters do not participate in merger events. Generally, freezing controls the nature of the aggregation process, as demonstrated by the final distribution of frozen clusters. The cluster mass distribution has a power-law tail, F k ∼k -γ , when the freezing process is sufficiently slow. Different exponents, γ = 1 and 3, are found for the constant and the product aggregation rates, respectively. For the latter case, the standard polymerization model, either no gels, or a single gel, or even multiple gels, may be produced

  2. [Social freezing - the male perspective]. (United States)

    Gromoll, J; Tüttelmann, F; Kliesch, S


    In Germany there is an emerging trend for postponing parenthood due to non-medical, sociocultural reasons. This clearly impacts on the reproductive success due to an age-dependent decrease in fertility. Thus, strategies and techniques are currently discussed which could preserve the female fertility status, among which social freezing (cryopreservation of oocytes) for later fertilization is the most realistic one; however, while there is an intensive discussion on the procedure and timing of oocyte cryopreservation, virtually no attention has been paid to the male side and the aging effects on the male germ cells. To evaluate the risk paternal age poses for the integrity of germ cells. For this review a literature search using PubMed, data from the Federal Statistical Office of Germany, the German in vitro fertilization (IVF) register as well as own data were used. Sperm cell integrity is clearly affected by age both at the genetic as well as at the epigenetic levels. The estimated mutation rate for spermatozoa doubles every 16.5 years. Monogenic and multifactorial diseases are strongly associated with paternal age. Men aged >40 years have an increased risk of passing age-related mutations to their children. Cryopreservation of spermatozoa is an option for men who postpone planning a family. Genetic counseling is recommended for couples undertaking social freezing and a male age of >40 years.

  3. Design of a blood-freezing system for leukemia research (United States)

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


    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.

  4. Accelerated storage testing of freeze-dried Pseudomonas ...

    African Journals Online (AJOL)


    storage and it would be useful to increase culture viability during storage. The viability of freeze-dried. P. fluorescens ... assessment of viable cells, while the residual water content is omitted (Hernandez et al., 2009). The ..... Effects of micronization on viability andthermotolerance of probiotic freeze-dried cultures. Int. Dairy J.

  5. Freezing and Food Safety (United States)

    ... Page ] Enzymes Enzyme activity can lead to the deterioration of food quality. Enzymes present in animals, vegetables, and fruit ... slows the enzyme activity that takes place in foods. It does not halt ... deterioration. This is called "blanching." For successful freezing, blanch ...

  6. Administrative Freezing of Capital Construction

    Directory of Open Access Journals (Sweden)

    Kropacheva A. V.


    Full Text Available The author estimates administrative freezing of capital construction as a type of punishment for violating laws in this sphere. The article provides the mechanism of freezing of capital construction as a serious alternative for fine sanctions

  7. Freeze-drying of lactic acid bacteria. (United States)

    Fonseca, Fernanda; Cenard, Stéphanie; Passot, Stéphanie


    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.

  8. Effects of freezing conditions on quality changes in blueberries. (United States)

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


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

  9. 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.


    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.

  10. Freezing-induced deformation of biomaterials in cryomedicine (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

  11. Freeze substitution in 3 hours or less. (United States)

    McDonald, K L; Webb, R I


    Freeze substitution is a process for low temperature dehydration and fixation of rapidly frozen cells that usually takes days to complete. New methods for freeze substitution have been developed that require only basic laboratory tools: a platform shaker, liquid nitrogen, a metal block with holes for cryotubes and an insulated container such as an ice bucket. With this equipment, excellent freeze substitution results can be obtained in as little as 90 min for cells of small volume such as bacteria and tissue culture cells. For cells of greater volume or that have significant diffusion barriers such as cuticles or thick cell walls, one can extend the time to 3 h or more with dry ice. The 3-h method works well for all manner of specimens, including plants and Caenorhabditis elegans as well as smaller samples. Here, we present the basics of the techniques and some results from Nicotiana leaves, C. elegans adult worms, Escherichia coli and baby hamster kidney tissue culture cells. © 2011 The Authors Journal of Microscopy © 2011 Royal Microscopical Society.

  12. Freezing of Coulomb liquids

    International Nuclear Information System (INIS)

    Tosi, M.P.


    Recent progress in the theory of liquid-solid coexistence as approached from the liquid phase in systems with Coulomb forces is reviewed. Main attention is given to (i) Wigner crystallization of the electron gas in the degenerate and classical limits, and (ii) localization of bond particles leading to freezing in a pseudoclassical liquid-state version of the bond-charge model for elemental semiconductors. These models serve to illustrate crystallization driven by pure Coulomb repulsions and crystallization resulting from the interplay of attraction and repulsions in multicomponent systems, respectively. (author). 29 refs, 4 figs

  13. Freeze-dried live attenuated smallpox vaccine prepared in cell culture "LC16-KAKETSUKEN": Post-marketing surveillance study on safety and efficacy compliant with Good Clinical Practice. (United States)

    Nishiyama, Yasumasa; Fujii, Tatsuya; Kanatani, Yasuhiro; Shinmura, Yasuhiko; Yokote, Hiroyuki; Hashizume, So


    In Japan, production of smallpox vaccine LC16m8 (named LC16-KAKETSUKEN) was restarted and was determined to be maintained as a national stockpile in March 2002. To conduct a post-marketing surveillance study of the vaccination of freeze-dried live attenuated smallpox vaccine prepared in cell culture LC16-KAKETSUKEN using attenuated vaccinia strain LC16m8. The study complied with Good Clinical Practice, focusing on a comparison between primary vaccinees and re-vaccinees. 268 personnel (261 males and 7 females) of the Japan Ground Self-Defense Force were inoculated with LC16-KAKETSUKEN and thereafter adverse events and efficacy were evaluated. Among 268 vaccinee participants, the following vaccinees showed adverse events, none serious: 53 of 196 primary vaccinees (without previous smallpox vaccination), 4 of 71 re-vaccinees (with previous smallpox vaccination) and 1 vaccinee with unknown previous vaccination history. A breakdown of adverse events observed in this study (total 268 vaccinees) showed the following minor or mild adverse events: 52 (19.4%) swelling of axillary lymph node, 4 (1.5%) fever, 2 (0.7%) fatigue, 1 (0.4%) of rash, 14 (5.2%) erythema at the inoculation site, 1 (0.4%) swelling at the inoculation site and 1 (0.4%) autoinoculation. The incidence of adverse events for primary vaccinees (53/196; 27.0%) was significantly higher than for re-vaccinees (4/71; 5.6%). However, the proportion of vaccine take was significantly higher for primary vaccinees (185/196; 94.4%) than for re-vaccinees (58/71; 81.7%). Although the proportion of vaccine take of re-vaccinees was significantly lower than for primary vaccinees due to preexisting immunity by previous vaccination, no significant difference was found in neutralizing antibody titers between primary vaccinees and re-vaccinees at 1, 4 and 7 months after LC16-KAKETSUKEN vaccination. The present post-marketing surveillance study compliant with Good Clinical Practice demonstrated the efficacy and safety of the

  14. Anhydrobiosis and Freezing-Tolerance

    DEFF Research Database (Denmark)

    McGill, Lorraine; Shannon, Adam; Pisani, Davide


    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...... 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...

  15. Freezing on a sphere (United States)

    Guerra, Rodrigo E.; Kelleher, Colm P.; Hollingsworth, Andrew D.; Chaikin, Paul M.


    The best understood crystal ordering transition is that of two-dimensional freezing, which proceeds by the rapid eradication of lattice defects as the temperature is lowered below a critical threshold. But crystals that assemble on closed surfaces are required by topology to have a minimum number of lattice defects, called disclinations, that act as conserved topological charges—consider the 12 pentagons on a football or the 12 pentamers on a viral capsid. Moreover, crystals assembled on curved surfaces can spontaneously develop additional lattice defects to alleviate the stress imposed by the curvature. It is therefore unclear how crystallization can proceed on a sphere, the simplest curved surface on which it is impossible to eliminate such defects. Here we show that freezing on the surface of a sphere proceeds by the formation of a single, encompassing crystalline ‘continent’, which forces defects into 12 isolated ‘seas’ with the same icosahedral symmetry as footballs and viruses. We use this broken symmetry—aligning the vertices of an icosahedron with the defect seas and unfolding the faces onto a plane—to construct a new order parameter that reveals the underlying long-range orientational order of the lattice. The effects of geometry on crystallization could be taken into account in the design of nanometre- and micrometre-scale structures in which mobile defects are sequestered into self-ordered arrays. Our results may also be relevant in understanding the properties and occurrence of natural icosahedral structures such as viruses.

  16. Freezing on a sphere. (United States)

    Guerra, Rodrigo E; Kelleher, Colm P; Hollingsworth, Andrew D; Chaikin, Paul M


    The best understood crystal ordering transition is that of two-dimensional freezing, which proceeds by the rapid eradication of lattice defects as the temperature is lowered below a critical threshold. But crystals that assemble on closed surfaces are required by topology to have a minimum number of lattice defects, called disclinations, that act as conserved topological charges-consider the 12 pentagons on a football or the 12 pentamers on a viral capsid. Moreover, crystals assembled on curved surfaces can spontaneously develop additional lattice defects to alleviate the stress imposed by the curvature. It is therefore unclear how crystallization can proceed on a sphere, the simplest curved surface on which it is impossible to eliminate such defects. Here we show that freezing on the surface of a sphere proceeds by the formation of a single, encompassing crystalline 'continent', which forces defects into 12 isolated 'seas' with the same icosahedral symmetry as footballs and viruses. We use this broken symmetry-aligning the vertices of an icosahedron with the defect seas and unfolding the faces onto a plane-to construct a new order parameter that reveals the underlying long-range orientational order of the lattice. The effects of geometry on crystallization could be taken into account in the design of nanometre- and micrometre-scale structures in which mobile defects are sequestered into self-ordered arrays. Our results may also be relevant in understanding the properties and occurrence of natural icosahedral structures such as viruses.

  17. 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 slot...

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

    Lifescience Database Archive (English)

    Full Text Available List Contact us DGBY Phenome data - Freeze-thaw stress Data detail Data name Phenome data - Freeze-thaw stress... DOI 10.18908/lsdba.nbdc00953-006 Description of data contents Yeasts used in bread making are exposed to freeze-thaw stress...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 closely related to freeze-thaw stress. The results showed that defects in the functions of va

  19. Calcium-dependent freezing tolerance in Arabidopsis involves membrane resealing via synaptotagmin SYT1. (United States)

    Yamazaki, Tomokazu; Kawamura, Yukio; Minami, Anzu; Uemura, Matsuo


    Plant freezing tolerance involves the prevention of lethal freeze-induced damage to the plasma membrane. We hypothesized that plant freezing tolerance involves membrane resealing, which, in animal cells, is accomplished by calcium-dependent exocytosis following mechanical disruption of the plasma membrane. In Arabidopsis thaliana protoplasts, extracellular calcium enhanced not only freezing tolerance but also tolerance to electroporation, which typically punctures the plasma membrane. However, calcium did not enhance survival when protoplasts were exposed to osmotic stress that mimicked freeze-induced dehydration. Calcium-dependent freezing tolerance was also detected with leaf sections in which ice crystals intruded into tissues. Interestingly, calcium-dependent freezing tolerance was inhibited by extracellular addition of an antibody against the cytosolic region of SYT1, a homolog of synaptotagmin known to be a calcium sensor that initiates exocytosis. This inhibition indicates that the puncture allowing the antibody to flow into the cytoplasm occurs during freeze/thawing. Thus, we propose that calcium-dependent freezing tolerance results from resealing of the punctured site. Protoplasts or leaf sections isolated from Arabidopsis SYT1-RNA interference (RNAi) plants lost calcium-dependent freezing tolerance, and intact SYT1-RNAi plants had lower freezing tolerance than control plants. Taken together, these findings suggest that calcium-dependent freezing tolerance results from membrane resealing and that this mechanism involves SYT1 function.

  20. Effects of various freezing containers for vitrification freezing on mouse oogenesis. (United States)

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


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

  1. Freeze-drying and related preparation techniques for biological microprobe analysis

    International Nuclear Information System (INIS)

    Wroblewski, R.; Wroblewski, J.; Anniko, M.; Edstroem, L.P.


    An X-ray microanalytical and morphological investigation has been carried out on rapidly frozen, freeze-dried or freeze-substituted tissues. A comparison was made between different embedding and polymerization procedures following freeze-substitution and freeze-drying. The investigation also included an analysis of specimens infiltrated, embedded and polymerized by ultraviolet irradiation at low temperatures with Lowicryl HM20. The morphological preservation of Lowicryl embedded tissue was adequate for the identification of different cell structures like nuclei, mitochondria, lysosomes and different types of endoplasmic reticulum. X-ray microanalytical investigation of low temperature embedded material displayed an elemental composition of cells and organelles similar to that found in freeze-dried cyosections. Compared with freeze-dried cryosections, low temperature embedded material could be sectioned for light microscopy and area of interest chosen for further thin sectioning. This is of great importance in work with tissues with complicated morphology and heterogenous cell populations

  2. Optoacoustic laser monitoring of cooling and freezing of tissues

    International Nuclear Information System (INIS)

    Larin, Kirill V; Larina, I V; Motamedi, M; Esenaliev, R O


    Real-time monitoring of cooling and freezing of tissues, cells, and other biological objects with a high spatial and time resolution, which is necessary for selective destruction of cancer and benign tumours during cryotherapy, as well as for preventing any damage to the structure and functioning of biological objects in cryobiology, is considered. The optoacoustic method, based on the measurement and analysis of acoustic waves induced by short laser pulses, is proposed for monitoring the cooling and freezing of the tissue. The effect of cooling and freezing on the amplitude and time profile of acoustic signals generated in real tissues and in a model object is studied. The experimental results indicate that the optoacoustic laser technique can be used for real-time monitoring of cooling and freezing of biological objects with a submillimeter spatial resolution and a high contrast. (laser biology and medicine)



    Soham Shukla


    Among the various methods of drying, this article has mentioned only one most important method, “Freeze drying”. This method is mainly used for the drying of thermo labile materials. This method works on the principle of sublimation. This method is divided into 3 steps for its better understanding; these are Freezing, Primary drying, and secondary drying. There are many advantages and disadvantages of this method, but still this is the most useful drying method nowadays.

  4. Cryo-scanning electron microscopy to study the freezing behavior of plant tissues. (United States)

    Fujikawa, Seizo; Endoh, Keita


    A cryo-scanning electron microscope (cryo-SEM) is a valuable tool for observing bulk frozen samples to monitor freezing responses of plant tissues and cells. Here, essential processes of a cryo-SEM to observe freezing behaviors of plant tissue cells are described.

  5. Effect of freezing on rabbit cultured chondrocytes

    Directory of Open Access Journals (Sweden)

    R.R Filgueiras


    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

  6. Fundamentals of freeze-drying. (United States)

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


    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

  7. Tissue Triage and Freezing for Models of Skeletal Muscle Disease (United States)

    Meng, Hui; Janssen, Paul M.L.; Grange, Robert W.; Yang, Lin; Beggs, Alan H.; Swanson, Lindsay C.; Cossette, Stacy A.; Frase, Alison; Childers, Martin K.; Granzier, Henk; Gussoni, Emanuela; Lawlor, Michael W.


    Skeletal muscle is a unique tissue because of its structure and function, which requires specific protocols for tissue collection to obtain optimal results from functional, cellular, molecular, and pathological evaluations. Due to the subtlety of some pathological abnormalities seen in congenital muscle disorders and the potential for fixation to interfere with the recognition of these features, pathological evaluation of frozen muscle is preferable to fixed muscle when evaluating skeletal muscle for congenital muscle disease. Additionally, the potential to produce severe freezing artifacts in muscle requires specific precautions when freezing skeletal muscle for histological examination that are not commonly used when freezing other tissues. This manuscript describes a protocol for rapid freezing of skeletal muscle using isopentane (2-methylbutane) cooled with liquid nitrogen to preserve optimal skeletal muscle morphology. This procedure is also effective for freezing tissue intended for genetic or protein expression studies. Furthermore, we have integrated our freezing protocol into a broader procedure that also describes preferred methods for the short term triage of tissue for (1) single fiber functional studies and (2) myoblast cell culture, with a focus on the minimum effort necessary to collect tissue and transport it to specialized research or reference labs to complete these studies. Overall, this manuscript provides an outline of how fresh tissue can be effectively distributed for a variety of phenotypic studies and thereby provides standard operating procedures (SOPs) for pathological studies related to congenital muscle disease. PMID:25078247

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

    Directory of Open Access Journals (Sweden)

    Gyoungju Nah

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

  9. Enhanced protein internalization and efficient endosomal escape using polyampholyte-modified liposomes and freeze concentration


    Ahmed, Sana; Fujita, Satoshi; Matsumura, Kazuaki


    Here we show a new strategy for efficient freeze concentration-mediated cytoplasmic delivery of proteins, obtained via the endosomal escape property of polyampholyte-modified liposomes. The freeze concentration method successfully induces the efficient internalization of proteins simply by freezing cells with protein and nanocarrier complexes. However, the mechanism of protein internalization remains unclear. Here, we designed a novel protein delivery carrier by modifying liposomes through in...

  10. A Theory of Immersion Freezing (United States)

    Barahona, Donifan


    Immersion freezing is likely involved in the initiation of precipitation and determines to large extent the phase partitioning in convective clouds. Theoretical models commonly used to describe immersion freezing in atmospheric models are based on the classical nucleation theory which however neglects important interactions near the immersed particle that may affect nucleation rates. This work introduces a new theory of immersion freezing based on two premises. First, immersion ice nucleation is mediated by the modification of the properties of water near the particle-liquid interface, rather than by the geometry of the ice germ. Second, the same mechanism that leads to the decrease in the work of germ formation also decreases the mobility of water molecules near the immersed particle. These two premises allow establishing general thermodynamic constraints to the ice nucleation rate. Analysis of the new theory shows that active sites likely trigger ice nucleation, but they do not control the overall nucleation rate nor the probability of freezing. It also suggests that materials with different ice nucleation efficiency may exhibit similar freezing temperatures under similar conditions but differ in their sensitivity to particle surface area and cooling rate. Predicted nucleation rates show good agreement with observations for a diverse set of materials including dust, black carbon and bacterial ice nucleating particles. The application of the new theory within the NASA Global Earth System Model (GEOS-5) is also discussed.

  11. Assessing storage of stability and mercury reduction of freeze-dried Pseudomonas putida within different types of lyoprotectant (United States)

    Azoddein, Abdul Aziz Mohd; Nuratri, Yana; Azli, Faten Ahada Mohd; Bustary, Ahmad Bazli


    Pseudomonas putida is a potential strain in biological treatment to remove mercury contained in the effluent of petrochemical industry due to its mercury reductase enzyme that able to reduce ionic mercury to elementary mercury. Freeze-dried P. putida allows easy, inexpensive shipping, handling and high stability of the product. This study was aimed to freeze dry P. putida cells with addition of lyoprotectant. Lyoprotectant was added into the cells suspension prior to freezing. Dried P. putida obtained was then mixed with synthetic mercury. Viability of recovery P. putida after freeze dry was significantly influenced by the type of lyoprotectant. Among the lyoprotectants, tween 80/ sucrose was found to be the best lyoprotectant. Sucrose was able to recover more than 78% (6.2E+09 CFU/ml) of the original cells (7.90E+09CFU/ml) after freeze dry and able to retain 5.40E+05 viable cells after 4 weeks storage at 4 °C without vacuum. Polyethylene glycol (PEG) pre-treated freeze dried cells and broth pre-treated freeze dried cells after the freeze-dry process recovered more than 64% (5.0 E+09CFU/ml) and >0.1% (5.60E+07CFU/ml). Freeze-dried P. putida cells in PEG and broth cannot survive after 4 weeks storage. Freeze dry also does not really change the pattern of growth P. putida but extension of lag time was found 1 hour after 3 weeks of storage. Additional time was required for freeze-dried P. putida cells to recover before introducing freeze-dried cells to more complicated condition such as mercury solution. The maximum mercury reduction of PEG pre-treated freeze-dried cells after freeze dry and after storage of 3 weeks was 17.91 %. The maximum of mercury reduction of tween 80/sucrose pre-treated freeze-dried cells after freeze dry and after storage 3 weeks was 25.03%. Freeze dried P. putida was found to have lower mercury reduction compare to the fresh P. putida that has been grown in agar. Result from this study may be beneficial and useful as initial reference before

  12. Freeze-in through portals

    CERN Document Server

    Blennow, Mattias; Zaldivar, Bryan


    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.

  13. A new cryosurgical device for controlled freezing. (United States)

    Rabin, Y; Coleman, R; Mordohovich, D; Ber, R; Shitzer, A


    A new cryosurgical device was developed in this study to facilitate examination of factors affecting the outcome of cryotreatment. Special emphasis was placed on the control of the cooling rate at the freezing front. In the new computer-controlled cryosurgical device, the controlling variable is the cryoprobe temperature, which is calculated to ensure prespecified cooling rates at the freezing front. Details of the new cryodevice, results of a validation test, and the system characteristics are presented in Part I of this study. In this part of the study initial results of 13 in vivo experimental cryotreatments, including histological observations, are presented. The in vivo pilot investigations include the normal, healthy skin and the underlying skeletal muscle of the thighs in rabbits. Using low cooling rate-controlled freezing, the new cryosurgical device is demonstrated here as an effective surgical tool. An in vivo temperature measurement technique is employed based on miniature thermocouples and X-ray images. Thermal analysis of the heat transfer in the cryotreated tissue is presented, based on the temperature measurements and on numerical heat transfer simulations. Cryotreated tissue was extracted either immediately or 4 or 7 days following the procedure. The histological observations on the skeletal muscle of the 4- and 7-day postcryoinjury were not substantially different. The effective penetration depth of the cryolesion was in the range of 5-15 mm, possibly extending up to 25 mm, depending on the specific area treated and operating parameters. The cryotreatment resulted in complete destruction of cells in the skin followed by rapid replacement by epithelial cells. Histological responses to cryotreatment of skeletal muscle were similar to those resulting from a range of traumatic episodes, e.g., crush damage. It was also found that most of the blood vessels in the cryotreated region remained intact without histological evidence of extravasation of

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


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

  15. 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


    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...

  16. Three-Dimensional Microstructure of Biological Tissues during Freezing and Thawing (United States)

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

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

  17. In vitro comparison of the efficacy of TGF-β1 and PDGF-BB in combination with freeze-dried bone allografts for induction of osteogenic differentiation in MG-63 osteoblast-like cells. (United States)

    Vahabi, Surena; Torshabi, Maryam; Esmaeil Nejad, Azadeh


    Predictable regeneration of alveolar bone defects has always been a challenge in implant dentistry. Bone allografts are widely used bone substitutes with controversial osteoinductive activity. This in vitro study aimed to assess the osteogenic potential of some commercially available freeze-dried bone allografts supplemented with human recombinant platelet-derived growth factor-BB and transforming growth factor beta-1. Cell viability, mineralization, and osteogenic gene expression of MG-63 osteoblast-like cells were compared among the allograft alone, allograft/platelet-derived growth factor-BB, allograft/transforming growth factor beta-1, and allograft/platelet-derived growth factor-BB/transforming growth factor beta-1 groups. The methyl thiazol tetrazolium assay, real-time quantitative reverse transcription polymerase chain reaction and alizarin red staining were performed, respectively, for assessment of cell viability, differentiation, and mineralization at 24-72 h post treatment. The allograft with greater cytotoxic effect on MG-63 cells caused the lowest differentiation among the groups. In comparison with allograft alone, allograft/transforming growth factor beta-1, and allograft/transforming growth factor beta-1/platelet-derived growth factor-BB caused significant upregulation of bone sialoprotein and osteocalcin osteogenic mid-late marker genes, and resulted in significantly higher amounts of calcified nodules especially in mineralized non-cytotoxic allograft group. Supplementation of platelet-derived growth factor-BB alone in 5 ng/mL concentration had no significant effect on differentiation or mineralization markers. According to the results, transforming growth factor beta-1 acts synergistically with bone allografts to enhance the osteogenic differentiation potential. Therefore, this combination may be useful for rapid transformation of undifferentiated cells into bone-forming cells for bone regeneration. However, platelet-derived growth factor

  18. The genetic characteristics in cytology and plant physiology of two wheat (Triticum aestivum) near isogenic lines with different freezing tolerances. (United States)

    Wang, Wenqiang; Hao, Qunqun; Wang, Wenlong; Li, Qinxue; Wang, Wei


    Freezing tolerance in taft plants relied more upon an ABA-independent- than an ABA-dependent antifreeze signaling pathway. Two wheat (Triticum aestivum) near isogenic lines (NIL) named tafs (freezing sensitivity) and taft (freezing tolerance) were isolated in the laboratory and their various cytological and physiological characteristics under freezing conditions were studied. Proplastid, cell membrane, and mitochondrial ultrastructure were less damaged by freezing treatment in taft than tafs plants. Chlorophyll, ATP, and thylakoid membrane protein contents were significantly higher, but malondialdehyde content was significantly lower in taft than tafs plants under freezing condition. Antioxidant capacity, as indicated by reactive oxygen species accumulation and antioxidant enzyme activity, and the relative gene expression were significantly greater in taft than tafs plants. Soluble sugars and abscisic acid (ABA) contents were significantly higher in taft plants than in tafs plants under both normal and freezing conditions. The upregulated expression levels of certain freezing tolerance-related genes were greater in taft than tafs plants under freezing treatment. The addition of sodium tungstate, an ABA synthesis inhibitor, led to only partial freezing tolerance inhibition in taft plants and the down-regulated expression of some ABA-dependent genes. Thus, both ABA-dependent and ABA-independent signaling pathways are involved in the freezing tolerance of taft plants. At the same time, freezing tolerance in taft plants relied more upon an ABA-independent- than an ABA-dependent antifreeze signaling pathway.

  19. Experimental Investigation of Thermal Conductivity of Meat During Freezing (United States)

    Shinbayeva, A.; Arkharov, I.; Aldiyarov, A.; Drobyshev, A.; Zhubaniyazova, M.; Kurnosov, V.


    The cryogenic technologies of processing and storage of agricultural products are becoming increasingly indispensable in the food industry as an important factor of ensuring food safety. One of such technologies is the shock freezing of meat, which provides a higher degree of preservation of the quality of frozen products in comparison with traditional technologies. The thermal conductivity of meat is an important parameter influencing the energy consumption in the freezing process. This paper presents the results of an experimental investigation of the temperature dependence of the thermal conductivity of beef. The measurements were taken by using a specially designed measurement cell, which allows covering the temperature range from 80 to 300 K.

  20. 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.


    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

  1. A tissue snap-freezing apparatus without sacrificial cryogens (United States)

    Vanapalli, Srinivas; Jagga, Sahil; Holland, Harry; ter Brake, Marcel


    Molecular technologies in cancer diagnosis require a fresh and frozen tissue, which is obtained by means of snap-freezing. Currently, coolants such as solid carbon dioxide and liquid nitrogen are used to preserve good morphology of the tissue. Using these coolants, snap freezing of tissues for diagnostic and research purposes is often time consuming, laborious, even hazardous and not user friendly. For that reason snap-freezing is not routinely applied at the location of biopsy acquisition. Furthermore, the influence of optimal cooling rate and cold sink temperature on the viability of the cells is not well known. In this paper, a snap-freezing apparatus powered by a small cryocooler is presented that will allow bio-medical research of tissue freezing methods and is safe to use in a hospital. To benchmark this apparatus, cooldown of a standard aluminum cryo-vial in liquid nitrogen is measured and the cooling rate is about -25 K/s between 295 K and 120 K. Sufficient cooling rate is obtained by a forced convective helium gas flow through a gap formed between the cryo-vial and a cold surface and is therefore chosen as the preferred cooling method. A conceptual design of the snap-apparatus with forced flow is discussed in this paper.

  2. 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.

  3. Influence of the freezing method on the changes that occur in grape samples after frozen storage. (United States)

    Santesteban, Luis G; Miranda, Carlos; Royo, José B


    Sample freezing is frequently used in oenological laboratories as a compromise solution to increase the number of samples that can be analysed, despite the fact that some grape characteristics are known to change after frozen storage. However, freezing is usually performed using standard freezers, which provide a slow freezing. The aim of this work was to evaluate whether blast freezing would decrease the impact of standard freezing on grape composition. Grape quality parameters were assessed in fresh and in frozen stored samples that had been frozen using three different procedures: standard freezing and blast freezing using either a blast freezer or an ultra-freezer. The implications of frozen storage in grape samples reported in earlier research were observed for the three freezing methods evaluated. Although blast freezing improved repeatability for the most problematic parameters (tartaric acidity, TarA; total phenolics, TP), the improvement was not important from a practical point of view. However, TarA and TP were relatively repeatable among the three freezing procedures, which suggests that freezing had an effect on these parameters independently of the method used . According to our results, the salification potential of the must is probably implied in the changes observed for TarA, whereas for TP the precipitation of protoanthocyanins after association with cell wall material is hypothesized to cause the lack of repeatability between fresh and frozen grapes. Blast freezing would not imply a great improvement if implemented in oenological laboratories, at least for the parameters included in this study. © 2013 Society of Chemical Industry.

  4. State transitions and physicochemical aspects of cryoprotection and stabilization in freeze-drying of Lactobacillus rhamnosus GG (LGG). (United States)

    Pehkonen, K S; Roos, Y H; Miao, S; Ross, R P; Stanton, C


    The frozen and dehydrated state transitions of lactose and trehalose were determined and studied as factors affecting the stability of probiotic bacteria to understand physicochemical aspects of protection against freezing and dehydration of probiotic cultures. Lactobacillus rhamnosus GG was frozen (-22 or -43 degrees C), freeze-dried and stored under controlled water vapour pressure (0%, 11%, 23% and 33% relative vapour pressure) conditions. Lactose, trehalose and their mixture (1 : 1) were used as protective media. These systems were confirmed to exhibit relatively similar state transition and water plasticization behaviour in freeze-concentrated and dehydrated states as determined by differential scanning calorimetry. Ice formation and dehydrated materials were studied using cold-stage microscopy and scanning electron microscopy. Trehalose and lactose-trehalose gave the most effective protection of cell viability as observed from colony forming units after freezing, dehydration and storage. Enhanced cell viability was observed when the freezing temperature was -43 degrees C. State transitions of protective media affect ice formation and cell viability in freeze-drying and storage. Formation of a maximally freeze-concentrated matrix with entrapped microbial cells is essential in freezing prior to freeze-drying. Freeze-drying must retain a solid amorphous state of protectant matrices. Freeze-dried matrices contain cells entrapped in the protective matrices in the freezing process. The retention of viability during storage seems to be controlled by water plasticization of the protectant matrix and possibly interactions of water with the dehydrated cells. Highest cell viability was obtained in glassy protective media. This study shows that physicochemical properties of protective media affect the stability of dehydrated cultures. Trehalose and lactose may be used in combination, which is particularly important for the stabilization of probiotic bacteria in dairy

  5. Thermodynamics of freezing and melting


    Pedersen, Ulf Rørbæk; Costigliola, Lorenzo; Bailey, Nicholas; Schrøder, Thomas; Dyre, Jeppe C.


    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 variatio...

  6. Medical and social egg freezing

    DEFF Research Database (Denmark)

    Lallemant, Camille; Vassard, Ditte; Andersen, Anders Nyboe


    INTRODUCTION: Until recently, limited options for preserving fertility in order to delay childbearing were available. Although egg freezing and successful thawing is now possible, it remains unclear to what extent women are aware of the availability of this technique, their attitudes towards its ...... planning. Reassurance regarding its efficacy appears more important than its potential adverse effects on their health or that of future children, or the costs of the procedure....

  7. Impregnation of leather during "freeze-drying"

    DEFF Research Database (Denmark)

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


    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...... suggest that the process which takes place within the leather during the freeze-drying in not actual freeze-drying, but rather a sophisticated way of distributing the impregnating agent. The pure ice phase freezes out, but the impregnating agent remains liquid as the temperature does not become low enough...

  8. PHBV/PLLA-based composite scaffolds fabricated using an emulsion freezing/freeze-drying technique for bone tissue engineering: surface modification and in vitro biological evaluation

    International Nuclear Information System (INIS)

    Sultana, Naznin; Wang Min


    Tissue engineering combines living cells with biodegradable materials and/or bioactive components. Composite scaffolds containing biodegradable polymers and nanosized osteoconductive bioceramic with suitable properties are promising for bone tissue regeneration. In this paper, based on blending two biodegradable and biocompatible polymers, namely poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and poly(l-lactic acid) (PLLA) with incorporated nano hydroxyapatite (HA), three-dimensional composite scaffolds with controlled microstructures and an interconnected porous structure, together with high porosity, were fabricated using an emulsion freezing/freeze-drying technique. The influence of various parameters involved in the emulsion freezing/freeze-drying technique was studied for the fabrication of good-quality polymer scaffolds based on PHBV polymers. The morphology, mechanical properties and crystallinity of PHBV/PLLA and HA in PHBV/PLLA composite scaffolds and PHBV polymer scaffolds were studied. The scaffolds were coated with collagen in order to improve wettability. During in vitro biological evaluation study, it was observed that SaOS-2 cells had high attachment on collagen-coated scaffolds. Significant improvement in cell proliferation and alkaline phosphatase activity for HA-incorporated composite scaffolds was observed due to the incorporation of HA. After 3 and 7 days of culture on all scaffolds, SaOS-2 cells also had normal morphology and growth. These results indicated that PHBV/PLLA-based scaffolds fabricated via an emulsion freezing/freeze-drying technique were favorable sites for osteoblastic cells and are promising for the applications of bone tissue engineering.

  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


    superoxide dismutase to compensate for the lack of the cytoplasmic enzyme. Free radicals generated as a result of freezing and thawing were detected in cells directly using electron paramagnetic resonance spectroscopy with either alpha-phenyl-N-tert-butylnitrone or 5, 5-dimethyl-1-pyrroline-N-oxide as spin...... 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. The Use of High Pressure Freezing and Freeze Substitution to Study Host-Pathogen Interactions in Fungal Diseases of Plants (United States)

    Mims, C. W.; Celio, Gail J.; Richardson, Elizabeth A.


    This article reports on the use of high pressure freezing followed by freeze substitution (HPF/FS) to study ultrastructural details of host pathogen interactions in fungal diseases of plants. The specific host pathogen systems discussed here include a powdery mildew infection of poinsettia and rust infections of daylily and Indian strawberry. The three pathogens considered here all attack the leaves of their hosts and produce specialized hyphal branches known as haustoria that invade individual host cells without killing them. We found that HPF/FS provided excellent preservation of both haustoria and host cells for all three host pathogen systems. Preservation of fungal and host cell membranes was particularly good and greatly facilitated the detailed study of host pathogen interfaces. In some instances, HPF/FS provided information that was not available in samples prepared for study using conventional chemical fixation. On the other hand, we did encounter various problems associated with the use of HPF/FS. Examples included freeze damage of samples, inconsistency of fixation in different samples, separation of plant cell cytoplasm from cell walls, breakage of cell walls and membranes, and splitting of thin sections. However, we believe that the outstanding preservation of ultrastructural details afforded by HPF/FS significantly outweighs these problems and we highly recommend the use of this fixation protocol for future studies of fungal host-plant interactions.

  11. Sequential Strangeness Freeze-out (United States)

    Bellwied, Rene


    I will describe the latest results from lattice QCD pertaining to a potential flavour hierarchy in the hadronic freeze-out from the QCD crossover region. I will compare these results to a variety of improved hadronic resonance gas calculations and to experimental data of fluctuations of net-charge, net-proton and net-kaon multiplicity distributions, which serve as a proxy for the susceptibilities of conserved quantum numbers on the lattice. I will conclude that there is intriguing evidence for a flavour dependent freezeout, and I will suggest expansions to the experimental program at RHIC and the LHC that could potentially demonstrate the impact of a flavour separation during hadronization.

  12. DNA comet assay to identify different freezing temperatures of irradiated liver chicken

    International Nuclear Information System (INIS)

    Duarte, Renato C.; Mozeika, Michel A.; Fanaro, Gustavo B.; Villavicencio, Anna L.C.H.; Marchioni, Eric


    The cold chain is a succession of steps which maintain the food at low temperature. The thawed food never be frozen again and the best solution being to consume it quickly to avoid the microorganism growth which causes decay and nutrients damage. One of most important point is that freezing process, unlike irradiation, do not destroy microorganisms, only inactive them as long as they remain in a frozen state. The Comet Assay is an original test used to detect irradiated foods that's recognize the DNA damage and can then be used to control the overall degradation of the food and in a certain extend to evaluate the damage caused by irradiation, different forms of freeze and storage time on liver chicken cells. Different freezing temperatures were used, deep freeze -196 deg C and slow freeze -10 deg C. Samples were irradiated in a 60 Co irradiator with 1.5, 3.0 and 4.5 kGy radiation doses. Fast freezing technique induces a low percent of DNA degradation comparing to slow freezing technique. This procedure could be a good choose to chicken freezing processing. (author)

  13. The vulnerability to freezing-induced xylem cavitation of Larrea tridentata (Zygophyllaceae) in the Chihuahuan desert. (United States)

    Martínez-Vilalta, Jordi; Pockman, William T


    The temperature dependence of freezing-induced xylem cavitation was studied in a Chihuahuan desert population of Larrea tridentata (Zygophyllaceae). Field measurements of wood temperature and xylem embolism were combined with anatomical studies and laboratory measurements of embolism in stem and root samples frozen under controlled conditions. Our laboratory experiments corroborated the previously observed relationship between minimum freezing temperature and embolism. The area of the low-temperature exotherms produced during the freezing treatments was correlated with the resulting embolism, suggesting that the freezing of water inside parenchyma cells is associated with the occurrence of xylem embolism. In the laboratory experiments, embolism in stems increased only at temperatures below -14°C. Although this meant that the studied population was more resistant to freezing-induced xylem embolism than a previously studied population from the Sonoran desert, the impact of freezing was nevertheless greater because of much lower environmental temperatures. This result suggests that dieback associated with periodic extreme freezes may contribute to limiting the present distribution of L. tridentata in central New Mexico. Although laboratory experiments showed that root xylem embolism increased after freezing to less negative minimum temperatures than stems (significant effects at T = -7°C), root embolism in the field was lower than shoot embolism in accordance with measured soil temperatures throughout the study.

  14. Well-plate freeze-drying

    DEFF Research Database (Denmark)

    Trnka, Hjalte; Rantanen, Jukka; Grohganz, Holger


    Abstract Context: Freeze-drying in presence of excipients is a common practice to stabilize biomacromolecular formulations. The composition of this formulation is known to affect the quality of the final product. Objective: The aim of this study was to evaluate freeze-drying in well...... no significant edge effect was found for any of the quality attributes analyzed. Conclusion: Freeze-drying in well-plates was found to be a suitable and representative high throughput platform for formulation screening....

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

  16. Freeze dehydration of milk using microwave energy

    International Nuclear Information System (INIS)

    Souda, K.B.; Akyel, C.; Bilgen, E.


    This paper presents the results of experimental studies on heat and mass transfer during a microwave freeze dehydration process. An experimental system and procedure was developed to freeze dry milk. A 2500-W microwave system with an appropriate wave guide was set up and instrumented, and a procedure was experimentally developed to obtain milk powder first by freezing milk and then dehydrating it at low pressure using microwave energy. An unsteady-state analysis was used to derive a one-dimensional mathematical model of the freeze dehydration process in a microwave electromagnetic field

  17. Freezing and thawing of processed meat in an industrial freezing tunnel


    Glaucio Antonio Marini; Eduarda Molardi Bainy; Marcelo Kaminski Lenzi; Marcos Lúcio Corazza


    Freezing is a commonly used preservation method in the meat industry. The understanding of the product behavior during the freezing process can assist in a better process management and quality control. This work reports the study of freezing and thawing of three types of processed meat in order to determine process parameters in an industrial forced‑air freezing tunnel at ‑30oC. Chicken sausages (frankfurter type), mortadela (bologna type) and mechanically deboned chicken meat (MDCM) were st...

  18. Mechanisms of deterioration of nutrients. [of freeze dried foods (United States)

    Karel, M.; Flink, J. M.


    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.

  19. The Design and Performance of a PEFC at a Temperature Below Freezing (United States)

    Hishinuma, Yukio; Chikahisa, Takemi; Kagami, Fumio; Ogawa, Tomohiro

    At temperatures below freezing, air humidity becomes lower and produced water at the cathode freezes on the surface of catalyst, and it is difficult to start a PEFC (Polymer Electrolyte Fuel Cell) at a cold district. The object of the work is to study the performance of the fuel cell below the freezing point by experiments and simulation. To investigate the characteristics of the starting of a temperature below freezing the performance of a single cell was measured at temperatures from -3 to -25°C and pressures from 1 to 2 atm. The results of the experiments and simulation indicate that the performance of a PEFC decreases at higher current densities and pressures, and lower cell temperatures because of ice more produced on the reactive area of the cathode. To maintain the cell performance below freezing point, it is effective to adjust the current densities and gas flow rate to balance the produced and removed water. However at -5°C, heat generated in the fuel cell is effective to warm the cell and make self-starting possible. These results shows that it is necessary to heat the cell with an additional heat source in order to start the fuel cell below -5°C.

  20. 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

  1. Anhydrobiosis and freezing-tolerance: adaptations that facilitate the establishment of Panagrolaimus nematodes in polar habitats. (United States)

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


    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

  2. Estimation of the freezing point of concentrated fruit juices for application in freeze concentration


    Auleda Amorós, Josep Maria; Raventós Santamaria, Mercè; Sánchez Machado, José; Hernández Yáñez, Eduard


    In freeze concentration operations the fluids remain at temperatures below 0 °C. For a good study of this concentration operation is very important to know the values of freezing point. The aim of this work was to establish a model that predicts the freezing point of fruit juices at various concentrations within the range of interest for freeze concentration (10–40 °Brix). The model proposed relates the freezing point of a juice with the concentrations of main sugars present in the juice: suc...

  3. Monitoring the Freezing Point of Buffalo Milk. (United States)

    Pesce, Antonella; Salzano, Caterina; De Felice, Anna; Garofalo, Francesca; Liguori, Salvatore; De Santo, Annunziata; Palermo, Pierpaolo; Guarino, Achille


    The aim of this study was to evaluate the basic freezing point of buffalo milk. Bulk milk samples were collected from buffalo and cattle farms in Caserta area from 2008 to 2014. The analysis involved a total of 1886 buffalo milk samples and 1711 bovine milk samples. These were also tested for fat, protein and lactose contents by means of infrared spectrometry. The freezing point was determined by means of a thermistor cryoscope. Data underwent statistical analysis. Our research showed an average freezing point of -0.528°C for buffalo milk and -0.522°C for bovine milk. Given the lack of data on the freezing point of buffalo milk, our study provides the first indication of a basic freezing point of the milk of this species in Italy.

  4. Transient freezing behavior in photophobic responses of Euglena gracilis investigated in a microfluidic device. (United States)

    Ozasa, Kazunari; Lee, Jeesoo; Song, Simon; Maeda, Mizuo


    We found that the transient freezing behavior in photophobic responses of Euglena gracilis is a good indicator of the metabolic status of the cells. The transient blue light photophobic responses of E. gracilis cells were investigated on-chip using a new measurement, 'trace momentum' (TM), to evaluate their swimming activity quantitatively in real time. When blue light of intensity >30 mW cm(-2) was repeatedly switched on and off, a large negative spike in the TM was observed at the onset of the 'blue-light-off' phase. Single-cell trace analysis at a blue light intensity of 40 mW cm(-2) showed that 48% (on average, n = 15) of tumbling Euglena cells ceased activity ('freezing') for 2-30 s at the onset of blue-light-off before commencing forward motion in a straight line (termed 'straightforward swimming'), while 45% smoothly commenced straightforward swimming without delay. The proportion of freezing Euglena cells depended on the blue light intensity (only 20% at 20 mW cm(-2)). When the cells were stimulated by four blue light pulses at the higher intensity, without pre-exposure, the transient freezing behavior was more prominent but, on repeating the stimuli after an 80 min interval in red light, the same cells did not freeze. This shows that the metabolism of the cells had changed to anti-freezing during the interval. The relationship between the interval time with/without light irradiation and the blue light adaptation was elucidated experimentally. The origin of the freezing behavior is considered to be a shortage of a metabolic substance that promotes smooth switching of flagellum movement from in situ rotation mode to a straightforward swimming mode. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email:

  5. Influence of dough freezing on Saccharomyces cerevisiae metabolism

    Directory of Open Access Journals (Sweden)

    Pejin Dušanka J.


    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.

  6. 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.


    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

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

  8. Initial boost release of transforming growth factor-β3 and chondrogenesis by freeze-dried bioactive polymer scaffolds. (United States)

    Krüger, Jan Philipp; Machens, Isabel; Lahner, Matthias; Endres, Michaela; Kaps, Christian


    In cartilage regeneration, bio-activated implants are used in stem and progenitor cell-based microfracture cartilage repair procedures. Our aim was to analyze the chondrogenic potential of freeze-dried resorbable polymer-based polyglycolic acid (PGA) scaffolds bio-activated with transforming growth factor-β3 (TGFB3) on human subchondral mesenchymal progenitor cells known from microfracture. Progenitor cells derived from femur heads were cultured in the presence of freeze-dried TGFB3 in high-density pellet culture and in freeze-dried TGFB3-PGA scaffolds for chondrogenic differentiation. Progenitor cell cultures in PGA scaffolds as well as pellet cultures with and without continuous application of TGFB3 served as controls. Release studies showed that freeze-dried TGFB3-PGA scaffolds facilitate a rapid, initial boost-like release of 71.5% of TGFB3 in the first 10 h. Gene expression analysis and histology showed induction of typical chondrogenic markers like type II collagen and formation of cartilaginous tissue in TGFB3-PGA scaffolds seeded with subchondral progenitor cells and in pellet cultures stimulated with freeze-dried TGFB3. Chondrogenic differentiation in freeze-dried TGFB3-PGA scaffolds was comparable to cultures receiving TGFB3 continuously, while non-stimulated controls did not show chondrogenesis during prolonged culture for 14 days. These results suggest that bio-activated, freeze-dried TGFB3-PGA scaffolds have chondrogenic potential and are a promising tool for stem cell-mediated cartilage regeneration.

  9. Protein freeze concentration and micro-segregation analysed in a temperature-controlled freeze container

    Directory of Open Access Journals (Sweden)

    Ulrich Roessl


    Full Text Available To examine effects of varied freezing conditions on the development of spatial heterogeneity in the frozen protein solution, macroscopic freeze concentration and micro-segregation of bovine serum albumin (BSA were investigated in a temperature-controlled 200-ml freeze container. Freezing to −40 °C promoted formation of protein concentration gradients (69–114 μg ml−1 in frozen samples taken from 12 different freezer positions, whereby slow freezing in 4 h or longer facilitated the evolution of strong spatial heterogeneities and caused local concentration increases by 1.15-fold relative to the initial protein concentration (100 μg ml−1. To visualize protein micro-segregation during phase separation, BSA was conjugated with fluorescein isothiocyanate and confocal laser scanning fluorescence microscopy was used to localize and size the freeze-concentrated protein regions. Slow freezing resulted in distinctly fewer and larger protein domains in the frozen bulk than fast freezing. Surface stress on the protein during freezing would therefore be minimized at low cooling rates; microscopic freeze concentration would however be highest under these conditions, potentially favoring protein aggregation.

  10. Hot big bang or slow freeze? (United States)

    Wetterich, C.


    We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze - a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple ;crossover model; without a big bang singularity. In the infinite past space-time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe.

  11. Study of the conditions necessary for propane-jet freezing of fresh biological tissues without detectable ice formation. (United States)

    Haggis, G H


    The performance of a commercial double-propane-jet freezer (Balzers QFD 101) has been assessed, for rapid freezing of fresh tissues in freeze-etch work. Samples of diaphragm muscle and intestinal villi were frozen between copper sheets, with a spacer to give 20-30 microns thickness of tissue. Fracture cuts were made with the Balzers BAF 400 freeze-etch microtome within 5-10 microns of a freezing face (i.e. a tissue face in contact with the copper sheets of the frozen sandwich). After some modifications to the QFD 101, replicas showing no evidence of ice were obtained of muscle cells, although for intestinal epithelial cells some evidence of ice formation was found. Infiltration with 5% glycerol or dimethylsulphoxide improves the depth of good freezing. Results and problems arising from such infiltration are briefly discussed.

  12. Effects of six substances on the growth and freeze-drying of Lactobacillus delbrueckii subsp. bulgaricus. (United States)

    Chen, He; Huang, Jie; Shi, Xiaoyu; Li, Yichao; Liu, Yu


    The efficacy of Lactobacillus delbrueckii subsp. bulgaricus as starter cultures for the dairy industry depends largely on the number of viable and active cells. Freeze-drying is the most convenient and successful method to preserve the bacterial cells. However, not all strains survived during freeze-drying. The effects of six substances including NaCl, sorbitol, mannitol, mannose, sodium glutamate, betaine added to the MRS medium on the growth and freeze-drying survival rate and viable counts of Lb. delbrueckii subsp. bulgaricus were studied through a single-factor test and Plackett-Burman design. Subsequently, the optimum freeze-drying conditions of Lb. delbrueckii subsp. bulgaricus were determined. Lb. delbrueckii subsp. bulgaricus survival rates were up to the maximum of 42.7%, 45.4%, 23.6%, while the concentrations of NaCl, sorbitol, sodium glutamate were 0.6%, 0.15%, 0.09%, respectively. In the optimum concentration, the viable counts in broth is 6.1, 6.9, 5.13 (×108 CFU/mL), respectively; the viable counts in freeze-drying power are 3.09, 5.2, 2.7 (×1010 CFU/g), respectively. Three antifreeze factors including NaCl, sorbitol, sodium glutamate have a positive effect on the growth and freeze-drying of Lb. delbrueckii subsp. bulgaricus. The results are beneficial for developing Lb. delbrueckii subsp. bulgaricus.

  13. Ultrasound assisted immersion freezing of broccoli (Brassica oleracea L. var. botrytis L.). (United States)

    Xin, Ying; Zhang, Min; Adhikari, Benu


    The aim of this study was to research the ultrasound-assisted freezing (UAF) of broccoli. CaCl2 solution was used as freezing medium. The comparative advantage of using UAF over normal freezing on the freezing time, cell-wall bound calcium to total calcium ratio, textural properties, color, drip loss and L-ascorbic acid contents was evaluated. The application of UAF at selected acoustic intensity with a range of 0.250-0.412 W/cm(2) decreased the freezing time and the loss of cell-wall bound calcium content. Compared to normal freezing, the values of textural properties, color, L-ascorbic acid content were better preserved and the drip loss was significantly minimized by the application of UAF. However, when outside that range of acoustic intensity, the quality of the ultrasound-assisted frozen broccoli was inferior compared to that of the normally frozen samples. Selected the appropriate acoustic intensity was very important for the application of UAF. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Breeding of Freeze-tolerant Yeast and the Mechanisms of Stress-tolerance (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. Tailorable Porous Ceramics via Freeze Casting (United States)

    National Aeronautics and Space Administration — Freeze casting of ceramics is a novel technique used to produce porous materials. The process involves solidifying a solvent in a ceramic slurry to produce a frozen...

  16. Freezing and refrigerated storage in fisheries

    National Research Council Canada - National Science Library

    Johnston, W. A


    ...; the factors affecting cold storage conditions, etc. In addition, the publication describes the methods used to calculate cold storage refrigeration loads as well as the costs of freezing and cold storage...

  17. Global Annual Freezing and Thawing Indices (United States)

    National Aeronautics and Space Administration — The total annual freezing and thawing indices are defined as the cumulative number of degree-days when air temperatures are below and above zero degrees Celsius. The...

  18. Freezing tolerance of conifer seeds and germinants. (United States)

    Hawkins, B J; Guest, H J; Kolotelo, D


    Survival after freezing was measured for seeds and germinants of four seedlots each of interior spruce (Picea glauca x engelmannii complex), lodgepole pine (Pinus contorta Dougl. ex Loud.), Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) and western red cedar (Thuja plicata Donn ex D. Donn). Effects of eight seed treatments on post-freezing survival of seeds and germinants were tested: dry, imbibed and stratified seed, and seed placed in a growth chamber for 2, 5, 10, 15, 20 or 30 days in a 16-h photoperiod and a 22/17 degrees C thermoperiod. Survival was related to the water content of seeds and germinants, germination rate and seedlot origin. After freezing for 3 h at -196 degrees C, dry seed of most seedlots of interior spruce, Douglas-fir and western red cedar had 84-96% germination, whereas lodgepole pine seedlots had 53-82% germination. Freezing tolerance declined significantly after imbibition in lodgepole pine, Douglas-fir and interior spruce seed (western red cedar was not tested), and mean LT50 of imbibed seed of these species was -30, -24.5 and -20 degrees C, respectively. Freezing tolerance continued to decline to a minimum LT50 of -4 to -7 degrees C after 10 days in a growth chamber for interior spruce, Douglas-fir and lodgepole pine, or after 15 days for western red cedar. Minimum freezing tolerance was reached at the stage of rapid hypocotyl elongation. In all species, a slight increase in freezing tolerance of germinants was observed once cotyledons emerged from the seed coat. The decrease in freezing tolerance during the transition from dry to germinating seed correlated with increases in seed water content. Changes in freezing tolerance between 10 and 30 days in the growth chamber were not correlated with seedling water content. Within a species, seedlots differed significantly in freezing tolerance after 2 or 5 days in the growth chamber. Because all seedlots of interior spruce and lodgepole pine germinated quickly, there was no correlation

  19. Freezing pattern and frost killing temperature of apple (Malus domestica) wood under controlled conditions and in nature. (United States)

    Pramsohler, Manuel; Hacker, Jürgen; Neuner, Gilbert


    The freezing pattern and frost killing temperatures of apple (Malus domestica Borkh.) xylem were determined by differential thermal analysis and infrared differential thermal analysis (IDTA). Results from detached or attached twigs in controlled freezing experiments and during natural field freezing of trees were compared. Non-lethal freezing of apoplastic water in apple xylem as monitored during natural winter frosts in the field occurred at -1.9 ± 0.4 °C and did not change seasonally. The pattern of whole tree freezing was variable and specific to the environmental conditions. On detached twigs high-temperature freezing exotherms (HTEs) occurred 2.8 K below the temperature observed under natural frosts in the field with a seasonal mean of -4.7 ± 0.5 °C. Microporous apple xylem showed freezing without a specific pattern within a few seconds in IDTA images during HTEs, which is in contrast to macroporous xylem where a 2D freezing pattern mirrors anatomical structures. The pith tissue always remained unfrozen. Increasing twig length increased ice nucleation temperature; for increased twig diameter the effect was not significant. In attached twigs frozen in field portable freezing chambers, HTEs were recorded at a similar mean temperature (-4.6 ± 1.0 °C) to those for detached twigs. Upon lethal intracellular freezing of apple xylem parenchyma cells (XPCs) low-temperature freezing exotherms (LTEs) can be recorded. Low-temperature freezing exotherms determined on detached twigs varied significantly between a winter minimum of -36.9 °C and a summer maximum -12.7 °C. Within the temperature range wherein LTEs were recorded by IDTA in summer (-12.7 ± 0.5 to -20.3 ± 1.1 °C) various tiny clearly separated discontinuous freezing events could be detected similar to that in other species with contrasting XPC anatomy. These freezing events appeared to be initially located in the primary and only later in the secondary xylem. During the LTE no

  20. Monitoring the Freezing Point of Buffalo Milk


    Pesce, Antonella; Salzano, Caterina; De Felice, Anna; Garofalo, Francesca; Liguori, Salvatore; De Santo, Annunziata; Palermo, Pierpaolo; Guarino, Achille


    The aim of this study was to evaluate the basic freezing point of buffalo milk. Bulk milk samples were collected from buffalo and cattle farms in Caserta area from 2008 to 2014. The analysis involved a total of 1886 buffalo milk samples and 1711 bovine milk samples. These were also tested for fat, protein and lactose contents by means of infrared spectrometry. The freezing point was determined by means of a thermistor cryoscope. Data underwent statistical analysis. Our research showed an aver...

  1. Bulk water freezing dynamics on superhydrophobic surfaces (United States)

    Chavan, S.; Carpenter, J.; Nallapaneni, M.; Chen, J. Y.; Miljkovic, N.


    In this study, we elucidate the mechanisms governing the heat-transfer mediated, non-thermodynamic limited, freezing delay on non-wetting surfaces for a variety of characteristic length scales, Lc (volume/surface area, 3 mm commercial superhydrophobic spray coatings, showing a monotonic increase in freezing time with coating thickness. The added thermal resistance of thicker coatings was much larger than that of the nanoscale superhydrophobic features, which reduced the droplet heat transfer and increased the total freezing time. Transient finite element method heat transfer simulations of the water slab freezing process were performed to calculate the overall heat transfer coefficient at the substrate-water/ice interface during freezing, and shown to be in the range of 1-2.5 kW/m2K for these experiments. The results shown here suggest that in order to exploit the heat-transfer mediated freezing delay, thicker superhydrophobic coatings must be deposited on the surface, where the coating resistance is comparable to the bulk water/ice conduction resistance.

  2. 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


    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.

  3. Freezing the Master Production Schedule Under Rolling Planning Horizons


    V. Sridharan; William L. Berry; V. Udayabhanu


    The stability of the Master Production Schedule (MPS) is a critical issue in managing production operations with a Material Requirements Planning System. One method of achieving stability is to freeze some portion or all of the MPS. While freezing the MPS can limit the number of schedule changes, it can also produce an increase in production and inventory costs. This paper examines three decision variables in freezing the MPS: the freezing method, the freeze interval length, and the planning ...

  4. Thermal analysis of ice and glass transitions in insects that do and do not survive freezing. (United States)

    Rozsypal, Jan; Moos, Martin; Šimek, Petr; Koštál, Vladimír


    Some insects rely on the strategy of freeze tolerance for winter survival. During freezing, extracellular body water transitions from the liquid to solid phase and cells undergo freeze-induced dehydration. Here we present results of a thermal analysis (from differential scanning calorimetry) of ice fraction dynamics during gradual cooling after inoculative freezing in variously acclimated larvae of two drosophilid flies, Drosophila melanogaster and Chymomyza costata. Although the species and variants ranged broadly between 0 and close to 100% survival of freezing, there were relatively small differences in ice fraction dynamics. For instance, the maximum ice fraction (IF max ) ranged between 67.9 and 77.7% total body water (TBW). The C. costata larvae showed statistically significant phenotypic shifts in parameters of ice fraction dynamics (melting point and IF max ) upon entry into diapause, cold-acclimation, and feeding on a proline-augmented diet. These differences were mostly driven by colligative effects of accumulated proline (ranging between 6 and 487 -1 TBW) and other metabolites. Our data suggest that these colligative effects per se do not represent a sufficient mechanistic explanation for high freeze tolerance observed in diapausing, cold-acclimated C. costata larvae. Instead, we hypothesize that accumulated proline exerts its protective role via a combination of mechanisms. Specifically, we found a tight association between proline-induced stimulation of glass transition in partially-frozen body liquids (vitrification) and survival of cryopreservation in liquid nitrogen. © 2018. Published by The Company of Biologists Ltd.

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

    International Nuclear Information System (INIS)

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


    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. - Highlights: • Silk fibroin/collagen was fabricated using 3D printing. • Physical characterization and Cell compatibility were compared. • 3D printed scaffold exhibited better overall performance.

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

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Kai [Tianjin First Center Hospital, No. 24 Fukang Road, Tianjin, TJ 300192 (China); Li, Ruixin [Institute of Medical Equipment, Academy of Military and Medical Sciences, No. 106, Wandong Street, Hedong District, Tianjin 300000 (China); Jiang, Wenxue, E-mail: [Tianjin First Center Hospital, No. 24 Fukang Road, Tianjin, TJ 300192 (China); Sun, Yufu [Tianjin First Center Hospital, No. 24 Fukang Road, Tianjin, TJ 300192 (China); Li, Hui [Tianjin Medical University General Hospital, No. 154 Anshan Road, Tianjin, TJ 300052 (China)


    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. - Highlights: • Silk fibroin/collagen was fabricated using 3D printing. • Physical characterization and Cell compatibility were compared. • 3D printed scaffold exhibited better overall performance.

  7. Evaluation of freeze-substitution and conventional embedding protocols for routine electron microscopic processing of eubacteria

    International Nuclear Information System (INIS)

    Graham, L.L.; Beveridge, T.J.


    Freeze-substitution and more conventional embedding protocols were evaluated for their accurate preservation of eubacterial ultrastructure. Radioisotopes were specifically incorporated into the RNA, DNA, peptidoglycan, and lipopolysaccharide of two isogenic derivatives of Escherichia coli K-12 as representative gram-negative eubacteria and into the RNA and peptidoglycan of Bacillus subtilis strains 168 and W23 as representative gram-positive eubacteria. Radiolabeled bacteria were processed for electron microscopy by conventional methods with glutaraldehyde fixation, osmium tetroxide postfixation, dehydration in either a graded acetone or ethanol series, and infiltration in either Spurr or Epon 812 resin. A second set of cells were simultaneously freeze-substituted by plunge-freezing in liquid propane, substituting in anhydrous acetone containing 2% (wt/vol) osmium tetroxide, and 2% (wt/vol) uranyl acetate, and infiltrating in Epon 812. Extraction of radiolabeled cell components was monitored by liquid scintillation counting at all stages of processing to indicate retention of cell labels. Electron microscopy was also used to visually confirm ultrastructural integrity. Radiolabeled nucleic acid and wall components were extracted by both methods. In conventionally embedded specimens, dehydration was particularly damaging, with ethanol-dehydrated cells losing significantly more radiolabeled material during dehydration and subsequent infiltration than acetone-treated cells. For freeze-substituted specimens, postsubstitution washes in acetone were the most deleterious step for gram-negative cells, while infiltration was more damaging for gram-positive cells

  8. SENSITIVE TO FREEZING2 Aides in Resilience to Salt and Drought in Freezing-Sensitive Tomato. (United States)

    Wang, Kun; Hersh, Hope Lynn; Benning, Christoph


    SENSITIVE TO FREEZING2 (SFR2) is crucial for protecting chloroplast membranes following freezing in Arabidopsis (Arabidopsis thaliana). It has been shown that SFR2 homologs are present in all land plants, including freezing-sensitive species, raising the question of SFR2 function beyond freezing tolerance. Similar to freezing, salt and drought can cause dehydration. Thus, it is hypothesized that in freezing-sensitive plants SFR2 may play roles in their resilience to salt or drought. To test this hypothesis, SlSFR2 RNAi lines were generated in the cold/freezing-sensitive species tomato (Solanum lycopersicum [M82 cv]). Hypersensitivity to salt and drought of SlSFR2-RNAi lines was observed. Higher tolerance of wild-type tomatoes was correlated with the production of trigalactosyldiacylglycerol, a product of SFR2 activity. Tomato SFR2 in vitro activity is Mg 2+ -dependent and its optimal pH is 7.5, similar to that of Arabidopsis SFR2, but the specific activity of tomato SFR2 in vitro is almost double that of Arabidopsis SFR2. When salt and drought stress were applied to Arabidopsis, no conditions could be identified at which SFR2 was induced prior to irreversibly impacting plant growth, suggesting that SFR2 protects Arabidopsis primarily against freezing. Discovery of tomato SFR2 function in drought and salt resilience provides further insights into general membrane lipid remodeling-based stress tolerance mechanisms and together with protection against freezing in freezing-resistant plants such as Arabidopsis, it adds lipid remodeling as a possible target for the engineering of abiotic stress-resilient crops. © 2016 American Society of Plant Biologists. All Rights Reserved.

  9. The big freeze may be over: a contracting universe for cryopreservation? (United States)

    Gale, Robert Peter; Ruiz-Argüelles, Guillermo J


    According to current cosmological theory, the universe will continue to expand indefinitely. If so, it should cool eventually reaching temperatures too cold to sustain life. This theory is commonly referred to as heat-death or the big freeze. Putting aside this potentially unpleasant scenario, unlikely in the lifetime of current readers (about 10 × E + 2500 years from now), freezing, in contrast, has played an important role in hematopoietic cell autotransplants for disease such as plasma cell myeloma and lymphomas. Let us consider how.

  10. Heat transfer coefficient of cryotop during freezing. (United States)

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


    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).

  11. Hot big bang or slow freeze?

    International Nuclear Information System (INIS)

    Wetterich, C.


    We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze — a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple “crossover model” without a big bang singularity. In the infinite past space–time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe

  12. Hot big bang or slow freeze?

    Energy Technology Data Exchange (ETDEWEB)

    Wetterich, C.


    We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze — a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple “crossover model” without a big bang singularity. In the infinite past space–time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe.

  13. Hot big bang or slow freeze?

    Directory of Open Access Journals (Sweden)

    C. Wetterich


    Full Text Available We confront the big bang for the beginning of the universe with an equivalent picture of a slow freeze — a very cold and slowly evolving universe. In the freeze picture the masses of elementary particles increase and the gravitational constant decreases with cosmic time, while the Newtonian attraction remains unchanged. The freeze and big bang pictures both describe the same observations or physical reality. We present a simple “crossover model” without a big bang singularity. In the infinite past space–time is flat. Our model is compatible with present observations, describing the generation of primordial density fluctuations during inflation as well as the present transition to a dark energy-dominated universe.

  14. Repeated freeze-thaw cycles reduce the survival rate of osteocytes in bone-tendon constructs without affecting the mechanical properties of tendons. (United States)

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


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

  15. 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


    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...... studies to describe freezing rates are appropriate for kaolinite aerosol particles. Mechanisms for contact freezing are briefly discussed....

  16. Cryoprotectant Production in Freeze-Tolerant Wood Frogs Is Augmented by Multiple Freeze-Thaw Cycles. (United States)

    Larson, Don J; Barnes, Brian M


    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.

  17. Drought increases freezing tolerance of both leaves and xylem of Larrea tridentata. (United States)

    Medeiros, Juliana S; Pockman, William T


    Drought and freezing are both known to limit desert plant distributions, but the interaction of these stressors is poorly understood. Drought may increase freezing tolerance in leaves while decreasing it in the xylem, potentially creating a mismatch between water supply and demand. To test this hypothesis, we subjected Larrea tridentata juveniles grown in a greenhouse under well-watered or drought conditions to minimum temperatures ranging from -8 to -24 °C. We measured survival, leaf retention, gas exchange, cell death, freezing point depression and leaf-specific xylem hydraulic conductance (k₁). Drought-exposed plants exhibited smaller decreases in gas exchange after exposure to -8 °C compared to well-watered plants. Drought also conferred a significant positive effect on leaf, xylem and whole-plant function following exposure to -15 °C; drought-exposed plants exhibited less cell death, greater leaf retention, higher k₁ and higher rates of gas exchange than well-watered plants. Both drought-exposed and well-watered plants experienced 100% mortality following exposure to -24 °C. By documenting the combined effects of drought and freezing stress, our data provide insight into the mechanisms determining plant survival and performance following freezing and the potential for shifts in L. tridentata abundance and range in the face of changing temperature and precipitation regimes. © 2010 Blackwell Publishing Ltd.

  18. Rheological behaviour, freezing curve, and density of coffee solutions at temperatures close to freezing


    Hernández Yáñez, Eduard; Moreno, Fabian Leonardo; Raventós Santamaria, Mercè; Santamaría, N.; Acosta, J.; Pirachican, Oscar; Torres, L.; Ruiz Pardo, Yolanda


    The physical properties of coffee solutions were determined for temperatures close to the freezing point. Rheological behaviour, freezing curve, density, and their relationship between coffee mass fraction and Brix degrees were determined for coffee mass fractions between 5 and 50% (wet basis) in the -6 to 20 degrees C temperature interval. Values of viscosity varied from 1.99 to 1037 mPa center dot s and values of density from 1000 to 1236 kg center dot m(-3). The freezing curve was generate...

  19. A comparative study of mouse embryo freeze-preservation including the examination of a thermoelectric freezing device. (United States)

    Schiewe, M C; Schmidt, P M; Wildt, D E


    In Study 1 over 2000 4- to 8-cell mouse embryos were randomly pooled and assigned to 1 of 12 treatment groups. A 2 X 2 X 3 factorial design was used to analyze two types of cryoprotectant/post-thaw (PT) dilutions (dimethyl sulfoxide [Me2SO]/stepwise dilution versus glycerol/sucrose dilution), two storage containers (glass ampoules versus plastic straws), and three cooling treatments. Two commercial, controlled-rate freezing machines were examined, employing either nitrogen gas (Planer) or thermoelectric (Glacier) cooling. Embryos were cooled slowly (0.5 degrees C/min) to -35 or -80 degrees C and then cooled rapidly by transfer into liquid nitrogen (LN2). Thawed embryos were cultured for 24 hr after which developmental stage, post-thaw survival (PTS), embryo degeneration rate (EDR), quality grade (QG), and fluorescein diacetate viability grade (VG) were assessed. Overall, PTS and EDR were similar (P greater than 0.05) among the three freezing unit/plunge temperature treatments. Cumulative results of container and cryoprotectant/PT dilution treatments consistently demonstrated greater PTS, QG, and VG ratings and lower EDR values when embryos were frozen in ampoules using glycerol/sucrose dilution. Embryos treated with Me2SO/stepwise dilution were particularly sensitive to freezing damage when stored in plastic straws and plunged into LN2 at -35 degrees C. Study 2 was directed at determining whether Study 1 methods for diluting Me2SO-protected embryos markedly affected PTS rates. Post-thaw culture percentages were no different (P greater than 0.05) for four- to eight-cell Me2SO-treated embryos frozen in ampoules (using the forced-LN2 device), thawed, and diluted either conventionally in reduced concentrations of Me2SO or in the sucrose treatment normally accorded glycerolated embryos.(ABSTRACT TRUNCATED AT 250 WORDS)

  20. The Arabidopsis RCC1 Family Protein TCF1 Regulates Freezing Tolerance and Cold Acclimation through Modulating Lignin Biosynthesis. (United States)

    Ji, Hongtao; Wang, Youning; Cloix, Catherine; Li, Kexue; Jenkins, Gareth I; Wang, Shuangfeng; Shang, Zhonglin; Shi, Yiting; Yang, Shuhua; Li, Xia


    Cell water permeability and cell wall properties are critical to survival of plant cells during freezing, however the underlying molecular mechanisms remain elusive. Here, we report that a specifically cold-induced nuclear protein, Tolerant to Chilling and Freezing 1 (TCF1), interacts with histones H3 and H4 and associates with chromatin containing a target gene, blue-copper-binding protein (BCB), encoding a glycosylphosphatidylinositol-anchored protein that regulates lignin biosynthesis. Loss of TCF1 function leads to reduced BCB transcription through affecting H3K4me2 and H3K27me3 levels within the BCB gene, resulting in reduced lignin content and enhanced freezing tolerance. Furthermore, plants with knocked-down BCB expression (amiRNA-BCB) under cold acclimation had reduced lignin accumulation and increased freezing tolerance. The pal1pal2 double mutant (lignin content reduced by 30% compared with WT) also showed the freezing tolerant phenotype, and TCF1 and BCB act upstream of PALs to regulate lignin content. In addition, TCF1 acts independently of the CBF (C-repeat binding factor) pathway. Our findings delineate a novel molecular pathway linking the TCF1-mediated cold-specific transcriptional program to lignin biosynthesis, thus achieving cell wall remodeling with increased freezing tolerance.

  1. The Arabidopsis RCC1 Family Protein TCF1 Regulates Freezing Tolerance and Cold Acclimation through Modulating Lignin Biosynthesis.

    Directory of Open Access Journals (Sweden)

    Hongtao Ji

    Full Text Available Cell water permeability and cell wall properties are critical to survival of plant cells during freezing, however the underlying molecular mechanisms remain elusive. Here, we report that a specifically cold-induced nuclear protein, Tolerant to Chilling and Freezing 1 (TCF1, interacts with histones H3 and H4 and associates with chromatin containing a target gene, blue-copper-binding protein (BCB, encoding a glycosylphosphatidylinositol-anchored protein that regulates lignin biosynthesis. Loss of TCF1 function leads to reduced BCB transcription through affecting H3K4me2 and H3K27me3 levels within the BCB gene, resulting in reduced lignin content and enhanced freezing tolerance. Furthermore, plants with knocked-down BCB expression (amiRNA-BCB under cold acclimation had reduced lignin accumulation and increased freezing tolerance. The pal1pal2 double mutant (lignin content reduced by 30% compared with WT also showed the freezing tolerant phenotype, and TCF1 and BCB act upstream of PALs to regulate lignin content. In addition, TCF1 acts independently of the CBF (C-repeat binding factor pathway. Our findings delineate a novel molecular pathway linking the TCF1-mediated cold-specific transcriptional program to lignin biosynthesis, thus achieving cell wall remodeling with increased freezing tolerance.

  2. Fibres as carriers for Lactobacillus rhamnosus during freeze-drying and storage in apple juice and chocolate-coated breakfast cereals. (United States)

    Saarela, Maria; Virkajärvi, Ilkka; Nohynek, Liisa; Vaari, Anu; Mättö, Jaana


    The capability of different fibre preparations to protect the viability and stability of Lactobacillus rhamnosus during freeze-drying, storage in freeze-dried form and after formulation into apple juice and chocolate-coated breakfast cereals was studied. In freeze-drying trials wheat dextrin and polydextrose proved to be promising carriers for the L. rhamnosus strains: both freeze-drying survival and storage stability at 37 degrees C were comparable to the control carrier (sucrose). Using apple fibre and inulin carriers resulted in powders with fairly good initial freeze-drying survival but with poor storage stability at 37 degrees C. When fresh L. rhamnosus cells were added into apple juice (pH 3.5) together with oat flour with 20% beta-glucan the survival of the cells was much better at 4 degrees C and at 20 degrees C than with sucrose, wheat dextrin and polydextrose, whereas with freeze-dried cells no protective effect of oat flour could be seen. The stability of freeze-dried L. rhamnosus cells at 20 degrees C was higher in chocolate-coated breakfast cereals compared to low pH apple juice. Similar to freeze-drying stability, wheat dextrin and polydextrose proved to be better carriers than oat flour in chocolate-coated breakfast cereals. Regardless of their differing capability to adhere to fibre preparations the two L. rhamnosus strains studied gave parallel results in the stability studies with different carriers.

  3. Device and method for determining freezing points (United States)

    Mathiprakasam, Balakrishnan (Inventor)


    A freezing point method and device (10) are disclosed. The method and device pertain to an inflection point technique for determining the freezing points of mixtures. In both the method and device (10), the mixture is cooled to a point below its anticipated freezing point and then warmed at a substantially linear rate. During the warming process, the rate of increase of temperature of the mixture is monitored by, for example, thermocouple (28) with the thermocouple output signal being amplified and differentiated by a differentiator (42). The rate of increase of temperature data are analyzed and a peak rate of increase of temperature is identified. In the preferred device (10) a computer (22) is utilized to analyze the rate of increase of temperature data following the warming process. Once the maximum rate of increase of temperature is identified, the corresponding temperature of the mixture is located and earmarked as being substantially equal to the freezing point of the mixture. In a preferred device (10), the computer (22), in addition to collecting the temperature and rate of change of temperature data, controls a programmable power supply (14) to provide a predetermined amount of cooling and warming current to thermoelectric modules (56).

  4. Liquid carbon: structure near the freezing line

    NARCIS (Netherlands)

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


    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

  5. Unitarity Constraints on Asymmetric Freeze-In

    Energy Technology Data Exchange (ETDEWEB)

    Hook, Anson; /SLAC


    This paper considers unitarity and CPT constraints on asymmetric freeze-in, the use of freeze-in to store baryon number in a dark sector. In this scenario, Sakharov's out of equilibrium condition is satisfied by placing the visible and hidden sectors at different temperatures while a net visible baryon number is produced by storing negative baryon number in a dark sector. It is shown that unitarity and CPT lead to unexpected cancellations. In particular, the transfer of baryon number cancels completely at leading order. This note has shown that if two sectors are in thermal equilibrium with themselves, but not with each other, then the leading effect transferring conserved quantities between the two sectors is of order the the weak coupling connecting them to the third power. When freeze-in is used to produce a net baryon number density, the leading order effect comes from {Omicron}({lambda}{sup 3}) diagrams where the intermediate state that goes on-shell has a different visible baryon number than the final state visible baryon number. Models in which the correct baryon number is generated with freeze-in as the dominant source of abundance, typically require {lambda} {approx}> 10{sup -6} and m{sub bath} {approx}> TeV. m{sub bath} is the mass of the visible particle which communicates with the hidden sector. The lower window is potentially observable at the LHC.

  6. Anomalous freezing behavior of nanoscale liposomes

    DEFF Research Database (Denmark)

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


    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...

  7. Susceptibility of blackberry flowers to freezing temperatures (United States)

    Injury of tight buds, open flowers and green fruit often occur in fruit crops during spring frost events. In this study, freezing tolerance of ‘Triple Crown’ blackberry flowers at different reproductive stages of development (tight bud to green drupe) was determined using two methods. One method i...

  8. Scaling-Up Eutectic Freeze Crystallization

    NARCIS (Netherlands)

    Genceli, F.E.


    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)

  9. Freeze block testing of buried waste lines

    International Nuclear Information System (INIS)

    Robbins, E.D.; Willi, J.C.


    An investigation was conducted to demonstrate application of freeze blocking in waste transfer lines such that a hydrostatic pressure test can be applied. A shop test was conducted on a 20-foot length, 3-inch schedule 40, carbon steel pipe using a coolant of dry ice and Freon. The positive results from these tests prompted a similar employment of the freeze block method in hydrostatic pressure testing the feed inlet leading to 241-S-101 Waste Tank. This pipeline is a 3-inch schedule 10, stainless steel pipe approximately 800 feet long. The freeze block was formed near the lower end of the pipe as it entered the 101-S Waste Tank and a pressure hold test was applied to this pipeline. This test proved the integrity of the pipeline in question, and demonstrated the validity of freeze blocking an open-ended pipeline which could not be hydrotested in other conventional ways. The field demonstration facility, costing $30,200 was completed late in 1975

  10. Sysnthesis of powders by freeze-drying

    International Nuclear Information System (INIS)

    Johnson, S.M.; Gusman, M.I.; Hildenbrand, D.L.


    The freeze-drying method of synthesizing powders of the superconducting oxide YBa 2 Cu 3 O 7 - δ is described. This process produces homogeneous, submicron powders of high purity. The effects of salt selection, solution concentration and pH on the process are described. Some evaluation of the sintering behavior and the effects on critical current density are included

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

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


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

  12. Water vapor movement in freezing aggregate base materials. (United States)


    The objectives of this research were to 1) measure the extent to which water vapor movement results in : water accumulation in freezing base materials; 2) evaluate the effect of soil stabilization on water vapor movement : in freezing base materials;...

  13. Static delectric behavior of charged fluids near freezing

    International Nuclear Information System (INIS)

    Fasolino, A.; Parrinello, M.; Tosi, M.P.


    The wavenumber-dependent, static dielectric function of classical charged fluids near freezing is obtained from structural data based on computer simulation or neutron diffraction, and its behavior is connected with the freezing process. (Auth.)

  14. Freeze-thaw performance testing of whole concrete railroad ties. (United States)


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

  15. Adaptive Control of Freeze-Form Extrusion Fabrication Processes (Preprint)

    National Research Council Canada - National Science Library

    Zhao, Xiyue; Landers, Robert G; Leu, Ming C


    Freeze-form Extrusion Fabrication (FEF) is an additive manufacturing process that extrudes high solids loading aqueous ceramic pastes in a layer-by-layer fashion below the paste freezing temperature for component fabrication...

  16. Canalization of freeze tolerance in an alpine grasshopper. (United States)

    Hawes, Timothy C


    In the Rock and Pillar Range, New Zealand, the alpine grasshopper, Sigaus australis Hutton, survives equilibrium freezing (EF) all-year round. A comparison of freeze tolerance (FT) in grasshoppers over four austral seasons for a 1 year period finds that: (a) the majority (>70%) of the sample population of grasshoppers survive single freeze-stress throughout the year; (b) exposure to increased freeze stress (multiple freeze-stress events) does not lead to a loss of freeze tolerance; and (c) responses to increased freeze stress reveal seasonal tuning of the FT adaptation to environmental temperatures. The Rock and Pillar sample population provides a clear example of the canalization of the FT adaptation. Seasonal variability in the extent of tolerance of multiple freezing events indicates that physiology is modulated to environmental temperatures by phenotypic plasticity - i.e. the FT adaptation is permanent and adjustable. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Nanomaterials for efficiently lowering the freezing point of anti-freeze coolants. (United States)

    Hong, Haiping; Zheng, Yingsong; Roy, Walter


    In this paper, we report, for the first time, the effect of the lowered freezing point in a 50% water/50% anti-freeze coolant (PAC) or 50% water/50% ethylene glycol (EG) solution by the addition of carbon nanotubes and other particles. The experimental results indicated that the nano materials are much more efficient (hundreds fold) in lowering the freezing point than the regular ionic materials (e.g., NaCl). The possible explanation for this interesting phenomenon is the colligative property of fluid and relative small size of nano material. It is quite certain that the carbon nanotubes and metal oxide nano particles could be a wonderful candidate for the nano coolant application because they could not only increase the thermal conductivity, but also efficiently lower the freezing point of traditional coolants.

  18. Effects of freezing stress on electrolyte leakage of Persian shallot (Allium altissimum Regel. under controlled condition

    Directory of Open Access Journals (Sweden)

    Shahram Rezvan beydokhti


    Full Text Available In order to evaluate the effect of freezing stress on electrolyte leakage of three Mooseers’ (Allium altissimum Regel. ecotypes (Shirvan, Kalat and Tandoureh, a factorial experiment based on completely randomized design with three replications was carried out under the controlled conditions in the Faculty of Agricultural, Ferdowsi University of Mashhad, Iran, during 2009. Plants were grown in natural conditions to acclimate with the growth condition, and then at two growth stages of emergence and seedling were transferred to the thermogradiant freezer with the six freezing temperatures (0,-4, -8,-12, -16 and -20 oC. Cell membrane integrity was measured by electrolyte leakage (EL index and the lethal temperature 50% (LT50 of samples were determined based on this index. The results showed that electrolyte leakage percentage and LT50el were significantly affected by experimental treatments. As the temperature decreased, EL (% of leaf, bulb and root of all ecotypes was significantly increased. In the all freezing temperatures, the highest and the lowest electrolyte leakage was observed in the root and leaf in the emergence stage. Shirvan ecotype at two growth stage (emergence and seedling showed the highest root electrolyte leakage and hence showed the lowest freezing tolerance in comparison with Kalat and Tandoreh ecotypes. The lowest and highest LT50el due to freezing were observed in Kalat, Tandoureh ecotype and Shirvan ecotype, respectively. At seedling stage, Shirvan ecotype’s root showed the highest LT50el which was more sensitive to freezing temperatures in comparison with leaf and bulb. Among all ecotypes, Kalat and Tandoreh ecotypes showed the lowest electrolyte leakage percentage, the lowest LT50el and the most tolerance to the freezing stress.

  19. Freeze-drying process design by manometric temperature measurement: design of a smart freeze-dryer. (United States)

    Tang, Xiaolin Charlie; Nail, Steven L; Pikal, Michael J


    To develop a procedure based on manometric temperature measurement (MTM) and an expert system for good practices in freeze drying that will allow development of an optimized freeze-drying process during a single laboratory freeze-drying experiment. Freeze drying was performed with a FTS Dura-Stop/Dura-Top freeze dryer with the manometric temperature measurement software installed. Five percent solutions of glycine, sucrose, or mannitol with 2 ml to 4 ml fill in 5 ml vials were used, with all vials loaded on one shelf. Details of freezing, optimization of chamber pressure, target product temperature, and some aspects of secondary drying are determined by the expert system algorithms. MTM measurements were used to select the optimum shelf temperature, to determine drying end points, and to evaluate residual moisture content in real-time. MTM measurements were made at 1 hour or half-hour intervals during primary drying and secondary drying, with a data collection frequency of 4 points per second. The improved MTM equations were fit to pressure-time data generated by the MTM procedure using Microcal Origin software to obtain product temperature and dry layer resistance. Using heat and mass transfer theory, the MTM results were used to evaluate mass and heat transfer rates and to estimate the shelf temperature required to maintain the target product temperature. MTM product dry layer resistance is accurate until about two-thirds of total primary drying time is over, and the MTM product temperature is normally accurate almost to the end of primary drying provided that effective thermal shielding is used in the freeze-drying process. The primary drying times can be accurately estimated from mass transfer rates calculated very early in the run, and we find the target product temperature can be achieved and maintained with only a few adjustments of shelf temperature. The freeze-dryer overload conditions can be estimated by calculation of heat/mass flow at the target product

  20. Deep-freezing of boar semen in plastic film 'cochettes'. (United States)

    Eriksson, B M; Rodriguez-Martinez, H


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

  1. Objective video quality assessment method for freeze distortion based on freeze aggregation (United States)

    Watanabe, Keishiro; Okamoto, Jun; Kurita, Takaaki


    With the development of the broadband network, video communications such as videophone, video distribution, and IPTV services are beginning to become common. In order to provide these services appropriately, we must manage them based on subjective video quality, in addition to designing a network system based on it. Currently, subjective quality assessment is the main method used to quantify video quality. However, it is time-consuming and expensive. Therefore, we need an objective quality assessment technology that can estimate video quality from video characteristics effectively. Video degradation can be categorized into two types: spatial and temporal. Objective quality assessment methods for spatial degradation have been studied extensively, but methods for temporal degradation have hardly been examined even though it occurs frequently due to network degradation and has a large impact on subjective quality. In this paper, we propose an objective quality assessment method for temporal degradation. Our approach is to aggregate multiple freeze distortions into an equivalent freeze distortion and then derive the objective video quality from the equivalent freeze distortion. Specifically, our method considers the total length of all freeze distortions in a video sequence as the length of the equivalent single freeze distortion. In addition, we propose a method using the perceptual characteristics of short freeze distortions. We verified that our method can estimate the objective video quality well within the deviation of subjective video quality.

  2. SLAPex Freeze/Thaw 2015: The First Dedicated Soil Freeze/Thaw Airborne Campaign (United States)

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


    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.

  3. 7 CFR 58.620 - Freezing and packaging rooms. (United States)


    ... 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 be...

  4. Effect of repeated freezing-thawing on the Achilles tendon of rabbits. (United States)

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


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

  5. Screening freeze-drying cryoprotectants for Saccharomyces boulardii by Plackett-Burman design

    Directory of Open Access Journals (Sweden)

    He CHEN


    Full Text Available As a lyophilized product, Saccharomyces boulardii has been commonly used to treat diarrhea in adults. However, there are few studies focusing on the preparation of its freeze-drying powder. This paper investigated the effect of lyoprotectants on the freeze-drying survival rate and the number of viable cells for vacuum freezedried powder of S. boulardii. Single factor experiment and Plackett-Burman design were conducted to obtain the optimal compound lyoprotectant formulations. The result showed that lactose, trehalose and sodium glutamate could significantly enhance the freeze-drying survival rate of S. boulardii. Meanwhile, all these three lyoprotectants showed positive effect on the freezedrying survival rate of the yeast, and the optimal lyoprotectant composition for S. boulardii was as follows: 18g/100mL lactose, 18g/100mL trehalose and 3g/100mL sodium glutamate.

  6. 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


    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...... temperature were less harmful to Salmonella than were low humidity and high temperature. Tolerance to adverse conditions was highest for Salmonella Infantis and one Salmonella Typhimurium U292 isolate and lowest for Salmonella Derby and one Salmonella Typhimurium DT170 isolate. Dehydration, in contrast...... to freezing, was differently tolerated by the Salmonella strains in this study, but tolerance to freezing and dehydration does not appear to contribute to the emergence of successful Salmonella clones....


    Directory of Open Access Journals (Sweden)

    Maria Bernadett SZMUTKU


    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.

  8. High ice nucleation activity located in blueberry stem bark is linked to primary freeze initiation and adaptive freezing behaviour of the bark (United States)

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


    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

  9. High ice nucleation activity located in blueberry stem bark is linked to primary freeze initiation and adaptive freezing behaviour of the bark. (United States)

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


    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. Published by Oxford University Press on behalf of the Annals of Botany Company.

  10. Fabrication of Chitosan Silk-based Tracheal Scaffold Using Freeze-Casting Method (United States)

    Nematollahi, Zeinab; Tafazzoli-Shadpour, Mohammad; Zamanian, Ali; Seyedsalehi, Amir; Mohammad-Behgam, Shadmehr; Ghorbani, Fariba; Mirahmadi, Fereshte


    Background: Since the treatments of long tracheal lesions are associated with some limitations, tissue engineered trachea is considered as an alternative option. This study aimed at preparing a composite scaffold, based on natural and synthetic materials for tracheal tissue engineering. Methods: Nine chitosan silk-based scaffolds were fabricated using three freezing rates (0.5, 1, and 2°C/min) and glutaraldehyde (GA) concentrations (0, 0.4, and 0.8 wt%). Samples were characterized, and scaffolds having mechanical properties compatible with those of human trachea and proper biodegradability were selected for chondrocyte cell seeding and subsequent biological assessments. Results: The pore sizes were highly influenced by the freezing rate and varied from 135.3×372.1 to 37.8×83.4 µm. Swelling and biodegradability behaviors were more affected by GA rather than freezing rate. Tensile strength raised from 120 kPa to 350 kPa by an increment of freezing rate and GA concentration. In addition, marked stiffening was demonstrated by increasing elastic modulus from 1.5 MPa to 12.2 MPa. Samples having 1 and 2°C/min of freezing rate and 0.8 wt% GA concentration made a non-toxic, porous structure with tensile strength and elastic modulus in the range of human trachea, facilitating the chondrocyte proliferation. The results of 21-day cell culture indicated that glycosaminoglycans content was significantly higher for the rate of 2°C/min (12.04 µg/min) rather than the other (9.6 µg/min). Conclusion: A homogenous porous structure was created by freeze drying. This allows the fabrication of a chitosan silk scaffold cross-linked by GA for cartilage tissue regeneration with application in tracheal regeneration. PMID:28131109

  11. Late Spring Freezes in Poland in Relation to Atmospheric Circulation

    Directory of Open Access Journals (Sweden)

    Ustrnul Zbigniew


    Full Text Available Late spring freeze events, a significant agroclimatic hazard, are investigated for Poland. Daily minimum air temperatures from 184 stations for the period 1951-2010 were used to analyze the frequency and conditional probability of late spring freezes. In addition, three classification schemes were employed to investigate the atmospheric circulation responsible for late spring freezes events. The findings suggest that knowledge of the airflow influencing late spring freezes can help to understand the complex historical trends and projected future changes in freeze risk for perennial crops

  12. 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)


    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)

  13. Study of freezing-point depression of selected food extracts

    Energy Technology Data Exchange (ETDEWEB)

    Tanaka, Fumihiko [Kagoshima Univ. (Japan). Dept. of Agricultural Systems Engineering; Murata, Satoshi; Habara, Kazuhiro; Amaratunga, K.S.P. [Kyushu Univ., Fukuoka (Japan). Dept. of Agricultural Engineering


    The phenomenon of freezing-point depression that accompanies the solute concentration of selected food extracts was investigated to reveal the characteristics of solid-liquid phase equilibrium. The freezing curves of various food extracts did not exhibit ideal solution behavior in the higher concentration range. The experimental data were fitted to new freezing-point depression equations by the method of nonlinear least squares, and the results clearly indicated that the calculated freezing points at various concentrations were in good agreement with the experimental data. Furthermore, by using the determined parameters, the freezing ratio and the activation coefficient were derived.

  14. Development and Design of Sludge Freezing Beds (United States)


    freezing often occurs in nature. A typical example is the desalin - ation of sea ice. Initially, brine is concentrated between ice crystal boundaries...Research, 67(3): 1085-1090. Cox, G.F.N. and W.F. Weeks (1975) Brine drainage and initial salt entrapment in sodium chloride ice. USA Cold Regions...Over a period of time, this brine drains out by gravity leaving relatively pure ice (Cox and Weeks 1975). Another example is the formation of pure ice


    Directory of Open Access Journals (Sweden)

    H. Sadeghi - Nejad


    Full Text Available (1 The secondary stage of freeze - drying, particularly the last day, is not important and I suggest the whole procedure is reduced to three days, the primary stage occupying two of these. (2 The mothod used obtains the satisfactory low level of residual moisture. RESUME (3 Experiments on reconstitution with saline and distilled water show that distilled water is more satisfactory and I suggest that saline should not he used.

  16. Microwave Augmented Freeze-Drying - Four Studies (United States)


    F.J. 1G76. Pigments in "Principles of Food Science". O.R. Fennema, ed., Marcel Dekkar, Inc., New York. Datta, A.K. and Hu, W. 1992. Optimization of...and Wilke, C.R 1967. The relationship between transport properties and rates of freeze-drying of poultry meat. AIChE J. 13:428. Slater, L.E. 1975

  17. 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


    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.

  18. 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. (United States)

    Tulha, Joana; Lima, Ana; Lucas, Cândida; Ferreira, Célia


    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. 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. 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.

  19. Sperm Preservation using Freeze-Drying Method

    Directory of Open Access Journals (Sweden)



    Full Text Available Since the discovery of cryopreservation method for bull semen, cryopreservation become an alternative method for maintaining gamet resources of certain animal which is threatened or near extinction. This technology was then applied to the preservation of embryo, oocyte, ovary and testis. The application of intracytoplasmic sperm injection (ICSI for which sperm motility is unnecessary had supported the effort to create simplified method such as freeze-drying for sperm preservation. Due to the benefit of ICSI over the conventional in vitro fertilization (IVF the spermatozoon could be mechanically driven to pass through the zona pellucida and entering the cytoplasm of oocytes prior to fertilization. The freeze-drying method is an alternative method in sperm preservation which ignored the motility of sperm. The sperm resulted from this technique is in drying state, therefore, it might be stored in room temperature or in refrigerator. Many reports have claimed that freeze-dried sperm which is not motile but has an intact DNA was able to fertilize oocytes, even produced offspring in mouse.

  20. Atmospheric freeze drying assisted by power ultrasound

    International Nuclear Information System (INIS)

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


    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.

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

    Directory of Open Access Journals (Sweden)

    Camila Figueiredo Borgognoni


    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

  2. Bulk specimen X-ray microanalysis of freeze-fractured, freeze-dried tissues in gerontological research

    International Nuclear Information System (INIS)

    Nagy, I.


    The rationale for choosing the freeze-fracture freeze-drying (FFFD) method of biological bulk specimen preparation as well as the theoretical and practical problems of this method are treated. FFFD specimens are suitable for quantitative X-ray microanalysis of biologically relevant elements. Although the spatial resolution of this analytical technique is low, the application of properly selected bulk standard crystals as well as the measurement of the intracellular water and dry mass content by means of another method developed in the same laboratory, allow us to obtain useful information about the age-dependent changes of ionic composition in the main intracellular compartments. The paper summarizes the problems with regard to specimen preparation, beam penetration and the quantitative analysis of FFFD specimens. The method has been applied so far mainly for the analysis of intranuclear and intracytoplasmic concentrations of Na, C1 and K in various types of cells and has resulted in a significant contribution to our understanding of the cellular mechanisms of aging. 84 references

  3. Response of New zealand mudsnails Potamopyrgus antipodarum to freezing and near freezing fluctuating water temperatures (United States)

    Moffitt, Christine M.; James, Christopher A.


    We explored the resilience of the invasive New Zealand mudsnail Potamopyrgus antipodarum to fluctuating winter freezing and near-freezing temperature cycles in laboratory tests. Our goal was to provide data to confirm field observations of mortality and presumed mortality in stream habitats with fluctuating freezing to near-freezing temperatures. We tested individuals from 2 locations with distinctly different thermal regimes and population densities. One location had low snail densities and water temperatures with strong diel and seasonal water variation. The other location had high snail densities and nearly constant water temperatures. Groups of individuals from both locations were tested in each of 3 laboratory-created diel thermal cycles around nominal temperatures of 0, 2, or 4°C. Mortality occurred in cycles around 0°C in both populations, and little to no mortality occurred at temperatures >0°C. Individuals from both sources held in diel 0°C cycles for 72 h showed 100% mortality. Our findings support observations from published field studies that survival was limited in infested habitats subject to freezing temperatures.

  4. 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


    superoxide dismutase to compensate for the lack of the cytoplasmic enzyme. Free radicals generated as a result of freezing and thawing were detected in cells directly using electron paramagnetic resonance spectroscopy with either alpha-phenyl-N-tert-butylnitrone or 5, 5-dimethyl-1-pyrroline-N-oxide as spin...... of superoxide radicals formed from oxygen and electrons leaked from the mitochondrial electron transport chain....

  5. Computational investigation of surface freezing in a molecular model of water. (United States)

    Haji-Akbari, Amir; Debenedetti, Pablo G


    Water freezes in a wide variety of low-temperature environments, from meteors and atmospheric clouds to soil and biological cells. In nature, ice usually nucleates at or near interfaces, because homogenous nucleation in the bulk can only be observed at deep supercoolings. Although the effect of proximal surfaces on freezing has been extensively studied, major gaps in understanding remain regarding freezing near vapor-liquid interfaces, with earlier experimental studies being mostly inconclusive. The question of how a vapor-liquid interface affects freezing in its vicinity is therefore still a major open question in ice physics. Here, we address this question computationally by using the forward-flux sampling algorithm to compute the nucleation rate in a freestanding nanofilm of supercooled water. We use the TIP4P/ice force field, one of the best existing molecular models of water, and observe that the nucleation rate in the film increases by seven orders of magnitude with respect to bulk at the same temperature. By analyzing the nucleation pathway, we conclude that freezing in the film initiates not at the surface, but within an interior region where the formation of double-diamond cages (DDCs) is favored in comparison with the bulk. This, in turn, facilitates freezing by favoring the formation of nuclei rich in cubic ice, which, as demonstrated by us earlier, are more likely to grow and overcome the nucleation barrier. The films considered here are ultrathin because their interior regions are not truly bulk-like, due to their subtle structural differences with the bulk.

  6. Urea loading enhances freezing survival and postfreeze recovery in a terrestrially hibernating frog. (United States)

    Costanzo, Jon P; Lee, Richard E


    We tested the hypothesis that urea, an osmolyte accumulated early in hibernation, functions as a cryoprotectant in the freeze-tolerant wood frog, Rana sylvatica. Relative to saline-treated, normouremic (10 micromol ml(-1)) frogs, individuals rendered hyperuremic (70 micromol ml(-1)) by administration of an aqueous urea solution exhibited significantly higher survival (100% versus 64%) following freezing at -4 degrees C, a potentially lethal temperature. Hyperuremic frogs also had lower plasma levels of intracellular proteins (lactate dehydrogenase, creatine kinase, hemoglobin), which presumably escaped from damaged cells, and more quickly recovered neurobehavioral functions following thawing. Experimental freezing-thawing did not alter tissue urea concentrations, but did elevate glucose levels in the blood and organs of all frogs. When measured 24 h after thawing commenced, glucose concentrations were markedly higher in urea-loaded frogs as compared to saline-treated ones, possibly because elevated urea retarded glucose clearance. Like other low-molecular-mass cryoprotectants, urea colligatively reduces both the amount of ice forming within the body and the osmotic dehydration of cells. In addition, by virtue of certain non-colligative properties, it may bestow additional protection from freeze-thaw damage not afforded by glucose.

  7. Response surface optimization of lyoprotectant for Lactobacillus bulgaricus during vacuum freeze-drying. (United States)

    Chen, He; Chen, Shiwei; Li, Chuanna; Shu, Guowei


    The individual and interactive effects of skimmed milk powder, lactose, and sodium ascorbate on the number of viable cells and freeze-drying survival for vacuum freeze-dried powder formulation of Lactobacillus bulgaricus were studied by response surface methodology, and the optimal compound lyoprotectant formulations were gained. It is shown that skim milk powder, lactose, and sodium ascorbate had a significant impact on variables and survival of cultures after freeze-drying. Also, their protective abilities could be enhanced significantly when using them as a mixture of 28% w/v skim milk, 24% w/v lactose, and 4.8% w/v sodium ascorbate. The optimal freeze-drying survival rate and the number of viable cells of Lactobacillus bulgaricus were observed to be (64.41±0.02)% and (3.22±0.02)×10(11) colony-forming units (CFU)/g using the optimal compound protectants, which were very close to the expected values 64.47% and 3.28×10(11) CFU/g.

  8. Recent developments in smart freezing technology applied to fresh foods. (United States)

    Xu, Ji-Cheng; Zhang, Min; Mujumdar, Arun S; Adhikari, Benu


    Due to the increased awareness of consumers in sensorial and nutritional quality of frozen foods, the freezing technology has to seek new and innovative technologies for better retaining the fresh like quality of foods. In this article, we review the recent developments in smart freezing technology applied to fresh foods. The application of these intelligent technologies and the associated underpinning concepts have greatly improved the quality of frozen foods and the freezing efficiency. These technologies are able to automatically collect the information in-line during freezing and help control the freezing process better. Smart freezing technology includes new and intelligent technologies and concepts applied to the pretreatment of the frozen product, freezing processes, cold chain logistics as well as warehouse management. These technologies enable real-time monitoring of quality during the freezing process and help improve product quality and freezing efficiency. We also provide a brief overview of several sensing technologies used to achieve automatic control of individual steps of freezing process. These sensing technologies include computer vision, electronic nose, electronic tongue, digital simulation, confocal laser, near infrared spectroscopy, nuclear magnetic resonance technology and ultrasound. Understanding of the mechanism of these new technologies will be helpful for applying them to improve the quality of frozen foods.

  9. Statistical analysis of intramembranous particles using freeze fracture specimens. (United States)

    Schladitz, Katja; Särkkä, Aila; Pavenstädt, Iris; Haferkamp, Otto; Mattfeldt, Torsten


    We studied the point processes of intramembranous particles of mitochondrial membranes from HeLa cells using the freeze fracture technique. Three groups - under normal conditions, after exposition with rotenone, and after exposition with sodium acid - were compared. First, we used several summary statistics in order to study the two-dimensional point patterns of intramembranous particles within each group. Then, we compared the patterns in different groups by bootstrap tests using the K-function and the nearest neighbour distance function G(r). Estimation of the G-function provided significant results but no significant differences between groups were found using the classical K-function; estimation of G(r) should therefore not be omitted when studying observed planar point patterns.

  10. Freeze Casting for Assembling Bioinspired Structural Materials. (United States)

    Cheng, Qunfeng; Huang, Chuanjin; Tomsia, Antoni P


    Nature is very successful in designing strong and tough, lightweight materials. Examples include seashells, bone, teeth, fish scales, wood, bamboo, silk, and many others. A distinctive feature of all these materials is that their properties are far superior to those of their constituent phases. Many of these natural materials are lamellar or layered in nature. With its "brick and mortar" structure, nacre is an example of a layered material that exhibits extraordinary physical properties. Finding inspiration in living organisms to create bioinspired materials is the subject of intensive research. Several processing techniques have been proposed to design materials mimicking natural materials, such as layer-by-layer deposition, self-assembly, electrophoretic deposition, hydrogel casting, doctor blading, and many others. Freeze casting, also known as ice-templating, is a technique that has received considerable attention in recent years to produce bioinspired bulk materials. Here, recent advances in the freeze-casting technique are reviewed for fabricating lamellar scaffolds by assembling different dimensional building blocks, including nanoparticles, polymer chains, nanofibers, and nanosheets. These lamellar scaffolds are often infiltrated by a second phase, typically a soft polymer matrix, a hard ceramic matrix, or a metal matrix. The unique architecture of the resultant bioinspired structural materials displays excellent mechanical properties. The challenges of the current research in using the freeze-casting technique to create materials large enough to be useful are also discussed, and the technique's promise for fabricating high-performance nacre-inspired structural materials in the future is reviewed. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Survival of freeze-dried microcapsules of α-galactosidase producing probiotics in a soy bar matrix. (United States)

    Chen, Mo; Mustapha, Azlin


    Soy oligosaccharides, mainly α-galactosides, are prevalently present in soy protein products, and can result in unfavorable digestive effects when consumed. The aim of this research was to investigate the efficiency of α-galactoside reduction by probiotic bacterial hydrolysis and if such bacteria could be maintained in a high number in a soy protein product in a microencapsulated and freeze-dried form. The probiotic bacterium, Lactobacillus acidophilus LA-2, when induced by raffinose, exhibited a high level of α-galactosidase activity at 5.0 U/mg. To preserve probiotics with high viability, cells were microencapsulated and freeze-dried. Optimization of microencapsulation presented that a combination of κ-carrageenan and inulin at a proportion of 1.9:0.1 (w:w) as capsule wall materials, significantly retained the viability of the probiotics through freeze-drying (P ≤ 0.05). Scanning electron microscopic images confirmed that the morphology of the microcapsules was well preserved after freeze-drying. Upon incorporation into soy protein bars, the freeze-dried microcapsules of L. acidophilus LA-2 remained in high numbers throughout 14 weeks of storage at 4 °C. Results of this work with the support of other studies on microencapsulation benefits indicate a promising use of freeze-dried α-galactosidase positive microencapsulated probiotics in a soy food. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Stability evaluation of freeze-dried Lactobacillus paracasei subsp. tolerance and Lactobacillus delbrueckii subsp. bulgaricus in oral capsules. (United States)

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


    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.

  13. Increasing viability, numbers, and motility of sperm in men with normal spermatogenesis exposed to saffron extract after freezing- thawing process

    Directory of Open Access Journals (Sweden)

    Y. Khazaei


    Full Text Available Background: Sperm freezing method is used frequently in assisted reproductive techniques, on the other hand in different studies negative effect of freezing have been shown on different sperm parameters. Objective: The aim of this study was to determine the effect of saffron extract as an antioxidant, on the different sperm parameters in men with normal spermatogenesis after freezing-thawing process. Methods: In this case-control study, collecting of samples was done in 2015 year from the Infertility Treatment Center, ACECR Branch of Qazvin, Qazvin, Iran. These men had normal spermatogenesis and their spouse had infertility problem. Semen samples was divided in two groups, control without saffron extract, and case with 50 mg/ml saffron extract. Then, samples freezed with snap freezing method. After two weeks, they were thawed and different sperm parameters were assessed. Data were analyzed by two-tail T test. Findings: Our results showed, mean percent of viability (72±0.99, motility (87±0.43, and the number of sperm cells (62.5±3.8 in treaded group was elevated significantly (P0.05. Conclusion: Our results showed that possibly antioxidant agents of saffron extract could scavenge free radicals and thus, optimize different sperm parameters (viability, motility, and number after freezing and thawing.

  14. Freeze-drying of live virus vaccines: A review. (United States)

    Hansen, L J J; Daoussi, R; Vervaet, C; Remon, J-P; De Beer, T R M


    Freeze-drying is the preferred method for stabilizing live, attenuated virus vaccines. After decades of research on several aspects of the process like the stabilization and destabilization mechanisms of the live, attenuated viruses during freeze-drying, the optimal formulation components and process settings are still matter of research. The molecular complexity of live, attenuated viruses, the multiple destabilization pathways and the lack of analytical techniques allowing the measurement of physicochemical changes in the antigen's structure during and after freeze-drying mean that they form a particular lyophilization challenge. The purpose of this review is to overview the available information on the development of the freeze-drying process of live, attenuated virus vaccines, herewith focusing on the freezing and drying stresses the viruses can undergo during processing as well as on the mechanisms and strategies (formulation and process) that are used to stabilize them during freeze-drying. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Physical Stability of Freeze-Dried Isomalt Diastereomer Mixtures

    DEFF Research Database (Denmark)

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


    Purpose 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. Methods Isomalt was freeze-dried in......Purpose 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. Methods Isomalt was freeze......-dried in four different diastereomer compositions and its physical stability was investigated with differential scanning calorimetry, Fourier-transform infrared and Raman spectroscopy, X-ray powder diffraction, Karl-Fischer titration and thermogravimetric analysis in order to verify the solid state form...

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

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


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

  17. Air-cooled steam condensers non-freeze warranties

    Energy Technology Data Exchange (ETDEWEB)

    Larinoff, M.W.


    What this paper is suggesting is the seller quote a condenser package with a LIMITED NON-FREEZE WARRANTY. Relieve the inexperienced buyer of the responsibility for selecting freeze protection design options. The seller cannot afford to over-design because of the added costs and the need for a competitive price. Yet he cannot under-design and allow the condenser tubes to freeze periodically and then pay the repair bills in accordance with the warranty.

  18. Evaluation of Factors Affecting Freezing Point of Milk


    Jelena Zagorska; Inga Ciprovica


    The freezing point of milk is in important indicator of the milk quality. The freezing point of milk is determined primarily to prove milk adulteration with water and to determine the amount of water in it. Chemical composition and properties of milk, thermal treatment and presence of any substance can influence freezing point of product. There are different substances, which can be added to milk with main purpose to prolong shelf-life of raw milk. There are detergent, pr...

  19. Anisotropic silk fibroin/gelatin scaffolds from unidirectional freezing

    Energy Technology Data Exchange (ETDEWEB)

    Asuncion, Maria Christine Tankeh, E-mail: [National University of Singapore, Department of Biomedical Engineering (Singapore); Goh, James Cho-Hong [National University of Singapore, Department of Biomedical Engineering (Singapore); National University of Singapore, Department of Orthopedic Surgery (Singapore); Toh, Siew-Lok [National University of Singapore, Department of Biomedical Engineering (Singapore); National University of Singapore, Department of Mechanical Engineering (Singapore)


    Recent studies have underlined the importance of matching scaffold properties to the biological milieu. Tissue, and thus scaffold, anisotropy is one such property that is important yet sometimes overlooked. Methods that have been used to achieve anisotropic scaffolds present challenges such as complicated fabrication steps, harsh processing conditions and toxic chemicals involved. In this study, unidirectional freezing was employed to fabricate anisotropic silk fibroin/gelatin scaffolds in a simple and mild manner. Morphological, mechanical, chemical and cellular compatibility properties were investigated, as well as the effect of the addition of gelatin to certain properties of the scaffold. It was shown that scaffold properties were suitable for cell proliferation and that mesenchymal stem cells were able to align themselves along the directed fibers. The fabricated scaffolds present a platform that can be used for anisotropic tissue engineering applications such as cardiac patches. - Highlights: • Silk/gelatin scaffolds with unidirectional alignment were fabricated using a simple and scalable process • Presence of gelatin in silk resulted to lesser shrinkage, better water retention and improved cell proliferation. • Mesenchymal stem cells were shown to align themselves according to the fiber alignment.

  20. Effect of Impurities on the Freezing Point of Zinc (United States)

    Sun, Jianping; Rudtsch, Steffen; Niu, Yalu; Zhang, Lin; Wang, Wei; Den, Xiaolong


    The knowledge of the liquidus slope of impurities in fixed-point metal defined by the International Temperature Scale of 1990 is important for the estimation of uncertainties and correction of fixed point with the sum of individual estimates method. Great attentions are paid to the effect of ultra-trace impurities on the freezing point of zinc in the National Institute of Metrology. In the present work, the liquidus slopes of Ga-Zn, Ge-Zn were measured with the slim fixed-point cell developed through the doping experiments, and the temperature characteristics of the phase diagram of Fe-Zn were furthermore investigated. A quasi-adiabatic Zn fixed-point cell was developed with the thermometer well surrounded by the crucible with the pure metal, and the temperature uniformity of less than 20 mK in the region where the metal is located was obtained. The previous doping experiment of Pb-Zn with slim fixed-point cell was checked with quasi-adiabatic Zn fixed-point cell, and the result supports the previous liquidus slope measured with the traditional fixed-point realization.

  1. Parameter Sensitivity of the Microdroplet Vacuum Freezing Process

    Directory of Open Access Journals (Sweden)

    Zhijun Zhang


    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.

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

    Directory of Open Access Journals (Sweden)

    Stephan G Anagnostaras


    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.

  3. Asset Freezing: Smart Sanction or Criminal Charge?

    Directory of Open Access Journals (Sweden)

    Melissa van den Broek


    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.

  4. Constraints for the thawing and freezing potentials (United States)

    Hara, Tetsuya; Suzuki, Anna; Saka, Shogo; Tanigawa, Takuma


    We study the accelerating present universe in terms of the time evolution of the equation of state w(z) (redshift z) due to thawing and freezing scalar potentials in the quintessence model. The values of dw/da and d^2w/da^2 at a scale factor of a = 1 are associated with two parameters of each potential. For five types of scalar potentials, the scalar fields Q and w as functions of time t and/or z are numerically calculated under the fixed boundary condition of w(z=0)=-1+Δ. The observational constraint w_obs (Planck Collaboration, arXiv:1502.01590) is imposed to test whether the numerical w(z) is in w_obs. Some solutions show thawing features in the freezing potentials. Mutually exclusive allowed regions in the dw/da vs. d^2w/da^2 diagram are obtained in order to identify the likely scalar potential and even the potential parameters for future observational tests.

  5. Beneficial effect of directional freezing on in vitro viability of cryopreserved sheep whole ovaries and ovarian cortical slices. (United States)

    Maffei, S; Pennarossa, G; Brevini, T A L; Arav, A; Gandolfi, F


    Does directional freezing improve the structural and functional integrity of ovarian fragments compared with conventional slow freezing and to whole ovary cryopreservation? Compared with slow freezing, the use of directional freezing significantly improves all structural and functional parameters of ovarian fragments assessed in vitro and, overall, whole ovaries were better preserved than ovarian fragments. Directional freezing has been developed to provide an alternative way to cryopreserve large biological samples and it is known to improve the structural and functional integrity of whole ovaries. Conventional slow freezing of ovarian fragments is the procedure more widely used in clinical settings but it causes substantial structural damage that limits the functional period after transfer back into the patient. We performed a 2 × 2 factorial design experiment on a total of 40 sheep ovaries, divided into four groups (n = 10 ovaries per group): (i) directional freezing of whole ovary (DFwo); (ii) directional freezing of ovarian fragments (DFof); (iii) conventional freezing of whole ovary (CFwo); (iv) conventional freezing of ovarian fragments (CFof). An additional eight ovaries were used as fresh controls. Ewe ovaries were randomly assigned to one of the experimental groups and frozen accordingly. Upon thawing, ovarian tissue was examined morphologically and cultured in vitro for 7 days. Samples were analyzed for cell proliferation and apoptosis, for DNA damage and repair activity, and for the presence of a panel of heat shock proteins (HSPs) by immunohistochemistry. Most studied parameters were significantly improved (P sheep ovaries, which are smaller than human ovaries and therefore may withstand the procedures better. Improved integrity of ovarian morphology may translate to improved outcomes after transplantation. Alternatively, the particularly good preservation of whole ovaries suggests they could provide a source of ovarian follicles for in vitro culture

  6. Effect of pre-fixation delay and freezing on mink testicular endpoints for environmental research.

    Directory of Open Access Journals (Sweden)

    Ellinor Spörndly-Nees

    Full Text Available There is growing interest in using wild animals to monitor the real-life cocktail effect of environmental chemicals on male reproduction. However, practical difficulties, such as long distances to the laboratory, generally prolong the time between euthanisation and specimen handling. For instance, tissue fixation is often performed on frozen material or on material where deterioration has started, which may affect tissue morphology. This study examined the effect of pre-fixation delay and freezing on mink testicular endpoints in order to determine robust endpoints in suboptimally handled specimens. Sexually mature farmed mink (n=30 selected at culling were divided into six groups and subjected to different time intervals between euthanisation and fixation or freezing: 0 hours (fixed immediately post mortem, 6 hours, 18 hours, 30 hours, 42 hours, or frozen 6 hours post mortem and thawed overnight. Unaffected endpoints when pre-fixation storage was extended to 30 hours included: area and diameter of the seminiferous tubules, length and weight of the testes, and acrosomes marked with Gata-4. Epithelial height, Sertoli cells marked with Gata-4 and cell morphology were affected endpoints after 6 hours of storage. Freezing the tissue prior to fixation severely altered cell morphology and reduced testicular weight, tubular diameter and area. Morphological changes seen after 6 hours included shredded germ cells and excess cytoplasm in seminiferous tubular lumen, chromatin rearrangements and increased germ cell death. Extended delay before fixation and freezing affected many endpoints in the mink testicular tissue. Some of these endpoints may mimic chemically induced effects, which is important to consider when evaluating specimens from wild animals for environmental toxicity.

  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


    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. Approaches to the preservation of human granulocytes by freezing

    Energy Technology Data Exchange (ETDEWEB)

    Frim, J; Mazur, P


    Because of its simplicity, the FDA assay can be used effectively as a screening test to eliminate procedures and treatments that are damaging to cells. In this context, a number of conclusions can be drawn from the data presented: (1) Exposure to 1 and 2 M glycerol at room temperature damages human granulocytes in a few minutes. Reducing the exposure temperature to 0/sup 0/C reduces the amount of injury substantially. (2) Human granulocytes respond to freezing and thawing in a manner typical of many mammalian cells in that they exhibit a maximum in survival at an optimum cooling rate slightly above 1/sup 0/C/min when combined with rapid warming. The use of rapid warming and a high (2 M) concentration of glycerol reduces the dependence of survival on cooling rate by broadening the range of rates over which survival is relatively high. (3) Human granulocytes show some sensitivity to dilution stresses since survival depends somewhat on the concentration of glycerol used and the severity of the dilution procedure. The reasons for the sharp decrease in cell viability following incubation of frozen-thawed granulocytes at 37/sup 0/C are not known. One possibility is that the phosphate buffered saline suspending medium used is not suitable for incubation at 37/sup 0/C. A second possibility is that some cell injury is not expressed at 0/sup 0/C and remains undetected by the FDA assay until the cells are incubated at 37/sup 0/C. There is also the possibility that lysosomal enzymes released by a few damaged cells in a sample will cause additional damage in other cells at 37/sup 0/C.

  9. Technological Development of Brewing in Domestic Refrigerator Using Freeze-Dried Raw Materials

    Directory of Open Access Journals (Sweden)

    Angelika-Ioanna Gialleli


    Full Text Available Development of a novel directly marketable beer brewed at low temperature in a domestic refrigerator combined with yeast immobilization technology is presented in this study. Separately, freeze-dried wort and immobilized cells of the cryotolerant yeast strain Saccharomyces cerevisiae AXAZ-1 on tubular cellulose were used in low-temperature fermentation (2, 5 and 7 °C. The positive eff ect of tubular cellulose during low-temperature brewing was examined, revealing that freeze-dried immobilized yeast cells on tubular cellulose signifi cantly reduced the fermentation rates in contrast to freeze-dried free cells, although they are recommended for home-made beer production. Immobilization also enhanced the yeast resistance at low-temperature fermentation, reducing the minimum brewing temperature value from 5 to 2 °C. In the case of high-quality beer production, the eff ect of temperature and initial sugar concentration on the fermentation kinetics were assessed. Sensory enrichment of the produced beer was confi rmed by the analysis of the fi nal products, revealing a low diacetyl concentration, together with improved polyphenol content, aroma profi le and clarity. The proposed process for beer production in a domestic refrigerator can easily be commercialized and applied by dissolving the content of two separate packages in tap water; one package containing dried wort and the other dried immobilized cells on tubular cellulose suspended in tap water.

  10. Application of the Quality by Design Approach to the Freezing Step of Freeze-Drying: Building the Design Space. (United States)

    Arsiccio, Andrea; Pisano, Roberto


    The present work shows a rational method for the development of the freezing step of a freeze-drying cycle. The current approach to the selection of freezing conditions is still empirical and non-systematic, thus resulting in poor robustness of control strategy. The final aim of this work is to fill this gap, describing a rational procedure, based on mathematical modelling, for properly choosing the freezing conditions. Mechanistic models are used for the prediction of temperature profiles during freezing and dimension of ice crystals being formed. Mathematical description of the drying phase of freeze-drying is also coupled with results obtained by freezing models, thus providing a comprehensive characterization of the lyophilization process. In this framework, deep understanding of the phenomena involved is required, and, according to the Quality by Design approach, this knowledge can be used to build the design space. The step by step procedure for building the design space for freezing is thus described and examples of applications are provided. The calculated design space is validated upon experimental data and we show that it allows easy control of the freezing process and fast selection of appropriate operating conditions. Copyright © 2018. Published by Elsevier Inc.

  11. Probability of freezing in the freeze-avoiding beetle larvae Cucujus clavipes puniceus (Coleoptera: Cucujidae) from interior Alaska. (United States)

    Sformo, T; McIntyre, J; Walters, K R; Barnes, B M; Duman, J


    Freeze-avoiding insects must resist freezing or die. A suite of adaptations to low temperatures, including the production of antifreeze proteins, colligative antifreezes (polyols), and dehydration allows most individuals to prevent freezing below the lowest ambient temperatures experienced in situ; however, there can be a wide variance in the minimum temperatures that individuals of freeze-avoiding species reach before freezing. We used logistic regression to explore factors that affect this variance and to estimate the probability of freezing in larvae of the freeze-avoiding beetle Cucujus clavipes puniceus. We hypothesized that water content ≤0.5 mg mg(-1) dry mass would lead to deep supercooling (avoidance of freezing below -58°C). We found a significant interaction between water content and ambient below-snow temperature and a significant difference between individuals collected from two locations in Alaska: Wiseman and Fairbanks. Individuals collected in Wiseman deep supercooled with greater water content and to a greater range of ambient temperatures than individuals collected in Fairbanks, leading to significantly different lethal water contents associated with 50% probability of freezing. Copyright © 2011 Elsevier Ltd. All rights reserved.

  12. Freezing cytorrhysis and critical temperature thresholds for photosystem II in the peat moss Sphagnum capillifolium. (United States)

    Buchner, Othmar; Neuner, Gilbert


    Leaflets of Sphagnum capillifolium were exposed to temperatures from -5 degrees C to +60 degrees C under controlled conditions while mounted on a microscope stage. The resultant cytological response to these temperature treatments was successfully monitored using a light and fluorescence microscope. In addition to the observable cytological changes during freezing cytorrhysis and heat exposure on the leaflets, the concomitant critical temperature thresholds for inactivation of photosystem II (PS II) were studied using a micro fibre optic and a chlorophyll fluorometer mounted to the microscope stage. Chlorophyllous cells of S. capillifolium showed extended freezing cytorrhysis immediately after ice nucleation at -1.1 degrees C in the water in which the leaflets were submersed during the measurement. The occurrence of freezing cytorrhysis, which was visually manifested by cell shrinkage, was highly dynamic and was completed within 2 s. A total reduction of the mean projected diameter of the chloroplast containing area during freezing cytorrhysis from 8.9 to 3.8 microm indicates a cell volume reduction of approximately -82%. Simultaneous measurement of chlorophyll fluorescence of PS II was possible even through the frozen water in which the leaf samples were submersed. Freezing cytorrhysis was accompanied by a sudden rise of basic chlorophyll fluorescence. The critical freezing temperature threshold of PS II was identical to the ice nucleation temperature (-1.1 degrees C). This is significantly above the temperature threshold at which frost damage to S. capillifolium leaflets occurs (-16.1 degrees C; LT(50)) which is higher than observed in most higher plants from the European Alps during summer. High temperature thresholds of PS II were 44.5 degrees C which is significantly below the heat tolerance of chlorophyllous cells (49.9 degrees C; LT(50)). It is demonstrated that light and fluorescence microscopic techniques combined with simultaneous chlorophyll fluorescence

  13. Effect of radiation and freezing on (/sup 3/H)DNA of Yersinia enterocolitica

    Energy Technology Data Exchange (ETDEWEB)

    Grecz, N.; El-zawahry, Y.A.


    Freezing of the enteropathogenic bacterium Yersinia enterocolitica to -18 and -75/sup 0/C caused 7 and 42% cell death, respectively, and 0.329 and 0.588 single-strand breaks per 10/sup 8/ daltons of DNA, respectively, while radiation to one D/sub 10/ dose (10% cell survival) combined with freezing to 2 to 0, -18 and -75/sup 0/C induces 0.05, 0.75, and 5.04 single-strand breaks, respectively. The increase in the effectiveness of radiation with respect to the yield of single-strand breaks at -18 to -75/sup 0/C is contrary to expectation and seems to be due to arrest of repair of single-strand breaks by these low temperatures. 27 references.

  14. Effect of radiation and freezing on [3H]DNA of Yersinia enterocolitica

    International Nuclear Information System (INIS)

    Grecz, N.; El-zawahry, Y.A.


    Freezing of the enteropathogenic bacterium Yersinia enterocolitica to -18 and -75 0 C caused 7 and 42% cell death, respectively, and 0.329 and 0.588 single-strand breaks per 10 8 daltons of DNA, respectively, while radiation to one D 10 dose (10% cell survival) combined with freezing to 2 to 0, -18 and -75 0 C induces 0.05, 0.75, and 5.04 single-strand breaks, respectively. The increase in the effectiveness of radiation with respect to the yield of single-strand breaks at -18 to -75 0 C is contrary to expectation and seems to be due to arrest of repair of single-strand breaks by these low temperatures. 27 references

  15. Microprocessor-controlled vs. "dump-freezing" platelet and lymphocyte cryopreservation: A quantitative and qualitative comparative study

    Directory of Open Access Journals (Sweden)

    Balint Bela


    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

  16. Accelerated storage testing of freeze-dried Pseudomonas ...

    African Journals Online (AJOL)

    The viability of freeze-dried P. fluorescens strains (BTP1, PI9 and BB2) was evaluated by using the Arrhenius model. This model was described by measuring the reaction rate constants (D or k) and temperature sensitivity of rate constant (z or Ea). The freeze-dried P. fluorescens strains were stored in glass tubes at 60, ...

  17. Effects of stress on bodily freezing in adolescents

    NARCIS (Netherlands)

    Niermann, H.C.M.; Figner, B.; Tyborowska, A.B.; Cillessen, A.H.N.; Roelofs, K.


    Freezing is a major defensive stress-response, characterized by reduced body-sway and heart rate. Exacerbated freezing in threatening situations has been associated with increased basal and stress-induced glucocorticoid levels and with long-lasting stress-related symptoms in animals. However, the

  18. Nanoclay Performance on Resistance of Clay under Freezing Cycles

    African Journals Online (AJOL)


    4 Department of Civil Engineering, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran KEYWORDS: Nanoclay, Resistance, Freezing, Sample, Uniaxial. ABSTRACT: This study aims to investigate effect of nanoclay on resistance of the exposed clay soils in freezing in vitro conditions.

  19. Applicable technical method for freeze-substitution of high pressure ...

    African Journals Online (AJOL)



    Nov 2, 2011 ... Cryo-fixation and freeze substitution followed by microscopy are commonly used sample preparation methods for visualizing the morphology of intracellular organelles. Freeze substitution is an especially important preparative step because it enables the preservation of intracellular structures in cryo-fixed.

  20. Prospective Primary School Teachers' Perceptions on Boiling and Freezing (United States)

    Senocak, Erdal


    The aim of this study was to investigate the perceptions of prospective primary school teachers on the physical state of water during the processes of boiling and freezing. There were three stages in the investigation: First, open-ended questions concerning the boiling and freezing of water were given to two groups of prospective primary school…

  1. A tissue snap-freezing apparatus without sacrificial cryogens

    NARCIS (Netherlands)

    Vanapalli, Srinivas; Jagga, Sahil; Holland, Harry; ter Brake, H.J.M.


    Molecular technologies in cancer diagnosis require a fresh and frozen tissue, which is obtained by means of snap-freezing. Currently, coolants such as solid carbon dioxide and liquid nitrogen are used to preserve good morphology of the tissue. Using these coolants, snap freezing of tissues for


    NARCIS (Netherlands)

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


    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

  3. Using infrared thermography to study freezing in plants (United States)

    Factors that determine when and to what extent a plant will freeze are complex. While thermocouples have served as the main method of monitoring the freezing process in plants, infrared thermography offers distinct advantages, and the use of this latter technology has provided new insights on the p...

  4. A subspecies of region crossing change, region freeze crossing change


    Inoue, Ayumu; Shimizu, Ryo


    We introduce a local move on a link diagram named a region freeze crossing change which is close to a region crossing change, but not the same. We study similarity and difference between region crossing change and region freeze crossing change.

  5. Savanna Tree Seedlings are Physiologically Tolerant to Nighttime Freeze Events (United States)

    O’Keefe, Kimberly; Nippert, Jesse B.; Swemmer, Anthony M.


    Freeze events can be important disturbances in savanna ecosystems, yet the interactive effect of freezing with other environmental drivers on plant functioning is unknown. Here, we investigated physiological responses of South African tree seedlings to interactions of water availability and freezing temperatures. We grew widely distributed South African tree species (Colophospermum mopane, Combretum apiculatum, Acacia nigrescens, and Cassia abbreviata) under well-watered and water-limited conditions and exposed individuals to nighttime freeze events. Of the four species studied here, C. mopane was the most tolerant of lower water availability. However, all species were similarly tolerant to nighttime freezing and recovered within one week following the last freezing event. We also show that water limitation somewhat increased freezing tolerance in one of the species (C. mopane). Therefore, water limitation, but not freezing temperatures, may restrict the distribution of these species, although the interactions of these stressors may have species-specific impacts on plant physiology. Ultimately, we show that unique physiologies can exist among dominant species within communities and that combined stresses may play a currently unidentified role in driving the function of certain species within southern Africa. PMID:26870065

  6. Solar desalination by freezing and distillation (United States)

    Kvajic, G.

    It is noted that among seawater desalination processes the absorption-freeze vapor compression processes based on the thermal heat pump, although untested commercially and still in the development stage, appears technically and economically an attractive application of low-grade (exergy) solar heat. The distillation processes proposed here may be conveniently powered by low-grade solar heat (from flat plate solar collectors). It is expected that the scaling problem will be insignificant in comparison with that encountered in the conventional multistage flash process. The novel feature here is the use of enlarged capacity for heat exchange between distillate and brine via latent heat of solid-liquid phase change of a suitable hydrophobic intermediate heat transfer material.

  7. Limonene encapsulation in freeze dried gellan systems. (United States)

    Evageliou, Vasiliki; Saliari, Dimitra


    The encapsulation of limonene in freeze-dried gellan systems was investigated. Surface and encapsulated limonene content was determined by measurement of the absorbance at 252nm. Gellan matrices were both gels and solutions. For a standard gellan concentration (0.5wt%) gelation was induced by potassium or calcium chloride. Furthermore, gellan solutions of varying concentrations (0.25-1wt%) were also studied. Limonene was added at two different concentrations (1 and 2mL/100g sample). Gellan gels encapsulated greater amounts of limonene than solutions. Among all gellan gels, the KCl gels had the greater encapsulated limonene content. However, when the concentration of limonene was doubled in these KCl gels, the encapsulated limonene decreased. The surface limonene content was significant, especially for gellan solutions. The experimental conditions and not the mechanical properties of the matrices were the dominant factor in the interpretation of the observed results. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Climatic variability and the evolution of insect freeze tolerance. (United States)

    Sinclair, Brent J; Addo-Bediako, A; Chown, Steven L


    Insects may survive subzero temperatures by two general strategies: Freeze-tolerant insects withstand the formation of internal ice, while freeze-avoiding insects die upon freezing. While it is widely recognized that these represent alternative strategies to survive low temperatures, and mechanistic understanding of the physical and molecular process of cold tolerance are becoming well elucidated, the reasons why one strategy or the other is adopted remain unclear. Freeze avoidance is clearly basal within the arthropod lineages, and it seems that freeze tolerance has evolved convergently at least six times among the insects (in the Blattaria, Orthoptera, Coleoptera, Hymenoptera, Diptera and Lepidoptera). Of the pterygote insect species whose cold-tolerance strategy has been reported in the literature, 29% (69 of 241 species studied) of those in the Northern Hemisphere, whereas 85 % (11 of 13 species) in the Southern Hemisphere exhibit freeze tolerance. A randomization test indicates that this predominance of freeze tolerance in the Southern Hemisphere is too great to be due to chance, and there is no evidence of a recent publication bias in favour of new reports of freeze-tolerant species. We conclude from this that the specific nature of cold insect habitats in the Southern Hemisphere, which are characterized by oceanic influence and climate variability must lead to strong selection in favour of freeze tolerance in this hemisphere. We envisage two main scenarios where it would prove advantageous for insects to be freeze tolerant. In the first, characteristic of cold continental habitats of the Northern Hemisphere, freeze tolerance allows insects to survive very low temperatures for long periods of time, and to avoid desiccation. These responses tend to be strongly seasonal, and insects in these habitats are only freeze tolerant for the overwintering period. By contrast, in mild and unpredictable environments, characteristic of habitats influenced by the Southern

  9. Structure-property-processing correlations in freeze-cast composite scaffolds. (United States)

    Hunger, Philipp M; Donius, Amalie E; Wegst, Ulrike G K


    Surprisingly few reports have been published, to date, on the structure-property-processing correlations observed in freeze-cast materials directionally solidified from polymer solutions, or ceramic or metal slurries. The studies that exist focus on properties of sintered ceramics, that is materials whose structure was altered by further processing. In this contribution, we report first results on correlations observed in alumina-chitosan-gelatin composites, which were chosen as a model system to test and compare the effect of particle size and processing parameters on their mechanical properties at a specific composition. Our study reveals that highly porous (>90%) hybrid materials can be manufactured by freeze casting, through the self-assembly of a polymer and a ceramic phase that occurs during directional solidification, without the need of additional processing steps such as sintering or infiltration. It further illustrates that the properties of freeze-cast hybrid materials can independently be tailored at two levels of their structural hierarchy, allowing for the simultaneous optimization of both mechanical and structural requirements. An increase in freezing rate resulted in decreases in lamellar spacing, cell wall thickness, pore aspect ratio and cross-sectional area, as well as increases in both Young's modulus and compressive yield strength. The mechanical properties of the composite scaffolds increased with an increasing particle size. The results show that both structure and mechanical properties of the freeze-cast composites can be custom-designed and that they are thus ideally suited for a large variety of applications that require high porosity at low or medium load-bearing capacity. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  10. Application of plackett-burman design in screening freeze drying cryoprotectants for Lactobacillus bulgaricus

    Directory of Open Access Journals (Sweden)

    Guowei SHU


    Full Text Available Lactobacillus bulgaricus is the bacteria commonly used in probiotic dairy product, including yogurt and cheese. The bacteria may be stored for long periods of time if it is freeze-dried. The cryoprotectant mixture for L. bulgaricus was optimized during the process of freeze-drying using a Plackett-Burman design and the steepest ascent test. In our initial tests, the cell survival rate and the number of viable cells were associated with the type of cyroprotectant used. Therefore, our optimization protocol focused on increasing survival rate. Substances that previously had a protective effect during freeze-drying were investigated, for example: sucrose, lactose, skim milk powder, sodium bicarbonate, sodium glutamate, magnesium sulfate, sodium ascorbate, yeast extract, vitamin B2, and phosphate buffer. We determined that the optimum cryoprotectant composition for L. bulgaricus consists of 28.0 g/100 mL skim milk powder, 24.0 g/100 mL lactose and 4.8 g/100 mL sodium ascorbate. The optimized cryoprotectant provides a 63.25% cell survival rate.

  11. Effects of freezing rates and cryoprotectant on thermal expansion of articular cartilage during freezing process. (United States)

    Xu, Y; Sun, H J; Lv, Y; Zou, J C; Lin, B L; Hua, T C


    The intact articular cartilage has not yet been successfully preserved at low temperature most likely due to the volume expansion from water to ice during freezing. The objective of this current study focuses on examining thermal expansion behavior of articular cartilage (AC) during freezing from 0 degree C to -100 degree C. Thermo Mechanical Analysis (TMA) was used to investigate the effects of different concentrations of dimethyl sulphoxide (DMSO) (0%, 10%, 30% and 60% v/v) and different freezing rates (1 C/min, 3 C/min and 5 C/min). The results showed that: (1) the inhomogeneous thermal expansion (or contraction) presents due to inhomogeneous water distributions in articular cartilage during freezing, which also may be the most likely reason that the matrix has been damaged in cryopreserved intact articular cartilage; (2) at the phase transition temperature range, the maximum thermal strain change value for 5C/min is approximately 1.45 times than that for 1 C/min, but the maximum thermal expansion coefficient of the later is about six times than that of the former; (3) the thermal expansion coefficient decreases with increasing cooling rate at the unfrozen temperature region, but some opposite results are obtained at the frozen temperature region; (4) the higher the DMSO concentration is, at the phase change temperature region, the smaller the thermal strain change as well as the maximum thermal expansion coefficient are, but DMSO concentration exhibits little effect on the thermal expansion coefficient at both unfrozen and frozen region. Once the DMSO concentration increasing enough, e.g. 60% v/v, the thermal strain decreases linearly and smoothly without any abrupt change due to little or no ice crystal forms (i.e. vitrification) in frozen articular cartilage. This study may improve our understanding of the thermal expansion (or contraction) behavior of cryopreserved articular cartilage and it may be useful for the future study on cryopreservation of intact

  12. Predictive modeling of freezing and thawing of frost-susceptible soils. (United States)


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

  13. Multiple Glass Transitions and Freezing Events of Aqueous Citric Acid (United States)


    Calorimetric and optical cryo-microscope measurements of 10–64 wt % citric acid (CA) solutions subjected to moderate (3 K/min) and slow (0.5 and 0.1 K/min) cooling/warming rates and also to quenching/moderate warming between 320 and 133 K are presented. Depending on solution concentration and cooling rate, the obtained thermograms show one freezing event and from one to three liquid–glass transitions upon cooling and from one to six liquid–glass and reverse glass–liquid transitions, one or two freezing events, and one melting event upon warming of frozen/glassy CA/H2O. The multiple freezing events and glass transitions pertain to the mother CA/H2O solution itself and two freeze-concentrated solution regions, FCS1 and FCS2, of different concentrations. The FCS1 and FCS2 (or FCS22) are formed during the freezing of CA/H2O upon cooling and/or during the freezing upon warming of partly glassy or entirely glassy mother CA/H2O. The formation of two FCS1 and FCS22 regions during the freezing upon warming to our best knowledge has never been reported before. Using an optical cryo-microscope, we are able to observe the formation of a continuous ice framework (IF) and its morphology and reciprocal distribution of IF/(FCS1 + FCS2). Our results provide a new look at the freezing and glass transition behavior of aqueous solutions and can be used for the optimization of lyophilization and freezing of foods and biopharmaceutical formulations, among many other applications where freezing plays a crucial role. PMID:25482069

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

    NARCIS (Netherlands)

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


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

  15. Effects of industrial pre-freezing processing and freezing handling on glucosinolates and antioxidant attributes in broccoli florets. (United States)

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


    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Initiation of urinary bladder carcinogenesis in rats by freeze ulceration with sodium saccharin promotion. (United States)

    Hasegawa, R; Greenfield, R E; Murasaki, G; Suzuki, T; Cohen, S M


    Sodium saccharin was previously shown to induce a significant incidence of transitional cell carcinoma of the bladder when administered to rats either immediately or beginning 2 weeks after ulceration of the bladder epithelium induced by freezing or cyclophosphamide injection. However, the marked regenerative hyperplasia following ulceration by either of these methods is not completely repaired until 3 to 4 weeks after ulceration. To determine whether initiation in this model was due to the ulceration and regenerative hyperplasia alone or if it was due to the administration of sodium saccharin acting on the hyperplastic epithelium, the effect of administering sodium saccharin at various times after ulceration was examined. Five-week-old F344 male rats were given sodium saccharin as 5% of the diet beginning either immediately (Group 1) or 2, 4, 6, or 18 weeks (Groups 2, 3, 4, or 5, respectively) after freezing of the bladder, and sacrificed 2 years after the start of the experiment. The incidences of rats with transitional cell carcinoma of the bladder were 11 of 36 rats (31%) in Group 1, 6 of 36 (17%) in Group 2, 12 of 40 (30%) in Group 3, 7 of 36 (19%) in Group 4, and 9 of 39 (23%) in Group 5. Sodium saccharin without prior ulceration induced a transitional cell papilloma in one rat, and freeze ulceration without subsequent sodium saccharin induced a transitional cell carcinoma in one rat. No bladder lesions were seen in the untreated control rats. Scanning electron microscopic examination of rats fed sodium saccharin after ulceration showed evidence of multifocal hyperplasia and significant surface changes either at Week 18 of the experiment (Groups 1 to 3) or 18 weeks after beginning sodium saccharin administration (Groups 4 and 5). These results indicate that freeze ulceration of the bladder induced irreversible changes in the epithelial cells related to bladder cancer initiation even though the regenerative hyperplasia is morphologically reversible, and that

  17. Physical Chemistry of the Freezing Process of Atmospheric Aqueous Drops. (United States)

    Bogdan, Anatoli; Molina, Mario J


    In supercooled aqueous solutions, ice nucleation is the initial stage of the freezing process. In this paper, we present experimental results that indicate that during the freezing of aqueous solutions, freeze-induced phase separation (FIPS) into pure ice and a freeze-concentrated solution (FCS) takes place. Our observations involve the use of an optical cryo-microscope (OC-M) to record images and movies. The results visually indicate for the first time that there are two freezing processes for (NH 4 ) 3 H(SO 4 ) 2 /H 2 O solutions: (i) contact freezing, as is the case for pure water drops, and (ii) the Wegener-Bergeron-Findeisen process, which is the growth of frozen drops (ice) at the expense of liquid ones. We also present OC-M images of frozen micrometer-scaled H 2 SO 4 /H 2 O drops that support our previous finding that freezing of these solutions generates mixed-phase particles, namely an ice core coated with a FCS. These results are relevant for atmospheric as well as for pharmaceutical sciences.

  18. Metabolic changes in Avena sativa crowns recovering from freezing. (United States)

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


    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.

  19. Freeze-thaw durability of air-entrained concrete. (United States)

    Shang, Huai-Shuai; Yi, Ting-Hua


    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.

  20. Freeze-Thaw Durability of Air-Entrained Concrete

    Directory of Open Access Journals (Sweden)

    Huai-Shuai Shang


    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.

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

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


    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.

  2. Universal pion freeze-out in heavy-ion collisions

    CERN Document Server

    Adamova, D; Appelshäuser, H; Belaga, V; Braun-Munzinger, P; Castillo, A; Cherlin, A; Damjanovic, S; Dietel, T; Dietrich, L; Drees, A; Esumi, S I; Filimonov, K; Fomenko, K; Fraenkel, Zeev; Garabatos, C; Glässel, P; Hering, G; Holeczek, J; Kushpil, V; Lenkeit, B C; Ludolphs, W; Maas, A; Marin, A; Milosevic, J; Milov, A; Miskowiec, D; Panebratsev, Yu A; Petchenova, O Yu; Petracek, V; Pfeiffer, A; Rak, J; Ravinovich, I; Rehak, P; Sako, H; Schükraft, Jürgen; Sedykh, S; Shimansky, S S; Slivova, J; Specht, H J; Stachel, J; Sumbera, M; Tilsner, H; Tserruya, Itzhak; Wessels, J P; Wienold, T; Windelband, B; Wurm, J P; Xie, W; Yurevich, S; Yurevich, V; Schmitz, W


    Based on an evaluation of recent systematic data on two-pion interferometry and on measured particle yields at mid-rapidity, we propose a universal condition for thermal freeze-out of pions in heavy-ion collisions. We show that freeze-out occurs when the mean free path of pions lambda_f reaches a value of approximately 2.5 fm, which is much smaller than the spatial extent of the system at freeze-out. This critical mean free path is independent of the centrality of the collision and its value is constant at all currently available beam energies from AGS to RHIC.

  3. Organic acid catalyzed carbon aerogels with freeze-drying (United States)

    Xu, Yuelong; Yan, Meifang; Liu, Zhenfa


    Carbon aerogels (CAs) were synthesized via a sol-gel process by condensation-polymerization of phloroglucinol, resorcinol and formaldehyde using 2,4-dihydroxybenzoic acid as catalyst with freeze-drying. The effects of the freeze-drying method on the texture and pore structure were studied. Meanwhile the structure of carbon aerogels was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and a surface-area analyzer. The results show that the freeze-drying method and acid catalyst were good for the specific surface area of carbon aerogel, up to 765m2 g-1, and pore size distribution.


    Directory of Open Access Journals (Sweden)



    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.

  5. X-ray microanalysis of freeze-dried and frozen-hydrated cryosections

    International Nuclear Information System (INIS)

    Zierold, K.


    The elemental composition and the ultrastructure of biological cells were studied by scanning transmission electron microscopy (STEM) combined with energy dispersive X-ray microanalysis. The preparation technique involves cryofixation, cryoultramicrotomy, cryotransfer, and freeze-drying of samples. Freeze-dried cryosections 100-nm thick appeared to be appropriate for measuring the distribution of diffusible elements and water in different compartments of the cells. The lateral analytical resolution was less than 50 nm, depending on ice crystal damage and section thickness. The detection limit was in the range of 10 mmol/kg dry weight for all elements with an atomic number higher than 12; for sodium and magnesium the detection limits were about 30 and 20 mmol/kg dry weight, respectively. The darkfield intensity in STEM is linearly related to the mass thickness. Thus, it becomes possible to measure the water content in intracellular compartments by using the darkfield signal of the dry mass remaining after freeze-drying. By combining the X-ray microanalytical data expressed as dry weight concentrations with the measurements of the water content, physiologically more meaningful wet weight concentrations of elements were determined. In comparison to freeze-dried cryosections frozen-hydrated sections showed poor contrast and were very sensitive against radiation damage, resulting in mass loss. The high electron exposure required for recording X-ray spectra made reproducible microanalysis of ultrathin (about 100-nm thick) frozen-hydrated sections impossible. The mass loss could be reduced by carbon coating; however, the improvement achieved thus far is still insufficient for applications in X-ray microanalysis. Therefore, at present only bulk specimens or at least 1-micron thick sections can be used for X-ray microanalysis of frozen-hydrated biological samples

  6. Estimating collision efficiencies from contact freezing experiments (United States)

    Nagare, B.; Marcolli, C.; Stetzer, O.; Lohmann, U.


    Interactions of atmospheric aerosols with clouds influence cloud properties and modify the aerosol life cycle. Aerosol particles act as cloud condensation nuclei and ice nucleating particles or become incorporated into cloud droplets by scavenging. For an accurate description of aerosol scavenging and ice nucleation in contact mode, collision efficiency between droplets and aerosol particles needs to be known. This study derives the collision rate from experimental contact freezing data obtained with the ETH Collision Ice Nucleation Chamber CLINCH. Freely falling 80 μm water droplets are exposed to an aerosol consisting of 200 nm diameter silver iodide particles of concentrations from 500-5000 cm-3, which act as ice nucleating particles in contact mode. The chamber is kept at ice saturation in the temperature range from 236-261 K leading to slow evaporation of water droplets giving rise to thermophoresis and diffusiophoresis. Droplets and particles bear charges inducing electrophoresis. The experimentally derived collision efficiency of 0.13 is around one order of magnitude higher than theoretical formulations which include Brownian diffusion, impaction, interception, thermophoretic, diffusiophoretic and electric forces. This discrepancy is most probably due to uncertainties and inaccuracies in the description of thermophoretic and diffusiophoretic processes acting together. This is to the authors knowledge the first dataset of collision efficiencies acquired below 273 K. More such experiments with different droplet and particle diameters are needed to improve our understanding of collision processes acting together.

  7. Adaptation to seasonality and the winter freeze

    Directory of Open Access Journals (Sweden)

    Jill Christine Preston


    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.

  8. GPR utilization in artificial freezing engineering

    International Nuclear Information System (INIS)

    Song, Lei; Yang, Weihao; Huang, Jiahui; Li, Haipeng; Zhang, Xiaojun


    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. Contamination of freezing soils: Australia's Antarctic opportunity

    International Nuclear Information System (INIS)

    Williams, P.


    Last month, the Federal government announced that millions of dollars were to be spent cleaning up Antarctica, for which Australia has special responsibilities. Australia's largesse is especially interesting in a world context. Antarctica, by international agreement, is free of any industrial development - mining, storage of wastes, or any other profit-making activity that would disturb the environment (tourism is allowed under increasingly controlled conditions). The importance of the more or less pristine frigid environment lies in the wide range of scientific research that is carried out there. Sophisticated techniques to improve environmental quality are evidently in the early development stage. That cold-loving organisms can thrive in frozen ground in Antarctica and the Arctic was a discovery so unexpected that few people could grasp its importance. Only later was it found that these bugs can eat up contaminants - and the discovery assumed enormous practical significance. Little is known about how to clean up contamination in freezing soils even though there is a pressing need to solve the growing problem with military, industrial and nuclear waste in the Northern Hemisphere

  10. Metabolic Process During the Repair of Freeze-Injury in Escherichia coli1 (United States)

    Ray, B.; Speck, M. L.


    After Escherichia coli was injured by freezing, the repair process was studied during incubation of the cells for 2 hr at 25 C in 0.5% K2HPO4 at pH 7.0 in the presence of specific metabolic inhibitors. The repair in K2HPO4 was not affected by inhibitors of the synthesis of protein, nucleic acids, and mucopeptide. These inhibitors prevented growth of the repaired cells in a minimal broth at 35 C for 24 hr (except actinomycin D and hydroxyurea). Several uncouplers of adenosine triphosphate (ATP) synthesis reduced the repair process in K2HPO4, but only cyanide and azide prevented growth in minimal medium. Data indicated that the cells synthesized energy in the form of ATP and probably utilized it for the repair process. Addition of ATP also facilitated the repair of injury. The freeze-injured cells showed extreme susceptibility to surface-active agents and lysozyme. The repaired cells, like the uninjured cells, became relatively resistant to these compounds. PMID:4564043

  11. Biological activity and mechanical stability of sol-gel-based biofilters using the freeze-gelation technique for immobilization of Rhodococcus ruber. (United States)

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


    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.

  12. Microwave Enhanced Freeze Drying of Solid Waste, Phase II (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...

  13. Natural Electrical Potentials That Arise When Soils Freeze. (United States)


    phenomena occurring during the freezing of dilute aqueous solutions and their possible relation- ship to thunderstorm electricity . Physical Review, 75(3): 2254-259 24 I. * - -- - , -U. - - S -- ~ v -~

  14. Microwave Enhanced Freeze Drying of Solid Waste, Phase I (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...

  15. Normal freezing of ideal ternary systems of the pseudobinary type (United States)

    Li, C. H.


    Perfect liquid mixing but no solid diffusion is assumed in normal freezing. In addition, the molar compositions of the freezing solid and remaining liquid, respectively, follow the solidus and liquidus curves of the constitutional diagram. For the linear case, in which both the liquidus and solidus are perfectly straight lines, the normal freezing equation giving the fraction solidified at each melt temperature and the solute concentration profile in the frozen solid was determined as early as 1902, and has since been repeatedly published. Corresponding equations for quadratic, cubic or higher-degree liquidus and solidus lines have also been obtained. The equation of normal freezing for ideal ternary liquid solutions solidified into ideal solid solutions of the pseudobinary type is given. Sample computations with the use of this new equation were made and are given for the Ga-Al-As system.

  16. Hyperbaric Oxygen as an Adjunct Treatment of Freezing Cold Injury

    National Research Council Canada - National Science Library

    Mekjavic, Igor B; Gorjanc, Jurij; Mekjavic, Polana J; Bajrovic, Fajko; Milcinski, Metka


    .... With the exception of freezing cold injury, hyperbaric oxygen therapy is now accepted as an adjunct therapy for the management of all other acute traumatic peripheral ischaemias, such as crush...

  17. Freeze-drying of silica nanoparticles: redispersibility toward nanomedicine applications. (United States)

    Picco, Agustin S; Ferreira, Larissa F; Liberato, Michelle S; Mondo, Gabriela B; Cardoso, Mateus B


    To study freeze-drying of silica nanoparticles (SiO 2 NPs) in order to find suitable conditions to produce lyophilized powders with no aggregation after resuspension and storage. SiO 2 NPs were synthesized using a Stöber-based procedure, and characterized by scanning electron microscopy, dynamic light scattering and nitrogen adsorption/desorption isotherms. SiO 2 NPs hydrodynamic diameters were compared prior and after freeze-drying in the presence/absence of carbohydrate protectants. Glucose was found to be the most suitable protectant against the detrimental effects of lyophilization. The minimum concentration of carbohydrate required to effectively protect SiO 2 NPs from aggregation during freeze-drying is influenced by the nanoparticle's size and texture. Negligible aggregation was observed during storage. Carbohydrates can be used during SiO 2 NPs freeze-drying process to obtain redispersable solids that maintain original sizes without residual aggregation.

  18. Cellular damage suffered by equine embryos after exposure to cryoprotectants or cryopreservation by slow-freezing or vitrification. (United States)

    Hendriks, W K; Roelen, B A J; Colenbrander, B; Stout, T A E


    Equine embryos are cryopreserved by slow-freezing or vitrification. While small embryos (cryopreservation reasonably well, larger embryos do not. It is not clear if slow-freezing or vitrification is less damaging to horse embryos. To compare the type and extent of cellular damage suffered by small and large embryos during cryopreservation by slow-freezing vs. vitrification. Sixty-three Day 6.5-7 embryos were subdivided by size and assigned to one of 5 treatments: control, exposure to slow-freezing or vitrification cryoprotectants (CPs), and cryopreservation by either technique. After thawing/CP removal, embryos were stained with fluorescent stains for various parameters of cellular integrity, and assessed by multiphoton microscopy. Exposing large embryos to vitrification CPs resulted in more dead cells (6.8 ± 1.3%: 95% confidence interval [CI], 3.1-10.4%) than exposure to slow-freezing media (0.3 ± 0.1%; 95% CI 0.0-0.6%: P = 0.001). Cryopreservation by either technique induced cell death and cytoskeleton disruption. Vitrification of small embryos resulted in a higher proportion of cells with fragmented or condensed (apoptotic) nuclei (P = 0.002) than slow-freezing (6.7 ± 1.5%, 95% CI 3.0-10.4% vs. 5.0 ± 2.1%, 95% CI 4.0-14.0%). Slow-freezing resulted in a higher incidence of disintegrated embryos (P = 0.01) than vitrification. Mitochondrial activity was low in control embryos, and was not differentially affected by cryopreservation technique, whereas vitrification changed mitochondrial distribution from a homogenous crystalline pattern in control embryos to a heterogeneous granulated distribution in vitrified embryos (P = 0.05). Cryopreservation caused more cellular damage to large embryos than smaller ones. While vitrification is more practical, it is not advisable for large embryos due to a higher incidence of dead cells. The choice is less obvious for small embryos, as vitrification led to occasionally very high percentages of dead or damaged

  19. Immunisation of Sheep with Bovine Viral Diarrhoea Virus, E2 Protein Using a Freeze-Dried Hollow Silica Mesoporous Nanoparticle Formulation.

    Directory of Open Access Journals (Sweden)

    Donna Mahony

    Full Text Available Bovine viral diarrhoea virus 1 (BVDV-1 is arguably the most important viral disease of cattle. It is associated with reproductive, respiratory and chronic diseases in cattle across the world. In this study we have investigated the capacity of the major immunological determinant of BVDV-1, the E2 protein combined with hollow type mesoporous silica nanoparticles with surface amino functionalisation (HMSA, to stimulate immune responses in sheep. The current work also investigated the immunogenicity of the E2 nanoformulation before and after freeze-drying processes. The optimal excipient formulation for freeze-drying of the E2 nanoformulation was determined to be 5% trehalose and 1% glycine. This excipient formulation preserved both the E2 protein integrity and HMSA particle structure. Sheep were immunised three times at three week intervals by subcutaneous injection with 500 μg E2 adsorbed to 6.2 mg HMSA as either a non-freeze-dried or freeze-dried nanoformulation. The capacity of both nanovaccine formulations to generate humoral (antibody and cell-mediated responses in sheep were compared to the responses in sheep immunisation with Opti-E2 (500 μg together with the conventional adjuvant Quil-A (1 mg, a saponin from the Molina tree (Quillaja saponira. The level of the antibody responses detected to both the non-freeze-dried and freeze-dried Opti-E2/HMSA nanoformulations were similar to those obtained for Opti-E2 plus Quil-A, demonstrating the E2 nanoformulations were immunogenic in a large animal, and freeze-drying did not affect the immunogenicity of the E2 antigen. Importantly, it was demonstrated that the long term cell-mediated immune responses were detectable up to four months after immunisation. The cell-mediated immune responses were consistently high in all sheep immunised with the freeze-dried Opti-E2/HMSA nanovaccine formulation (>2,290 SFU/million cells compared to the non-freeze-dried nanovaccine formulation (213-500 SFU/million cells

  20. Large p/sub t/ enhancement from freeze out

    CERN Document Server

    Magas, V K; Csernai, László P; Grassi, Frédérique; Greiner, W; Hama, Y; Kodama, T; Lázár, Z I; Stöcker, H


    Freeze out of particles across three dimensional space-time hypersurface is discussed in a simple kinetic model. The final momentum distribution of emitted particles, for freeze out surfaces with space-like normal, shows a $9 non-exponential transverse momentum spectrum. The slope parameter of the p/sub t/ distribution increases with increasing p/sub t/, in agreement with recently measured SPS pion and h/sup -/ spectra. (11 refs).

  1. Ice, Ice, Baby? A Sociological Exploration of Social Egg Freezing


    Baldwin, K.


    Social egg freezing is a fertility preservation strategy which enables women to preserve a number of healthy unfertilised eggs for potential future use when faced with the threat of age-related fertility decline. The overall aim of this thesis was to explore how women understand, construct and experience social egg freezing in the context of debates surrounding reproductive ‘choice’ and ‘delayed motherhood’. The study sought to provide insights into how women perceive the risks and benefi...

  2. Boundary lubrication properties of materials with expansive freezing


    Jagla, E. A.


    We have performed molecular dynamics simulations of solid-solid contacts lubricated by a model fluid displaying many of the properties of water, particularly its expansive freezing. Near the region where expansive freezing occurs, the lubricating film remains fluid, and the friction force decreases linearly as the shear velocity is reduced. No sign of stick-slip motion is observed even at the lowest velocities. We give a simple interpretation of these results, and suggest that in general good...

  3. On the calculation of the electrolyte aqueous solution freezing temperature

    International Nuclear Information System (INIS)

    Ptitsyn, A.B.


    Analytical expression of the dependence of freezing temperature of electrolyte (CdCl 2 , CdSO 4 , BeSO 4 ) aqueous solutions on their composition was found. Three different methods were suggested were suggested for calculation of water activity in multicomponent solution. It was established that sublinear extrapolation of freezing temperature of salt aqueous solutions was possible not up to -5 deg C, but up to -15 deg C, independently of their composition. 6 refs.; 2 figs


    Directory of Open Access Journals (Sweden)

    V. M. Stefanovskiy


    Full Text Available The concept of «ideal product» is proposed for the study of mass transfer during partial freezing of food products by freezing plate. The ideal product is a product, in which number of factors affecting the «real product» (meat are excluded. These factors include chemical composition of meat, quality grade of raw material (NOR, DFD, PSE, cryoscopic temperature that determines the degree of water transformation into ice, the phenomenon of osmosis, rate of freezing, etc. By using the concept of «ideal product» and its implementation in a physical experiment, it is proved that the “piston effect” causing the migration of moisture is due to frozen crust formation during partial freezing of the body. During partial freezing of the product by freezing plate, «ideal» and «real» food environment is transformed from closed system into open one with inflow of moisture to unfrozen part of the body. In the «ideal product», there is an expulsion of unfrozen moisture from freezing front, so the water appears on the body surface. Thus, the displacement of moisture increases by the same law, according to which the thickness (weight of frozen layer increases. During partial freezing of ground meat, moisture does not appear on the surface of the product, but hydrates the unfrozen part of meat. The reason of this phenomenon is the expulsion of water during formation of frozen crust and water-binding capacity of meat.

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

    Energy Technology Data Exchange (ETDEWEB)

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


    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.

  6. Acceleration of oxidation-reduction reactions in freezing solution


    Takenaka,Norimichi; Ueda,Akihiro; Maeda,Yasuaki


    In general, most chemical reactions are accelerated at higher temperature, while reactions of which activation energy is negative are accelerated at lower temperature. The reaction rates, however, become much slower in the ice phase because molecule cannot move easily. It was found that a few reactions of which activation energies were positive were accelerated in freezing aqueous solution. The reaction rates in the freezing process were 10 to (10)^5 times faster than those in solution at 25℃...

  7. Simulation Study on Freeze-drying Characteristics of Mashed Beef


    Tambunan, Armansyah H; Solahudin, M; Rahajeng, Estri


    Drying characteristic of a particular product is important in analyzing the appropriateness of the drying method for the product. This is especially important for freeze drying, which is known as the most expensive drying method, asideji-om its good drying quality. The objectives of this experiment are to develop a computer simulation program using a retreating drying-frontmodel for predicting freeze drying characteristics of mashed nteat, especially for the influence of sublimation temperatu...

  8. Freeze-drying wet digital prints: An option for salvage?

    International Nuclear Information System (INIS)

    Juergens, M C; Schempp, N


    On the occasion of the collapse of the Historical Archive of the City of Cologne in March 2009 and the ensuing salvage effort, questions were raised about the use of freeze-drying for soaked digital prints, a technique that has not yet been evaluated for these materials. This study examines the effects of immersion, air-drying, drying in a blotter stack, freezing and freeze-drying on 35 samples of major digital printing processes. The samples were examined visually before, during and after testing; evaluation of the results was qualitative. Results show that some prints were already damaged by immersion alone (e.g. bleeding inks and soluble coatings) to the extent that the subsequent choice of drying method made no significant difference any more. For those samples that did survive immersion, air-drying proved to be crucial for water-sensitive prints, since any contact with the wet surface caused serious damage. Less water-sensitive prints showed no damage throughout the entire procedure, regardless of drying method. Some prints on coated media suffered from minor surface disruption up to total delamination of the surface coating due to the formation of ice crystals during shock-freezing. With few exceptions, freeze-drying did not cause additional damage to any of the prints that hadn't already been damaged by freezing. It became clear that an understanding of the process and materials is important for choosing an appropriate drying method.

  9. Mechanisms of deterioration of nutrients. [improved quality of freeze-dried foods (United States)

    Karel, M.; Flink, J. M.


    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.

  10. Freeze-drying microscopy in mathematical modeling of a biomaterial freeze-drying

    Directory of Open Access Journals (Sweden)

    Camila Figueiredo Borgognoni


    Full Text Available Transplantation brings hope for many patients. A multidisciplinary approach on this field aims at creating biologically functional tissues to be used as implants and prostheses. The freeze-drying process allows the fundamental properties of these materials to be preserved, making future manipulation and storage easier. Optimizing a freeze-drying cycle is of great importance since it aims at reducing process costs while increasing product quality of this time-and-energy-consuming process. Mathematical modeling comes as a tool to help a better understanding of the process variables behavior and consequently it helps optimization studies. Freeze-drying microscopy is a technique usually applied to determine critical temperatures of liquid formulations. It has been used in this work to determine the sublimation rates of a biological tissue freeze-drying. The sublimation rates were measured from the speed of the moving interface between the dried and the frozen layer under 21.33, 42.66 and 63.99 Pa. The studied variables were used in a theoretical model to simulate various temperature profiles of the freeze-drying process. Good agreement between the experimental and the simulated results was found.A prática da transplantação traz esperança para muitos pacientes. Uma visão multidisciplinar nessa área visa à produção de tecidos biológicos para serem utilizados como implantes e próteses. A liofilização é um processo de secagem que preserva características essenciais desses materiais, facilitando sua manipulação e armazenamento. A liofilização é um processo que requer muito tempo e energia e sua otimização é muito importante, pois permite reduzir custos de processo melhorando a qualidade do produto. A modelagem matemática é uma ferramenta que permite descrever o comportamento do produto durante o processo e, consequentemente, auxilia os estudos de otimização. Microscopia óptica acoplada à liofilização, uma t

  11. Seasonal Variation in the Hepatoproteome of the Dehydration- and Freeze-Tolerant Wood Frog, Rana sylvatica

    Directory of Open Access Journals (Sweden)

    Jon P. Costanzo


    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.

  12. Evaluation of 3D-Printed Polycaprolactone Scaffolds Coated with Freeze-Dried Platelet-Rich Plasma for Bone Regeneration

    Directory of Open Access Journals (Sweden)

    Junda Li


    Full Text Available Three-dimensional printing is one of the most promising techniques for the manufacturing of scaffolds for bone tissue engineering. However, a pure scaffold is limited by its biological properties. Platelet-rich plasma (PRP has been shown to have the potential to improve the osteogenic effect. In this study, we improved the biological properties of scaffolds by coating 3D-printed polycaprolactone (PCL scaffolds with freeze-dried and traditionally prepared PRP, and we evaluated these scaffolds through in vitro and in vivo experiments. In vitro, we evaluated the interaction between dental pulp stem cells (DPSCs and the scaffolds by measuring cell proliferation, alkaline phosphatase (ALP activity, and osteogenic differentiation. The results showed that freeze-dried PRP significantly enhanced ALP activity and the mRNA expression levels of osteogenic genes (ALP, RUNX2 (runt-related gene-2, OCN (osteocalcin, OPN (osteopontin of DPSCs (p < 0.05. In vivo, 5 mm calvarial defects were created, and the PRP-PCL scaffolds were implanted. The data showed that compared with traditional PRP-PCL scaffolds or bare PCL scaffolds, the freeze-dried PRP-PCL scaffolds induced significantly greater bone formation (p < 0.05. All these data suggest that coating 3D-printed PCL scaffolds with freeze-dried PRP can promote greater osteogenic differentiation of DPSCs and induce more bone formation, which may have great potential in future clinical applications.

  13. A Robust, Cell-free Production System for On-Demand Protein Synthesis in Space (United States)

    National Aeronautics and Space Administration — We will develop a new cell-free expression system that functions after rehydrating from a freeze-dried condition. Freeze-dried powder that can be stored or...

  14. Reproducing Black’s experiments: freezing point depression and supercooling of water


    Güémez, Júlio; Fiolhais, Carlos; Fiolhais, Manuel


    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,...

  15. Effects of Pressure-shift Freezing on the Structural and Physical Properties of Gelatin Hydrogel Matrices


    Kim, Byeongsoo; Gil, Hyung Bae; Min, Sang-Gi; Lee, Si-Kyung; Choi, Mi-Jung


    This study investigates the effects of the gelatin concentration (10-40%, w/v), freezing temperatures (from -20? to -50?) and freezing methods on the structural and physical properties of gelatin matrices. To freeze gelatin, the pressure-shift freezing (PSF) is being applied at 0.1 (under atmospheric control), 50 and 100 MPa, respectively. The freezing point of gelatin solutions decrease with increasing gelatin concentrations, from -0.2? (10% gelatin) to -6.7? (40% gelatin), while the extent ...

  16. Freezing of gait: Promising avenues for future treatment. (United States)

    Gilat, Moran; Lígia Silva de Lima, Ana; Bloem, Bastiaan R; Shine, James M; Nonnekes, Jorik; Lewis, Simon J G


    Freezing of gait is a devastating symptom of Parkinson's disease and other forms of parkinsonism. It poses a major burden on both patients and their families, as freezing often leads to falls, fall-related injuries and a loss of independence. Treating freezing of gait is difficult for a variety of reasons: it has a paroxysmal and unpredictable nature; a multifaceted pathophysiology, with an interplay between motor elements (disturbed stepping mechanisms) and non-motor elements (cognitive decline, anxiety); and a complex (and likely heterogeneous) underlying neural substrate, involving multiple failing neural networks. In recent years, advances in translational neuroscience have offered new insights into the pathophysiology underlying freezing. Furthermore, the mechanisms behind the effectiveness of available treatments (or lack thereof) are better understood. Driven by these concepts, researchers and clinicians have begun to improve currently available treatment options, and develop new and better treatment methods. Here, we evaluate the range of pharmacological (i.e. closed-looped approaches), surgical (i.e. multi-target and adaptive deep brain and spinal cord stimulation) and behavioural (i.e. biofeedback and cueing on demand) treatment options that are under development, and propose novel avenues that are likely to play a crucial role in the clinical management of freezing of gait in the near future. The outcomes of this review suggest that the successful future management of freezing of gait will require individualized treatments that can be implemented in an on-demand manner in response to imminent freezing. With this review we hope to guide much-needed advances in treating this devastating symptom of Parkinson's disease. Copyright © 2018 Elsevier Ltd. All rights reserved.

  17. X-ray microanalysis of high-pressure frozen, freeze-substituted, Ni hyperaccumulating plants

    International Nuclear Information System (INIS)

    Bidwell, S.D.; Crawford, S.A.; Woodrow, I.E.; Sommer-Knudsen, J.; Marshall, A.T.


    Full text: The preparation of plant material for the x-ray microanalysis of diffusible elements is notoriously difficult, especially where cells are highly vacuolated. It is shown that leaves of a Ni hyperaccumulator, Hybanthus floribundus, are well preserved at the ultrastructural level by high-pressure freezing followed by an anhydrous freeze-substitution protocol (Marshall, 1980) Dry cut sections were subjected to x-ray microanalysis in a Jeol 1200EX STEM to produce quantitative element distribution maps. Element distribution maps showed that Ni was predominantly localised in the vacuoles of epidermal cells in the leaves. Quantitation of Ni revealed concentrations up to 275 mmol kg -1 (embedded tissue) in some epidermal vacuoles. The accumulation of Ni in these cells was associated with a decrease in the concentration of Na and K. There was no indication that Ni was associated with P, S or Cl in the vacuoles Ni also concentrated on the outside of cell walls throughout the leaf section, indicating that apoplastic compartmentation is also involved in Ni tolerance and accumulation in this plant. Copyright (2002) Australian Society for Electron Microscopy Inc

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

    NARCIS (Netherlands)

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


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

  19. The impact of freeze-drying infant fecal samples on measures of their bacterial community profiles and milk-derived oligosaccharide content. (United States)

    Lewis, Zachery T; Davis, Jasmine C C; Smilowitz, Jennifer T; German, J Bruce; Lebrilla, Carlito B; Mills, David A


    Infant fecal samples are commonly studied to investigate the impacts of breastfeeding on the development of the microbiota and subsequent health effects. Comparisons of infants living in different geographic regions and environmental contexts are needed to aid our understanding of evolutionarily-selected milk adaptations. However, the preservation of fecal samples from individuals in remote locales until they can be processed can be a challenge. Freeze-drying (lyophilization) offers a cost-effective way to preserve some biological samples for transport and analysis at a later date. Currently, it is unknown what, if any, biases are introduced into various analyses by the freeze-drying process. Here, we investigated how freeze-drying affected analysis of two relevant and intertwined aspects of infant fecal samples, marker gene amplicon sequencing of the bacterial community and the fecal oligosaccharide profile (undigested human milk oligosaccharides). No differences were discovered between the fecal oligosaccharide profiles of wet and freeze-dried samples. The marker gene sequencing data showed an increase in proportional representation of Bacteriodes and a decrease in detection of bifidobacteria and members of class Bacilli after freeze-drying. This sample treatment bias may possibly be related to the cell morphology of these different taxa (Gram status). However, these effects did not overwhelm the natural variation among individuals, as the community data still strongly grouped by subject and not by freeze-drying status. We also found that compensating for sample concentration during freeze-drying, while not necessary, was also not detrimental. Freeze-drying may therefore be an acceptable method of sample preservation and mass reduction for some studies of microbial ecology and milk glycan analysis.

  20. The impact of freeze-drying infant fecal samples on measures of their bacterial community profiles and milk-derived oligosaccharide content

    Directory of Open Access Journals (Sweden)

    Zachery T. Lewis


    Full Text Available Infant fecal samples are commonly studied to investigate the impacts of breastfeeding on the development of the microbiota and subsequent health effects. Comparisons of infants living in different geographic regions and environmental contexts are needed to aid our understanding of evolutionarily-selected milk adaptations. However, the preservation of fecal samples from individuals in remote locales until they can be processed can be a challenge. Freeze-drying (lyophilization offers a cost-effective way to preserve some biological samples for transport and analysis at a later date. Currently, it is unknown what, if any, biases are introduced into various analyses by the freeze-drying process. Here, we investigated how freeze-drying affected analysis of two relevant and intertwined aspects of infant fecal samples, marker gene amplicon sequencing of the bacterial community and the fecal oligosaccharide profile (undigested human milk oligosaccharides. No differences were discovered between the fecal oligosaccharide profiles of wet and freeze-dried samples. The marker gene sequencing data showed an increase in proportional representation of Bacteriodes and a decrease in detection of bifidobacteria and members of class Bacilli after freeze-drying. This sample treatment bias may possibly be related to the cell morphology of these different taxa (Gram status. However, these effects did not overwhelm the natural variation among individuals, as the community data still strongly grouped by subject and not by freeze-drying status. We also found that compensating for sample concentration during freeze-drying, while not necessary, was also not detrimental. Freeze-drying may therefore be an acceptable method of sample preservation and mass reduction for some studies of microbial ecology and milk glycan analysis.

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

    Directory of Open Access Journals (Sweden)

    Inmaculada Aranaz


    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.

  2. Immersion freezing of ambient dust using WISDOM setup (United States)

    Rudich, Y.; Reicher, N.


    A small subset of the atmospheric particles has the ability to induce ice formation. Among them are mineral dust particles that originate from arid regions. Mineral dust particles are internally mixed with various types of minerals such as kaolinite and illite from the clay minerals, quartz and feldspar. The mineral composition of the dust particles determine their freezing efficiency. Much attention was given to the clay group, as they are the most common minerals transported in the atmosphere. Recently, much focus has been directed to the feldspars, since its ice efficiency is higher at warmer temperatures, and as such is may dominate freezing in mixed phase clouds. Moreover, it was found that samples that contained higher content of feldspar had higher nucleation activity. In this study, we examine the immersion freezing of ambient dust particles that were collected in Rehovot, Israel (31.9N, 34.8E about 80m AMSL), during dust storms from the Sahara and the Syrian deserts. The size-segregated dust particles were collected on cyclopore polycarbonate filters using a Micro-orifice Uniform deposit Impactor (MOUDI). Freezing experiments were done using the WeIzmann Supercooled Droplets Observation on Microarray set (WISDOM). The particles were extracted from the filters by sonication and subsequently immersed in 100μm droplets that were cooled in a rate of 1°CPM to -37°C (homogenous freezing threshold). Investigation of the particles mineralogy was also performed. We observed freezing onset at 253K for particles of different diameters (0.3, 1.0, 1.8 and 3.2 μm). Most of the droplets were completely frozen by 243K. The number of active sites ranged from 108 to 1012 per m-2. Droplets that contained larger particles (higher surface area) froze at slightly warmer temperatures and contained slightly higher number of active sites. The freezing behavior fits well with measurements of K-feldspar particles and this may suggest that the feldspar dominated the dust freezing

  3. Responses of Lithium-Modified Bath to a Shift in Heat Input/Output Balance and Observation of Freeze-Lining Formation During the Heat Balance Shift (United States)

    Liu, Jingjing; Taylor, Mark; Dorreen, Mark


    In the aluminum electrolysis process, new industrial aluminum/electricity power markets demand a new cell technology to extend the cell heat balance and amperage operating window of smelters by shifting the steady states. The current work investigates the responses of lithium-modified bath system when the input/output balance is shifted in a laboratory analogue to the industrial heat balance shift. Li2CO3 is added to the cryolite-AlF3-CaF2-Al2O3 system as a bath modifier. A freeze deposit is formed on a `cold finger' dipped into the bath and investigated by X-ray diffraction analysis and electron probe X-ray microanalysis. The macro- and micro-structure of the freeze lining varies with the bath superheat (bath temperature minus bath liquidus temperature) and an open crystalline layer with entrapped liquid dominates the freeze thickness. Compared with the cryolite-AlF3-CaF2-Al2O3 bath system, the lithium-modified bath freeze is more sensitive to the heat balance shift. This freeze investigation provides primary information to understand the variation of the side ledge in an industrial cell when the lithium-modified bath system is used.

  4. A theoretical extension of the soil freezing curve paradigm (United States)

    Amiri, Erfan A.; Craig, James R.; Kurylyk, Barret L.


    Numerical models of permafrost evolution in porous media typically rely upon a smooth continuous relation between pore ice saturation and sub-freezing temperature, rather than the abrupt phase change that occurs in pure media. Soil scientists have known for decades that this function, known as the soil freezing curve (SFC), is related to the soil water characteristic curve (SWCC) for unfrozen soils due to the analogous capillary and sorptive effects experienced during both soil freezing and drying. Herein we demonstrate that other factors beyond the SFC-SWCC relationship can influence the potential range over which pore water phase change occurs. In particular, we provide a theoretical extension for the functional form of the SFC based upon the presence of spatial heterogeneity in both soil thermal conductivity and the freezing point depression of water. We infer the functional form of the SFC from many abrupt-interface 1-D numerical simulations of heterogeneous systems with prescribed statistical distributions of water and soil properties. The proposed SFC paradigm extension has the appealing features that it (1) is determinable from measurable soil and water properties, (2) collapses into an abrupt phase transition for homogeneous media, (3) describes a wide range of heterogeneity within a single functional expression, and (4) replicates the observed hysteretic behavior of freeze-thaw cycles in soils.

  5. 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)


    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)

  6. 'Banking time': egg freezing and the negotiation of future fertility. (United States)

    Waldby, Catherine


    This paper examines the relatively recent practice of non-medical egg freezing, in which women bank their eggs for later use in conceiving a child. Non-medical egg freezing has only been available for about the last five years, as new vitrification techniques have made the success rates for actual conception more reliable than the earlier method of slow freezing. I draw on interviews with both clinicians and women who have banked their eggs to consider how this novel practice articulates with broader issues about the relationship between sexuality, reproduction and the political economy of household formation. Non-medical egg-freezing provides a technical solution to a number of different problems women face with regard to the elongation of the life course, the extension of education, the cost of household establishment and the iterative nature of relationship formation, thematised by the ubiquity of internet dating among the interviewees. I focus on the ways women used egg freezing to manage and reconcile different forms of time.

  7. Apparatus for freeze drying of biologic and sediment samples

    International Nuclear Information System (INIS)



    Freeze drying to obtain water from individual samples, though not complicated, usually requires considerable effort to maintain the cold traps on a 24-hr basis. In addition, the transfer of a sample from sample containers to freeze-dry flasks is usually made with some risk of contamination to the sample. If samples are large, 300 g to 600 g, usually several days are required to dry the samples. The use of an unattended system greatly improves personnel and drying efficiency. Commercial freeze dryers are not readily applicable to the problems of collecting water from individual samples, and lab-designed collectors required sample transfer and continual replenishment of the dry ice. A freeze-dry apparatus for collecting water from individual sediment and/or biological samples was constructed to determine the tritium concentrations in fish for dose calcaluations and the tritium distribution in sediment cores for water movement studies. The freeze, dry apparatus, which can handle eight samples simultaneously and conveniently, is set up for unattended 24-hr operation and is designed to avoid sample transfer problems

  8. Cryoprotectant-free freezing of the goat epididymal sperm. (United States)

    Katanbafzadeh, H; Barati, F; Tabandeh, M


    Cryoprotectant free approach successfully removed the impact of physical and chemical damages in preserving human sperm in a vitrification protocol. There is no any report on this technology in farm animal sperm freezing. The aim of the present study was to find the efficacy of cholesterol-loaded cyclodextrin (CLC; 1 mg per 60 million) and sucrose (0.1 and 0.2 M) on freezing of the goat epididymal sperm. Caudal epididymides (n=5 pairs) were dissected, incised and incubated in the Tris-BSA solution for 15 min, followed by swim-up at room temperature. Sperm was loaded in 0.25 mL French straws and cooled on nitrogen vapor for 3 min then immersed in liquid nitrogen and remained for 48 h. Then the straws thawed by immersing in 37 degree C waterbath for 30 sec and analyzed. The results showed the impact of freezing on the goat epididymal sperm motility, viability and DNA fragmentation that were improved by incorporation of CLC and sucrose (0.2 M). In conclusion, the goat epididymal sperm was frozen in a cryoprotectant-free freezing model. CLC and 0.4 M sucrose protected the goat epididymal sperm against freezing-induced damages.

  9. The effect of sublethal injury by heating, freezing, drying and gamma-radiation on the duration of the lag phase of Salmonella typhimurium

    International Nuclear Information System (INIS)

    Mackey, B.M.; Derrick, C.M.


    The duration of the lag phase of Salmonella typhimurium surviving heat, freezing, drying and gamma-radiation was used to indicate the time needed to repair sublethal injury. Following equivalent lethal treatments, heat and freeze-injured cells needed longer to repair than those injured by drying or gamma-radiation. Measurement of repair on membrane filters showed that in a heat-injured population having a lag time of 9 h, some individual cells needed up to 14 h to recover maximum tolerance to 3% NaCl. (author)

  10. 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)


    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

  11. Effect of rapid freezing-thawing techniques on the sperm parameters and ultrastructure of Chinese Taihang black goat spermatozoa. (United States)

    Shi, Liguang; Ren, Youshe; Zhou, Hanlin; Hou, Guanyu; Xun, Wenjuan; Yue, Wenbin; Zhang, Chunxiang; Yang, Rujie


    Supercooling sperm in liquid nitrogen vapour is a feasible and economic technique for the practical production. The study aimed to reveal the negative effects of this rapid freezing and thawing processes on Taihang black goat spermatozoa and to find out the changing of spermatozoa motility and ultrastructure by using CASA and TEM. Qualified semen samples, which collected from twenty Chinese Taihang black goats using artificial vagina were pooled and investigated the kinematics parameters and ultrastructural morphology. The results showed that freezing-thawing caused a significant reduction in the spermatozoon total motility (P<0.001), in rapid and medium cell numbers (P<0.001) and motility parameters (VAP, VSL, VCL, ALH and BCF) (P<0.01). Immotile spermatozoa number was increased significantly after freezing-thawing (P<0.001). In the ultrastructural analysis, the shape with a sperm nucleus characterized by ruptures, bend and deformity was observed. The plasma membranes were broken, and nucleoplasm erupted. The mitochondria in the middle piece were disturbed by partial absence or additional accumulations. Swelling, coiling, vacuolization and structural disorganization of mitochondria were also observed. In conclusion, Freezing-thawing procedure has a detrimental effect on motility, membrane integrity and mitochondria of goat spermatozoa. Transmission electron microscopy provides an intuitive observation to investigate deformity spermatozoa. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. High-freezing-point fuels used for aviation turbine engines (United States)

    Friedman, R.


    Broadened-specification aviation fuels could be produced from a greater fraction of crude source material with improvements in fuel supply and price. These fuels, particularly those with increased final boiling temperatures, would have higher freezing temperatures than current aviation turbine fuels. The higher-freezing-point fuels can be substituted in the majority of present commercial flights, since temperature data indicate that in-flight fuel temperatures are relatively mild. For the small but significant fraction of commercial flights where low fuel temperatures make higher freezing-point fuel use unacceptable, adaptations to the fuel or fuel system may be made to accommodate this fuel. Several techniques are discussed. Fuel heating is the most promising concept. One simple system design uses existing heat rejection from the fuel-lubricating oil cooler, another uses an engine-driven generator for electrical heating. Both systems offer advantages that outweigh the obvious penalties.

  13. Positron probe to study the freezing of nanodroplets

    International Nuclear Information System (INIS)

    Pujari, P.K.


    Positron is an excellent in situ probe to study the phase behavior of fluid confined in nanodomains. The study of phase behavior (freezing/melting) of nano confined fluid or nanodroplet has great relevance in fundamental research as well as applications in nano-tribology, nanofabrication, membrane separation, interfacial adhesion and lubrication. It is seen that the properties of freezing/melting of nanodroplets are different from their bulk behavior due to the combined effects of finite size, surface force, surface anisotropy, pore disorder and reduced dimensionality. We have used positron annihilation spectroscopy (PAS) to study the freezing/melting behavior of different organic liquids like benzene, ethylene glycol and isopropanol confined in nanopores of ZSM5 zeolite and silica gel

  14. Encapsulation and subsequent freeze-drying of Lactobacillus reuteri CRL 1324 for its potential inclusion in vaginal probiotic formulations. (United States)

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


    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. Copyright © 2015 Elsevier B.V. All rights reserved.

  15. Contact freezing observed with a high speed video camera (United States)

    Hoffmann, Nadine; Koch, Michael; Kiselev, Alexei; Leisner, Thomas


    Freezing of supercooled cloud droplets on collision with ice nucleating particle (INP) has been considered as one of the most effective heterogeneous freezing mechanisms. Potentially, it could play an important role in rapid glaciation of a mixed phase cloud especially if coupled with ice multiplication mechanism active at moderate subzero temperatures. The necessary condition for such coupling would be, among others, the presence of very efficient INPs capable of inducing ice nucleation of the supercooled drizzle droplets in the temperature range of -5°C to -20°C. Some mineral dust particles (K-feldspar) and biogenic INPs (pseudomonas bacteria, birch pollen) have been recently identified as such very efficient INPs. However, if observed with a high speed video (HSV) camera, the contact nucleation induced by these two classes of INPs exhibits a very different behavior. Whereas bacterial INPs can induce freezing within a millisecond after initial contact with supercooled water, birch pollen need much more time to initiate freezing. The mineral dust particles seem to induce ice nucleation faster than birch pollen but slower than bacterial INPs. In this contribution we show the HSV records of individual supercooled droplets suspended in an electrodynamic balance and colliding with airborne INPs of various types. The HSV camera is coupled with a long-working-distance microscope, allowing us to observe the contact nucleation of ice at very high spatial and temporal resolution. The average time needed to initiate freezing has been measured depending on the INP species. This time do not necessarily correlate with the contact freezing efficiency of the ice nucleating particles. We discuss possible mechanisms explaining this behavior and potential implications for future ice nucleation research.

  16. Effects of freezing on the bactericidal activity of human milk. (United States)

    Takci, Sahin; Gulmez, Dolunay; Yigit, Sule; Dogan, Ozlem; Dik, Kezban; Hascelik, Gulsen


    Storage of human milk by freezing has been recommended for long-term storage. The present study analyzed the bactericidal activity of human milk on Escherichia coli and Pseudomonas aeruginosa and determined the changes in bactericidal activity following freezing at -20°C and -80°C for 1 month and 3 months. Forty-eight milk samples were collected from 48 lactating mothers. Each sample was divided into 10 aliquots. Two of the samples were processed immediately and the others were stored at both -20°C and -80°C until analysis after 1 month and 3 months of freezing. All of the fresh milk samples showed bactericidal activity against E coli and P aeruginosa. Freezing at -20°C for 1 month did not cause statistically significant alteration in bactericidal activity (P > 0.017), whereas storage for 3 months lowered the degree of bactericidal activity significantly (P Bactericidal activity was protected when the samples were stored at -80°C. There was no statistically significant difference in the bactericidal activity of human milk against E coli between freezing at -20°C and -80°C for 1 month (P > 0.017); however, when milk was stored for 3 months, -80°C was significantly more protective (P bactericidal activity against P aeruginosa (P > 0.05). Storage by freezing at -80°C is more appropriate to keep bactericidal capacity of stored human milk >1 month if affordable and available, especially in intensive care settings.

  17. Ultrasonic emissions from conifer xylem exposed to repeated freezing. (United States)

    Mayr, Stefan; Zublasing, Verena


    Ultrasonic emission measurements enable the analysis of xylem cavitation induced by drought and freeze-thaw events. Several studies have indicated that ultrasonic acoustic emissions (UAE) in conifers occur upon freezing and not upon thawing, although classical theory has postulated gas bubble formation during freezing and cavitation during thawing. We analyzed the pattern and quality of freeze-thaw-induced UAE in seven conifers (Abies alba, Larix decidua, Juniperus communis, Picea abies, Pinus cembra, Pinus mugo, Pinus sylvestris). Axes samples dehydrated to different water potentials were exposed to repeated frost cycles. The number, amplitude and energy of UAE signals were registered and related to water potential, temperature course and wood characteristics (wood density, tracheid diameter). For P. abies, ultrasonic emission analysis was also performed on bark samples, xylem samples without bark, as well as on stems of young potted trees. In all conifers, UAE were registered in water-stressed samples but not in saturated or dehydrated samples. No signals were emitted by the bark of P. abies. Ultrasonic activity occurred only upon freezing, and identical patterns were observed in axes samples and stems of potted P. abies trees. A weak positive relationship between tracheid diameter and UAE energy was observed, indicating wide tracheids to emit signals with higher energy. The classical bubble formation hypothesis cannot sufficiently explain the occurrence of UAE during freezing and upon repeated temperature cycles, as demonstrated in this study. We suggest that the low water potential of ice induces air-seeding near the ice-water interface, and consequently, causes UAE.

  18. The effect of dryer load on freeze drying process design. (United States)

    Patel, Sajal M; Jameel, Feroz; Pikal, Michael J


    Freeze-drying using a partial load is a common occurrence during the early manufacturing stages when insufficient amounts of active pharmaceutical ingredient (API) are available. In such cases, the immediate production needs are met by performing lyophilization with less than a full freeze dryer load. However, it is not obvious at what fractional load significant deviations from full load behavior begin. The objective of this research was to systematically study the effects of variation in product load on freeze drying behavior in laboratory, pilot and clinical scale freeze-dryers. Experiments were conducted with 5% mannitol (high heat and mass flux) and 5% sucrose (low heat and mass flux) at different product loads (100%, 50%, 10%, and 2%). Product temperature was measured in edge as well as center vials with thermocouples. Specific surface area (SSA) was measured by BET gas adsorption analysis and residual moisture was measured by Karl Fischer. In the lab scale freeze-dryer, the molar flux of inert gas was determined by direct flow measurement using a flowmeter and the molar flux of water vapor was determined by manometric temperature measurement (MTM) and tunable diode laser absorption spectroscopy (TDLAS) techniques. Comparative pressure measurement (capacitance manometer vs. Pirani) was used to determine primary drying time. For both 5% mannitol and 5% sucrose, primary drying time decreases and product temperature increases as the load on the shelves decreases. No systematic variation was observed in residual moisture and vapor composition as load decreased. Further, SSA data suggests that there are no significant freezing differences under different load conditions. Independent of dryer scale, among all the effects, variation in radiation heat transfer from the chamber walls to the product seems to be the dominant effect resulting in shorter primary drying time as the load on the shelf decreases (i.e., the fraction of edge vials increases).

  19. FY 2017 Status of Sodium Freezing and Remelting Tests

    Energy Technology Data Exchange (ETDEWEB)

    Lv, Q. [Argonne National Lab. (ANL), Argonne, IL (United States); Boron, E. [Argonne National Lab. (ANL), Argonne, IL (United States); Momozaki, Y. [Argonne National Lab. (ANL), Argonne, IL (United States); Chojnowski, D. B. [Argonne National Lab. (ANL), Argonne, IL (United States); Sienicki, J. J. [Argonne National Lab. (ANL), Argonne, IL (United States); Reed, C. B. [Argonne National Lab. (ANL), Argonne, IL (United States)


    The Sodium Freezing and Remelting experiment facility at Argonne National Laboratory has been significantly modified and improved. The main improvement was replacement of the two original stainless steel test sections that had strain gages limited by their bonds to the stainless steel to maximum temperatures of 350°C with a single new test section with strain gages that can be utilized up to 980°C and a thin wall to enhance measured strains. Wetting of stainless steel by sodium within a practical time of one to a few days is expected to require temperatures of 450°C or greater. Thus, the higher temperature strain gages enable wetting in a short time of a few days. Wetting below 350°C would have required an impractically long time of at least weeks. Other improvements included upgrading of the loop configuration, incorporation of a cold finger to purify sodium, a new data acquisition system, and reinstallation of the many heaters, heater controllers, and thermocouples. After the loop had been heated to 400°C for about two hours, an initial sodium freezing test was conducted. It is thought that the sodium might have at least partially wetted the stainless steel wall under these conditions. The strain gage measurements indicate that an incremental step inward deformation of the test section thin wall occurred as the temperature decreased through the sodium freezing temperature. This behavior is consistent with sodium initially adhering to the stainless steel inner wall but breaking away from the wall as the freezing sodium contracted. Conduct of additional sodium freezing tests under well wetted conditions was prevented as a result of stoppage of all electrical work at Argonne by the Laboratory Director on July 25, 2017. A pathway to resuming electrical work is now in place at Argonne and additional sodium freezing testing will resume next fiscal year.

  20. Impact of fermentation pH and temperature on freeze-drying survival and membrane lipid composition of Lactobacillus coryniformis Si3. (United States)

    Schoug, Asa; Fischer, Janett; Heipieper, Hermann J; Schnürer, Johan; Håkansson, Sebastian


    During the industrial stabilization process, lactic acid bacteria are subjected to several stressful conditions. Tolerance to dehydration differs among lactic acid bacteria and the determining factors remain largely unknown. Lactobacillus coryniformis Si3 prevents spoilage by mold due to production of acids and specific antifungal compounds. This strain could be added as a biopreservative in feed systems, e.g. silage. We studied the survival of Lb. coryniformis Si3 after freeze-drying in a 10% skim milk and 5% sucrose formulation following different fermentation pH values and temperatures. Initially, a response surface methodology was employed to optimize final cell density and growth rate. At optimal pH and temperature (pH 5.5 and 34 degrees C), the freeze-drying survival of Lb. coryniformis Si3 was 67% (+/-6%). The influence of temperature or pH stress in late logarithmic phase was dependent upon the nature of the stress applied. Heat stress (42 degrees C) did not influence freeze-drying survival, whereas mild cold- (26 degrees C), base- (pH 6.5), and acid- (pH 4.5) stress significantly reduced survival. Freeze-drying survival rates varied fourfold, with the lowest survival following mild cold stress (26 degrees C) prior to freeze-drying and the highest survival after optimal growth or after mild heat (42 degrees C) stress. Levels of different membrane fatty acids were analyzed to determine the adaptive response in this strain. Fatty acids changed with altered fermentation conditions and the degree of membrane lipid saturation decreased when the cells were subjected to stress. This study shows the importance of selecting appropriate fermentation conditions to maximize freeze-drying viability of Lb. coryniformis as well as the effects of various unfavorable conditions during growth on freeze-drying survival.

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

    Directory of Open Access Journals (Sweden)

    Yalong Dang


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

  2. High freezing point fuels used for aviation turbine engines (United States)

    Friedman, R.


    Broadened-specification aviation fuels could be produced from a greater fraction of crude source material with improvements in fuel supply and price. These fuels, particularly those with increased final boiling temperatures, would have higher freezing temperatures than current aviation turbine fuels. For the small but significant fraction of commercial flights where low fuel temperatures make higher freezing-point fuel use unacceptable, adaptations to the fuel or fuel system may be made to accommodate this fuel. Several techniques are discussed. Fuel heating is the most promising concept. One simple design uses existing heat rejection from the fuel-lubricating oil cooler, another uses an engine-driven generator for electrical heating.

  3. Maximum freeze-out baryon density in nuclear collisions

    Energy Technology Data Exchange (ETDEWEB)

    Randrup, Jorgen; Cleymans, Jean


    Using simple parametrizations of the thermodynamicfreeze-out parameters vextracted from the data over a wide beam-energyrange, we reexpress the hadronic freeze-out line in terms of theunderlying dynamical quantities, the net baryon density rhoB and theenergy density epsilon, which are subject to local conservation laws.This analysis makes it apparent that rhoB exhibits a maximum as thecollision energy is decreased. This maximum freeze-out density hasmu=400-500 MeV, which is above the critical value, and it is reached fora fixed-target bombarding energy of 20-30 A GeV.

  4. Experimental and analytical study of thermal stresses during pipe freezing


    Keary, A.C.; Syngellakis, S.; Bowen, R.J.


    A review of experimental investigations on stress development during the blockage of a water-filled pipe by freezing was undertaken with the parallel development of an effective finite element thermal stress model. A wide spread of measured stress values was noted as well as a degree of uncertainty in the cases when the gauge output did not return to zero at the end of the freezing cycle. A methodical examination of stress- and temperature-time histories showed that it is possible to divide a...

  5. Accelerated redox reaction between chromate and phenolic pollutants during freezing

    Energy Technology Data Exchange (ETDEWEB)

    Ju, Jinjung; Kim, Jaesung [Department of Environmental Sciences and Biotechnology, Hallym University, Chuncheon, Gangwon-do 24252 (Korea, Republic of); Vetráková, Ľubica [Department of Chemistry and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno (Czech Republic); Seo, Jiwon [School of Urban and Environmental Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919 (Korea, Republic of); Heger, Dominik [Department of Chemistry and Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Kamenice 5, 625 00 Brno (Czech Republic); Lee, Changha [School of Urban and Environmental Engineering, Ulsan National Institute of Science and Technology (UNIST), Ulsan 44919 (Korea, Republic of); Yoon, Ho-Il [Korea Polar Research Institute (KOPRI), Incheon 21990 (Korea, Republic of); Kim, Kitae, E-mail: [Korea Polar Research Institute (KOPRI), Incheon 21990 (Korea, Republic of); Kim, Jungwon, E-mail: [Department of Environmental Sciences and Biotechnology, Hallym University, Chuncheon, Gangwon-do 24252 (Korea, Republic of)


    Highlights: • Redox conversion of 4-CP/Cr(VI) was significantly accelerated during freezing. • Accelerated redox conversion in ice is ascribed to the freeze concentration effect. • 4-CP, Cr(VI), and protons are concentrated in the liquid brine by freezing. • Redox conversions of various phenolic pollutants/Cr(VI) were significant in ice. • Freezing-accelerated redox conversion was observed in real polluted water. - Abstract: The redox reaction between 4-chlorophenol (4-CP) and chromate (Cr(VI)) (i.e., the simultaneous oxidation of 4-CP by Cr(VI) and reduction of Cr(VI) by 4-CP) in ice (i.e., at −20 °C) was compared with the corresponding reaction in water (i.e., at 25 °C). The redox conversion of 4-CP/Cr(VI), which was negligible in water, was significantly accelerated in ice. This accelerated redox conversion of 4-CP/Cr(VI) in ice is ascribed to the freeze concentration effect occurring during freezing, which excludes solutes (i.e., 4-CP and Cr(VI)) and protons from the ice crystals and subsequently concentrates them in the liquid brine. The concentrations of Cr(VI) and protons in the liquid brine were confirmed by measuring the optical image and the UV–vis absorption spectra of cresol red (CR) as a pH indicator of frozen solution. The redox conversion of 4-CP/Cr(VI) was observed in water when the concentrations of 4-CP/protons or Cr(VI)/protons increased by 100/1000-fold. These results corroborate the freeze concentration effect as the reason for the accelerated redox conversion of 4-CP/Cr(VI) in ice. The redox conversion of various phenolic pollutants/Cr(VI) and 4-CP/Cr(VI) in real wastewater was successfully achieved in ice, which verifies the environmental relevance and importance of freezing-accelerated redox conversion of phenolic pollutants/Cr(VI) in cold regions.

  6. Thermal Conductivity of Human Bone in Cryoprobe Freezing as Related to Density. (United States)

    Walker, Kyle E; Baldini, Todd; Lindeque, Bennie G


    Cryoprobes create localized cell destruction through freezing. Bone is resistant to temperature flow but is susceptible to freezing necrosis at warmer temperatures than tumor cells. Few studies have determined the thermal conductivity of human bone. No studies have examined conductivity as related to density. The study goal was to examine thermal conductivity in human bone while comparing differences between cancellous and cortical bone. An additional goal was to establish a relationship between bone density and thermal conductivity. Six knee joints from 5 cadavers were obtained. The epiphyseal region was sliced in half coronally prior to inserting an argon-circulating cryoprobe directed away from the joint line. Thermistor thermometers were placed perpendicularly at measured increments, and the freezing cycle was recorded until steady-state conditions were achieved. For 2 cortical samples, the probe was placed intramedullary in metaphyseal samples and measurements were performed radially from the central axis of each sample. Conductivity was calculated using Fournier's Law and then plotted against measured density of each sample. Across samples, density of cancellous bone ranged from 0.86 to 1.38 g/mL and average thermal conductivity ranged between 0.404 and 0.55 W/mK. Comparatively, cortical bone had a density of 1.70 to 1.86 g/mL and thermal conductivity of 0.0742 to 0.109 W/mK. A strong 2-degree polynomial correlation was seen (R 2 =0.8226, P<.001). Bone is highly resistant to temperature flow. This resistance varies and inversely correlates strongly with density. This information is clinically relevant to maximize tumor ablation while minimizing morbidity through unnecessary bone loss and damage to surrounding structures. [Orthopedics. 2017; 40(2):90-94.]. Copyright 2016, SLACK Incorporated.

  7. Cyclopropanation of unsaturated fatty acids and membrane rigidification improve the freeze-drying resistance of Lactococcus lactis subsp. lactis TOMSC161. (United States)

    Velly, H; Bouix, M; Passot, S; Penicaud, C; Beinsteiner, H; Ghorbal, S; Lieben, P; Fonseca, F


    This work aimed at characterizing the biochemical and biophysical properties of the membrane of Lactococcus lactis TOMSC161 cells during fermentation at different temperatures, in relation to their freeze-drying and storage resistance. Cells were cultivated at two different temperatures (22 and 30 °C) and were harvested at different growth phases (from the middle exponential phase to the late stationary phase). Bacterial membranes were characterized by determining the fatty acid composition, the lipid phase transition, and the membrane fluidity. Cultivability and acidification activity losses of L. lactis were quantified after freezing, drying, and 3 months of storage. The direct measurement of membrane fluidity by fluorescence anisotropy was linked to lipid composition, and it was established that the cyclopropanation of unsaturated fatty acids with concomitant membrane rigidification during growth led to an increase in the freeze-drying and storage resistance of L. lactis. As expected, cultivating cells at a lower fermentation temperature than the optimum growth temperature induced a homeoviscous adaptation that was demonstrated by a lowered lipid phase transition temperature but that was not related to any improvement in freeze-drying resistance. L. lactis TOMSC161 was therefore able to develop a combined biochemical and biophysical response at the membrane level during fermentation. The ratio of cyclic fatty acids to unsaturated fatty acids (CFA/UFA) appeared to be the most relevant parameter associated with membrane rigidification and cell resistance to freeze-drying and storage. This study increased our knowledge about the physiological mechanisms that explain the resistance of lactic acid bacteria (LAB) to freeze-drying and storage stresses and demonstrated the relevance of complementary methods of membrane characterization.

  8. Freeze-fracture of infected plant leaves in ethanol for scanning electron microscopic study of fungal pathogens. (United States)

    Moore, Jayma A; Payne, Scott A


    Fungi often are found within plant tissues where they cannot be visualized with the scanning electron microscope (SEM). We present a simple way to reveal cell interiors while avoiding many common causes of artifact. Freeze-fracture of leaf tissue using liquid nitrogen during the 100% ethanol step of the dehydration process just before critical point drying is useful in exposing intracellular fungi to the SEM.

  9. The Freezing Point Depression Law in Physical Chemistry. (United States)

    Franzen, Hugo F.


    Suggests a change in physical chemistry courses to use a slightly more complicated but significantly more useful generalization of the simple freezing point depression law. Lists reasons for the change and presents the treatment of solid-liquid equilibria where solid-solution is allowed. Provides a mathematical treatment. (MVL)

  10. Sperm vacuoles are not modified by freezing--thawing procedures. (United States)

    Gatimel, Nicolas; Leandri, Roger; Parinaud, Jean


    Since the development of the motile sperm organellar morphology examination (MSOME) in 2001 for observing the cephalic vacuoles at high magnification, no study as yet assessed the effect of cryopreservation on these vacuoles, although sperm freezing-thawing procedures are known to affect sperm quality. Examination of the vacuoles before and after freezing-thawing would indicate whether the same normality criteria can be applied for frozen as for fresh spermatozoa when performing intracytoplasmic morphologically selected sperm injection. In 27 sperm samples from fertile men, analysis of conventional sperm parameters (motility, vitality, percentage of normal forms) and a morphological analysis at high magnification (×6000) using image analysis software was performed before freezing and after thawing. Whereas there were expected decreases in motility (Pvacuole area, total vacuole area, vacuole area in the anterior, median and basal parts of the head, percentage of spermatozoa with a vacuole area ≤6.5% and percentage of spermatozoa with a vacuole area >13%). Freezing-thawing procedures have no effect on human sperm vacuoles. Copyright © 2012 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  11. [Super sweet corn hybrids adaptability for industrial processing. I freezing]. (United States)

    Alfonzo, Braunnier; Camacho, Candelario; Ortiz de Bertorelli, Ligia; De Venanzi, Frank


    With the purpose of evaluating adaptability to the freezing process of super sweet corn sh2 hybrids Krispy King, Victor and 324, 100 cobs of each type were frozen at -18 degrees C. After 120 days of storage, their chemical, microbiological and sensorial characteristics were compared with a sweet corn su. Industrial quality of the process of freezing and length and number of rows in cobs were also determined. Results revealed yields above 60% in frozen corns. Length and number of rows in cobs were acceptable. Most of the chemical characteristics of super sweet hybrids were not different from the sweet corn assayed at the 5% significance level. Moisture content and soluble solids of hybrid Victor, as well as total sugars of hybrid 324 were statistically different. All sh2 corns had higher pH values. During freezing, soluble solids concentration, sugars and acids decreased whereas pH increased. Frozen cobs exhibited acceptable microbiological rank, with low activities of mesophiles and total coliforms, absence of psychrophiles and fecal coliforms, and an appreciable amount of molds. In conclusion, sh2 hybrids adapted with no problems to the freezing process, they had lower contents of soluble solids and higher contents of total sugars, which almost doubled the amount of su corn; flavor, texture, sweetness and appearance of kernels were also better. Hybrid Victor was preferred by the evaluating panel and had an outstanding performance due to its yield and sensorial characteristics.

  12. Freezing tolerance of wheat cultivars at the early growing season ...

    African Journals Online (AJOL)

    Cold stress is a worldwide abiotic stress in temperate regions that affects plant development and yield of winter wheat (Triticum aestivum L.) cultivars and other winter crops. This study was conducted to evaluate the effect of freezing stress at the early growing season on survival and also the relationship between resistances ...

  13. Dynamic Melting of Freezing Droplets on Ultraslippery Superhydrophobic Surfaces. (United States)

    Chu, Fuqiang; Wu, Xiaomin; Wang, Lingli


    Condensed droplet freezing and freezing droplet melting phenomena on the prepared ultraslippery superhydrophobic surface were observed and discussed in this study. Although the freezing delay performance of the surface is common, the melting of the freezing droplets on the surface is quite interesting. Three self-propelled movements of the melting droplets (ice- water mixture) were found including the droplet rotating, the droplet jumping, and the droplet sliding. The melting droplet rotating, which means that the melting droplet rotates spontaneously on the superhydrophobic surface like a spinning top, is first reported in this study and may have some potential applications in various engineering fields. The melting droplet jumping and sliding are similar to those occurring during condensation but have larger size scale and motion scale, as the melting droplets have extra-large specific surface area with much more surface energy available. These self-propelled movements make all the melting droplets on the superhydrophobic surface dynamic, easily removed, which may be promising for the anti-icing/frosting applications.

  14. Biomass, thermal inertia, and radiative freeze occurrence in leafless forests (United States)

    Brian E. Potter; John C. Zasada


    Using field measurements of air temperature, wind, and relative humidity from a clear-cut site and two wooded sites in northern Wisconsin, we used a radiative transfer model to simulate temperatures on seven calm, clear nights similar to those on which freezes typically occur. Each night was simulated twice for the wooded sites. One simulation ignored the presence of...

  15. Nanoclay Performance on Resistance of Clay under Freezing Cycles

    African Journals Online (AJOL)

    This study aims to investigate effect of nanoclay on resistance of the exposed clay soils in freezing in vitro conditions. The clay sample was selected from the considered land. Then they were mixed on water in 1.5, 3, 4.5 and 6 percents as well as without nanoclay. There were prepared cylinders with 3.81 cm diameter and ...

  16. Physiological and environmental causes of freezing injury in red spruce (United States)

    Paul G. Schaberg; Donald H. DeHayes


    For many, concerns about the implications of "environmental change" conjure up scenarios of forest responses to global warming, enrichment of greenhouse gases, such as carbon dioxide and methane, and the northward migration of maladapted forests. From that perspective, the primary focus of this chapter, that is, causes of freezing injury to red spruce (

  17. Freeze-Spray Processing of Layered Ceramic Composites (Preprint) (United States)


    used to spray thin layers of slurries on a cryogenically cooled metallic plate . The slurries freeze almost instantly, forming solid structures. The...34Optimization of FGM TBC and Their Thermal Cycling Stability," Mater. Sci Forum, 492,9-14 (2005). K . An, K. Ravichandran, R. Dutton, and S. Semiatin

  18. Standardinng initial cooling of sheep semen before freezing

    African Journals Online (AJOL)

    sheep semen before freezing. C. Kemp. Animal and Dairy Science Research Institute, Private Bag X2,. Irene, 1675 Republic of South Africa. Received 6 June 1985. A practical and repeatable method for the cooling, during the processing phase, of sheep semen, with the aim of minimizing inter-experiment variation is ...

  19. Antifreeze proteins enable plants to survive in freezing conditions

    Indian Academy of Sciences (India)

    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 ...

  20. Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification. (United States)

    Pradiee, J; Esteso, M C; Castaño, C; Toledano-Díaz, A; Lopez-Sebastián, A; Guerra, R; Santiago-Moreno, J


    This work examines the effectiveness of a TCG (Tris, citric acid, glucose, 6% egg yolk and 5% glycerol) and a TEST (TES, Tris, glucose, 6% egg yolk and 5% glycerol) sperm extender in the freezing of mouflon spermatozoa at slow cooling rates, using different pre-freezing equilibration times (2-3 hr). It also examines the tolerance of mouflon spermatozoa to different concentrations of cryoprotectants (5, 10, 20% glycerol; 5%, 10%, 20% dimethyl sulfoxide; 6% polyvinylpyrrolidone) and/or sucrose (100, 300, 500 mm). The highest quality (p spermatozoa were obtained when using the TEST extender and an equilibration time of 3 hr. Sperm motility and membrane integrity were strongly reduced when using rapid freezing rates (60-85°C min -1 ), independent of the concentration of cryoprotectants. The lowest sucrose concentration (100 mm) provided the highest (p spermatozoa and live spermatozoa with an intact acrosome. Vitrified-warmed sperm variables were at their best when the spermatozoa was diluted in TCG-6% egg yolk + 100 mm sucrose and warmed at 60°C. Slow warming at 37°C strongly reduced (p < .05) sperm motility and viability. However, sperm vitrification returned lower fertility, sperm motility and sperm viability values than conventional sperm freezing. © 2016 Blackwell Verlag GmbH.

  1. 7 CFR 58.638 - Freezing the mix. (United States)


    ... 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 THE...

  2. Structural damages in adsorbed vaccines affected by freezing. (United States)

    Kurzątkowski, Wiesław; Kartoğlu, Ümit; Staniszewska, Monika; Górska, Paulina; Krause, Aleksandra; Wysocki, Mirosław Jan


    This study was planned to evaluate structural damages in adsorbed vaccines affected by freezing using scanning electron microscopy and X-ray analysis of the elements. Randomly selected 42 vials of eight different types of WHO pre-qualified adsorbed freeze-sensitive vaccines from 10 manufacturers were included in the study. Vaccines were kept at 5 °C. Selected numbers of vials from each type were then exposed to -25 °C for 24 h periods. All samples were evaluated for their structure using scanning electron microscopy, X-ray analysis of the elements and precipitation time. Scanning electron microscopy of vaccines affected by freezing showed either smooth or rough surfaced conglomerates associated with phosphate content of the precipitate. These vaccines precipitated 2-15 times faster compared to non-frozen samples. Non-frozen samples showed uniform flocculent structure either dense or dispersed. X-ray analysis of precipitates in frozen samples confirmed that the precipitate is mainly aluminium clutters. Scanning electron microscopy confirmed that the lattice structure of bonds between adsorbent and the antigen is broken and aluminium forms conglomerates that grow in size and weight. The precipitation time of vaccines affected by freezing is 4.5 times faster on average compared to non-frozen samples. These facts form the basis of the "shake test". Copyright © 2012 World Health Organization. Published by Elsevier Ltd.. All rights reserved.

  3. Freeze/thaw phenomena in concrete at low temperatures

    DEFF Research Database (Denmark)

    Johannesson, Björn


    , 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...

  4. A heat equation for freezing processes with phase change

    DEFF Research Database (Denmark)

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


    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...

  5. Lattice QCD results on chemical freeze-out

    Directory of Open Access Journals (Sweden)

    Ratti Claudia


    Full Text Available The latest lattice QCD results on fluctuations of conserved charges at finite temperature and density are reviewed, and connected to the experimental moments of their net-multiplicity distributions, in order to describe the chemical freeze-out from first principles.

  6. Freezing of ionic melts into a superionic phase

    International Nuclear Information System (INIS)

    March, N.H.; Tosi, M.P.


    With the specific case of BaCl 2 in mind, for which neutron diffraction data is available, we have studied the theory of freezing of an ionic melt into a superionic phase. The idea underlying the theory presented here is that the rather highly ordered cation assembly in molten BaCl 2 revealed by the neutron experiment, leads to cation sub-lattice freezing with the usual volume change associated with the first-order phase transition. This has two effects on the anions: (i) it imposes a volume change and (ii) it modulates the number density of the anions from its homogeneous form above the freezing temperature. Explicitly, this modulation is calculated in linear response approximation in terms of (i) the Fourier components of the density in the cation sub-lattice and (ii) the liquid direct correlation functions csub(Ba-Cl) and csub(Cl-Cl). This treatment shows that the freezing transition is a balance between the free energy increase associated with the ordering of the singlet densities of anions and cations and the decrease due to the volume contraction. Taking the volume change from experiment, the modulation of the singlet densities at the first reciprocal lattice vector can be estimated. (author)

  7. Seeing fearful body language rapidly freezes the observer's motor cortex

    NARCIS (Netherlands)

    Borgomaneri, Sara; Vitale, Francesca; Gazzola, V.; Avenanti, Alessio

    Fearful body language is a salient signal alerting the observer to the presence of a potential threat in the surrounding environment. Although detecting potential threats may trigger an immediate reduction of motor output in animals (i.e., freezing behavior), it is unclear at what point in time

  8. Seeing fearful body language rapidly freezes the observer's motor cortex

    NARCIS (Netherlands)

    Borgomaneri, S.; Vitale, F.; Gazzola, V.; Avenanti, A.


    Fearful body language is a salient signal alerting the observer to the presence of a potential threat in the surrounding environment. Although detecting potential threats may trigger an immediate reduction of motor output in animals (i.e., freezing behavior), it is unclear at what point in time

  9. Freezing Point Depressions of Phase Change CO2 Solvents

    DEFF Research Database (Denmark)

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


    . The experimental values indicate that the DEEA–water interaction is almost similar to that of MEA–water interaction. MAPA has shown a stronger nonideal behavior compared to DEEA. A correlation for the freezing points as a function of solution composition was formulated for the unloaded binary and ternary systems......., 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...

  10. Phase separation during freezing upon warming of aqueous solutions (United States)

    Bogdan, A.; Loerting, T.


    Using differential scanning calorimetry, we show that the addition of solute(s) to emulsified water lowers the freezing temperature to nanodrops adsorbed on fumed silica resemble bulk water more than water confined in nanoscaled confinement and also more than nanoscaled water domains in aqueous solution.

  11. Anatomy of a Tuition Freeze: The Case of Ontario (United States)

    Rexe, Deanna


    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…

  12. CFD modelling of condensers for freeze-drying processes

    Indian Academy of Sciences (India)

    ... 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.

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

    NARCIS (Netherlands)

    Enriquez Paz y Puente, O.R.


    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

  14. Large tundra methane burst during onset of freezing

    DEFF Research Database (Denmark)

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


    of global atmospheric methane concentrations indicate that the observed early winter emission burst improves the agreement between the simulated seasonal cycle and atmospheric data from latitudes north of 60N. Our findings suggest that permafrost-associated freeze-in bursts of methane missions from tundra...

  15. Freeze desalination of seawater using LNG cold energy. (United States)

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


    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. CFD modelling of condensers for freeze-drying processes

    Indian Academy of Sciences (India)

    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 consid- ered and the influence exerted by the inert gas as well as other parameters has been.

  17. Freeze desalination of seawater using LNG cold energy

    KAUST Repository

    Chang, Jian


    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. Resistance to freezing and thawing of mortar specimens made from ...

    Indian Academy of Sciences (India)



    Jun 3, 2000 ... Abstract. Resistance to freezing and thawing of mortar specimens made from sulphoaluminate–belite cement. (M–SAB) is compared with that of mortars made from portland cement (M–PC). The results suggest that lar- ger median radius of the pores and total porosity of M–SAB compared to those of M–PC ...

  19. Preparation of superconducting powders by freeze-drying

    International Nuclear Information System (INIS)

    Johnson, S.M.; Gusman, M.I.; Rowcliffe, D.J.; Geballe, T.H.; Sun, J.Z.


    A method of preparing superconducting powders by freeze-drying is described. Powders produced by this method are homogeneous, have high purities, and are very reactive. Materials sintered from these powders have densities up to 89% of the theoretical density, and exhibit very sharp resistivity drops and large Meissner effects. The microstructure of the materials is very sensitive to the sintering temperature

  20. Freeze-drying of filamentous fungi and yeasts

    NARCIS (Netherlands)

    Tan, C.S.


    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

  1. Freezing of gait: moving forward on a mysterious clinical phenomenon

    NARCIS (Netherlands)

    Nutt, J.G.; Bloem, B.R.; Giladi, N.; Hallett, M.; Horak, F.B.; Nieuwboer, A.


    Freezing of gait (FoG) is a unique and disabling clinical phenomenon characterised by brief episodes of inability to step or by extremely short steps that typically occur on initiating gait or on turning while walking. Patients with FoG, which is a feature of parkinsonian syndromes, show variability

  2. Monte Carlo simulation of the OCP freezing transition

    International Nuclear Information System (INIS)

    DeWitt, H.E.; Slattery, W.L.; Yang, Juxing


    The One Component Plasma (OCP) in three dimensions is a system of classical point charges moving in a fixed uniform neutralizing background. In nature the OCP is a rough approximation of the conditions in a white dwarf star in which one has fully ionized nuclei such as carbon, oxygen, and smaller amounts of heavier elements up to iron all moving in a nearly uniform background provided by relativistically degenerate electrons. The OCP is also a mathematical limiting model for a non-neutral plasma of ions in a Penning trap and cooled to strongly coupled conditions. Similarly, a collection of charge colloidal suspensions in water can exhibit the Coulomb freezing behavior of the OCP. A single dimensionless parameter, Γ is sufficient to describe the system. For very weak coupling, Γ much-lt 1, the thermodynamic properties of the OCP are given rigorously by the Debye-Huckel theory. This paper reports on Monte Carlo simulation of the freezing of the OCP from a random start for particle numbers ranging from 500 to 2000. In one case the authors obtained a perfect bcc lattice, but in most cases the final state would be an imperfect crystal or two different microcrystals, fcc and bcc, growing into each other. With a cluster analysis program the authors looked at the formation of nucleating clusters, and followed the actual freezing process. Roughly 80 particles are needed in a cluster before it starts to grow rapidly and freeze

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

    International Nuclear Information System (INIS)

    Burkhardt, Anja; Warmer, Martin; Panneerselvam, Saravanan; Wagner, Armin; Zouni, Athina; Glöckner, Carina; Reimer, Rudolph; Hohenberg, Heinrich; Meents, Alke


    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

  4. Xylem diameter changes during osmotic stress, desiccation and freezing in Pinus sylvestris and Populus tremula. (United States)

    Lintunen, Anna; Lindfors, Lauri; Nikinmaa, Eero; Hölttä, Teemu


    Trees experience low apoplastic water potential frequently in most environments. Low apoplastic water potential increases the risk of embolism formation in xylem conduits and creates dehydration stress for the living cells. We studied the magnitude and rate of xylem diameter change in response to decreasing apoplastic water potential and the role of living parenchyma cells in it to better understand xylem diameter changes in different environmental conditions. We compared responses of control and heat-injured xylem of Pinus sylvestris (L.) and Populus tremula (L.) branches to decreasing apoplastic water potential created by osmotic stress, desiccation and freezing. It was shown that xylem in control branches shrank more in response to decreasing apoplastic water potential in comparison with the samples that were preheated to damage living xylem parenchyma. By manipulating the osmotic pressure of the xylem sap, we observed xylem shrinkage due to decreasing apoplastic water potential even in the absence of water tension within the conduits. These results indicate that decreasing apoplastic water potential led to withdrawal of intracellular water from the xylem parenchyma, causing tissue shrinkage. The amount of xylem shrinkage per decrease in apoplastic water potential was higher during osmotic stress or desiccation compared with freezing. During desiccation, xylem diameter shrinkage involved both dehydration-related shrinkage of xylem parenchyma and water tension-induced shrinkage of conduits, whereas dehydration-related shrinkage of xylem parenchyma was accompanied by swelling of apoplastic ice during freezing. It was also shown that the exchange of water between symplast and apoplast within xylem is clearly faster than previously reported between the phloem and the xylem. Time constant of xylem shrinkage was 40 and 2 times higher during osmotic stress than during freezing stress in P. sylvestris and P. tremula, respectively. Finally, it was concluded that the

  5. Inactivation of Kudoa septempunctata in olive flounder meat by liquid freezing. (United States)

    Ohnishi, Takahiro; Akuzawa, Sayuri; Furusawa, Hiroko; Yoshinari, Tomoya; Kamata, Yoichi; Sugita-Konishi, Yoshiko


    Kudoa septempunctata in olive flounder meat was inactivated using 3 distinct freezing methods:liquid freezing for 5 min, air blast freezing at -30℃ for 5 h, and -80℃ for 1 h. The fracture curve of olive flounder meat subjected to liquid freezing resembled that of meat stored at 4℃, indicating that the structure of olive flounder muscle was well preserved. In contrast, air blast freezing induced the disappearance of the fracture point in the fracture curve, indicating that there was deterioration in the meat quality. Liquid freezing preserved the transparency of olive flounder meat to the same degree as that of meat stored at 4°C. However, air blast freezing induced meat cloudiness. These results indicate that liquid freezing can be used for K. septempunctata inactivation without affecting the meat quality.

  6. Experimental research of "microcable in a microconduct" system stability to effect of freezing water (United States)

    Andreev, Vladimir A.; Burdin, Vladimir A.; Nikulina, Tatiana G.; Alekhin, Ivan N.; Gavryushin, Sergey A.; Nikulin, Aleksey G.; Praporshchikov, Denis E.


    Results of experimental researches of "optical microcable in a microduct" system stability to effect of freezing water are presented. It is shown this system is steadier to water freezing in comparison to lighten optical cable in protective polymer tube.

  7. Cryomicroscopy: a method for visualisation of freezing processes; Kryomikroskopie: Eine Methode zur Visualisierung von Gefrierprozessen

    Energy Technology Data Exchange (ETDEWEB)

    Spindler, Ralf; Glasmacher, Birgit [Leibniz Univ. Hannover (Germany). Inst. fuer Mehrphasenprozesse


    Cryomicroscopy allows the investigation of a variety of freezing processes under controlled thermal conditions. For cryopreservation the thermal effects in biological materials can be observed. Fundamental phenomena can be investigated and freezing protocols can be optimized. (orig.)

  8. Effects of freezing and cold acclimation on the plasma membrane of isolated protoplasts. [Annual report], May 16, 1993--January 29, 1994

    Energy Technology Data Exchange (ETDEWEB)

    Steponkus, P.L.


    Our aim is to provide a mechanistic understanding of the cellular and molecular aspects of freezing injury and cold acclimation from a perspective of the structural and functional integrity of the plasma membrane-the primary site of freezing injury in winter cereals. We established that destabilization of the plasma membrane of winter rye, the most freezing-tolerant winter cereal, can result from several different lesions: expansion induced lysis, lamellar-to-hexagonal II phase transitions, and the fracture-jump lesion. The occurrence and incidence of these various lesions, depends on the freeze/thaw protocol and the stage of cold acclimation. In non-acclimated leaves and protoplasts, expansion-induced lysis is the predominant lesion at temperatures between {minus}2 and {minus}5{degree}C, whereas freeze-induced formation of the H{sub II} phase is the predominant lesion at temperatures below {minus}10{degree}C. We investigated whether the difference in freezing tolerance and the threshold temperatures at which the lesions occur in rye and oat are a consequence of differences in the lipid composition of the plasma membrane. There are substantial differences between rye and oat cell membranes both before and after cold acclimation. The plasma membrane of oat contains greater proportions of acylated sterylglucosides and cerebrosides than that of rye, and there is little change in these two lipid classes during cold acclimation. The lyotropic phase behavior of lipid mixtures that resemble the plasma membrane of rye and oat was studied. The differences in lipid composition of rye and oat are of mechanistic significance because of their influence on the hydration characteristics of the plasma membrane, the propensity for dehydration-induced lipid-lipid demixing, and the intrinsic curvature of the lipid monolayers. These studies suggest that strategies for improving the freezing tolerance of winter cereals should include approaches to modify membrane lipid composition.

  9. Freezing of liquid alkali metals as screened ionic plasmas

    International Nuclear Information System (INIS)

    Badirkhan, Z.; Rovere, M.; Tosi, M.P.


    The relationship between Wigner crystallization of the classical ionic plasma and the liquid-solid transition of alkali metals is examined within the density wave theory of freezing. Freezing of the classical plasma on a rigid neutralizing background into the bcc structure is first re-evaluated, in view of recent progress in the determination of its thermodynamic functions by simulation and of the known difficulties of the theory relating to the order parameter at the (200) star of reciprocal lattice vectors. Freezing into the fcc structure is also considered in this context and found to be unfavoured. On allowing for long-wavelength deformability of the background, the ensuing appearance of a volume change on freezing into the bcc structure is accompanied by reduced stability of the fluid phase and by an increase in the entropy of melting. Freezing of alkali metals into the bcc structure is next evaluated, taking their ionic pair structure as that of an ionic plasma reference fluid screened by conduction electrons and asking that the correct ionic coupling strength at liquid-solid coexistence should be approximately reproduced. The ensuring values of the volume and entropy changes across the phase transition, as estimated from the theory by two alternative routes, are in reasonable agreement with experiment. The order parameters of the phase transition, excepting the (200) one, conform rather closely to a Gaussian behaviour and yield a Lindemann ratio in reasonable agreement with the empirical value for melting of bcc crystals. It is suggested that ionic ordering at the (200) star in the metal may be (i) assisted by medium range ordering in the conduction electrons, as indicated by differences in X-ray and neutron diffraction intensities from the liquid, and/or (ii) quite small in the hot bcc solid. Such a possible premelting behaviour of bcc metals should be worth testing experimentally by diffraction. (author). 48 refs, 1 fig., 1 tab

  10. Freezing shortens the lifetime of DNA molecules under tension. (United States)

    Chung, Wei-Ju; Cui, Yujia; Chen, Chi-Shuo; Wei, Wesley H; Chang, Rong-Shing; Shu, Wun-Yi; Hsu, Ian C


    DNA samples are commonly frozen for storage. However, freezing can compromise the integrity of DNA molecules. Considering the wide applications of DNA molecules in nanotechnology, changes to DNA integrity at the molecular level may cause undesirable outcomes. However, the effects of freezing on DNA integrity have not been fully explored. To investigate the impact of freezing on DNA integrity, samples of frozen and non-frozen bacteriophage lambda DNA were studied using optical tweezers. Tension (5-35 pN) was applied to DNA molecules to mimic mechanical interactions between DNA and other biomolecules. The integrity of the DNA molecules was evaluated by measuring the time taken for single DNA molecules to break under tension. Mean lifetimes were determined by maximum likelihood estimates and variances were obtained through bootstrapping simulations. Under 5 pN of force, the mean lifetime of frozen samples is 44.3 min with 95% confidence interval (CI) between 36.7 min and 53.6 min while the mean lifetime of non-frozen samples is 133.2 min (95% CI: 97.8-190.1 min). Under 15 pN of force, the mean lifetimes are 10.8 min (95% CI: 7.6-12.6 min) and 78.5 min (95% CI: 58.1-108.9 min). The lifetimes of frozen DNA molecules are significantly reduced, implying that freezing compromises DNA integrity. Moreover, we found that the reduced DNA structural integrity cannot be restored using regular ligation process. These results indicate that freezing can alter the structural integrity of the DNA molecules.

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

    Directory of Open Access Journals (Sweden)

    Slavica Golc Teger


    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.





    Ice rnorphology (size and shape) influence decisively in sensory appreciation, texture and quality of rnany frozen foods. Ice rnorphology is also irnportant in sorne technological processes such as freeze drying and freeze concentration, which influences the efficiency ofthese processes. The overall objective of this thesis was to increase our knowledge about the control on rnorphology of the ice phase in freezing food and related processes such as freeze concentration. Freezin...

  13. Evaluation of Freezing Tolerance of Three Ajowan (Trachyspermum ammi (Linn. Sprague Ecotypes in Controlled Conditions

    Directory of Open Access Journals (Sweden)

    Z Boroumand Rezazadeh


    Full Text Available Ajowan is one of the endemic plants in Khorasan province, and there is a little information on its tolerance to cold stress. In order to study freezing tolerance of ajowan, an experiment was conducted in faculty of agriculture, Ferdowsi University of Mashhad, based on factorial-completely randomized design with three replications and three ecotypes of ajowan (Neishabour, Birjand and Torbat-e-Heidarieh were imposed on eight freezing temperatures (0 (control, -1.5, -3, -4.5, -6,-7.5, -9 and -10.5 °C. Plants were grown in natural environment till 4-5 leaf stage, then for freezing treatments transferred to thermo-gradient freezer. The cell membrane stability was evaluated by electrolyte leakage index (EL and temperature for killing 50% of samples according to the electrolyte leakage (LT50el was determined. Furthermore, survival percentage, leaf number and dry weight, temperature for killing 50% of samples according to survival (LT50su and reduced dry matter temperature 50 (RDMT50 were determined after three weeks recovery in the glasshouse. Response of ajowan ecotypes for electrolyte leakage was different and birjand ecotype had the lowest %EL, whereas the slope of %EL in mentioned ecotype was lower than two other ecotypes. However there were no significant differences among ajowan ecotypes on LT50su. Decreasing temperature to -7.5 °C reduced survival percentage of Neishabour and Torbat-e-Heidarieh ecotypes to lower than 20 percent, whiles in this temperature Birjand’s survival percentage was about 60 percent. It seems that Birjand ecotype with the lowest electrolyte leakage, the highest survival and dry matter and the lowest LT50su was more tolerant than two other ecotypes.

  14. 14 CFR 25.1455 - Draining of fluids subject to freezing. (United States)


    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Draining of fluids subject to freezing. 25.1455 Section 25.1455 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF... § 25.1455 Draining of fluids subject to freezing. If fluids subject to freezing may be drained...

  15. Effect of non-volatile solute on the freezing point of malonic acid ...

    African Journals Online (AJOL)

    The mixture of malonic acid and potassium chloride were obtained in various proportions. The freezing point of pure malonic acid was obtained in the range 134 and 135oC which is in agreement with the literature value of 135 OC. the freezing point of all the mixtures were below the literature value of 135 OC. the freezing ...

  16. 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


    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

  17. Cryogenic freezing of fresh date fruits for quality preservation during frozen storage

    Directory of Open Access Journals (Sweden)

    Abdullah Alhamdan


    Full Text Available Fresh date fruits, especially Barhi cultivar, are favored and widely consumed at the Khalal maturity stage (first color edible stage. These fruits are seasonal and perishable and there is a need for extending their shelf life. This study evaluates two different freezing methods, namely cryogenic freezing using liquid nitrogen and conventional deep freezing on preserving the quality and stability of date fruits (cv. Barhi at Khalal maturity stage. Fresh date fruits (cv. Barhi at Khalal stage were frozen utilizing the two methods. The produced frozen dates were stored under frozen storage conditions for nine months (at −20 °C and −40 °C for the conventional and cryogenic freezing, respectively. Color values, textural properties (hardness, elasticity, chewiness and resilience, and nutrition attributes (enzymes and sugars for fresh dates before freezing and for the frozen dates were measured every three months during the frozen storage. Color values of the frozen dates were affected by the freezing method and the frozen storage period. There are substantial differences in the quality of the frozen fruits in favor of cryogenic freezing compared to the conventional slow freezing. The results revealed a large disparity between the times of freezing of the two methods. The freezing time accounted to 10 min in the cryogenic freezing method, whereas it was 1800 min for the conventional slow freezing system.

  18. Gelatin-Filtered Consomme: A Practical Demonstration of the Freezing and Thawing Processes (United States)

    Lahne, Jacob B.; Schmidt, Shelly J.


    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…

  19. Sintering of a freeze-dried 10 mol% Y2O3-stabilized zirconia

    International Nuclear Information System (INIS)

    Rakotoson, A.; Paulus, M.


    After presenting the results of freeze drying a sulfate solution, the authors describe a preparation process in which the freeze-drying technique by addition of a suspension of stabilized zirconia in the liquid solution before freeze-drying. This process breaks the polymeric chains, increases the green density of the compact, and decreases the sintering temperature. The mechanisms involved are discussed

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

    Tellinghuisen, Joel


    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…

  1. The glycocalyx and stereociliary interconnections of the vestibular sensory epithelia of the guinea pig - A freeze-fracture, low-voltage cryo-SEM, SEM and TEM study

    NARCIS (Netherlands)

    Valk, WL; Oei, MLYM; Segenhout, JM; Dijk, F; Stokroos, [No Value; Albers, FWJ


    In this study freeze fracture, low-voltage cryo-SEM, SEM and TEM were used to characterise the glycocalyx and stereociliary interconnections in the hair cell bundle of the vestibular sensory epithelia of the guinea pig. The glycocalyx resembles a shell-like structure separately surrounding each

  2. Screening of freeze-dried protective agents for the formulation of biocontrol strains, Bacillus cereus AR156, Burkholderia vietnamiensis B418 and Pantoea agglomerans 2Re40. (United States)

    Zhan, Y; Xu, Q; Yang, M-M; Yang, H-T; Liu, H-X; Wang, Y-P; Guo, J-H


    The effects of different freeze-drying protective agents on the viabilities of biocontrol strains Bacillus cereus AR156, Burkholderia vietnamiensis B418 and Pantoea agglomerans 2Re40 were investigated. Several concentrations of protective and rehydration media were tested to improve the survival of biocontrol agents after freeze-drying. The subsequent survival rates during storage and rehydration media of freeze-dried biocontrol strains were also examined. The results indicated that cellobiose (5%) and d-galactose (5%) gave maximum viability of strains Bu. vietnamiensis B418 and P. agglomerans 2Re40 (98 and 54·3% respectively) while the perfect one (100%) of strain B. cereus AR156 was obtained with sucrose (5%) during freeze-drying, and the highest survival of the three strains was reached when they were rehydrated with 10% nonfat skim milk. In the following storage, the survival rates showed that B. cereus AR156 could still reach 50% after 12 months. This study showed that freeze-drying could be used to stabilize cells of these three biocontrol strains. Further studies should focus on the scale-up possibilities and formulation development. © Nanjing Agricultural University. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

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

    Gupta, Siddhi; Goswami, Sudipta; Sinha, Arvind


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

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

    International Nuclear Information System (INIS)

    Liu, Yang; Ren, Li; Wang, Yingjun


    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

  5. 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: [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: [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)


    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.

  6. Distribution of coniferin in freeze-fixed stem of Ginkgo biloba L. by cryo-TOF-SIMS/SEM (United States)

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


    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.

  7. Summer Freezing Resistance: A Critical Filter for Plant Community Assemblies in Mediterranean High Mountains. (United States)

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


    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

  8. Evaluation of Freezing Tolerance of Hexaploid Triticale Genotypes under Controlled Conditions

    Directory of Open Access Journals (Sweden)

    Ahmad, NEZAMI


    Full Text Available In order to evaluate freezing tolerance of different triticale (X Triticosecale Wittmack genotypes, an experiment was carried out under controlled conditions in 2007 and 2008 at college of agriculture, Ferdowsi University of Mashhad. In this study seeven triticale genotypes (Juanilo-92, ET-82-15, ET-82-8, ET-83-20, ET-83-19, ET-83-18 and ET-79-17, across six temperatures (0C, -4C, -8C, -12C, -16C and -20C were evaluated within a factorial-completely randomized design with three replications. Plants were kept until 2 leaf stage in chamber with temperature of 20/15C (day/night and 12.5 h photoperiod. At the end of this stage, plants were under acclimation for three weeks. After exposing to acclimation freezing the cell membrane integrity was measured through electrolyte leakage (EL and the lethal temperature (LT50 of samples was measured. After the exposure to freezing temperatures the samples were transferred to the greenhouse. Survival percentage, plant height, leaf area and number, chlorophyll content, and plant dry weight were determined after 3 weeks. Results showed that the effect of different freezing temperature and genotypes were significant on all plant characteristics. As temperature decreased, %EL of all genotypes was increased. Minimum and Maximum EL % in leaf and crown were observed at 0C (21% and -20C (88.5%. ET-79-17 and Juanilo-92 genotypes showed the highest EL% (55.5% and 44.8% and ET-83-20 the lowest EL% (47.3% and 41.2% in leaf and crown. Dry weight and leaf area decreased by 48% and 42% respectively compared to non frozen control plants. ET-79-17 and ET-82-15 genotypes showed the highest dry weight (83.8 mg and highest leaf area (14.3 cm2 respectively and ET-83-20 cultivar showed the lowest dry weight and leaf area (58.2 mg and 8.7 cm2.

  9. Cold tolerance and freeze-induced glucose accumulation in three terrestrial slugs

    DEFF Research Database (Denmark)

    Slotsbo, Stine; Hansen, Lars Monrad; Jordaens, Kurt


    in their habitat. Slugs spontaneously froze at about -4 °C when cooled under dry conditions, but freezing of body fluids was readily induced at -1 °C when in contact with external ice crystals. All three species survived freezing for 2 days at -1 °C, and some A. rufus and A. lusitanicus also survived freezing....... 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....

  10. Effect of Freezing on Spermatozoa from Tigaie Rams Belonging to the Mountain Ecotype

    Directory of Open Access Journals (Sweden)

    Vasile Miclea


    Full Text Available Our aim was to study the influence of freezing on the viability and frequency of abnormalities in frozen ram spermatozoa. Sperm was collected form 20 rams belonging to the mountain ecotype of the Tigaie breed using the artificial vagina technique and volume and motility were assessed. Afterward it was diluted with Tryladil (1:4 supplemented with 20% egg yolk and heated at 37°C. Subsequently the temperature decreased at a rate of 0.2°C/minute until reaching 4°C and an equilibration time of 2 hours followed. During this time the diluted sperm was packaged in 0.25 ml straws. After sealing these were kept 6 cm above liquid nitrogen level for 13 minutes (- 120°C and then plunged into nitrogen. Volume, motility and concentration were assessed before freezing. After thawing sperm morphology was assessed using Hancock’s method and at the same time the endurance (at 10, 30 and 60 minutes and HOST tests were performed. The highest motility (0.40 was graded at 30 minutes. It could be correlated with the increased percentage of HOST positive spermatozoa, 27.78%. The percentage of abnormal spermatozoa was also high (47.89%, 38.44% of them having acrosome flaws. Cryopreservation has a negative effect on the characteristics of sperm cells from Tigaie rams belonging to the mountain ecotype.

  11. Freeze-drying of HI-6-loaded recombinant human serum albumin nanoparticles for improved storage stability. (United States)

    Dadparvar, Miriam; Wagner, Sylvia; Wien, Sascha; Worek, Franz; von Briesen, Hagen; Kreuter, Jörg


    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. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Demineralized Freeze-Dried Bovine Cortical Bone: Its Potential for Guided Bone Regeneration Membrane

    Directory of Open Access Journals (Sweden)

    David B. Kamadjaja


    Full Text Available Background. Bovine pericardium collagen membrane (BPCM had been widely used in guided bone regeneration (GBR whose manufacturing process usually required chemical cross-linking to prolong its biodegradation. However, cross-linking of collagen fibrils was associated with poorer tissue integration and delayed vascular invasion. Objective. This study evaluated the potential of bovine cortical bone collagen membrane for GBR by evaluating its antigenicity potential, cytotoxicity, immune and tissue response, and biodegradation behaviors. Material and Methods. Antigenicity potential of demineralized freeze-dried bovine cortical bone membrane (DFDBCBM was done with histology-based anticellularity evaluation, while cytotoxicity was analyzed using MTT Assay. Evaluation of immune response, tissue response, and biodegradation was done by randomly implanting DFDBCBM and BPCM in rat’s subcutaneous dorsum. Samples were collected at 2, 5, and 7 days and 7, 14, 21, and 28 days for biocompatibility and tissue response-biodegradation study, respectively. Result. DFDBCBM, histologically, showed no retained cells; however, it showed some level of in vitro cytotoxicity. In vivo study exhibited increased immune response to DFDBCBM in early healing phase; however, normal tissue response and degradation rate were observed up to 4 weeks after DFDBCBM implantation. Conclusion. Demineralized freeze-dried bovine cortical bone membrane showed potential for clinical application; however, it needs to be optimized in its biocompatibility to fulfill all requirements for GBR membrane.

  13. Chemical freeze-out in relativistic heavy-ion collisions

    Directory of Open Access Journals (Sweden)

    Jun Xu


    Full Text Available One surprising result in relativistic heavy-ion collisions is that the abundance of various particles measured in experiments is consistent with the picture that they reach chemical equilibrium at a temperature much higher than the temperature they freeze out kinetically. Using a multiphase transport model to study particle production in these collisions, we find, as an example, that the effective pion to nucleon ratio, which includes those from resonance decays, indeed changes very little during the evolution of the hadronic matter from the chemical to the kinetic freeze-out, and it is also accompanied by an almost constant specific entropy. We further use a hadron resonance gas model to illustrate the results from the transport model study.


    Directory of Open Access Journals (Sweden)

    Maria Bernadett SZMUTKU


    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. Conception following insemination with a freeze-preserved retrograde ejaculate. (United States)

    Kapetanakis, E; Rao, R; Dmowski, W P; Scommegna, A


    Retrograde ejaculation, an infrequent cause of male infertility, may be the sequela of prostate or bladder neck surgery or the result of interruption in the sympathetic innervation to the bladder neck. The diagnosis is established by history and examination of the urine. In infertile couples artificial insemination homologous (AIH) using retrograde ejaculate recovered from the bladder has been successfully accomplished, but conception has occurred only in isolated cases. A literature review of the subject is presented. The technique of semen recovery from the bladder is time-consuming and uncomfortable to the patient, while the quality of the semen recovered is variable and the risk of iatrogenic urinary tract infection is increased. To avoid the above problems, and to have sperm available for AIH at the time of induced ovulation in an anovulatory wife, we were able to freeze-preserve ejaculate recovered from the bladder for future use. A case of conception following AIH with freeze-preserved retrograde ejaculate is presented.

  16. Heterogeneous ice nucleation: bridging stochastic and singular freezing behavior (United States)

    Niedermeier, D.; Shaw, R. A.; Hartmann, S.; Wex, H.; Clauss, T.; Voigtländer, J.; Stratmann, F.


    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.

  17. Heat and Mass Transfer Model in Freeze-Dried Medium (United States)

    Alfat, Sayahdin; Purqon, Acep


    There are big problems in agriculture sector every year. One of the major problems is abundance of agricultural product during the peak of harvest season that is not matched by an increase in demand of agricultural product by consumers, this causes a wasted agricultural products. Alternative way was food preservation by freeze dried method. This method was already using heat transfer through conduction and convection to reduce water quality in the food. The main objective of this research was to design a model heat and mass transfer in freeze-dried medium. We had two steps in this research, the first step was design of medium as the heat injection site and the second was simulate heat and mass transfer of the product. During simulation process, we use physical property of some agriculture product. The result will show how temperature and moisture distribution every second. The method of research use finite element method (FEM) and will be illustrated in three dimensional.

  18. Different freezing behavior of millimeter- and micrometer-scaled (NH₄)₂SO₄/H₂O droplets. (United States)

    Bogdan, A; Molina, M J; Tenhu, H; Mayer, E; Bertel, E; Loerting, T


    Although the freezing of aqueous solutions is important for nature and different branches of science and freeze-applications, our understanding of the freezing process is not complete. For example, numerous measurements of micrometer-scaled (NH(4))(2)SO(4)/H(2)O droplets report one freezing event below the eutectic point. However, measurements of larger millimeter-scaled droplets reveal two freezing events: the freezing out of ice and subsequent freezing of a residual freeze-concentrated solution. To resolve this apparent contradiction we performed numerous calorimetric measurements which indicate that the freezing of a residual solution of millimeter-scaled 5-38 wt% (NH(4))(2)SO(4) droplets occurs mainly between ∼ 210 and 225 K. We also find that micrometer-scaled droplets produce one freezing event which is within or in the vicinity of the ∼ 210-225 K region. This fact and the analysis of thermograms suggest that the residual solution of micrometer-scaled droplets may partly crystallize simultaneously with ice and partly transform to glass at T(g)≈172 K. Our results suggest for the first time that the size of (NH(4))(2)SO(4)/H(2)O droplets may affect the number of freezing events below the eutectic point.

  19. Optimization Of Freeze-Dried Starter For Yogurt By Full Factorial Experimental Design

    Directory of Open Access Journals (Sweden)

    Chen He


    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.

  20. Freezing of body fluids induces metallothionein gene expression in earthworms (Dendrobaena octaedra). (United States)

    Fisker, Karina Vincents; Holmstrup, Martin; Sørensen, Jesper Givskov


    The molecular mechanisms activated by environmental contaminants and natural stressors such as freezing need to be investigated in order to better understand the mechanisms of interaction and potential effects that combined stressors may have on organisms. Using the freeze-tolerant earthworm Dendrobaena octaedra as model species, we exposed worms to freezing and exposure to sublethal copper in a factorial design and investigated the transcription of candidate genes for metal and cold stress. We hypothesised that both freezing and copper would induce transcription of genes coding for heat shock proteins (hsp10 and hsp70), metallothioneins (mt1 and mt2), and glutathione-S-transferase (gst), and that the combined effects of these two stressors would be additive. The gene transcripts hsp10, hsp70, and gst were significantly upregulated by freezing, but only hsp10 was upregulated by copper. We found that copper at the time of sampling had no effect on transcription of two metallothionein genes whereas transcription was strongly upregulated by freezing. Moreover, there was a significant interaction causing more than additive transcription rates of mt1 in the copper/freezing treatment suggesting that freeze-induced cellular dehydration increases the concentration of free copper ions in the cytosol. This metallothionein response to freezing is likely adaptive and possibly provides protection against freeze-induced elevated metal concentrations in the cytosol and excess ROS levels due to hypoxia during freezing. Copyright © 2015 Elsevier Inc. All rights reserved.

  1. Effects of Pressure-shift Freezing on the Structural and Physical Properties of Gelatin Hydrogel Matrices. (United States)

    Kim, Byeongsoo; Gil, Hyung Bae; Min, Sang-Gi; Lee, Si-Kyung; Choi, Mi-Jung


    This study investigates the effects of the gelatin concentration (10-40%, w/v), freezing temperatures (from -20℃ to -50℃) and freezing methods on the structural and physical properties of gelatin matrices. To freeze gelatin, the pressure-shift freezing (PSF) is being applied at 0.1 (under atmospheric control), 50 and 100 MPa, respectively. The freezing point of gelatin solutions decrease with increasing gelatin concentrations, from -0.2℃ (10% gelatin) to -6.7℃ (40% gelatin), while the extent of supercooling did not show any specific trends. The rheological properties of the gelatin indicate that both the storage (G') and loss (G") moduli were steady in the strain amplitude range of 0.1-10%. To characterize gelatin matrices formed by the various freezing methods, the ice crystal sizes which were being determined by the scanning electron microscopy (SEM) are affected by the gelatin concentrations. The ice crystal sizes are affected by gelatin concentrations and freezing temperature, while the size distributions of ice crystals depend on the freezing methods. Smaller ice crystals are being formed with PSF rather than under the atmospheric control where the freezing temperature is above -40℃. Thus, the results of this study indicate that the PSF processing at a very low freezing temperature (-50℃) offers a potential advantage over commercial atmospheric freezing points for the formation of small ice crystals.

  2. Variation in seedling freezing response is associated with climate in Larrea. (United States)

    Medeiros, Juliana S; Marshall, Diane L; Maherali, Hafiz; Pockman, William T


    Variation in freezing severity is hypothesized to have influenced the distribution and evolution of the warm desert evergreen genus Larrea. If this hypothesis is correct, performance and survival of species and populations should vary predictably along gradients of freezing severity. If freezing environment changes in the future, the ability of Larrea to adapt will depend on the structure of variation for freezing resistance within populations. To test whether freezing responses vary among and within Larrea populations, we grew maternal families of seedlings from high and low latitude L. divaricata and high latitude L. tridentata populations in a common garden. We measured survival, projected plant area and dark-adapted chlorophyll fluorescence (F (v) /F (m)) before and after cold acclimation and for 2 weeks following a single freeze. We detected significant variation in freezing resistance among species and populations. Maternal family lines differed significantly in their responses to cold acclimation and/or freezing for two out of the three populations: among L. tridentata maternal families and among low latitude L. divaricata maternal families. There were no significant differences across maternal families of high latitude L. divaricata. Our results indicate that increased freezing resistance in high latitude populations likely facilitated historical population expansion of both species into colder climates, but this may have occurred to a greater extent for L. tridentata than for L. divaricata. Differences in the structure of variation for cold acclimation and freezing responses among populations suggest potential differences in their ability to evolve in response to future changes in freezing severity.

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

    Czech Academy of Sciences Publication Activity Database

    Nunes, Cristiana Lara; Slížková, Zuzana


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

  4. Compression Freezing Kinetics of Water to Ice VII (United States)

    Gleason, A. E.; Bolme, C. A.; Galtier, E.; Lee, H. J.; Granados, E.; Dolan, D. H.; Seagle, C. T.; Ao, T.; Ali, S.; Lazicki, A.; Swift, D.; Celliers, P.; Mao, W. L.


    Time-resolved x-ray diffraction (XRD) of compressed liquid water shows transformation to ice VII in 6 nsec, revealing crystallization rather than amorphous solidification during compression freezing. Application of classical nucleation theory indicates heterogeneous nucleation and one-dimensional (e.g., needlelike) growth. These first XRD data demonstrate rapid growth kinetics of ice VII with implications for fundamental physics of diffusion-mediated crystallization and thermodynamic modeling of collision or impact events on ice-rich planetary bodies.

  5. Molecular dynamics simulations of freezing of water and salt solutions

    Czech Academy of Sciences Publication Activity Database

    Vrbka, Luboš; Jungwirth, Pavel


    Roč. 134, č. 1 (2007), s. 64-70 ISSN 0167-7322 R&D Projects: GA MŠk LC512; GA ČR(CZ) GD203/05/H001 Institutional research plan: CEZ:AV0Z40550506 Keywords : ice freezing * salt ions * molecular dynamics Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 0.982, year: 2007

  6. Synthesis and structural evaluation of freeze-cast porous alumina

    Energy Technology Data Exchange (ETDEWEB)

    Souza, Douglas F., E-mail: [Department of Metallurgical and Materials Engineering, Federal University of Minas Gerais — UFMG, Avenida Presidente Antônio Carlos, 6627, Campus UFMG, Belo Horizonte, MG CEP: 31270-901, Escola de Engenharia, bloco 2, sala 2230 (Brazil); Nunes, Eduardo H.M., E-mail: [Department of Metallurgical and Materials Engineering, Federal University of Minas Gerais — UFMG, Avenida Presidente Antônio Carlos, 6627, Campus UFMG, Belo Horizonte, MG CEP: 31270-901, Escola de Engenharia, bloco 2, sala 2230 (Brazil); Pimenta, Daiana S.; Vasconcelos, Daniela C.L. [Department of Metallurgical and Materials Engineering, Federal University of Minas Gerais — UFMG, Avenida Presidente Antônio Carlos, 6627, Campus UFMG, Belo Horizonte, MG CEP: 31270-901, Escola de Engenharia, bloco 2, sala 2230 (Brazil); Nascimento, Jailton F.; Grava, Wilson [Petrobras/CENPES, Avenida Horácio Macedo 950, Cidade Universitária, Ilha do Fundão, Rio de Janeiro, RJ CEP:21941-915 (Brazil); Houmard, Manuel [Department of Materials Engineering and Civil Construction, Federal University of Minas Gerais — UFMG, Avenida Presidente Antônio Carlos, 6627, Campus UFMG, Belo Horizonte, MG CEP: 31270-901, Escola de Engenharia, bloco 1, sala 3304 (Brazil); Vasconcelos, Wander L., E-mail: [Department of Metallurgical and Materials Engineering, Federal University of Minas Gerais — UFMG, Avenida Presidente Antônio Carlos, 6627, Campus UFMG, Belo Horizonte, MG CEP: 31270-901, Escola de Engenharia, bloco 2, sala 2230 (Brazil)


    In this work we fabricated alumina samples by the freeze-casting technique using tert-butanol as the solvent. The prepared materials were examined by scanning electron microscopy and X-ray microtomography. Next, they were coated with sol–gel silica films by dip-coating. Permeability tests were carried out in order to assess the permeation behavior of the materials processed in this study. We observed that the sintering time and alumina loading showed a remarkable effect on both the structural properties and flexural strength of the freeze-cast samples. Nitrogen adsorption tests revealed that the silica prepared in this study exhibited a microporous structure. It was observed that the presence of silica coatings on the alumina surface decreased the CO{sub 2} permeance by about one order of magnitude. Because of the similar kinetic diameters of nitrogen and carbon dioxide, the CO{sub 2}/N{sub 2} system showed a separation efficiency that was lower than that observed for the He/CO{sub 2} and He/N{sub 2} systems. We noticed that increasing the feed pressure improved the separation capacity of the obtained materials. - Highlights: • Porous alumina samples obtained by the freeze-casting technique • Microporous silica coating prepared by a simple sol–gel dip-coating methodology • Samples examined by SEM, μ-CT, and nitrogen sorption tests • Mechanical tests were carried out in the freeze-cast samples. • The presence of silica coatings on the alumina surface decreased the CO{sub 2} permeance.

  7. Entree Production Guides for Cook/Freeze Systems (United States)


    Mein 245 Turkey Tetrazzini 248 209 .. . .-- - __".. ___, _ . .. ... . . ..______ . ......__•__, * .% ,-. - 7 ARROZ CON POLLO C-1 Yield: 100 Portions...and freeze. 210 • . 2 ARROZ CON POLLO Ingredients Percent Pounds Grams Procedure Paprika 0.13 0.08 34 Garlic, dehydrated, 0.05 0.03 13 granulated...raw weight to cooked edible (boned) meat. Edible weight of chicken equals 11.7 lb (5307 g). 211 ARROZ CON POLLO Ingradients Meet 1. Bacon, NSN-8905

  8. Isolation of mutations affecting the development of freezing tolerance in Arabidopsis thaliana (L.) Heynh. (United States)

    Warren, G; McKown, R; Marin, A L; Teutonico, R


    We screened for mutations deleterious to the freezing tolerance of Arabidopsis thaliana (L.) Heynh. ecotype Columbia. Tolerance was assayed by the vigor and regrowth of intact plants after cold acclimation and freezing. From a chemically mutagenized population, we obtained 13 lines of mutants with highly penetrant phenotypes. In 5 of these, freezing sensitivity was attributable to chilling injury sustained during cold acclimation, but in the remaining 8 lines, the absence of injury prior to freezing suggested that they were affected specifically in the development of freezing tolerance. In backcrosses, freezing sensitivity from each line segregated as a single nuclear mutation. Complementation tests indicated that the 8 lines contained mutations in 7 different genes. The mutants' freezing sensitivity was also detectable in the leakage of electrolytes from frozen leaves. However, 1 mutant line that displayed a strong phenotype at the whole-plant level showed a relatively weak phenotype by the electrolyte leakage assay.

  9. Preparation and Characterization of Directionally Freeze-cast Copper Foams

    Directory of Open Access Journals (Sweden)

    Aurelia I. Cuba Ramos


    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.

  10. Sensitivity of liquid clouds to homogenous freezing parameterizations. (United States)

    Herbert, Ross J; Murray, Benjamin J; Dobbie, Steven J; Koop, Thomas


    Water droplets in some clouds can supercool to temperatures where homogeneous ice nucleation becomes the dominant freezing mechanism. In many cloud resolving and mesoscale models, it is assumed that homogeneous ice nucleation in water droplets only occurs below some threshold temperature typically set at -40°C. However, laboratory measurements show that there is a finite rate of nucleation at warmer temperatures. In this study we use a parcel model with detailed microphysics to show that cloud properties can be sensitive to homogeneous ice nucleation as warm as -30°C. Thus, homogeneous ice nucleation may be more important for cloud development, precipitation rates, and key cloud radiative parameters than is often assumed. Furthermore, we show that cloud development is particularly sensitive to the temperature dependence of the nucleation rate. In order to better constrain the parameterization of homogeneous ice nucleation laboratory measurements are needed at both high (>-35°C) and low (<-38°C) temperatures. Homogeneous freezing may be significant as warm as -30°CHomogeneous freezing should not be represented by a threshold approximationThere is a need for an improved parameterization of homogeneous ice nucleation.

  11. Warm and cold fermionic dark matter via freeze-in

    International Nuclear Information System (INIS)

    Klasen, Michael; Yaguna, Carlos E.


    The freeze-in mechanism of dark matter production provides a simple and intriguing alternative to the WIMP paradigm. In this paper, we analyze whether freeze-in can be used to account for the dark matter in the so-called singlet fermionic model. In it, the SM is extended with only two additional fields, a singlet scalar that mixes with the Higgs boson, and the dark matter particle, a fermion assumed to be odd under a Z 2 symmetry. After numerically studying the generation of dark matter, we analyze the dependence of the relic density with respect to all the free parameters of the model. These results are then used to obtain the regions of the parameter space that are compatible with the dark matter constraint. We demonstrate that the observed dark matter abundance can be explained via freeze-in over a wide range of masses extending down to the keV range. As a result, warm and cold dark matter can be obtained in this model. It is also possible to have dark matter masses well above the unitarity bound for WIMPs

  12. Freeze concentration of proteins in Antarctic krill wash water (United States)

    Qi, Xiangming; Xu, Jing; Zhao, Kuo; Guo, Hui; Ma, Lei


    Water-washing removes fluoride from Antarctic krill but produces large volumes of wash water containing water- soluble proteins and fluoride. The freeze concentration method was tested to determine if it could be used to recover water-soluble proteins while leaving the fluoride in solution. After freezing and thawing the wash water, protein and fluoride contents of the thawed fractions were determined to explore the melting regularity of components in the wash water. The highest concentration factors of protein and fluoride were obtained after 80 min of thawing, such as 1.48 ± 0.06 and 1.35 ± 0.04 times, respectively. The free amino-nitrogen (FAN) content and sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern results revealed that the highest concentrations of all ingredients were obtained after 80 min of the process. The degree of hydrolysis of all fractions from the thawing process fluctuated in a narrow range around 12% during the entire process, indicating that the thawing order did not change with various proteins or time during the entire thawing course. These results demonstrate that the freeze concentration method can be used to concentrate protein solutions, even those with fluoride. It was concluded that condensation was achieved and no ingredient could be separated, regardless of fluoride, amino acids, or different proteins in the water.

  13. Studies of heterogeneous freezing by three different desert dust samples

    Directory of Open Access Journals (Sweden)

    P. J. Connolly


    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

  14. Immersion freezing of ice nucleation active protein complexes

    Directory of Open Access Journals (Sweden)

    S. Hartmann


    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

  15. A practical approach to obtain the soil freezing characteristic curve and the freezing/melting point of a soil-water system

    NARCIS (Netherlands)

    Aukenthaler, Manuel; Brinkgreve, R.B.J.; Haxaire, A


    Knowing that extensive field tests and laboratory tests are time-consuming and expensive, this paper describes a practical approach to obtain crucial properties of frozen soil such as the soil freezing characteristic curve (SFCC), the freezing/melting point of a soil-water system and its hydraulic

  16. Effects of sugar alcohol and proteins on the survival of Lactobacillus bulgaricus LB6 during freeze drying. (United States)

    Chen, He; Chen, Shiwei; Chen, Hongli; Wu, Yanyan; Shu, Guowei


    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. 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. 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). 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.

  17. Effect of physical properties on the stability of Lactobacillus bulgaricus in a freeze-dried galacto-oligosaccharides matrix. (United States)

    Tymczyszyn, E Elizabeth; Sosa, Natalia; Gerbino, Esteban; Hugo, Ayelen; Gómez-Zavaglia, Andrea; Schebor, Carolina


    The ability of galacto-oligosaccharides (GOS) to protect Lactobacillus delbrueckii subsp. bulgaricus upon freeze drying was analyzed on the basis of their capacity to form glassy structures. Glass transition temperatures (T(g)) of a GOS matrix at various relative humidities (RH) were determined by DSC. Survival of L. bulgaricus in a glassy GOS matrix was investigated after freezing, freeze drying, equilibration at different RHs and storage at different temperatures. At 32 °C, a drastic viability loss was observed. At 20 °C, the survival was affected by the water content, having the samples stored at lower RHs, the highest survival percentages. At 4°C, no decay in the cells count was observed after 45 days of storage. The correlation between molecular mobility [as measured by Proton nuclear magnetic resonance (¹H NMR)] and loss of viability explained the efficiency of GOS as cryoprotectants. The preservation of microorganisms was improved at low molecular mobility and this condition was obtained at low water contents and low storage temperatures. These results are important in the developing of new functional foods containing pre and probiotics. Copyright © 2012 Elsevier B.V. All rights reserved.

  18. 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))


    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.

  19. The performance of five fruit-derived and freeze-dried potentially probiotic Lactobacillus strains in apple, orange and grape juices. (United States)

    Garcia, Estefânia Fernandes; de Oliveira Araújo, Amanda; Luciano, Winnie Alencar; de Albuquerque, Thatyane Mariano Rodrigues; de Oliveira Arcanjo, Narciza Maria; Madruga, Marta Suely; Dos Santos Lima, Marcos; Magnani, Marciane; Saarela, Maria; de Souza, Evandro Leite


    This study assessed the survival of the fruit-derived and freeze-dried L. plantarum 49, L. brevis 59, L. paracasei 108, L. fermentum 111 and L. pentosus 129 strains during frozen storage and when incorporated into apple, orange and grape juice stored under refrigeration. Physicochemical parameters of juices containing the freeze-dried Lactobacillus strains and the survival of the test strains in the fruit juices during in vitro digestion were also evaluated. No decreases in survival rates (log N/log N0) of the freeze-dried cells were observed up to 1 month of storage. The survival rates of the freeze-dried strains L. plantarum 49 and L. paracasei 108 were >0.75 up to 4 months of storage. All freeze-dried strains exhibited survival rates of >0.75 up to 2 weeks of storage in apple juice; only L. plantarum 49 and L. paracasei 108 showed similar survival rates in orange and grape juices up to 2 weeks of storage. The contents of the monitored organic acids or sugars during storage varied depending on the added strain and the type of fruit juice. At the end of the in vitro digestion, L. brevis 59, L. paracasei 108 and L. fermentum 111 showed survival rates of >0.80 in apple juice. Apple juice was as the best substrate to the survival of the tested freeze-dried Lactobacillus strains over time. L. paracasei 108 and L. plantarum 49 as the strains presenting the best performance for incorporation in potentially probiotic fruit juices. This article is protected by copyright. All rights reserved.

  20. Safety assessment of freeze-dried powdered Cassiae Semen: evaluation of chronic toxicity (26-week in Sprague-Dawley rats

    Directory of Open Access Journals (Sweden)

    Yanyu Pei

    Full Text Available There is a lack of safety assessment data regarding the long-term consumption of Cassiae Semen (Leguminosae, the seeds of Cassia obtusifolia L. and Cassia tora L.. Thus, we evaluated the toxicity of freeze-dried powdered Cassiae Semen in male and female Sprague-Dawley rats. Rats were intragastrically administered freeze-dried powdered Cassiae Semen at a dose of 0.5, 2.2, or 10.0 g/kg body weight/day for 26 weeks; several variables were assessed after 13 and 26 weeks as well as after a 4-week recovery period. No mortality was observed in the treated animals, and body weight increased in a dose-dependent manner. The total bilirubin (TBIL levels also displayed a dose-dependent relationship. In males, at 26 weeks, there were significant increases in relative kidney weights in the 2.2 and 10.0 g/kg groups compared with that in the negative control group (p < 0.05 or p < 0.01. Pigment deposition in the epithelial cells of the renal proximal convoluted tubules and atrophy or regeneration of renal tubules were observed in the 10.0 g/kg group after 26 weeks, and these changes were not fully reversed after the 4-week recovery period. Under the studied conditions, the primary toxicity organs for freeze-dried powdered Cassiae Semen in the 10.0 g/kg group were the kidneys. Keywords: Cassiae Semen, Freeze-dried powder, Chronic toxicity, Toxicity organs

  1. 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.


    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.

  2. Robustness testing in pharmaceutical freeze-drying: inter-relation of process conditions and product quality attributes studied for a vaccine formulation. (United States)

    Schneid, Stefan C; Stärtzel, Peter M; Lettner, Patrick; Gieseler, Henning


    The recent US Food and Drug Administration (FDA) legislation has introduced the evaluation of the Design Space of critical process parameters in manufacturing processes. In freeze-drying, a "formulation" is expected to be robust when minor deviations of the product temperature do not negatively affect the final product quality attributes. To evaluate "formulation" robustness by investigating the effect of elevated product temperature on product quality using a bacterial vaccine solution. The vaccine solution was characterized by freeze-dry microscopy to determine the critical formulation temperature. A conservative cycle was developed using the SMART™ mode of a Lyostar II freeze dryer. Product temperature was elevated to imitate intermediate and aggressive cycle conditions. The final product was analyzed using X-ray powder diffraction (XRPD), scanning electron microscopy (SEM), Karl Fischer, and modulated differential scanning calorimetry (MDSC), and the life cell count (LCC) during accelerated stability testing. The cakes processed at intermediate and aggressive conditions displayed larger pores with microcollapse of walls and stronger loss in LCC than the conservatively processed product, especially during stability testing. For all process conditions, a loss of the majority of cells was observed during storage. For freeze-drying of life bacterial vaccine solutions, the product temperature profile during primary drying appeared to be inter-related to product quality attributes.

  3. Novel real-time diagnosis of the freezing process using an ultrasonic transducer. (United States)

    Tseng, Yen-Hsiang; Cheng, Chin-Chi; Cheng, Hong-Ping; Lee, Dasheng


    The freezing stage governs several critical parameters of the freeze drying process and the quality of the resulting lyophilized products. This paper presents an integrated ultrasonic transducer (UT) in a stainless steel bottle and its application to real-time diagnostics of the water freezing process. The sensor was directly deposited onto the stainless steel bottle using a sol-gel spray technique. It could operate at temperature range from -100 to 400 °C and uses an ultrasonic pulse-echo technique. The progression of the freezing process, including water-in, freezing point and final phase change of water, were all clearly observed using ultrasound. The ultrasonic signals could indicate the three stages of the freezing process and evaluate the cooling and freezing periods under various processing conditions. The temperature was also adopted for evaluating the cooling and freezing periods. These periods increased with water volume and decreased with shelf temperature (i.e., speed of freezing). This study demonstrates the effectiveness of the ultrasonic sensor and technology for diagnosing and optimizing the process of water freezing to save energy.

  4. Repeated freeze-thaw cycles induce embolism in drought stressed conifers (Norway spruce, stone pine). (United States)

    Mayr, Stefan; Gruber, Andreas; Bauer, Helmut


    Freezing and thawing lead to xylem embolism when gas bubbles caused by ice formation expand during the thaw process. However, previous experimental studies indicated that conifers are resistant to freezing-induced embolism, unless xylem pressure becomes very negative during the freezing. In this study, we show that conifers experienced freezing-induced embolism when exposed to repeated freeze-thaw cycles and simultaneously to drought. Simulating conditions at the alpine timberline (128 days with freeze-thaw events and thawing rates of up to 9.5 K h(-1) in the xylem of exposed twigs during winter), young trees of Norway spruce [Picea abies (L.) Karst.] and stone pine (Pinus cembra L.) were exposed to 50 and 100 freeze-thaw cycles. This treatment caused a significant increase in embolism rates in drought-stressed samples. Upon 100 freeze-thaw cycles, vulnerability thresholds (50% loss of conductivity) were shifted 1.8 MPa (Norway spruce) and 0.8 MPa (stone pine) towards less negative water potentials. The results demonstrate that freeze-thaw cycles are a possible reason for winter-embolism in conifers observed in several field studies. Freezing-induced embolism may contribute to the altitudinal limits of conifers.

  5. Thermal properties of ration components as affected by moisture content and water activity during freezing. (United States)

    Li, J; Chinachoti, P; Wang, D; Hallberg, L M; Sun, X S


    Beef roast with vegetables is an example of a meal, ready-to-eat (MRE) ration entrée. It is a mixture of meat, potato, mushroom, and carrot with a gravy sauce. The thermal properties of each component were characterized in terms of freezing point, latent heat, freezable and unfreezable water contents, and enthalpy during freezing using differential scanning calorimetry. Freezing and thawing curves and the effect of freezing and thawing cycles on thermal properties were also evaluated. The freezing points of beef, potato, mushroom, and sauce were all in the range of -5.1 to -5.6 degrees C, but moisture content, water activity, latent heat, freezable and unfreezable water contents, and enthalpy varied among these components. Freezing temperature greatly affected the unfrozen water fraction. The unfreezable water content (unfrozen water fraction at -50 degrees C) of ration components was in the range of 8.2% to 9.7%. The freezing and thawing curves of vegetables with sauce differed from those of beef but took similar time to freeze or thaw. Freezing and thawing cycles did not greatly affect the thermal properties of each component. Freezing point and latent heat were reduced by decreasing moisture content and water activity of each component. Water activity was proportionally linear to freezing point at a(w) > 0.88, and moisture content was proportionally linear to freezable water content in all ration components. Water was not available for freezing when moisture content was reduced to 28.8% or less. This study indicates that moisture content and water activity are critical factors affecting thermal behavior of ration components during freezing.

  6. 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. (United States)

    Peiren, Jindrich; Hellemans, Ann; De Vos, Paul


    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.

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

    Energy Technology Data Exchange (ETDEWEB)

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


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

  8. Freeze-fracture studies of photoreceptor membranes: new observations bearing upon the distribution of cholesterol. (United States)

    Andrews, L D; Cohen, A I


    We performed electron microscopy of replicas from freeze-fractured retinas exposed during or after fixation to the cholesterol-binding antibiotic, filipin. We observed characteristic filipin-induced perturbations throughout the disk and plasma membranes of retinal rod outer segments of various species. It is evident that a prolonged exposure to filipin in fixative enhances rather than reduces presumptive cholesterol detection in the vertebrate photoreceptor cell. In agreement with the pattern seen in our previous study (Andrews, L.D., and A. I. Cohen, 1979, J. Cell Biol., 81:215-228), filipin-binding in membranes exhibiting particle-free patches seemed largely confined to these patches. Favorably fractured photoreceptors exhibited marked filipin-binding in apical inner segment plasma membrane topologically confluent with and proximate to the outer segment plasma membrane, which was comparatively free of filipin binding. A possible boundary between these differing membrane domains was suggested in a number of replicas exhibiting lower filipin binding to the apical plasma membrane of the inner segment in the area surrounding the cilium. This area contains a structure (Andrews, L. D., 1982, Freeze-fracture studies of vertebrate photoreceptors, In Structure of the Eye, J. G. Hollyfield and E. Acosta Vidrio, editors, Elsevier/North-Holland, New York, 11-23) that resembles the active zones of the nerve terminals for the frog neuromuscular junction. These observations lead us to hypothesize that these structures may function to direct vesicle fusion to occur near them, in a domain of membrane more closely resembling outer than inner segment plasma membrane. The above evidence supports the views that (a) all disk membranes contain cholesterol, but the particle-free patches present in some disks trap cholesterol from contiguous particulate membrane regions; (b) contiguous inner and outer segment membranes may greatly differ in cholesterol content; and (c) the suggested

  9. Freeze-drying behaviour of pasteurized whole egg

    International Nuclear Information System (INIS)

    Melike Sakin; Merve Samli; Gizem Kor, A.; Figen Kaymak-Ertekin


    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 (a w ), 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)

  10. Equilibrium theory of molecular fluids: Structure and freezing transitions (United States)

    Ram, Jokhan


    In this article we review equilibrium theory of molecular fluids which includes structure and freezing transitions. The application of the theory to evaluate the pair correlation functions using Integral Equation methods and Computer Simulations have been discussed. Freezing of classical complex fluids based on the density functional approach is also discussed and compare a variety of its versions. Transitions discussed are sensitive to the value of direct correlation functions of the effective liquid which is required as an input information in the theory. Accurate evaluation of pair correlation functions is emphasized. Calculation of these correlation functions which pose problems in the case of ordered phases is discussed. The pair correlation functions of the ordered phase, which are supposed to be made up of two contributions, one that preserves the symmetry of the isotropic phase and a second that breaks it, are discussed. A new free-energy functional developed for an inhomogeneous system that contains both symmetry conserved and symmetry broken parts of the direct pair correlation function is discussed. The most useful three dimensional reference interaction site model (3D-RISM) and its extension done recently by many workers is discussed. Application of this theory to a large variety of complex systems in combination with the density functional theory method implemented in the Amsterdam density functional software package is discussed. Coupling of the 3D-RISM salvation theory with molecular dynamics in the Amber molecular dynamics package is also given. The importance of the density functional theory for the study of the structure and phase behaviour of hard polyhedral is also discussed. The dynamical density functional and its generalized form applied for many important class of problems such as binary mixture, anisotropic particles dynamics of freezing and wetting, colloidal samples, particle self diffusion in complex environment, colloidal sedimentation

  11. Modelling of the granular products vacuum freeze-dried process

    Directory of Open Access Journals (Sweden)

    S. T. Antipov


    Full Text Available In the work reviewed and simulated the process of vacuum freeze-drying of granular products with the destruction of the dried layer. As development of this direction serves the method based on removal of the dried product layer from a surface of a granule, formed in the form of spherical bodies and placed in the punched drum. After process of preliminary freezing of a granule get to the punched drum located in the vacuum drying chamber. In case of fixed hashing of granules of a product due to their friction among themselves and about drum walls a dry part of a product undergoes destruction and regularly separates from the refrigerated remaining balance. Process comes to an end when all product in a granule turns into dry powder. For creation of adequate model of process of drying, it is reasonable to consider a separate granule, but not a layer of granules in general, but taking into account influence of granules at each other in the course of drying. For this purpose, the scheme and mathematical description of radiation heat flux of the individual granules of the product. To account for the intermittent nature of exposure to granules suggested ratio and its two alternative approaches to its definition. As well as an algorithm for determining the coefficient that takes into account the one-sided irradiation of the pellets. Formulated mathematical model of the process the vacuum - freeze-drying of granular products, which is a one-dimensional parabolic boundary value problem with moving boundary. It includes the unsteady heat conduction equation, initial condition, boundary conditions and defining a movable boundary conditions. The sought quantities are the temperature field and the free boundary. Explained the origin of the functions of internal heat sources present in the equations of heat conduction.

  12. Investigation of the Influence of Heat Balance Shifts on the Freeze Microstructure and Composition in Aluminum Smelting Bath System: Cryolite-CaF2-AlF3-Al2O3 (United States)

    Liu, Jingjing; Fallah-Mehrjardi, Ata; Shishin, Denis; Jak, Evgueni; Dorreen, Mark; Taylor, Mark


    In an aluminum electrolysis cell, the side ledge forms on side walls to protect it from the corrosive cryolitic bath. In this study, a series of laboratory analogue experiments have been carried out to investigate the microstructure and composition of side ledge (freeze linings) at different heat balance steady states. Three distinct layers are found in the freeze linings formed in the designed Cryolite-CaF2-AlF3-Al2O3 electrolyte system: a closed (columnar) crystalline layer, an open crystalline layer, and a sealing layer. This layered structure changes when the heat balance is shifted between different steady states, by melting or freezing the open crystalline layer. Phase chemistry of the freeze lining is studied in this paper to understand the side ledge formation process upon heat balance shifts. Electron probe X-ray microanalysis (EPMA) is used to characterize the microstructure and compositions of distinct phases existing in the freeze linings, which are identified as cryolite, chiolite, Ca-cryolite, and alumina. A freeze formation mechanism is further developed based on these microstructural/compositional investigations and also thermodynamic calculations through the software—FactSage. It is found that entrapped liquid channels exist in the open crystalline layer, assisting with the mass transfer between solidified crystals and bulk molten bath.

  13. Biofilm-like Lactobacillus rhamnosus probiotics encapsulated in alginate and carrageenan microcapsules exhibiting enhanced thermotolerance and freeze-drying resistance. (United States)

    Cheow, Wean Sin; Hadinoto, Kunn


    Microcapsules containing high-density biofilm-like Lactobacillus rhamnosus probiotics, in place of planktonic cells, are developed in order to enhance the cell viability upon exposures to stresses commonly encountered during food lifecycle (i.e., heating, freeze-drying, refrigerated storage, and acid). The high-density (HD) capsules are prepared by in situ cultivation of the planktonic cells in the confined space of polysaccharide-based capsules (i.e., chitosan-coated alginate and carrageenan capsules). Compared to their planktonic counterparts, the HD capsules exhibit higher freeze-drying resistance (~40×) and higher thermotolerance upon prolonged wet heat exposures at 60 and 70 °C (~12-8000×), but not at higher temperatures even for short exposures (i.e., 80 and 100 °C). The enhanced viability of the HD capsules, however, is not observed during the refrigerated storage and exposure to the simulated gastric juice. The alginate capsules are superior to carrageenan owed to their better cell release profile in the simulated intestinal juice and storage viability.

  14. Experimental research on durability of recycled aggregate concrete under freeze- thaw cycles (United States)

    Cheng, Yanqiu; Shang, Xiaoyu; Zhang, Youjia


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

  15. 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


    -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......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...... 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...

  16. Research on strength attenuation law of concrete in freezing - thawing environment (United States)

    Xiao, qianhui; Cao, zhiyuan; Li, qiang


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

  17. Do freeze events create a demographic bottleneck for Colophospermum mopane?

    CSIR Research Space (South Africa)

    Whitecross, MA


    Full Text Available reference: SAJB 815 P-authorquery-v11 Page 1 of 1 UNCORRECTED P R OO F 1 Highlights 2 South African Journal of Botany xxx (2012) xxx? xxx 4 5 Do freeze events create a demographic bottleneck for Colophospermum mopane? 6 7 M.A. Whitecrossa, S... at South African Journal of Botany xx (2012) xxx SAJB-00815; No of Pages 1 1 demographi 2 2Q1 3 e Witwatersrand 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 Keywords...

  18. Combined electrohydrodynamic (EHD) and vacuum freeze drying of shrimp

    International Nuclear Information System (INIS)

    Hu, Yucai; Huang, Qiang; Bai, Yaxiang


    To improve the drying qualities of shrimp, a combination of electrohydrodynamic (EHD) and vacuum freeze drying (FD) is examined. The drying rate, the shrinkage, the rehydration ratio, and the sensory properties including the color and trimness of the dried products under different drying methods (including combination drying of EHD and FD, EHD drying and FD drying) are measured. Compared with FD and EHD drying alone, the combined process consumes less drying time, and the product processed by combined drying displays lower shrinkage, higher rehydration rate and better sensory qualities.

  19. Deformed liquid marbles: Freezing drop oscillations with powders

    KAUST Repository

    Marston, Jeremy


    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.

  20. Electrochemical properties of carbon aerogels with freeze - drying (United States)

    Xu, Yuelong; Yan, Meifang; Liua, Zhenfa


    Carbon aerogels (CAs) were prepared via a sol-gel process by polymerization of phloroglucinol, resorcinol and formaldehyde using 2,4-dihydroxybenzoic acid as catalyst with freeze-drying. The electrochemical properties were characterized using cyclic voltammetry, galvanostatic charge-discharge measurements and electrochemical impedance spectroscopy (EIS). The specific capacitance of corresponding CAs was up to 131 F g-1 and 105 F g-1 at the density of 0.5 A g-1 and 1.0 A g-1, respectively.



    FÃtima Revia Granja Lima


    Three experiments were carried out aiming to evaluate the influence of different factors on parameter of post-thawing semen motility and pregnancy index of rams. The first one showed the influence of extender type (skim milk, HEPES and TRIS), freezing temperature (-79oC, -90oC and -120oC) and final concentration of sperm (600 x 106, 800 x 106 and 1000 x 106 sperm per mL) on semen motility parameters of Santa Ines rams. The second one experiment evaluated the influence of egg yolk typ...

  2. Numerical simulation of the effect of groundwater salinity on artificial freezing wall in coastal area (United States)

    Hu, Rui; Liu, Quan


    During the engineering projects with artificial ground freezing (AFG) techniques in coastal area, the freezing effect is affected by groundwater salinity. Based on the theories of artificially frozen soil and heat transfer in porous material, and with the assumption that only the variations of total dissolved solids (TDS) impact on freezing point and thermal conductivity, a numerical model of an AFG project in a saline aquifer was established and validated by comparing the simulated temperature field with the calculated temperature based on the analytic solution of rupak (reference) for single-pipe freezing temperature field T. The formation and development of freezing wall were simulated with various TDS. The results showed that the variety of TDS caused the larger temperature difference near the frozen front. With increasing TDS in the saline aquifer (1 35g/L), the average thickness of freezing wall decreased linearly and the total formation time of the freezing wall increased linearly. Compared with of the scenario of fresh-water (<1g/L), the average thickness of frozen wall decreased by 6% and the total formation time of the freezing wall increased by 8% with each increasing TDS of 7g/L. Key words: total dissolved solids, freezing point, thermal conductivity, freezing wall, numerical simulation Reference D.J.Pringel, H.Eicken, H.J.Trodahl, etc. Thermal conductivity of landfast Antarctic and Arctic sea ice[J]. Journal of Geophysical Research, 2007, 112: 1-13. Lukas U.Arenson, Dave C.Sego. The effect of salinity on the freezing of coarse- grained sand[J]. Canadian Geotechnical Journal, 2006, 43: 325-337. Hui Bing, Wei Ma. Laboratory investigation of the freezing point of saline soil[J]. Cold Regions Science and Technology, 2011, 67: 79-88.

  3. Impact of nucleon mass shift on the freeze-out process

    International Nuclear Information System (INIS)

    Zschocke, Sven; Csernai, Laszlo Pal; Molnar, Etele; Nyiri, Agnes; Manninen, Jaakko


    The freeze-out of a massive nucleon gas through a finite layer with a timelike normal is studied. The impact of the in-medium nucleon mass shift on the freeze-out process is investigated. A considerable modification of the thermodynamic variables of temperature, flow velocity, energy density, and particle density has been found. Because of the nucleon mass shift the freeze-out particle distribution functions are changed noticeably in comparison with the evaluations, which use the vacuum nucleon mass

  4. Practical limitations of ITS-90 from the mercury triple point to the silver freeze point

    International Nuclear Information System (INIS)

    Tavener, J. P.; Tavener, S. J.; Tavener, I. F.; Davies, N.


    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

  5. Effects of Artificial Supercooling Followed by Slow Freezing on the Microstructure and Qualities of Pork Loin. (United States)

    Kim, Yiseul; Hong, Geun-Pyo


    This study investigated the effects of artificial supercooling followed by still air freezing (SSF) on the qualities of pork loin. The qualities of pork frozen by SSF were compared with the fresh control (CT, stored at 4℃ for 24 h), slow freezing (SAF, still air freezing) and rapid freezing (EIF, ethanol immersion freezing) treatments. Compared with no supercooling phenomena of SAF and EIF, the extent of supercooling obtained by SSF treatment was 1.4℃. Despite that SSF was conducted with the same method with SAF, application of artificial supercooling accelerated the phase transition (traverse from -0.6℃ to -5℃) from 3.07 h (SAF) to 2.23 h (SSF). The observation of a microstructure indicated that the SSF prevented tissue damage caused by ice crystallization and maintained the structural integrity. The estimated quality parameters reflected that SSF exhibited superior meat quality compared with slow freezing (SAF). SSF showed better water-holding capacity (lower thawing loss, cooking loss and expressible moisture) and tenderness than SAF, and these quality parameters of SSF were not significantly different with ultra-fast freezing treatment (EIF). Consequently, the results demonstrated that the generation of supercooling followed by conventional freezing potentially had the advantage of minimizing the quality deterioration caused by the slow freezing of meat.

  6. Estimation of Freezing Point of Hydrocarbon and Hydrofluorocarbon Mixtures for Mixed Refrigerant jt Cryocooler (United States)

    Hwang, G.; Lee, J.; Jeong, S.


    Estimating the freezing point of refrigerant is an essential part in designing an MR JT (Mixed refrigerant Joule-Thomson) cryocooler to prevent itself from clogging and to operate with stability. There were researches on estimating freezing point, but some of them resulted in the wrong prediction of clogging. In this paper, the freezing point of the MR is precisely estimated with caution of clogging. The solubility of HC (hydrocarbon) and HFC (hydrofluorocarbon) mixture components are obtained with their activity coefficients, which represent the molecular interaction among the components. The freezing points of the MR JT cryocooler are systematically investigated in the operating temperature range from 70 K to 90 K.

  7. Successful long-term preservation of rat sperm by freeze-drying.

    Directory of Open Access Journals (Sweden)

    Takehito Kaneko

    Full Text Available BACKGROUND: Freeze-drying sperm has been developed as a new preservation method where liquid nitrogen is no longer necessary. An advantage of freeze-drying sperm is that it can be stored at 4 °C and transported at room temperature. Although the successful freeze-drying of sperm has been reported in a number of animals, the possibility of long-term preservation using this method has not yet been studied. METHODOLOGY/PRINCIPAL FINDINGS: Offspring were obtained from oocytes fertilized with rat epididymal sperm freeze-dried using a solution containing 10 mM Tris and 1 mM EDTA adjusted to pH 8.0. Tolerance of testicular sperm to freeze-drying was increased by pre-treatment with diamide. Offspring with normal fertility were obtained from oocytes fertilized with freeze-dried epididymal sperm stored at 4 °C for 5 years. CONCLUSIONS AND SIGNIFICANCE: Sperm with -SS- cross-linking in the thiol-disulfide of their protamine were highly tolerant to freeze-drying, and the fertility of freeze-dried sperm was maintained for 5 years without deterioration. This is the first report to demonstrate the successful freeze-drying of sperm using a new and simple method for long-term preservation.

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

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

    Directory of Open Access Journals (Sweden)

    Bo Ke


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

  10. A Numerical and Experimental Investigation of the Internal Flow of a Freezing Water Droplet


    Karlsson, Linn


    The overarching aim of this work is to study the freezing process of a single water droplet freezing on a cold surface, which is an interesting and important phenomenon with possible applications in many areas. Understanding the freezing process of a single water droplet is for example an important step when preventing unwanted icing, e.g. in the case of airplane wings and propellers, wind turbine rotor blades, and road surfaces.As a step in understanding the freezing process, the study speci...

  11. A New Mechanism of Canopy Effect in Unsaturated Freezing Soils

    Directory of Open Access Journals (Sweden)

    Teng Jidong


    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.

  12. Thermalization, Freeze-out, and Noise: Deciphering Experimental Quantum Annealers (United States)

    Marshall, Jeffrey; Rieffel, Eleanor G.; Hen, Itay


    By contrasting the performance of two quantum annealers operating at different temperatures, we address recent questions related to the role of temperature in these devices and their function as "Boltzmann samplers." Using a method to reliably calculate the degeneracies of the energy levels of large-scale spin-glass instances, we are able to estimate the instance-dependent effective temperature from the output of annealing runs. Our results corroborate the "freeze-out" picture which posits two regimes, one in which the final state corresponds to a Boltzmann distribution of the final Hamiltonian with a well-defined "effective temperature" determined at a freeze-out point late in the annealing schedule, and another regime in which such a distribution is not necessarily expected. We find that the output distributions of the annealers do not, in general, correspond to a classical Boltzmann distribution for the final Hamiltonian. We also find that the effective temperatures at different programing cycles fluctuate greatly, with the effect worsening with problem size. We discuss the implications of our results for the design of future quantum annealers to act as more-effective Boltzmann samplers and for the programing of such annealers.

  13. Freezing Point Determination of Water–Ionic Liquid Mixtures

    DEFF Research Database (Denmark)

    Liu, Yanrong; Meyer, Anne S.; Nie, Yi


    of water. On the basis of the cryoscopicbehavior recorded, the solid phases formed at higher IL contents werepresumed to be hydrates of the form IL·nH2O. The HOEtpyBr+H2O and HOEtmimBr+H2O systems formed simple eutectic systems. The eutectic points werefound to be at a water mole fraction of 0.617 and 219.......841 K in thefirst system and at a water mole fraction of 0.657 and 202.565 K inthe second system. Water activities in aqueous IL solutions were predictedby COSMO-RS and COSMO-SAC and compared to water activities derivedfrom the experimentally determined freezing points. The COSMO-RS predictionswere...... closer to the experimental water activities than the COSMO-SACpredictions. The experimental results indicate that the freezing pointsof IL+H2O systems are affected by the nature of both cationsand anions. However, according to the COSMO-RS excess enthalpy predictionresults, the anions have a relatively...

  14. Effect of freezing on the ram sperm morphometric parameters

    Directory of Open Access Journals (Sweden)

    Choquepuma W


    Full Text Available The objective of the study was to evaluate the effect of freezing on the morphometric parameters of ram spermatozoa. For this purpose, the semen of 2 young rams of the Corriedale breed of 2 and 2.5 years old was used, with a body condition of 3.5. The volume, motility, concentration, viability, membrane integrity and sperm morphometry were evaluated in the samples. All morphometric variables were determined using a CASA system (ISAS, Proiser. The freezing process was performed with TRIS based diluent with 3% soy lecithin and glycerol as cryoprotectant. A total of 12 morphometric parameters were analyzed (length, with, area, perimeter, ellipticity, elongation, regularity and roughness of head spermatozoa, acrosome percentage, and the width, distance and insertion angle mid piece ram sperm. A population of 2518 spermatozoa was evaluated in fresh, refrigerated and thawed semen. Our results indicate that some cryopreserved sperm morphometric parameters (length, width, area and perimeter of the head are smaller (p <0.05 than that found in fresh and refrigerated semen.

  15. Immersion Freezing of Total Ambient Aerosols and Ice Residuals

    Directory of Open Access Journals (Sweden)

    Gourihar Kulkarni


    Full Text Available This laboratory study evaluates an experimental set-up to study the immersion freezing properties of ice residuals (IRs at a temperature ranging from −26 to −34 °C using two continuous-flow diffusion chamber-style ice nucleation chambers coupled with a virtual impactor and heat exchanger. Ice was nucleated on the total ambient aerosol through an immersion freezing mechanism in an ice nucleation chamber (chamber 1. The larger ice crystals formed in chamber 1 were separated and sublimated to obtain IRs, and the frozen fraction of these IRs was investigated in a second ice nucleation chamber (chamber 2. The ambient aerosol was sampled from a sampling site located in the Columbia Plateau region, WA, USA, which is subjected to frequent windblown dust events, and only particles less than 1.5 μm in diameter were investigated. Single-particle elemental composition analyses of the total ambient aerosols showed that the majority of the particles are dust particles coated with organic matter. This study demonstrated a capability to investigate the ice nucleation properties of IRs to better understand the nature of Ice Nucleating Particles (INPs in the ambient atmosphere.

  16. Optimization of a protective medium for enhancing the viability of freeze-dried Bacillus amyloliquefaciens B1408 based on response surface methodology. (United States)

    Han, Lingjuan; Pu, Taixun; Wang, Xi; Liu, Beiling; Wang, Yonghong; Feng, Juntao; Zhang, Xing


    Response surface methodology (RSM) is a commonly used system to optimize cryoprotectants of biocontrol strains when they are subjected to preparations. Various kinds of cryoprotectants and centrifugal conditions were tested to improve the survival of biocontrol agents after freeze-drying. To determine the optimum levels of incorporation of three cryoprotectants (glucose, trehalose and xylitol) in the freeze-drying process of strain Bacillus amyloliquefaciens B1408, a range of experiments based on Box-Behnken Design (BBD) were conducted. The results indicated that the suitable centrifugation conditions were 5000 r/min,10 min and the optimum concentrations of cryoprotectants were glucose 1.00%, trehalose 4.74% and xylitol 1.45%. The proven survival rate of cells after freeze-drying was 91.24%. These results convincingly demonstrated that freeze-drying could be used to preparation of biocontrol strain B1408. This study provides a theoretical basis for commercial possibilities and formulation development. Copyright © 2018. Published by Elsevier Inc.

  17. Improvement of a dry formulation of Pseudomonas fluorescens EPS62e for fire blight disease biocontrol by combination of culture osmoadaptation with a freeze-drying lyoprotectant. (United States)

    Cabrefiga, J; Francés, J; Montesinos, E; Bonaterra, A


    To study the effect of lyoprotectants and osmoadaptation on viability of Pseudomonas fluorescens EPS62e during freeze-drying and storage and to evaluate the formulation in terms of efficacy in biocontrol and fitness on pear flowers. A wettable powder formulation of a biocontrol agent of fire blight was optimized by means of lyoprotectants and culture osmoadaptation. Freeze-drying was used to obtain dehydrated cells, and the best viability (70% of survival) was obtained using lactose as lyoprotectant. Survival during lyophilization was additionally improved using physiological adaptation of cells during cultivation under salt-amended medium (osmoadaptation). The procedure increased the survival of cells after freeze-drying attaining viability values close to a 100% in the lactose-formulated product (3 × 10(11) CFU g(-1) ), and through the storage period of 1 year at 4°C. The dry formulation showed also an improved biocontrol efficacy and survival of EPS62e on pear flowers under low relative humidity conditions. Cell viability after freeze-drying was improved using lactose as lyoprotectant combined with a procedure of osmoadaptation during cultivation. The powder-formulated product remained active for 12 months and retained biocontrol levels similar to that of fresh cells. The formulation showed an improved survival of EPS62e on flowers and an increase of the efficacy of biocontrol of fire blight at low relative humidity. The results have a potential value for commercial application in biocontrol agents not only of fire blight but also of other plant diseases. © 2014 The Society for Applied Microbiology.

  18. [Freeze-etch study on the brush border structure of Cysticercus cellulosae]. (United States)

    Meng, X Q; Wang, S S; Zhou, W Q; Wang, B X; Zhang, Y Y; Liu, G S


    The images of C. cellulosae detected by freeze-itch technique were exactly similar to those observed in the thin section. The scolex microtriches might be divided into two parts, a thick proximal portion and a thinner spinal distal end, the former measuring about 0.2-0.4 micron in length, the latter measuring about 1-2 microns in length. On the extracellular surface of the proximal portion of microthrix, 1 or 2 particles could occasionally be seen, while numerous particles appeared on the protoplasmic fracture face (PF face) of the proximal portion. The PF face were closely packed and irregularly distributed with membranous particles, which were spherical or subspherical in shape, measured 0.01-0.02 micron in diameter. The extracellular fracture face (EF face), of proximal portion and all four membranous faces of spinal end were smooth. The tegumentary plasmic membrane was 0.02 micron in thickness, with smooth ES face. The PF face of this membrane packed closely with numerous particles, which were similar with those on the PF face of the microthrix. Numerous globular or discoidal vesicles scattered at random in the tegumentary matrix. Most mitochondria were near the basement membrane of the matrix. In the parenchymal layer, the parenchymal cells, calcareous corpuscles and flame-cells could easily be seen. Most of the calcareous corpuscles and flame-cells were located closely beneath the muscle layer. The flame-cell of C. cellulosae mainly consisted of three portions, cell body, a bundle of cilia and a filtered cell lumen. The flame-cell body had a large nucleus and relatively few cytoplasm with several mitochondria.(ABSTRACT TRUNCATED AT 250 WORDS)

  19. Distinction by concanavalin A agglutination between ulceration and repair of rat bladder epithelium induced by freezing or cyclophosphamide and the effect of sodium saccharin. (United States)

    Suzuki, T; Hasegawa, R; Murasaki, G; Cohen, S M


    Agglutination of rat urinary bladder epithelial cells by concanavalin A (Con A) has been reported to be an early marker of bladder carcinogenesis. Ulceration of the bladder, induced by cyclophosphamide (CP) or freezing, followed by sodium saccharin in the diet results in the induction of bladder cancer. In the present studies, the agglutination of rat urinary bladder epithelial cells by Con A was shown to be increased during the regenerative hyperplasia following ulceration induced by i.p. CP injection, but it returned to normal levels by Day 21 when the preparative process was nearly complete. This effect correlated quantitatively with the dose of CP. However, if CP administration was followed by sodium saccharin in the diet beginning 14 days after the injection, the agglutinability of bladder cells by Con A persisted. In contrast, agglutination of bladder cells by Con A during regenerative hyperplasia following ulceration induced by freezing was not increased whether sodium saccharin was fed or not. These results indicate that Con A agglutination distinguishes between the regenerative hyperplasia induced by CP or freezing, even though either method followed by sodium saccharin in the diet results in bladder cancer in the rat.

  20. Freeze/thaw-induced embolism: probability of critical bubble formation depends on speed of ice formation

    Directory of Open Access Journals (Sweden)

    Sanna eSevanto


    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.

  1. The Effect of Freezing Stress on Percentage of Electrolytes Leakage and Survival of Flixweed (Descurainia sophia L. Seedlings

    Directory of Open Access Journals (Sweden)

    E Izadi-Darbandi


    Full Text Available Introduction Flixweed (Descurainia sophia L. is a medicinal plant from Brassicaceae family which also known as a weed for winter cereals and oil seed rape. Low temperatures are one of the most important abiotic stresses that threat Flixweed growth and productivity. Therefore it is important to recognize the freeze tolerance of Flixweed for successful planting and utilization in cold regions such as Mashhad in Khorasan Razavi Province (Iran’s north. east. Among many laboratory methods which have been developed to estimate and to evaluate plants response or their tolerance to freez¬ing temperatures, electrolyte leakage (EL test is widely used. This test is based on this principle that damage to the cell membranes results in enhanced leakage of solutes into the apoplastic water, hence recording the amount of leakage after stress treatments provides an estimation of tissue injury. Indeed continuing integration of plasma membrane is one important factor for survival of plants under freezing stress and any disturbance in membrane structure can lead to damage and death. So determination of LT50 point or critical temperature for electrolytes leakage and survival of plant is the most reliable, quantitative and simple methods for evaluating the cold tolerance of plants. The aim of this trial was to determine the LT50 according to the EL and SU% for Flixweed ecotypes. Materials and Methods In order to evaluate freeze tolerance in Flixweed, a factorial experiment based on completely randomized design with three replications was carried out in college of agriculture, Ferdowsi University of Mashhad. Experimental factors included five ecotypes of Flixweed (Eghlid, Sabzewar, Hamedan, Torbat-e-Jam and Neyshabour and 10 freezing temperature levels (0, -2,-4, -6, -8,-10,-12,-14,-16 and -18°C. Flixweed seeds were cultivated in pots in autumn of 2008 and were grown until 5-7 leaf stage under natural weather conditions for acclimation. Then to apply freezing

  2. 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

  3. Dietary feeding of freeze-dried whole cranberry inhibits intestinal tumor development in Apcmin/+ mice (United States)

    Dong, Wenxiao; Zhang, Yujie; Wang, Sinan; Xie, Runxiang; Wang, Bangmao; Cao, Hailong


    It is increasingly perceived that dietary components have been linked with the prevention of intestinal cancer. Cranberry is a rich source of phenolic constituents and non-digestible fermentable dietary fiber, which shows anti-proliferation effect in colorectal cancer cells. Herein, we investigated the efficacy of long-term cranberry diet on intestinal adenoma formation in Apcmin/+ mice. Apcmin/+ mice were fed a basal diet or a diet containing 20% (w/w) freeze-dried whole cranberry powder for 12 weeks, and the number and size of tumors were recorded after sacrifice. Our results showed that cranberry strongly prevented the growth of intestinal tumors by 33.1%. Decreased cell proliferation and increased apoptosis were observed in tumors of cranberry-fed mice. Cranberry diet reduced the expression profile of colonic inflammatory cytokines (IFN-γ, IL-1β and TNF-α) accompanied with increased levels of anti-inflammatory cytokines (IL-4 and IL-10). Moreover, the number of colonic goblet cells and MUC2 production were increased, and the intestinal barrier function was also improved. In addition, cranberry diet increased caecal short chain fatty acids concentrations, and down-regulated epidermal growth factor receptor signaling pathway. These data firstly show the efficacy and associated mechanisms of cranberry diet on intestinal tumor growth in Apcmin/+ mice, suggesting its chemopreventive potential against intestinal cancer. PMID:29228651


    Directory of Open Access Journals (Sweden)

    V. V. Turov


    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.

  5. Projecting potential adoption of genetically engineered freeze-tolerant Eucalyptus in the United States (United States)

    David N. Wear; Ernest Dixon IV; Robert C. Abt; Navinder Singh


    Development of commercial Eucalyptus plantations has been limited in the United States because of the species’ sensitivity to freezing temperatures. Recently developed genetically engineered clones of a Eucalyptus hybrid, which confer freeze tolerance, could expand the range of commercial plantations. This study explores how...

  6. Freezing Point of Milk: A Natural Way to Understand Colligative Properties (United States)

    Novo, Mercedes; Reija, Belen; Al-Soufi, Wajih


    A laboratory experiment is presented in which the freezing point depression is analyzed using milk as solution. The nature of milk as a mixture of different solutes makes it a suitable probe to learn about colligative properties. The first part of the experiment illustrates the analytical use of freezing point measurements to control milk quality,…

  7. Effect of the rehydration medium on the recovery of freeze-dried lactic acid bacteria. (United States)

    de Valdez, G F; de Giori, G S; de Ruiz Holgado, A P; Oliver, G


    Sixteen cultures of lactic acid bacteria were freeze-dried in 10% nonfat skim milk plus 0.75 M adonitol and rehydrated by using different rehydration media. Marked variations in their capacity to repair cellular damage after freeze-drying were observed among the species and strains under consideration. PMID:3911901

  8. Freeze injury to roots of southern pine seedlings in the USA | South ...

    African Journals Online (AJOL)

    Roots of pine seedlings can be injured by freezing temperatures and the degree of injury is affected by genotype and stage of acclimation. Local sources of Pinus echinata and P. virginiana that were acclimated by cold temperatures were relatively freeze tolerant when grown in nurseries located where average minimum ...

  9. The effect of freezing rate on the quality of Striped Bass Spermatozoa (United States)

    Several studies have been conducted in an attempt to determine the optimal freezing rate for the cryopreservation of striped bass (Morone saxatilis) spermatozoa. In this study, the effects of freezing rate (-10, -15, -20 and -40oC/minute) on gamete quality including, viability, motion characteristi...

  10. Preliminary report on a novel straw semen freezing method for ram ...

    African Journals Online (AJOL)

    A straw (0.25ml French) semen freezing method for ram spermatozoa using a temperartre controlled stainless steel plate is described to possibly replace the conventional static liquid nitrogen vapour freezing method. 'n Strooitjie (0.25ml Franse) semenvriesmetode vir ramspermatosoa deur gebruikmaking van 'n ...

  11. Gait-related cerebral alterations in patients with Parkinson's disease with freezing of gait

    NARCIS (Netherlands)

    Snijders, A.H.; Leunissen, H.P.; Bakker, M.; Overeem, S.; Helmich, R.C.G.; Bloem, B.R.; Toni, I.


    Freezing of gait is a common, debilitating feature of Parkinson’s disease. We have studied gait planning in patients with freezing of gait, using motor imagery of walking in combination with functional magnetic resonance imaging. This approach exploits the large neural overlap that exists between

  12. Gait-related cerebral alterations in patients with Parkinson's disease with freezing of gait

    NARCIS (Netherlands)

    Snijders, A.H.; Leunissen, I.; Bakker, M.; Overeem, S.; Helmich, R.C.G.; Bloem, B.R.; Toni, I.


    Freezing of gait is a common, debilitating feature of Parkinson's disease. We have studied gait planning in patients with freezing of gait, using motor imagery of walking in combination with functional magnetic resonance imaging. This approach exploits the large neural overlap that exists between

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

    DEFF Research Database (Denmark)

    Johannesson, Björn


    Samples of concrete at different water-to-cement ratios and air contents subjected to freeze/thaw cycles with the lowest temperature at about -80 degrees C are investigated. By adopting a novel technique, a scanning calorimeter is used to obtain data from which the ice contents at different freeze...

  14. Charge-charge liquid structure factor and the freezing of alkali halides

    International Nuclear Information System (INIS)

    March, N.H.; Tosi, M.P.


    The peak height of the charge-charge liquid structure factor Ssub(QQ) in molten alkali halides is proposed as a criterion for freezing. Available data on molten alkali chlorides, when extrapolated to the freezing point suggests Ssub(QQ)sup(max) approximately 5. (author)


    NARCIS (Netherlands)

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


    Fumigation of winter wheat with H2S during low temperature acclimation substantially reduced the development of freezing tolerance of the leaves. After 6 weeks of low temperature exposure (3-degrees-C), the freezing tolerance was increased by 6 and 2-degrees-C at 0 and 0.25-mu-l l-1 H2S,

  16. Degradation of ATP and glycogen in cod ( Gadus morhua ) muscle during freezing

    DEFF Research Database (Denmark)

    Cappeln, Gertrud; Jessen, Flemming


    , 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...

  17. Heat and Mass Transfer of Droplet Vacuum Freezing Process Based on Dynamic Mesh

    Directory of Open Access Journals (Sweden)

    Lili Zhao


    Full Text Available A numerical simulation using dynamic mesh method by COMSOL has been developed to model heat and mass transfer during vacuum freezing by evaporation of a single droplet. The initial droplet diameter, initial droplet temperature, and vacuum chamber pressure effect are studied. The surface and center temperature curve was predicted to show the effect. The mass transfer rate and radius displacement were also calculated. The results show the dynamic mesh shows well the freezing process with the radius reduction of droplet. The initial droplet diameter, initial droplet temperature, and vacuum pressure have obvious effect on freezing process. The total freezing time is about 200 s, 300 s, and 400 s for droplet diameter 7.5 mm, 10.5 mm, and 12.5 mm, respectively. The vacuum pressure less than 200 Pa is enough for the less time to freezing the droplet, that is, the key point in freezing time. The initial droplet temperature has obvious effect on freezing but little effect on freezing temperature.

  18. Control of crystal growth in water purification by directional freeze crystallization (United States)

    Conlon, William M. (Inventor)


    A Directional Freeze Crystallization system employs an indirect contact heat exchanger to freeze a fraction of liquid to be purified. The unfrozen fraction is drained away and the purified frozen fraction is melted. The heat exchanger must be designed in accordance with a Growth Habit Index to achieve efficient separation of contaminants. If gases are dissolved in the liquid, the system must be pressurized.

  19. Novel Foams Based on Freeze-Dried Renewable Vital Wheat Gluten

    DEFF Research Database (Denmark)

    Blomfeldt, Thomas O.J.; Olsson, Richard T.; Menon, Mohan


    A new way of producing rigid or semi-rigid foams from vital wheat gluten using a freeze-drying process is reported. Water/gluten-based mixtures were frozen and freeze-dried. Different foam structures were obtained by varying the mixing process and wheat gluten concentration, or by adding glycerol...

  20. A Dynamic Design Space for Primary Drying During Batch Freeze-Drying

    DEFF Research Database (Denmark)

    Mortier, Séverine Thérèse F C; Van Bockstal, Pieter Jan; Nopens, Ingmar


    Biopharmaceutical products are emerging within the pharmaceutical industry. However, biopharmaceuticals are often unstable in aqueous solution. Freeze-drying (lyophilisation) is the preferred method to achieve a stable product with an increased shelf-life. During batch freeze-drying, there are only...

  1. Direct comparative analysis of conventional and directional freezing for the cryopreservation of whole ovaries

    NARCIS (Netherlands)

    Maffei, S.; Hanenberg, M.; Pennarossa, G.; Roberto V. Silva, J.; Tiziana, A.; Brevini, L.; Pharm, D.; Arav, A.; Gandolfi, F.


    INTERVENTION(S): Eighty-one ovaries were randomly assigned to fresh control, conventional freezing (CF), and directional freezing (DF) group. Ovaries of CF and DF groups were perfused via the ovarian artery with Leibovitz L-15 medium, 10% fetal bovine serum, and 1.5 M dimethyl sulfoxide for 5

  2. Cognitive Contributions to Freezing of Gait in Parkinson Disease: Implications for Physical Rehabilitation. (United States)

    Peterson, Daniel S; King, Laurie A; Cohen, Rajal G; Horak, Fay B


    People with Parkinson disease (PD) who show freezing of gait also have dysfunction in cognitive domains that interact with mobility. Specifically, freezing of gait is associated with executive dysfunction involving response inhibition, divided attention or switching attention, and visuospatial function. The neural control impairments leading to freezing of gait have recently been attributed to higher-level, executive and attentional cortical processes involved in coordinating posture and gait rather than to lower-level, sensorimotor impairments. To date, rehabilitation for freezing of gait primarily has focused on compensatory mobility training to overcome freezing events, such as sensory cueing and voluntary step planning. Recently, a few interventions have focused on restitutive, rather than compensatory, therapy. Given the documented impairments in executive function specific to patients with PD who freeze and increasing evidence of overlap between cognitive and motor function, incorporating cognitive challenges with mobility training may have important benefits for patients with freezing of gait. Thus, a novel theoretical framework is proposed for exercise interventions that jointly address both the specific cognitive and mobility challenges of people with PD who freeze. © 2016 American Physical Therapy Association.

  3. Low-Temperature Failure Mode for Nickel-Hydrogen Cells

    National Research Council Canada - National Science Library

    Zimmerman, A. H


    .... It has been shown that there are chemical processes that can occur within the operating nickel-hydrogen cell that can raise the electrolyte freezing point in some cell designs up to the -5 to -10...

  4. Bilateral maxillary sinus floor augmentation with tissue-engineered autologous osteoblasts and demineralized freeze-dried bone

    Directory of Open Access Journals (Sweden)

    Aashish Deshmukh


    Full Text Available The pneumatization of the maxillary sinus often results in a lack of sufficient alveolar bone for implant placement. In the last decades, maxillary sinus lift has become a very popular procedure with predictable results. Sinus floor augmentation procedures are generally carried out using autologous bone grafts, bone substitutes, or composites of bone and bone substitutes. However, the inherent limitations associated with each of these, have directed the attention of investigators to new technologies like bone tissue engineering. Bone marrow stromal cells have been regarded as multi-potent cells residing in bone marrow. These cells can be harvested from a person, multiplied outside his body using bioengineering principles and technologies and later introduced into a tissue defect. We present a case where tissue-engineered autologous osteoblasts were used along with demineralized freeze-dried bone for sinus floor augmentation.

  5. Spacecraft Radiator Freeze Protection Using a Regenerative Heat Exchanger (United States)

    Ungar, Eugene K.; Schunk, Richard G.


    An active thermal control system architecture has been modified to include a regenerative heat exchanger (regenerator) inboard of the radiator. Rather than using a radiator bypass valve a regenerative heat exchanger is placed inboard of the radiators. A regenerator cold side bypass valve is used to set the return temperature. During operation, the regenerator bypass flow is varied, mixing cold radiator return fluid and warm regenerator outlet fluid to maintain the system setpoint. At the lowest heat load for stable operation, the bypass flow is closed off, sending all of the flow through the regenerator. This lowers the radiator inlet temperature well below the system set-point while maintaining full flow through the radiators. By using a regenerator bypass flow control to maintain system setpoint, the required minimum heat load to avoid radiator freezing can be reduced by more than half compared to a radiator bypass system.

  6. Freeze-Casting of Porous Biomaterials: Structure, Properties and Opportunities

    Directory of Open Access Journals (Sweden)

    Sylvain Deville


    Full Text Available The freeze-casting of porous materials has received a great deal of attention during the past few years. This simple process, where a material suspension is simply frozen and then sublimated, provides materials with unique porous architectures, where the porosity is almost a direct replica of the frozen solvent crystals. This review focuses on the recent results on the process and the derived porous structures with regards to the biomaterials applications. Of particular interest is the architecture of the materials and the versatility of the process, which can be readily controlled and applied to biomaterials applications. A careful control of the starting formulation and processing conditions is required to control the integrity of the structure and resulting properties. Further in vitro and in vivo investigations are required to validate the potential of this new class of porous materials.

  7. Freezing heat transfer within water-saturated porous media

    International Nuclear Information System (INIS)

    Sasaki, Akira; Aiba, Shinya; Fukusako, Shoichiro.


    In the present study, analytical and experimental investigations were performed so as to clarify the characteristics of freezing heat transfer in porous media saturated with water in a vertical rectangular cavity. In order to establish the momentum equation, the law of conservation of momentum was applied to the fluid in our control volume, and the equation took into account Forchheimer's extension as the resistance to flow in the porous media. Three different sizes of glass, iron and copper beads were used as the porous media in this study. The temperature of the cold wall was kept at -10degC, while that of the hot wall was varied from 2degC to 22 degC. Comparisons between the analytical results and the experimental ones show good agreement with the exception of the copper bead results. (author)

  8. Diagnosing and curing system freeze-ups: Part II

    Energy Technology Data Exchange (ETDEWEB)

    Frey, R. [Heating and Plumbing Service, Sparta, NJ (United States)


    In our last article, we discussed determination and possible causes leading to frozen pipes in the heating system. We now turn to the matter of dealing with such freeze-ups. There are two major categories of frozen heating systems. The first and worst we can label the {open_quotes}Floridian Flew{close_quotes} type, when the usual occupants have taken off for warmer climates. The second is the {open_quotes}Howcudit {open_quote}B{close_quote} virus,{close_quotes} which occurs to normal people under an unusual Arctic Attack right while they are living there. Regarding the first, I never cease to be amazed how folks can just up and leave their house for three days to three months without making proper arrangements for monitoring the temperature of their home during their absence. There are a frightening number of humanoids out there, who either lack a minimal presence of common sense, or who are terrible gamblers. The amazing thing is that some of these folks wouldn`t even gamble a postage stamp on a magazine sweepstakes, but still will gamble the contents of their home by driving off for two weeks at Disneyland in the middle of January without ever making any provision for assuring that their house doesn`t freeze up while they are away. The {open_quotes}Howcudit {open_quote}B{close_quote}{close_quotes} type is not nearly as devasting as the {open_quotes}Floridian Flew.{close_quotes} Imagine coming home to an icy cold house after flying in from the sunny beaches of some tropical paradise. The oilburner switch is on. The thermostate is set at 60 where we left it, but the needle is buried somewhere down behind the cover, like it is cowering from guilt. {open_quotes}Oh Man! I just checked the bathroom; there is an icicle hanging from the vanity faucet and the toilet bowl is a block of ice.{close_quotes}

  9. A Numerical Analysis on Freezing Behavior of Flowing Water inside a Pipe Cooled from Surroundings (United States)

    Chiba, Ryoichi; Izumi, Masaaki

    A freezing phenomenon in forced convectional flow inside a pipe is investigated numerically in this paper .The numerical analysis is carried out to assess the transient freezing behavior of flowing water inside a pipe cooled from surroundings in consideration of pressure drop caused by the freezing. The finite element technique is applied to solve the equations of motion and energy transport for laminar flow. The numerical model attempts to capture the solid-fluid interface on a fixed computational grid. The correlations among cooling conditions of pipe, velocity and temperature of water at the inlet, and location at which the freezing starts are examined to show the critical velocity to avoid freezing. In addition, under the condition that pressure remains constant at the inlet, the period in which the pipe is not blockaded by ice is calculated. The period is illustrated with some dimensionless parameters to predict the conditions under which blockage occurs within a given time.

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

    Directory of Open Access Journals (Sweden)

    Xinbao Yu


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

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

    DEFF Research Database (Denmark)

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


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

  12. Limited Impact of Subglacial Supercooling Freeze-on for Greenland Ice Sheet Stratigraphy (United States)

    Dow, Christine F.; Karlsson, Nanna B.; Werder, Mauro A.


    Large units of disrupted radiostratigraphy (UDR) are visible in many radio-echo sounding data sets from the Greenland Ice Sheet. This study investigates whether supercooling freeze-on rates at the bed can cause the observed UDR. We use a subglacial hydrology model to calculate both freezing and melting rates at the base of the ice sheet in a distributed sheet and within basal channels. We find that while supercooling freeze-on is a phenomenon that occurs in many areas of the ice sheet, there is no discernible correlation with the occurrence of UDR. The supercooling freeze-on rates are so low that it would require tens of thousands of years with minimal downstream ice motion to form the hundreds of meters of disrupted radiostratigraphy. Overall, the melt rates at the base of the ice sheet greatly overwhelm the freeze-on rates, which has implications for mass balance calculations of Greenland ice.

  13. Microstructure study of a material on the basis of YSZ obtained be freeze-drying

    International Nuclear Information System (INIS)

    Rizea, A.; Abrudeanu, M.; Petot, C.; Petot Ervas, G.


    Freeze-drying is a dehydration proceeding of the products in a frozen state, which is based on the ice sublimation process. It is a method, which leads to a very good homogeneity of the products and it allows obtaining very fine powders, which directs to reducing the sintering temperature. Freeze drying always supposes three stages: - freezing, sublimation and absorption of the residual water. The preparation of ZrO 20.91 Y 2 O 30.09 samples proceeds through the following stages: - a. solution preparation; b. solution spraying (into small droplets in liquid nitrogen); c. freeze drying processing; d. calcination of the freeze dried powder; e. powder compacting; f. sintering at four different temperature. The different structure of samples with different density are characterized on basis of micrographs. The results of these analyses are presented, discussed and explained through the chemical composition of the samples

  14. Freeze-drying-induced changes in the properties of graphene oxides

    International Nuclear Information System (INIS)

    Ham, Heon; Van Khai, Tran; Gil Na, Han; Jung Kwon, Yong; Yeon Cho, Hong; Woo Kim, Hyoun; Park, No-Hyung; So, Dae Sup; Lee, Joon-Woo


    We have characterized and evaluated changes in graphene oxide (GO) induced by means of freeze-drying. In order to evaluate these changes, we investigated the effects of freeze-drying and chemical reduction processes on the structure, morphology, chemical composition, and Raman properties of GO and reduced GO. The freeze-dried GO had a pore structure, maintaining a pored morphology even after thermal annealing. The freeze-dried samples were composed of a single folded nanosheet or a few nanosheets stacked and folded. The oxygen-containing functional groups were removed not only during the freeze-drying but also during the reduction processes, with an accompanying decrease in the average size of the sp 2 carbon domain (i.e. an increase in the I D /I G value). (papers)


    Directory of Open Access Journals (Sweden)

    O. L. Lukoyanova


    Full Text Available Evaluation of prolonged storage of breast milk safety for nutrition of newborns remains actual problem of modern nutritiology. Objectives: to evaluate an influence of freezing and prolonged storage of expressed breast milk on its nutritive, biological values and microbiological safety. Methods: samples of expressed breast milk (native samples and after 1 and 3 months of storage in containers Philips AVENT in t -18°С were analyzed; levels of secretory IgA, lysozyme, transforming growth factor (TGF β1, potassium, calcium, magnesium, pH, buffer capacity and bacterial cells were detected. Results: there is no statistically significant influence of low temperature and prolonged storage of expressed breast milk on levels of secretory IgA, lysozyme, TGF β1 and bacterial cells. Potassium and calcium levels significantly decreased (on about 10% and 20% accordingly, pH increased, and buffer capacity of milk lowered after freezing during 3 months. Conclusion: children’s feeding with native and expressed breast milk defrosted after 3 months of storage is equal in nutritive and biological values and microbiological safety.

  16. Exploring the Possibility of Cryopreservation of Feline and Canine Erythrocytes by Rapid Freezing with Penetrating and Non-Penetrating Cryoprotectants.

    Directory of Open Access Journals (Sweden)

    Denys Pogozhykh

    Full Text Available Efficient application of veterinary blood transfusion approaches for small companion animals requires readily available supply of the donor material. This can be achieved by developing of effective biobanking technologies allowing long-term storage of donor blood components via cryopreservation. Transfusion of an erythrocyte concentrate allows the successful correction of various hematological pathologies, severe bleeding, and etc. While in the past there were several approaches to cryopreserve red blood cells of dogs, to our knowledge there is virtually no data on cryopreservation of feline erythrocytes. In this paper, we performed a comprehensive parameter optimization for low temperature storage of RBCs of both species. Here, the efficiency of single-component and multicomponent cryoprotective media as well as necessary time of pre-incubation with penetrating and non-penetrating cryoprotectants prior to rapid freezing is analyzed. This study showed that glycerol was not sufficient for cryopreservation of red blood cells of the studied species under the investigated conditions. Application of 10% (v/v ME2SO allowed for a significant reduction of canine and feline erythrocytes hemolysis after thawing. 17.5% hydroxyethyl starch demonstrated the highest cryoprotective activity for both species. It was found that dog RBCs should be incubated in cryoprotective media for 30 min at 22°C prior to freezing, while for cat RBCs 20 min is sufficient. Combination of CPAs was less effective. Presented data may be considered in further studies in veterinary transfusion and blood banking optimization.

  17. The freezing point of raw and heat treated sheep milk and its variation during lactation

    Directory of Open Access Journals (Sweden)

    Bohumíra Janštová


    Full Text Available The freezing point of milk is an important indicator of the adulteration of the milk with water, but heat treatment may also affect its value. The aim of this study was determine freezing point of raw and heat treated sheep milk and its variation during lactation. The freezing point was determined in 42 bulk tank raw sheep milk samples and 42 pasteurized milk samples collected during lactation of sheep at one ecofarm in Moravian Walachia (Valašsko in the Czech Republic. The freezing point was determined in accordance with the standard ČSN 57 0538 using a thermistor cryoscope. The average freezing point of raw milk was -0.617 ± 0.052 °C, with a range from -0.560 to -0.875 °C. The freezing point was lower in the first months of lactation and increased at the end of lactation. The freezing point correlated (r = 0.8967 with the content of total non-fat solids. The average freezing point of sheep milk pasteurized at 65 °C for 30 min was -0.614 ± 0.053 °C, with a range from -0.564 to -0.702 °C. The median of freezing point differences between raw and pasteurized milk was 0.004 °C. Our study extends data about physico-chemical properties of sheep milk and registers for the first time specific changes in the freezing point value of sheep milk by heating.

  18. Cryoprotectant-free ultra-rapid freezing of human spermatozoa in cryogenic vials. (United States)

    Zhu, J; Jin, R-T; Wu, L-M; Johansson, L; Guo, T-H; Liu, Y-S; Tong, X-H


    The objective of this study was to investigate the effect of ultra-rapid freezing (direct immersion in liquid nitrogen) on human spermatozoa in cryogenic vials (≥0.5 ml) at different concentrations of sucrose. After swim-up, the sperm suspensions (N = 58) were diluted with sperm preparation medium and divided into six aliquots: swim-up (fresh), conventional freezing group (slow freezing) and four ultra-rapid freezing groups containing sucrose at different concentrations (0.15 m, 0.20 m, 0.25 m and 0.30 m). Sperm motility, progressive motility, plasma membrane integrity, DNA stability and acrosome integrity of fresh and cooled-warmed spermatozoa were analysed. The progressive motility, plasma membrane and acrosome integrity of spermatozoa in the 0.20 m sucrose group were significantly higher than those of the slow freezing group (47.5 ± 6.8% versus 36.4 ± 8.7%, 73.2 ± 6.9% versus 63.9 ± 6.3%, 53.7 ± 10.0% versus 35.9 ± 9.7% respectively, P 0.05 respectively) between the 0.20 m sucrose and the slow freezing group. No differences were found between the ultra-rapid and slow freezing group at the other concentrations of sucrose. Our findings suggest that the method of ultra-rapid freezing of human spermatozoa in cryogenic vials with a solution containing 0.20 m sucrose results in recovery of spermatozoon of superior qualities. In contrast to slow freezing, the ultra-rapid freezing technique of human spermatozoa seems to reduce cryoinjuries and maintain important physiological characteristics of the spermatozoa after warming. © 2013 Blackwell Verlag GmbH.

  19. Chlorophyll fluorescence imaging accurately quantifies freezing damage and cold acclimation responses in Arabidopsis leaves

    Directory of Open Access Journals (Sweden)

    Hincha Dirk K


    Full Text Available Abstract Background Freezing tolerance is an important factor in the geographical distribution of plants and strongly influences crop yield. Many plants increase their freezing tolerance during exposure to low, nonfreezing temperatures in a process termed cold acclimation. There is considerable natural variation in the cold acclimation capacity of Arabidopsis that has been used to study the molecular basis of this trait. Accurate methods for the quantitation of freezing damage in leaves that include spatial information about the distribution of damage and the possibility to screen large populations of plants are necessary, but currently not available. In addition, currently used standard methods such as electrolyte leakage assays are very laborious and therefore not easily applicable for large-scale screening purposes. Results We have performed freezing experiments with the Arabidopsis accessions C24 and Tenela, which differ strongly in their freezing tolerance, both before and after cold acclimation. Freezing tolerance of detached leaves was investigated using the well established electrolyte leakage assay as a reference. Chlorophyll fluorescence imaging was used as an alternative method that provides spatial resolution of freezing damage over the leaf area. With both methods, LT50 values (i.e. temperature where 50% damage occurred could be derived as quantitative measures of leaf freezing tolerance. Both methods revealed the expected differences between acclimated and nonacclimated plants and between the two accessions and LT50 values were tightly correlated. However, electrolyte leakage assays consistently yielded higher LT50 values than chlorophyll fluorescence imaging. This was to a large part due to the incubation of leaves for electrolyte leakage measurements in distilled water, which apparently led to secondary damage, while this pre-incubation was not necessary for the chlorophyll fluorescence measurements. Conclusion Chlorophyll

  20. Critical Radius of Supercooled Water Droplets: On the Transition toward Dendritic Freezing. (United States)

    Buttersack, Tillmann; Bauerecker, Sigurd


    The freezing of freely suspended supercooled water droplets with a diameter of bigger than a few micrometers splits into two rather different freezing stages. Within the first very fast dendritic freezing stage a spongy network ice with an ice portion of less than one-third forms and more than two-thirds of liquid water remain. In the present work the distribution of the ice portion in the droplet directly after the dendritic freezing phase as well as the evolution of the ice and temperature distribution has been investigated in dependence of the most relevant parameters as droplet diameter, dendritic freezing velocity (which correlates with the supercooling) and heat transfer coefficient to the surroundings (which correlates with the relative droplet velocity compared to the ambient air and with the droplet size). For this purpose on the experimental side acoustically levitated droplets in climate chambers have been investigated in combination with high-speed cameras. The obtained results have been used for finite element method (FEM) simulations of the dendritic freezing phase under consideration of the beginning second, much slower heat-transfer dominated freezing phase. A theoretical model covering 30 layers and 5 shells of the droplet has been developed which allows one to describe the evolution of both freezing phases at the same time. The simulated results are in good agreement with experimental as well as with calculated results exploiting the heat balance equation. The most striking result of this work is the critical radius of the droplet which describes the transition of one-stage freezing of the supercooled water droplet toward the thermodynamically forced dendritical two-stage freezing in which the droplet cannot sufficiently get rid of the formation heat anymore. Depending on the parameters named above this critical radius was found to be in the range of 0.1 to 1 μm by FEM simulation.