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Sample records for thawed frozen human

  1. Thawing of Frozen Tuna Meat

    Science.gov (United States)

    Tanaka, Takeo; Nishiwaki, Kôji; Kakuda, Kitonari; Tomimatsu, Takao

    Frozen southern bluefin tuna meat discolors easily and sometimes contracts when thawed caused by thaw rigor. These phenomenon often become problematic in the transaction or handling of this kind of frozen tuna. Frozen meat blocks of southern Bluefin tuna were thawed separately by air thawing, running water thawing and microwave thawing. Changes occurring during thawing were checked for meat color by met-myoglobin ratio determination and for contract by microscopic observation. Results are as follows : (1) Discoloration scarcely occurred in the process of running water thawing (at 10°C for 50 min, or at 0°C for 6 hr). (2) No contraction was observed during thawing with running water described above and air thawing (at 18-20°C for 6 hr). (3) Discoloration and contraction seemed to be minimized, as to latently contractile blocks, when meat temperature passed through rapidly between -10°C and -5°C, and slowly (for 5-6 hr) between -5°C and -1°C. When the block was originally not contractile, discloration was minimized by rising meat temperature rapidly from -10°C to -l°C.

  2. Breed differences of bull frozen-thawed semen.

    Science.gov (United States)

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

    2016-12-01

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

  3. Transfer of human frozen-thawed embryos with further cleavage during culture increases pregnancy rates

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    Bharat V Joshi

    2010-01-01

    Full Text Available Aim: To compare the pregnancy rate following transfer of frozen-thawed embryos with or without overnight culture after thawing. Settings and Design: This is a retrospective analysis of frozen-thawed embryo transfer (FET cycles performed between January 2006 and December 2008. Materials and Methods: Out of 518 thaw cycles, 504 resulted in embryo transfers (ETs. Of the total FET cycles, 415 were performed after an overnight culture of embryos (group A; and in 89 cycles, ET was performed within 2 hours of embryo thawing (group B. Statistical Analysis: The data were statistically analyzed using chi-square test. Results: We observed that with FET, women ≤30 years of age had a significantly higher (P=0.003 pregnancy rate (PR=28.9% as compared to women >30 years of age (17.5%. A significantly higher (P<0.001FNx08 pregnancy rate was also observed in women receiving 3 frozen-thawed embryos (29% as compared to those who received less than 3 embryos (10.7%. The difference in PR between group A (PR=24.3% and group B (PR=20.3% was not statistically significant. However, within group A, ET with cleaved embryos showed significantly ( P≤0.01 higher pregnancy rate compared to the uncleaved embryos, depending on the number of cleaved embryos transferred. Conclusion: No significant difference was noticed between FETs made with transfer of embryos with overnight culture and those without culture. However, within the cultured group, transfer of embryos cleaved during overnight culture gave significantly higher PR than transfers without any cleavage.

  4. In vitro development of donated frozen-thawed human embryos in a prototype static microfluidic device: a randomized controlled trial

    NARCIS (Netherlands)

    Kieslinger, Dorit C.; Hao, Zhenxia; Vergouw, Carlijn G.; Kostelijk, Elisabeth H.; Lambalk, Cornelis B.; le Gac, Severine

    Objective: To compare the development of human embryos in microfluidic devices with culture in standard microdrop dishes, both under static conditions. Design: Prospective randomized controlled trial. Setting: In vitro fertilization laboratory. Patient(s): One hundred eighteen donated frozen-thawed

  5. Cooking frozen and thawed roasts: beef, pork, and lamb cuts.

    Science.gov (United States)

    Fulton, C; Davis, C

    1975-09-01

    Cooking time, yield, and palatability of paired beef, pork, and lamb roasts cooked from the frozen and thawed states were compared. Cooking time for all roasts averaged from 3 to 22 min. per pound longer for meat cooked from the frozen state. The longer cooking time from the frozen state. The longer cooking time from the frozen state was greater for roasts with a large amount of bone and for cuts cooked by braising than for less bony roasts and cuts cooked by roasting. Except for thawed beef rump roasts, which had a higher yield of cooked lean meat, yield of cooked lean meat from the various cuts of beef, pork, and lamb was not affected by the state at the start of cooking. Collectively, all pork roasts had a higher yield of cooked lean meat when cooked from the frozen state. Juiciness and natural flavor of the roasts were not affected by the state at the start of cooking. Lamb leg and rib roasts were more tender when cooked from the thawed state.

  6. Successful artificial insemination in the Asian elephant (Elephas maximus using chilled and frozen-thawed semen

    Directory of Open Access Journals (Sweden)

    Wongkalasin Warut

    2009-07-01

    Full Text Available Abstract Background Artificial insemination (AI using frozen-thawed semen is well established and routinely used for breeding in various mammalian species. However, there is no report of the birth of elephant calves following AI with frozen-thawed semen. The objective of the present study was to investigate the fertilizing ability of chilled and frozen-thawed semen in the Asian elephant following artificial insemination (AI. Methods Semen samples were collected by from 8 bulls (age range, 12-to 42-years by manual stimulation. Semen with high quality were either cooled to 4°C or frozen in liquid nitrogen (-196°C before being used for AI. Blood samples collected from ten elephant females (age range, 12-to 52-years were assessed for estrus cycle and elephants with normal cycling were used for AI. Artificial insemination series were conducted during 2003 to 2008; 55 and 2 AI trials were conducted using frozen-thawed and chilled semen, respectively. Pregnancy was detected using transrectal ultrasonography and serum progestagen measurement. Results One female (Khod inseminated with chilled semen became pregnant and gave birth in 2007. The gestation length was 663 days and the sex of the elephant calf was male. One female (Sao inseminated with frozen-thawed semen showed signs of pregnancy by increasing progestagen levels and a fetus was observed for 5 months by transrectal ultrasonography. Conclusion This is the first report showing pregnancy following AI with frozen-thawed semen in the Asian elephant. Successful AI in the Asian elephant using either chilled or frozen-thawed semen is a stepping stone towards applying this technology for genetic improvement of the elephant population.

  7. Evaluation of the ovarian reserve in women transplanted with frozen and thawed ovarian cortical tissue

    DEFF Research Database (Denmark)

    Greve, Tine; Schmidt, Kirsten Tryde; Kristensen, Stine Gry

    2012-01-01

    To investigate ovarian reserve and ovarian function in women transplanted with frozen/thawed ovarian tissue.......To investigate ovarian reserve and ovarian function in women transplanted with frozen/thawed ovarian tissue....

  8. Thawing and storage system for frozen food in the food service industries

    Energy Technology Data Exchange (ETDEWEB)

    Komada, Hitoshi; Tamura, Toshiyuki

    1988-02-01

    Thawing and storage system for frozen food in the food service industries was studied. The air flowing system was adequate for thawing owing to the convenience and quality retaining. Thawing time could not be reduced at 98% humidity and over 5/sup 0/C, even if the thawing temperature was elevated, and was lengthened at 5/sup 0/C or less. Further the thawing time could not be reduced by increasing the flowing speed at 1m/s or more. When the humidity was 65%, the thawing time was lengthened by 15%. The free drip was increased linearly with the increase of thawing temperature, so that a little higher thawing temperature than the frozen one was preferable. After the completion of thawing, the food was cooled suddenly in the pressing process to uniform the thawing and prevent the surface drip. Considering the structure decomposition and change, the storage temperature of -1/sup 0/C and the variation of +0.5/sup 0/C were desirable. The newly developed high humidity thawing cabinet resulted the better recovery of cell structure, less dripping amount and high quality thawing. A Super Chiller employing an inverter compressor resulted lower temperature variation and was effective for beef curing. (25 figs, 7 tabs, 4 refs)

  9. Comparison of enterotomy leak pressure among fresh, cooled, and frozen-thawed porcine jejunal segments.

    Science.gov (United States)

    Aeschlimann, Kimberly A; Mann, F A; Middleton, John R; Belter, Rebecca C

    2018-05-01

    OBJECTIVE To determine whether stored (cooled or frozen-thawed) jejunal segments can be used to obtain dependable leak pressure data after enterotomy closure. SAMPLE 36 jejunal segments from 3 juvenile pigs. PROCEDURES Jejunal segments were harvested from euthanized pigs and assigned to 1 of 3 treatment groups (n = 12 segments/group) as follows: fresh (used within 4 hours after collection), cooled (stored overnight at 5°C before use), and frozen-thawed (frozen at -12°C for 8 days and thawed at room temperature [23°C] for 1 hour before use). Jejunal segments were suspended and 2-cm enterotomy incisions were made on the antimesenteric border. Enterotomies were closed with a simple continuous suture pattern. Lactated Ringer solution was infused into each segment until failure at the suture line was detected. Leak pressure was measured by use of a digital transducer. RESULTS Mean ± SD leak pressure for fresh, cooled, and frozen-thawed segments was 68.3 ± 23.7 mm Hg, 55.3 ± 28.1 mm Hg, and 14.4 ± 14.8 mm Hg, respectively. Overall, there were no significant differences in mean leak pressure among pigs, but a significant difference in mean leak pressure was detected among treatment groups. Mean leak pressure was significantly lower for frozen-thawed segments than for fresh or cooled segments, but mean leak pressure did not differ significantly between fresh and cooled segments. CONCLUSIONS AND CLINICAL RELEVANCE Fresh porcine jejunal segments or segments cooled overnight may be used for determining intestinal leak pressure, but frozen-thawed segments should not be used.

  10. Resumption of mitosis in frozen-thawed embryos is not related to the chromosomal constitution

    DEFF Research Database (Denmark)

    Agerholm, Inge E; Kølvrå, Steen; Crüger, Dorthe G

    2007-01-01

    OBJECTIVE: To study the relation between the resumption of mitosis after thaw and chromosomal constitution in frozen-thawed embryos. In addition, to evaluate the correlation among the three parameters of resumption of mitosis after thaw, postthaw blastomere loss, and multinucleation. DESIGN: Frozen......(S): Forty IVF and/or intracytoplasmic sperm injection patients. INTERVENTION(S): Embryo thawing, morphological evaluation, and fluorescence in situ hybridization analysis for aneuploidy screening. MAIN OUTCOME MEASURE(S): Resumption of mitosis, blastomere loss, multinucleation, and chromosome enumeration....... RESULT(S): No difference was observed in the chromosomal constitution of embryos with and without resumption of mitosis. Neither was the postthaw blastomere loss connected to the chromosomal constitution. The resumption of mitosis was not associated with postthaw loss of blastomeres...

  11. Frozen-Thawed Embryo Transfer Cycles Have a Lower Incidence of Ectopic Pregnancy Compared With Fresh Embryo Transfer Cycles.

    Science.gov (United States)

    Zhang, Xinyu; Ma, Caihong; Wu, Zhangxin; Tao, Liyuan; Li, Rong; Liu, Ping; Qiao, Jie

    2017-01-01

    To evaluate the risk of ectopic pregnancy of embryo transfer. A retrospective cohort study on the incidence of ectopic pregnancy in fresh and frozen-thawed embryo transfer cycles from January 1 st , 2010, to January 1 st , 2015. Infertile women undergoing frozen-thawed transfer cycles or fresh transfer cycles. In-vitro fertilization, fresh embryo transfer, frozen-thawed embryo transfer, ectopic pregnancy. Ectopic pregnancy rate and clinical pregnancy rate. A total of 69 756 in vitro fertilization-embryo transfer cycles from 2010 to 2015 were analyzed, including 45 960 (65.9%) fresh and 23 796 (34.1%) frozen-thawed embryo transfer cycles. The clinical pregnancy rate per embryo transfer was slightly lower in fresh embryo transfer cycles compared with frozen-thawed embryo transfer cycles (40.8% vs 43.1%, P cycles, blastocyst transfer shows a significantly lower incidence of ectopic pregnancy (0.8% vs 1.8%, P = .002) in comparison with day 3 cleavage embryo transfer. The risk of ectopic pregnancy is lower in frozen-thawed embryo transfer cycles than fresh embryo transfer cycles, and blastocyst transfer could further decrease the ectopic pregnancy rate in frozen-thawed embryo transfer cycles.

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

    Science.gov (United States)

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

    2017-02-01

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

  13. Basic study on promotion of thawing frozen soil by shock loading

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    Toshiaki WATANABE

    2008-06-01

    Full Text Available The aim of study is to confirm a new technique that can crush the frozen soil and/or ice block using underwater shock wave generated by the underwater explosion of explosive. This technique can lead to the earlier sowing, which can have the larger harvest because the duration of sunshine increases. Especially, in Hokkaido prefecture, Japan, if the sowing is carried out in April, we can expect to have 150% of harvest in the ordinary season. In the case of small processing area such as road repairing, frozen soil is thawed by using the heat of gas burner and/or the electric heater. It is not a suitable plan to apply these heating methods to agriculture, from the point of view enormous amount of processing area. Thawing technique for frozen soil is effective against the cold regions, for example, Russia, Norway, and Sweden, etc. At first, we carried out experiments using a detonating fuse and ice block. The propagation process of shock wave into the ice block was observed by means of a high-speed camera. In order to check about that influence we tried to give an actual frozen soil a shock wave. We could get a result that existence of water layer serves an important role in promotion of thawing by the shock loading to the frozen soil.

  14. Triggering of final oocyte maturation with gonadotropin-releasing hormone agonist or human chorionic gonadotropin. Live birth after frozen-thawed embryo replacement cycles

    DEFF Research Database (Denmark)

    Griesinger, Georg; Kolibianakis, E M; Papanikolaou, E G

    2007-01-01

    OBJECTIVE: To report the outcome of frozen-thawed embryo replacement cycles after GnRH-agonist triggering of final oocyte maturation in the collecting cycle with GnRH-antagonist. DESIGN: Prospective, observational, multicentric clinical study. SETTING: Tertiary university-affiliated IVF centers...... a total of 228 participants. Surplus embryos or oocytes at the pronuclear stage were cryopreserved in 53 patients after hCG administration and 32 patients after GnRH-agonist administration on the basis of patient choice, pronuclear/embryo availability, and local laws. INTERVENTION(S): Transfer of frozen......-thawed embryos. MAIN OUTCOME MEASURE(S): Live birth rate. RESULT(S): Thirty-one and 23 patients after administration of hCG and GnRH-agonist, respectively, started a frozen-embryo replacement cycle by September 2005, with 25 and 16 patients eventually undergoing at least one frozen-thawed ET. Live birth rate per...

  15. Use of frozen-thawed semen in equine artificial insemination programmes

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    Alonso A

    2016-12-01

    Full Text Available The use of equine frozen semen has variable results. Pregnancy rates per cycle may be between 10 to 70%. Although an inseminating dose of 280 a 320 millones progressive motile sperm (PMS has been recommended, under experimental conditions it is posible to obtain acceptable results with doses of 50 to 100 millones PMS. Deep intrauterine insemination is prefered when using frozen thawed semen, which consists in depositing the dose at the tip of the horn, ipsiltateral to ovulation. Young mares usually have better uterine condition and that increase their chance of become pregnant. It is important to make a correct follicular monitoring, in order to determine the right time to induce ovulation. Ovulation may be induced with Human Chorionic Gonadotrophin (hCG or deslorelin. It posible to perform a fixed time insemination or to choose an ultrasound follicular following protocol, inseminating within 6 h of ovulation. This option offers the best prenancy rates. Our laboratory manages a protocol which includes ultrasound monitoring every 6 hours, beginning 24 hous after induction of ovulation; this protocol had a first insemination pregnancy rate of 73%. If we consider that one of the main disadvantages of frozen-thawed semen is decreased fertility, these results are encouraging, since they are similar to those obtained with fresh or cooled semen, so it may represent an advantage when having stored semen of stallions that are dead, or have been sold or even castrated.

  16. In vitro development of donated frozen-thawed human embryos in a prototype static microfluidic device: a randomized controlled trial.

    Science.gov (United States)

    Kieslinger, Dorit C; Hao, Zhenxia; Vergouw, Carlijn G; Kostelijk, Elisabeth H; Lambalk, Cornelis B; Le Gac, Séverine

    2015-03-01

    To compare the development of human embryos in microfluidic devices with culture in standard microdrop dishes, both under static conditions. Prospective randomized controlled trial. In vitro fertilization laboratory. One hundred eighteen donated frozen-thawed human day-4 embryos. Random allocation of embryos that fulfilled the inclusion criteria to single-embryo culture in a microfluidics device (n = 58) or standard microdrop dish (n = 60). Blastocyst formation rate and quality after 24, 28, 48, and 72 hours of culture. The percentage of frozen-thawed day-4 embryos that developed to the blastocyst stage did not differ significantly in the standard microdrop dishes and microfluidic devices after 28 hours of culture (53.3% vs. 58.6%) or at any of the other time points. The proportion of embryos that would have been suitable for embryo transfer was comparable after 28 hours of culture in the control dishes and microfluidic devices (90.0% vs. 93.1%). Furthermore, blastocyst quality was similar in the two study groups. This study shows that a microfluidic device can successfully support human blastocyst development in vitro under static culture conditions. Future studies need to clarify whether earlier stage embryos will benefit from the culture in microfluidic devices more than the tested day-4 embryos because many important steps in the development of human embryos already take place before day 4. Further improvements of the microfluidic device will include parallel culture of single embryos, application of medium refreshment, and built-in sensors. NTR3867. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  17. Increasing vaginal progesterone gel supplementation after frozen-thawed embryo transfer significantly increases the delivery rate

    DEFF Research Database (Denmark)

    Alsbjerg, Birgit; Polyzos, Nikolaos P; Elbaek, Helle Olesen

    2013-01-01

    The aim of this study was to evaluate the reproductive outcome in patients receiving frozen-thawed embryo transfer before and after doubling of the vaginal progesterone gel supplementation. The study was a retrospective study performed in The Fertility Clinic, Skive Regional Hospital, Denmark....... A total of 346 infertility patients with oligoamenorrhoea undergoing frozen-thawed embryo transfer after priming with oestradiol and vaginal progesterone gel were included. The vaginal progesterone dose was changed from 90mg (Crinone) once a day to twice a day and the reproductive outcome during the two...... rate (8.7% versus 20.5%, respectively; P=0.002). Doubling of the vaginal progesterone gel supplementation during frozen-thawed embryo transfer cycles decreased the early pregnancy loss rate, resulting in a significantly higher delivery rate. This study evaluated the reproductive outcome of 346 women...

  18. Effect of semen extenders on frozen-thawed boar sperm characteristics and distribution in the female genital tract after deep intrauterine insemination in sows.

    Science.gov (United States)

    Noguchi, Michiko; Yoshioka, Koji; Hikono, Hirokazu; Suzuki, Chie; Kikuchi, Kazuhiro

    2015-12-01

    We compared the effects of extenders of frozen-thawed semen on post-thaw sperm characteristics and the distribution of frozen-thawed spermatozoa in the female genital tract after fixed-timed deep intrauterine insemination (DIUI) in sows. Frozen semen samples were thawed and diluted in either modified Modena solution (mMS) or porcine fertilization medium (PFM) containing theophylline, adenosine and cysteine. Sperm quality, assessed in vitro based on motility using a computer-assisted sperm analyzer and the integrity of the plasma and acrosomal membranes using flow cytometry, was evaluated at 0.5, 1.5, 3 and 6h after thawing. Progressive motility and the percentage of spermatozoa with damaged acrosomal membranes in PFM were significantly better than in mMS throughout the 6h. Sows with estrus synchronized using prostaglandin F2 alpha, equine chorionic gonadotropin and human chorionic gonadotropin (hCG) were inseminated once with mMS- or PFM-diluted 5 × 10(8) frozen-thawed spermatozoa by DIUI at 34 h after the hCG injection. At 4h after DIUI, reproductive tracts were recovered from 30 sows. There were significantly fewer polymorphonuclear leukocytes (PMNs) and more spermatozoa outside PMNs in the uterine horn after PFM treatment than with mMS. When 22 sows were administered DIUI with 10 × 10(8) frozen-thawed spermatozoa at 36 h after hCG, the pregnancy rates did not differ significantly between the mMS- (36%) and PFM- (64%) treated groups. Thus, PFM enhanced progressive sperm motility but increased sperm membrane damage compared with mMS; it also suppressed the migration of PMNs into the uterine lumen. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2012-09-01

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

  20. Viability of bovine demi embryo after splitting of fresh and frozen thawed embryo derived from in vitro embryo production

    Directory of Open Access Journals (Sweden)

    M Imron

    2007-06-01

    Full Text Available In vivo embryo production was limited by number of donor, wide variability respond due to superovulation program and also immunoactifity of superovulation hormone (FSH. Splitting technology could be an alternative to increase the number of transferrable embryos into recipien cows. Splitting is done with cutting embryo becoming two equal pieces (called demi embrio base on ICM orientation. The objective of this research was to determine the viability of demi embryo obtained from embryo splitting of fresh and frozen thawed embryo. The results showed that demi embryos which performed blastocoel reexpansion 3 hours after embryo splitting using fresh and frozen thawed embryos were 76.9 and 76.2% respectively. Base on existention of inner cell mass (ICM, the number of demi embryos developed with ICM from fresh and frozen thawed embryos were not significantly different (90.6 and 85.7% respectively. The cell number of demi embryo from fresh embryos splitting was not different compared with those from frozen thawed embryos (36.1 and 35.9 respectively. These finding indicated that embryo splitting can be applied to frozen thawed embryos with certain condition as well as fresh embryos.

  1. Perinatal outcomes of children born after frozen-thawed embryo transfer: a Nordic cohort study from the CoNARTaS group

    DEFF Research Database (Denmark)

    Wennerholm, UB; Henningsen, Anna-Karina Aaris; Romundstad, LB

    2013-01-01

    What are the risks of adverse outcomes in singletons born after frozen-thawed embryo transfer (FET)?......What are the risks of adverse outcomes in singletons born after frozen-thawed embryo transfer (FET)?...

  2. Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis.

    Science.gov (United States)

    Anand, Mukul; Yadav, Sarvajeet

    2016-02-01

    The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis. A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck). Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process. Significantly (plive percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (pvitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats.

  3. Isolation of total ribonucleic acid from fresh and frozen-thawed boar ...

    African Journals Online (AJOL)

    RNA) from raw fresh semen and frozen-thawed boar semen, using a protocol comprising the conventional TRIzol assay and a membrane-based technique, the PureLink RNA mini kit. Bioanalyzer profile revealed that the sperm RNA size ...

  4. Effect of heterologous seminal plasma and semen extenders on motility of frozen-thawed ram spermatozoa

    Directory of Open Access Journals (Sweden)

    G.A. Mataveia

    2010-05-01

    Full Text Available Ram seminal plasma increases the fertility of frozen-thawed ram spermatozoa deposited into the cervix. The aim of the current study was to compare the effect of ram seminal plasma to that of bull seminal plasma, dog prostatic fluid, protein-free TALP, TrilEq (Triladyl with 0.5 mℓ of Equex STM paste added to each 100 mℓ and heat-treated skim milk on longevity and percentages of progressively motile and aberrantly motile frozen-thawed ram spermatozoa. Three ejaculates from each of 6 rams were extended in TrilEq, pooled and frozen in straws as a single batch per ram. One hundred and eight straws (3 straws from each ram for each fluid were thawed in random order. Once thawed, a straw was emptied into a tube with 0.85mℓ of the appropriate fluid at 37 °C and kept at that temperature for 6 h. Motility was assessed at x200 magnification immediately (time zero and 2, 4 and 6 h after thawing. Progressive motility decreased from each time to the next (P < 0.05 and was 39.0% (0 h, 26.0% (2 h, 19.6% (4 h and 12.6% (6 h; SEM 1.24, n=108 for each group. Ram seminal plasma resulted in higher progressive motility than bull seminal plasma, lower than milk, and similar to the other fluids. Ram seminal plasma resulted in lower aberrant motility than protein-free TALP and similar aberrant motility to other fluids. The effect of ram seminal plasma and dog prostatic fluid was very similar. The effect of ram seminal plasma on the fertility of frozen-thawed ram spermatozoa deposited into the cervix is not due an exceptionally beneficial effect on the motility of spermatozoa.

  5. Physiological responses of planting frozen and thawed Douglas-fir seedlings

    Science.gov (United States)

    M. Anisul Islam; Kent G. Apostol; Douglass F. Jacobs; R. Kasten Dumroese

    2008-01-01

    We studied the short-term (7-day) physiological responses of planting thawed and frozen root plugs of Douglas-fir (Pseudotsuga menziesii) seedlings in 2 separate experiments under cool-moist and warm-dry growing conditions, respectively. Our results showed that shoot water potential, root hydraulic conductance, net photosynthesis (A), and...

  6. Effects of trehalose supplementation on cell viability and oxidative stress variables in frozen-thawed bovine calf testicular tissue.

    Science.gov (United States)

    Zhang, Xiao-Gang; Wang, Yan-Hua; Han, Cong; Hu, Shan; Wang, Li-Qiang; Hu, Jian-Hong

    2015-06-01

    Trehalose is widely used for cryopreservation of various cells and tissues. Until now, the effect of trehalose supplementation on cell viability and antioxidant enzyme activity in frozen-thawed bovine calf testicular tissue remains unexplored. The objective of the present study was to compare the effect of varying doses of trehalose in cryomedia on cell viability and key antioxidant enzymes activities in frozen-thawed bovine calf testicular tissue. Bovine calf testicular tissue samples were collected and cryopreserved in the cryomedias containing varying doses (0, 5, 10, 15, 20 and 25%; v/v) of trehalose, respectively. Cell viability, total antioxidant capacity (T-AOC) activity, catalase (CAT) activity, superoxide dismutase (SOD) activity, glutathione (GSH) content and malondialdehyde (MDA) content were measured and analyzed. The results showed that cell viability, T-AOC activity, SOD activity, CAT activity and GSH content of frozen-thawed bovine calf testicular tissue was decreased compared with that of fresh group (Pcell viability and antioxidant enzyme activity (SOD and CAT) among frozen-thawed groups (P0.05). In conclusion, the cryomedia added 15% trehalose reduced the oxidative stress and improved the cryoprotective effect of bovine calf testicular tissue. Further studies are required to obtain more concrete results on the determination of antioxidant capacity of trehalose in frozen-thawed bovine calf testicular tissue. Copyright © 2015 Elsevier Inc. All rights reserved.

  7. Two successful pregnancies following autotransplantation of frozen/thawed ovarian tissue

    DEFF Research Database (Denmark)

    Andersen, Claus Yding; Rosendahl, M.; Byskov, A.G.

    2008-01-01

    of frozen/thawed ovarian tissue. METHODS: One complete ovary was cryopreserved from each of six patients who were 26-35 years old prior to treatment. Tissue from three of the patients was transported 4-5 h on ice prior to cryopreservation. After a period of 17-32 months, orthotopic autotransplantation...

  8. Physical and mathematical modeling of process of frozen ground thawing under hot tank

    Science.gov (United States)

    Zemenkova, M. Y.; Shastunova, U.; Shabarov, A.; Kislitsyn, A.; Shuvaev, A.

    2018-05-01

    A description of a new non-stationary thermophysical model in the “hot tank-frozen ground” system is given, taking into account mass transfer of pore moisture. The results of calculated and experimental data are presented, and the position of the thawing front is shown to be in good agreement with the convective heat transfer due to moisture migration in the thawed ground.

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

    Science.gov (United States)

    Kolhe, Parag; Badkar, Advait

    2011-01-01

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

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

    Science.gov (United States)

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

    2015-04-01

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

  11. Assessment of motion and kinematic characteristics of frozen-thawed Sirohi goat semen using computer-assisted semen analysis

    Directory of Open Access Journals (Sweden)

    Mukul Anand

    2016-02-01

    Full Text Available Aim: The aim was to determine the motion and kinematics characteristic of frozen-thawed spermatozoa in Sirohi goat using computer-assisted semen analysis. Materials and Methods: A study was carried out in Sirohi buck. Semen collection was made biweekly from each buck with the help of artificial vagina. A total of 12 ejaculates were collected from two bucks (six ejaculates from each buck. Freshly collected semen was pooled and later evaluated. The pooled semen sample was extended with standard glycerolated egg yolk tris extender and later subjected to a process of cryopreservation. The motion and kinematic characteristics of spermatozoa were studied during freez-thawing process. Results: Significantly (p<0.01 higher value of live percent, hypo-osmotic swelling test, and acrosomal integrity were recorded in neat semen followed by diluted and frozen thaw semen. The proportion of spermatozoa showing slow progression were the highest in the neat and diluted semen followed by rapid and non-progressively motile, while a reverse pattern was observed in the frozen thaw semen where the proportion of non-progressively motile spermatozoa were significantly (p<0.01 higher followed by slow and rapid progression. Conclusion: This study showed that the best results for motion, vitality, plasma membrane integrity, and acrosome status were obtained in the neat semen followed by diluted and frozen thaw semen. Further, the process of cryopreservation results in a shift of motility from slow to non-progressive in the post-thaw semen with a significant decrease in the path velocities when compared to neat and diluted semen. Hence, it can be concluded that freezing-thawing process reduces the motility and kinematic characters spermatozoa and may be an important factor affecting the fertilizing ability of spermatozoa resulting in poor conception rate after insemination in goats.

  12. Effects of addition of sodium lauryl sulfate on frozen-thawed canine spermatozoa.

    Science.gov (United States)

    Hori, Tatsuya; Kaseki, Hanae; Fukuhara, Youko; Oba, Hiromichi; Mizutani, Tatsuji; Kawakami, Eiichi; Tsutsui, Toshihiko

    2006-10-01

    The addition of Orvus ES paste (OEP) to extender may be essential for preparing frozen dog semen. The major ingredient of OEP is sodium lauryl sulfate (SLS). In this study, we compared the effect of SLS on frozen dog semen with that of OEP. There were no significant differences between the 2-mg/ml SLS group and OEP group concerning sperm motility, viability and the percentage of viable sperm with intact acrosomes after freeze-thawing. These results suggest that the effectiveness of frozen dog semen extender containing 2 mg/ml of SLS is similar effective to that demonstrated for OEP.

  13. The effect of elevated progesterone levels before HCG triggering in modified natural cycle frozen-thawed embryo transfer cycles

    DEFF Research Database (Denmark)

    Groenewoud, Eva R; Macklon, Nick S; Cohlen, Ben J

    2017-01-01

    follicular phase progesterone levels may occur in unstimulated cycles before frozen-thawed embryo transfer, or what affect they may have on outcomes. In this cohort study, 271 patients randomized to the modified natural cycle arm of a randomized controlled trial comparing two endometrial preparation regimens....... Whether monitoring of progesterone and LH in natural cycle frozen-thawed embryo transfer has added clinical value should studied further....

  14. Fertility and flow cytometric evaluations of frozen-thawed rooster semen in cryopreservation medium containing low-density lipoprotein.

    Science.gov (United States)

    Shahverdi, A; Sharafi, M; Gourabi, H; Yekta, A Amiri; Esmaeili, V; Sharbatoghli, M; Janzamin, E; Hajnasrollahi, M; Mostafayi, F

    2015-01-01

    Frozen-thawed rooster semen is not reliable for use in artificial insemination in commercial stocks. Low-density lipoprotein (LDL) has been assessed for effectiveness as a cryoprotectant in the extender to improve the quality of frozen-thawed rooster semen. Although LDL has been evaluated in a few studies in other species for semen cryopreservation, so far no study has been conducted to examine this cryoprotectant for cryopreservation of fowl semen. Thus, this study aims to analyze the effects of different concentrations of LDL (0%, 2%, 4%, 6%, and 8%) in a Beltsville extender for cryopreservation of rooster spermatozoa. In experiment 1, motion parameters, membrane integrity, acrosome integrity, apoptosis status, and mitochondria activity were assessed after freeze-thawing. The highest quality frozen-thawed semen was selected to be used for evaluation of the fertility rate in experiment 2. Semen was collected from six roosters, twice weekly, then extended in a Beltsville extender that contained different concentrations of LDL as follows: 0% (control), 1% (Beltsville plus 1% LDL [BLDL1]), 2% (BLDL2), 4% (BLDL4), 6% (BLDL6), and 8% (BLDL8). Supplementation of the Beltsville extender with 4% LDL produced the most significant percentage of motility (43.1 ± 1.3), membrane integrity (59.4 ± 2.1),mitochondria activity (49.1 ± 1.19), and viable spermatozoa (45 ± 2.28) compared with the control treatment with the results of 22.7 ± 1.3 (motility), 38.4 ± 2.1 (membrane integrity), 40.25 ± 1.19 (mitochondrial activity), and 37.8 ± 2.28 (viability). In experiment 2, a significantly higher percentage of fertility rate was observed for frozen-thawed semen in the extender supplemented with 4% LDL (49.5 ± 1.6) compared with the control (29.2 ± 2.9). Progressive motility and acrosome integrity were not affected by LDL levels in the extenders. The results revealed that supplementation of the Beltsville extender with 4% LDL resulted in higher quality of frozen-thawed rooster

  15. The effect of extender, method of thawing, and duration of storage on in vitro fertility measures of frozen-thawed boar sperm.

    Science.gov (United States)

    Knox, R V; Ringwelski, J M; McNamara, K A; Aardsma, M; Bojko, M

    2015-08-01

    Frozen-thawed boar sperm (FTS) has reduced in vitro and in vivo life span compared to liquid semen. Experiments tested whether extenders, thawing procedures, and storage temperatures could extend the fertile life span of FTS. Experiment 1 tested the effect of six extenders on postthaw motility (MOT) and viability (VIA). Straws from boars (n = 6) were thawed, diluted into each extender, and evaluated at 20, 60, and 120 minutes. There was a trend (P = 0.08) for an extender-by-time interaction for MOT and effect of extender and time for MOT (P extender (P = 0.10) and time (P boar ejaculates (n = 15) were thawed at 50 °C for 10, 20, or 30 seconds or at 70 °C for 5, 10, or 20 seconds and evaluated at 5, 30, and 60 minutes. There was an effect of thawing treatment on MOT, VIA, and ACR (viable sperm with intact acrosomes, P extenders, thawing, and storage. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Acrosin release and acrosin activity during incubation in capacitating media using fresh and frozen-thawed dog sperm

    Directory of Open Access Journals (Sweden)

    Mónica de los Reyes

    2011-01-01

    Full Text Available We evaluated the effect of time and temperature on acrosin release from the acrosomal cap and the activity of this enzyme during in vitro capacitation in fresh and frozen/thawed dog sperm. Sperm-rich fractions of six ejaculates from three dogs were processed as fresh and frozen samples. Each sperm sample was incubated in canine capacitation medium (CCM for 0, 1, 2 and 3 h at 20°C and at 37°C. After incubation, the samples were assessed by the indirect immunofluorescent staining technique. The probability of having unlabeled sperm (PUS, indicating acrosin loss, was modelled by a binomial distribution using logistic regression. There was a linear relationship between PUS and time at both temperatures (p<0.001; however, a major percentage of unlabeled sperm was observed in frozen/thawed samples soon after incubation, indicating that the release of acrosin was affected by capacitation time, mainly in frozen samples. Temperature influenced acrosin release only in cryopreserved sperm (p<0.05. Acrosin activity was measured by digestion halos on slides coated with gelatin-substrate film during each time period; a significant increase in the number of large halos was observed in fresh samples throughout the experiment, whereas frozen/thawed sperm showed a decreased rate of halo diameters during culture. Thus, there appears to differences between fresh and frozen dog sperm in terms of acrosin release and the level of acrosin activity in the course of in vitro capacitation.

  17. Analytical Method for Differentiation of Chilled and Frozen-Thawed Chicken Meat

    Directory of Open Access Journals (Sweden)

    Ivanova Ivelina

    2014-12-01

    Full Text Available : Chilled and frozen chicken breast and thigh meat were stored at +4±1 oC and - 20±1 oC, respectively. Maillard reaction between ribose and meat proteins of the chicken samples was initiated. The changes in the ribose-induced Maillard reaction rate during chilled and frozen storage of chicken meat were evaluated on the bases of corrected absorbance values (A420* and bovine melanoidin equivalent values (mg BME/g. Application of BME as a measure of ribose-induced Maillard reaction rate enables comparability of the data obtained by different spectrophotometers. It was found that the BME values of chicken meat frozen stored for more than 15 days were significantly (P<0.05 lower than BME values of chilled-stored samples. According to the suggested threshold limit values the chicken thigh and breast meat with BME values lower than 30 mg BME/g and 51 mg BME/g, respectively could be classified as frozen-thawed.

  18. Conception rate and litter size in multiparous sows after intrauterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand.

    Science.gov (United States)

    Chanapiwat, Panida; Olanratmanee, Em-On; Kaeoket, Kampon; Tummaruk, Padet

    2014-10-01

    The aim of the present study was to determine the conception rate and litter size in sows after fixed time intra-uterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand. Sixty-nine Landrace multiparous sows were randomly allocated into two groups, including control (n=36) and treatment (n=33). The control sows were inseminated with extended fresh semen (3 × 10(9) motile sperm/dose, 100 ml) at 24, 36 and 48 hr after the onset of estrus. The treatment sows were inseminated with frozen-thawed semen (2 × 10(9) motile sperm/dose, 20 ml) at 24 and 36 hr after induction of ovulation by human chorionic gonadotropin. All inseminations were carried out by using an intra-uterine insemination technique. The time of ovulation was determined by using transrectal real-time B-mode ultrasonography. The conception rate, farrowing rate, total number of piglets born/litter (TB) and number of piglets born alive/litter (BA) were evaluated. The sows inseminated with extended fresh semen yield a higher TB (10.8 versus 9.0 piglets/l, P=0.015) and tended to have a higher conception rate (88.9% versus 75.8%, P=0.150) than sows inseminated with frozen-thawed semen. In conclusion, insemination using frozen-thawed boar semen can be practiced with convinced fertility under field conditions by fixed-time intrauterine insemination with 2 × 10(9) sperm/ dose of 20 ml at 24 and 36 hr after the onset of estrus.

  19. Conception Rate and Litter Size in Multiparous Sows after Intrauterine Insemination Using Frozen-Thawed Boar Semen in a Commercial Swine Herd in Thailand

    Science.gov (United States)

    CHANAPIWAT, Panida; OLANRATMANEE, Em-On; KAEOKET, Kampon; TUMMARUK, Padet

    2014-01-01

    ABSTRACT The aim of the present study was to determine the conception rate and litter size in sows after fixed time intra-uterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand. Sixty-nine Landrace multiparous sows were randomly allocated into two groups, including control (n=36) and treatment (n=33). The control sows were inseminated with extended fresh semen (3 × 109 motile sperm/dose, 100 ml) at 24, 36 and 48 hr after the onset of estrus. The treatment sows were inseminated with frozen-thawed semen (2 × 109 motile sperm/dose, 20 ml) at 24 and 36 hr after induction of ovulation by human chorionic gonadotropin. All inseminations were carried out by using an intra-uterine insemination technique. The time of ovulation was determined by using transrectal real-time B-mode ultrasonography. The conception rate, farrowing rate, total number of piglets born/litter (TB) and number of piglets born alive/litter (BA) were evaluated. The sows inseminated with extended fresh semen yield a higher TB (10.8 versus 9.0 piglets/l, P=0.015) and tended to have a higher conception rate (88.9% versus 75.8%, P=0.150) than sows inseminated with frozen-thawed semen. In conclusion, insemination using frozen-thawed boar semen can be practiced with convinced fertility under field conditions by fixed-time intrauterine insemination with 2 × 109 sperm/ dose of 20 ml at 24 and 36 hr after the onset of estrus. PMID:24954517

  20. Transplantation of frozen-thawed ovarian tissue

    DEFF Research Database (Denmark)

    Gellert, S E; Pors, S E; Kristensen, Stine Gry

    2018-01-01

    . CONCLUSION: This study found no indications of sufficient numbers of malignant cells present in the ovarian tissue to cause recurrence of cancer after OTT. Further, it is unlikely that OTC affects the well-being of children born. OTC is now an established method of fertility preservation in Denmark......PURPOSE: The purpose of the study is to review all peer-reviewed published reports of women receiving ovarian tissue transplantation (OTT) with frozen/thawed tissue (OTC) with respect to age, diagnosis, transplantation site, fertility outcome, and potential side effects, including data from all...... women in the Danish program. METHODS: A systematic review of the literature was performed in PubMed combined with results from all patients who had received OTT in Denmark up to December 2017. RESULTS: OTT has been reported from 21 different countries comprising a total of 360 OTT procedures in 318...

  1. Evaluation of motility, membrane status and DNA integrity of frozen-thawed bottlenose dolphin (Tursiops truncatus) spermatozoa after sex-sorting and recryopreservation.

    Science.gov (United States)

    Montano, G A; Kraemer, D C; Love, C C; Robeck, T R; O'Brien, J K

    2012-06-01

    Artificial insemination (AI) with sex-sorted frozen-thawed spermatozoa has led to enhanced management of ex situ bottlenose dolphin populations. Extended distance of animals from the sorting facility can be overcome by the use of frozen-thawed, sorted and recryopreserved spermatozoa. Although one bottlenose dolphin calf had been born using sexed frozen-thawed spermatozoa derived from frozen semen, a critical evaluation of in vitro sperm quality is needed to justify the routine use of such samples in AI programs. Sperm motility parameters and plasma membrane integrity were influenced by stage of the sex-sorting process, sperm type (non-sorted and sorted) and freezing method (straw and directional) (P0.05) at 24 h. The viability of sorted spermatozoa was higher (Pdolphin spermatozoa undergoing cryopreservation, sorting and recryopreservation are of adequate quality for use in AI.

  2. Human chorionic gonadotropin-administered natural cycle versus spontaneous ovulatory cycle in patients undergoing two pronuclear zygote frozen-thawed embryo transfer.

    Science.gov (United States)

    Lee, You-Jung; Kim, Chung-Hoon; Kim, Do-Young; Ahn, Jun-Woo; Kim, Sung-Hoon; Chae, Hee-Dong; Kang, Byung-Moon

    2018-03-01

    To compare human chorionic gonadotropin (HCG)-administered natural cycle with spontaneous ovulatory cycle in patients undergoing frozen-thawed embryo transfer (FTET) in natural cycles. In this retrospective cohort study, we analyzed the clinical outcome of a total of 166 consecutive FTET cycles that were performed in either natural cycle controlled by HCG for ovulation triggering (HCG group, n=110) or natural cycle with spontaneous ovulation (control group, n=56) in 166 infertile patients between January 2009 and November 2013. There were no differences in patients' characteristics between the 2 groups. The numbers of oocytes retrieved, mature oocytes, fertilized oocytes, grade I or II embryos and frozen embryos in the previous in vitro fertilization (IVF) cycle in which embryos were frozen were comparable between the HCG and control groups. Significant differences were not also observed between the 2 groups in clinical pregnancy rate (CPR), embryo implantation rate, miscarriage rate, live birth rate and multiple CPR. However, the number of hospital visits for follicular monitoring was significantly fewer in the HCG group than in the control group ( P cycle reduces the number of hospital visits for follicular monitoring without any detrimental effect on FTET outcome when compared with spontaneous ovulatory cycles in infertile patients undergoing FTET in natural ovulatory cycles.

  3. Influence of cryoprotectants glycerol and amides, combined with antioxidants on quality of frozen-thawed boar sperm.

    Science.gov (United States)

    Buranaamnuay, K; Grossfeld, R; Struckmann, C; Rath, D

    2011-08-01

    The present study was undertaken to examine whether the cooling and freezing extenders containing a mixture of antioxidants (AOs) catalase, Na-pyruvate and mercaptoethanol and one of three types of cryoprotectants (CPs) would be able to improve the quality of frozen-thawed boar sperm. The collected semen, only the sperm-rich fraction, was diluted 1:1 with Androhep plus™ extender, stored at 15°C for 2 h and centrifuged. The centrifuged sperm pellet was re-suspended in lactose-egg yolk extender and divided into four groups for mixing with freezing extenders containing different kinds of CPs at 5°C: (I) glycerol (GLY) as control; (II) GLY with AOs; (III) dimethyl formamide (DMF) with AOs and (IV) dimethyl acetamide (DMA) with AOs. Processed sperm were packaged in 0.25-mL straws and frozen using a controlled rate freezer. After thawing, the diluted thawed sperm were incubated at 38°C for 10 min and was assessed for motility by CASA, membrane/acrosome integrity by FITC-PNA/PI and DNA integrity (DFI) by SCSA. All sperm parameters evaluated, except DFI, were negatively affected (P0.05). There was no difference in DFI among the studied groups (P>0.05). In conclusion, based on the present results, addition of AOs to cooling and freezing extenders and/or replacement of GLY with DMF or DMA could not improve quality of frozen-thawed boar sperm. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Enriching the captive elephant population genetic pool through artificial insemination with frozen-thawed semen collected in the wild.

    Science.gov (United States)

    Hildebrandt, T B; Hermes, R; Saragusty, J; Potier, R; Schwammer, H M; Balfanz, F; Vielgrader, H; Baker, B; Bartels, P; Göritz, F

    2012-10-01

    The first successful AI in an elephant was reported in 1998, using fresh semen. Since then almost 40 calves have been produced through AI in both Asian and African elephants worldwide. Following these successes, with the objective of enriching the captive population with genetic material from the wild, we evaluated the possibility of using frozen-thawed semen collected from wild bulls for AI in captivity. Semen, collected from a 36-yr-old wild African savanna elephant (Loxodonta africana) in South Africa was frozen using the directional freezing technique. This frozen-thawed semen was used for four inseminations over two consecutive days, two before and two after ovulation, in a 26-yr-old female African savanna elephant in Austria. Insemination dose of 1200 × 10(6) cells per AI with 61% motility resulted in pregnancy, which was confirmed through ultrasound examination 75, 110 and 141 days after the AI procedure. This represents the first successful AI using wild bull frozen-thawed semen in elephants. The incorporation of AI with frozen-thawed semen into the assisted reproduction toolbox opens the way to preserve and transport semen between distant individuals in captivity or, as was done in this study, between wild and captive populations, without the need to transport stressed or potentially disease-carrying animals or to remove animals from the wild. In addition, cryopreserved spermatozoa, in combination with AI, are useful methods to extend the reproductive lifespan of individuals beyond their biological lifespan and an important tool for genetic diversity management and phenotype selection in these endangered mammals. Copyright © 2012 Elsevier Inc. All rights reserved.

  5. Theoretical study of heat transfer with moving phase-change interface in thawing of frozen food

    International Nuclear Information System (INIS)

    Leung, M; Ching, W H; Leung, D Y C; Lam, G C K

    2005-01-01

    A theoretical solution was obtained for a transient phase-change heat transfer problem in thawing of frozen food. In the physical model, a sphere originally at a uniform temperature below the phase-change temperature is suddenly immersed in a fluid at a temperature above the phase-change temperature. As the body temperature increases, the phase-change interface will be first formed on the surface. Subsequently, the interface will absorb the latent heat and move towards the centre until the whole body undergoes complete phase change. In the mathematical formulation, the nonhomogeneous problem arises from the moving phase-change interface. The solution in terms of the time-dependent temperature field was obtained by use of Green's function. A one-step Newton-Raphson method was specially designed to solve for the position of the moving interface to satisfy the interface condition. The theoretical results were compared with numerical results generated by a finite difference model and experimental measurements collected from a cold water thawing process. As a good agreement was found, the theoretical solution developed in this study was verified numerically and experimentally. Besides thawing of frozen food, there are many other practical applications of the theoretical solution, such as food freezing, soil freezing/thawing, metal casting and bath quenching heat treatment, among others

  6. Theoretical study of heat transfer with moving phase-change interface in thawing of frozen food

    Science.gov (United States)

    Leung, M.; Ching, W. H.; Leung, D. Y. C.; Lam, G. C. K.

    2005-02-01

    A theoretical solution was obtained for a transient phase-change heat transfer problem in thawing of frozen food. In the physical model, a sphere originally at a uniform temperature below the phase-change temperature is suddenly immersed in a fluid at a temperature above the phase-change temperature. As the body temperature increases, the phase-change interface will be first formed on the surface. Subsequently, the interface will absorb the latent heat and move towards the centre until the whole body undergoes complete phase change. In the mathematical formulation, the nonhomogeneous problem arises from the moving phase-change interface. The solution in terms of the time-dependent temperature field was obtained by use of Green's function. A one-step Newton-Raphson method was specially designed to solve for the position of the moving interface to satisfy the interface condition. The theoretical results were compared with numerical results generated by a finite difference model and experimental measurements collected from a cold water thawing process. As a good agreement was found, the theoretical solution developed in this study was verified numerically and experimentally. Besides thawing of frozen food, there are many other practical applications of the theoretical solution, such as food freezing, soil freezing/thawing, metal casting and bath quenching heat treatment, among others.

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

    Science.gov (United States)

    Benli, Hakan

    2016-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Hakan Benli

    2016-01-01

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

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

    Science.gov (United States)

    Benli, Hakan

    2016-01-01

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

  10. THE INFLUENCE OF SLOW THAWING ON EVOLUTION OF SOME BIOCHEMICAL COMPOUNDS IN FROZEN FISHES

    Directory of Open Access Journals (Sweden)

    Marcel AVRAMIUC

    2017-06-01

    Full Text Available The aim of this work was to study the evolution of pH, amino nitrogen and nitrogen from aminoacids in four fish species, during 48 hours of slow thawing, in order to assess the fish spoilage speed in these keeping conditions. The biological material was represented by frozen fishes (carp, catfish, mackerel and hake which were subjected to slow thawing at room temperature (+20..+22°C, by analysing, at certain time intervals, pH, amino nitrogen - AN (mg %, and nitrogen from aminoacids - NAA (g %. The pH was determined with a digital pH-meter type Hanna, and the nitrogen from aminoacids according to Sörensen method. The amino nitrogen was determined by the difference between the nitrogen content of volatile bases and the nitrogen content of the ammonia and primary amines. As compared to frozen samples, both pH and the amino nitrogen values of all fish samples showed constant and significant increases up to the end of the analyzed period, while the nitrogen from aminoacids only in the first 30 hours of thawing. The amino nitrogen and the nitrogen from amino acids values have indicated the highest spoilage speed in catfish and hake, and the least speed in mackerel.

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

    Science.gov (United States)

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

    2014-03-01

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

  12. Modified natural cycle for embryo transfer using frozen-thawed blastocysts: A satisfactory option.

    Science.gov (United States)

    Le, Quoc V; Abhari, Sina; Abuzeid, Omar M; DeAnna, Jennifer; Satti, Mohamed A; Abozaid, Tarek; Khan, Iqbal; Abuzeid, Mostafa I

    2017-06-01

    To describe pregnancy outcomes of frozen-thawed blastocysts cycles using modified natural cycle frozen embryo transfers (NC-FET) and down-regulated hormonally controlled frozen embryo transfers (HC-FET) protocols. This retrospective cohort study included all patients undergoing either modified NC-FET or down-regulated HC-FET using frozen-thawed day 5 embryos. Cycles with donor blastocysts were excluded. Four hundred twenty eight patients underwent a total of 493 FET cycles. Patients with regular menses and evidence of ovulation underwent modified NC-FET. These patients were given hCG 10,000 IU IM on the day of LH-surge. Vaginal progesterone (P4) was started two days later and blastocyst transfer was planned seven days after detecting the LH surge. Anovulatory patients and some ovulatory patients underwent down-regulated HC-FET. These patients were placed on medroxy-progesterone acetate (10mg) for 10days to bring on menses and were also given a half-dose of GnRH-agonist (GnRH-a) on the third day of medroxy-progesterone acetate. Exogenous estradiol was initiated on the third day of menses. Once serum E2 levels reached >500pg/mL and endometrial lining reached >8mm, intramuscular (IM) P4 in oil was administered. Blastocyst FET was planned 6days after initiating P4. The primary outcomes included clinical pregnancy and delivery rates. There were 197 patients in the modified NC-FET protocol and 181 in the down-regulated HC-FET protocol. Mean age (years), day-3 FSH levels (mIU/mL) and percentage of patients with male factor infertility were significantly higher and mean BMI (kg/m 2 ) was significantly lower in modified NC-FET compared to HC-FET, respectively. Analysis of the first cycle pregnancy outcomes revealed no significant differences in clinical pregnancy rate (54.3% vs. 52.5%) and delivery rate (47.2% vs. 43.6%) between modified NC-FET and HC-FET. Logistic regression analysis showed age (OR=0.939, 95% CI 0.894-0.989, p=0.011), number of blastocysts transferred (OR

  13. 86 successful births and 9 ongoing pregnancies worldwide in women transplanted with frozen-thawed ovarian tissue

    DEFF Research Database (Denmark)

    Jensen, Annette Klüver; Macklon, Kirsten Tryde; Fedder, Jens

    2017-01-01

    Purpose: This study aims to make an account of the children born following transplantation of frozen-thawed ovarian tissue worldwide with specific focus on the perinatal outcome of the children. Furthermore, perinatal outcome of seven deliveries (nine children) from Denmark is reported. Methods...

  14. In vitro fertilization of in vitro matured canine oocytes using frozen-thawed dog semen.

    Science.gov (United States)

    De los Reyes, M; Carrion, R; Barros, C

    2006-10-01

    Experiments were conducted to evaluate in vitro fertilization (IVF) of in vitro matured (IVM) bitch oocytes using dog spermatozoa frozen in three different extenders. Sperm-rich fraction from eight ejaculates of five dogs was frozen in each one of three egg yolk Tris extenders with additional: (A) 1.4 g citric acid and 0.8 g glucose; (B) 0.7 g citric acid and 3.5 g glucose; or (C) 1.4 g citric acid and 0.8 g fructose (all with 5% glycerol in 100 mL milliQ water). Thawed sperm were co-incubated with IVM bitch oocytes for 6 h. Oocytes were fixed and evaluated under an epifluorescence microscope; penetrated oocytes were defined as those having sperm heads in the perivitelline space or in the oocyte cytoplasm. Higher penetration rates (P < 0.05) were obtained in oocytes cultured with spermatozoa frozen in extenders B and C than those frozen in extender A (33.1, 34.2 and 26.4%, respectively).

  15. A frozen-thawed in vitro-matured bovine oocyte derived calf with normal growth and fertility.

    Science.gov (United States)

    Otoi, T; Yamamoto, K; Koyama, N; Tachikawa, S; Suzuki, T

    1996-08-01

    The growth and fertility of a female calf obtained from a frozen-thawed bovine oocyte was assessed. The birth weight of the calf was lower than the mean birth weight of calves from in vitro fertilized embryos (IVF-controls) and calves obtained by artificial insemination (AI-controls). The growth rate of the calf up to 6 months was slower than that of the IVF-controls, but similar to that of the AI-controls. When the calf developed into a heifer (200 kg), she was inseminated with frozen semen and 280 days later delivered a male calf. The chromosoms of this cow were normal. These findings suggest that the growth and fertility of the calf derived from the frozen oocyte are normal.

  16. Intravaginal artificial insemination in bitches using frozen/thawed semen after dilution in powdered coconut water (ACP-106c).

    Science.gov (United States)

    Uchoa, D C; Silva, T F P; Mota Filho, A C; Silva, L D M

    2012-12-01

    The aim of this study was to evaluate powdered coconut water extender (ACP-106c; ACP Serviços Tecnológicos Ltda, ACP Biotecnologia, Fortaleza, Ceará, Brazil) as a diluent for freezing dog semen and the fertility after vaginal insemination of semen frozen therein. Ten ejaculates were collected from five dogs, evaluated fresh, diluted in ACP-106c, 10% egg yolk and 6% glycerol, cooled and frozen. In the first phase of the study, straws with frozen semen were thawed and immediately subjected to the same analysis as the fresh semen and, in addition, to Computer-Assisted Semen Analysis (CASA). In phase 2, 10 bitches that had been subjected to natural breeding during a preceding oestrous cycle were vaginally inseminated with thawed semen that had been re-diluted in ACP-106c. After thawing, a mean of 77% sperm motility was obtained through subjective analysis and 77.3% through CASA. Following artificial insemination, a 60% pregnancy rate was observed, resulting in a 50% parturition rate and a mean litter size of 3.4 (SEM 0.6), with 47.1% males and 52.9% females. ACP-106c can be successfully used for freezing canine semen, and vaginal deposition of such semen yields similar pregnancy rates to those reported in other studies. © 2012 Blackwell Verlag GmbH.

  17. Singleton Pregnancy Outcomes after In Vitro Fertilization with Fresh or Frozen-Thawed Embryo Transfer and Incidence of Placenta Praevia

    Directory of Open Access Journals (Sweden)

    Sara Korosec

    2014-01-01

    Full Text Available The aim of the study was to compare the single pregnancy and neonate outcome after fresh and frozen-thawed embryo transfer in the in vitro fertilization programme (IVF. The study focused on clinical and laboratory factors affecting the abnormal placentation, especially placenta praevia, in patients conceiving in the IVF programme. The results confirm that neonates born after frozen-thawed embryo transfer had significantly higher mean birth weight than after fresh embryo transfer (ET. Moreover, the birth weight distribution in singletons was found to shift towards “large for gestation” (LGA after frozen-thawed ET. On the other hand, the pregnancies after fresh ET were characterized by a higher incidence of placenta praevia and 3rd trimester bleeding. Placenta praevia was more common in IVF patients with fresh ET in a stimulated cycle than in patients with ET in a spontaneous cycle. It occurred more frequently in patients with transfer of 2 embryos. From this point of view, single ET and ET in a spontaneous cycle should be encouraged in good prognosis patients in the future with more than two good quality embryos developed. An important issue arose of how the ovarian hormonal stimulation relates to abnormal placentation and if the serum hormone levels interfere with in the IVF treatment results.

  18. Frozen allogeneic human epidermal cultured sheets for the cure of complicated leg ulcers.

    Science.gov (United States)

    Bolívar-Flores, Y J; Kuri-Harcuch, W

    1999-08-01

    Skin ulcers due to venous stasis or diabetes are common among the elderly and are difficult to treat. Repeated applications of cell-based products have been reported to result in cure or improvement of leg ulcers of small size in a fraction of patients. To examine the effects of frozen human allogeneic epidermal cultures for the treatment of acute and chronic ulcers. We treated a series of 10 consecutive patients with leg ulcers of different etiology and duration with frozen human allogeneic epidermal cultures stored frozen and thawed for 5-10 minutes at room temperature before application. Three patients had ulcers with exposed Achilles or extensor tendon. The ulcers treated were as large as 160 cm2 in area and of up to 20-years' duration. After preliminary preparation of the wounds by debridement to remove necrotic tissue and application of silver sulfadiazine to control infection, thawed cultures were applied biweekly from 2 to 15 times depending on the size and complexity of the ulcer. All ulcers healed, including those with tendon exposure. After the first few applications, granulation tissue formed in the ulcer bed and on exposed tendons, and epidermal healing took place through proliferation and migration of cells from the margins of the wound. The time required for complete healing ranged from 1 to 31 weeks after the first application. The use of frozen human allogeneic epidermal cultures is a safe and effective treatment for venous or diabetic ulcers, even those with tendon exposure. It seems possible that any leg ulcer will be amenable to successful treatment by this method.

  19. Observations on procedures for thawing and spit-roasting frozen dressed chickens, and post-cooking care and storage: with particular reference to food-poisoning bacteria

    Science.gov (United States)

    Roberts, Diane

    1972-01-01

    A comparison was made of four methods of thawing frozen chickens and an average thaw time for each method was determined. Fully and partially thawed chickens, inoculated with salmonellas, Clostridium welchii and Staphylococcus aureus were cooked in a spit-roasting oven at different temperatures for different lengths of time. The chickens were examined freshly cooked and after storage under various conditions. Spit roasting fully thawed chickens until the outer skin was golden brown was sufficient heat-treatment to kill salmonellas and Staph. aureus but Cl. welchii could survive. Salmonellas could also survive if the chickens were not fully thawed before cooking. Incorrect storage after cooking was shown to encourage the growth of pathogens. The incidence of intestinal pathogens in frozen dressed chickens and environmental hazards in spit-roasting establishments were also studied. Of raw chickens examined 35% contained salmonellas (9 serotypes), 63% contained Cl. welchii and 63% Staph. aureus. PMID:4342001

  20. The first woman to give birth to two children following transplantation of frozen/thawed ovarian tissue

    DEFF Research Database (Denmark)

    Ernst, Erik; Bergholdt, Stinne; Jørgensen, Jan Stener

    2010-01-01

    Worldwide eight children have been born as a result of transplanting frozen/thawed ovarian tissue. Two of these children were born in Denmark following transport of the ovarian tissue for a period of 5 h prior to cryopreservation. One of these women, who was originally transplanted with six pieces...

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

    Science.gov (United States)

    Eriksson, B M; Rodriguez-Martinez, H

    2000-11-01

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

  2. Effects of diluents and plasma on honey bee (Apis mellifera L.) drone frozen-thawed semen fertility.

    Science.gov (United States)

    Gül, Aziz; Şahinler, Nuray; Onal, Ali G; Hopkins, Brandon K; Sheppard, Walter S

    2017-10-01

    Cryopreservation is an advanced method used to protect germplasm in liquid nitrogen. Honey bees are of special interest to protect because of their pollination activity and critical role in agriculture. There has been important progress in the cryopreservation of honey bee germplasm in recent years, leading to practical recovery of genetic material for breeding purposes following freezing. However, there remains room for improvement and the goal of the present study was to evaluate the effect of different "extenders" added post-thaw on the fertilization rate of cryopreserved honey bee semen. The purpose of adding extender post-thaw was to dilute the cryoprotectant to remove chemicals after centrifugation because of potential adverse effects. The control consisted of frozen-thawed semen without the addition of an extender; treatment groups included the addition of one of the following extenders: glucose solution, fresh ram semen plasma, fresh honey bee semen plasma, extender solution. All of the above treatments and frozen-thawed control were compared to fresh semen. For each group, 15 virgin queens were instrumentally inseminated with the semen-diluent solution and introduced into nucleus colonies to determine the brood patterns of the queens. Percentages of worker brood produced in the fresh semen, frozen-thawed semen control, glucose, fresh ram semen plasma, fresh honey bee semen plasma, and extender solution supplemented groups were 98.±1.1%, 47.0 ± 0.9%, 3.0 ± 0.8%, 0.3 ± 0.1%, 48.1 ± 4.1% and 40.3 ± 2.4%, respectively. Similiarly, spermatozoa numbers in the spermathecae of the same treatment groups were 3.6 × 10 6 , 1.6 × 10 6 , 7.3 × 10 5 , 4.7 × 10 5 , 8.1 × 10 5 , and 4.6 × 10 5 spermatozoa for the same treatment, respectively. The differences in both worker brood percentage and sperm count in the spermatheca were statistically significant (P drone semen plasma group. We found a positive correlation between sperm count in

  3. Fertility response of artificial insemination methods in sheep with fresh and frozen-thawed semen.

    Science.gov (United States)

    Masoudi, Reza; Zare Shahneh, Ahmad; Towhidi, Armin; Kohram, Hamid; Akbarisharif, Abbas; Sharafi, Mohsen

    2017-02-01

    The aim of this study was to evaluate the fertility response of artificial insemination (AI) methods with fresh and frozen sperm in sheep. In experiment 1, one hundred and fifty fat tailed Zandi ewes were assigned into 3 equal groups and inseminated with three AI methods consisting of vaginal, laparoscopic and trans-cervical AI with fresh semen. In experiment 2, a factorial study (3 AI methods × 2 extenders) was used to analyze the effects of three AI methods and two freezing extenders containing soybean lecithin (SL) or Egg yolk (EY) on reproductive performance of 300 fat tailed Zandi ewes. Also, total motility, progressive motility, viability and lipid peroxidation of semen were evaluated after freeze-thawing in two extenders. In result, there was no significant difference among three AI methods when fresh semen was used. In experiment 2, the highest percentage of pregnancy rate, parturition rate and lambing rate were obtained in laparoscopic AI group (P semen, trans-cervical AI was more efficient than vaginal method when frozen-thawed semen was used, but its efficiency was not as high as laparoscopic method. Also, SL extender can be an efficient alternative extender to preserve ram sperm during cryopreservation procedure without adverse effects of EY. Copyright © 2016. Published by Elsevier Inc.

  4. Relationships among frozen-thawed semen fertility, physical parameters, certain routine sperm characteristics and testosterone in breeding Murrah buffalo (Bubalus bubalis bulls

    Directory of Open Access Journals (Sweden)

    A. K. Singh

    2014-09-01

    Full Text Available Aim: The present study was carried out to examine the relationships among frozen-thawed semen fertility, physical parameters, seminal quality, and testosterone concentration in Murrah buffalo bulls. Materials and Methods: A total of 30 breeding Murrah buffalo bulls (either progeny tested or under progeny testing program were randomly selected from two government bull farms in Punjab. None of the bulls selected for this study had any preceding physical abnormality. A field fertility trial was conducted to determine the first service conception rate (FSCR. The number of females inseminated per bull semen was 10. All the bulls were inspected for structural soundness, measurement of scrotal circumference, testicular biometry, and internal pelvic area (IPA. Frozen-thawed semen was evaluated for total motility, progressive motility, viability, concentration, abnormality, and hypo-osmotic swelling test (HOST. Testosterone was estimated in blood plasma, seminal plasma as well as frozen-thawed semen extracts for establishing relationship. Results: The FSCR was 48% in the bulls having a scrotal circumference of ≥44 cm, although, there was no significant correlation between FSCR and scrotal circumference. Similarly, no consistent relationship existed between sperm concentration and scrotal circumference. A positive correlation was observed between IPA and FSCR (r=0.294. Of the six post-thaw seminal components (total motility, progressive motility, viability, HOST (%, total abnormality and concentration only total motility had a high significant (p<0.01 correlation with FSCR (r=0.694. Varied correlations existed between other seminal parameters and fertility. Using a simple regression analysis, the post-thaw motility, IPA, prepuce length and testosterone (independent variables combined to explain approximately 62% of the variation in the FSCR (dependent variable. Conclusion: The present study indicated that despite low to high correlations between seminal

  5. Effect of different concentrations of egg yolk and virgin coconut oil in Tris-based extenders on chilled and frozen-thawed bull semen.

    Science.gov (United States)

    Tarig, A A; Wahid, H; Rosnina, Y; Yimer, N; Goh, Y M; Baiee, F H; Khumran, A M; Salman, H; Ebrahimi, M

    2017-07-01

    The aim of this study was to evaluate the effects of 8% virgin coconut oil (VCO) combined with different percentages of egg yolk in Tris extender on the quality of chilled and frozen-thawed bull semen. A total of 24 ejaculates from four bulls were collected using an electroejaculator. Semen samples were diluted with 8% VCO in Tris extender which contained different concentrations 0% (control), 4%, 8%, 12%, 16% and 20% egg yolk. The diluted semen samples were divided into two fractions: one was chilled and stored at 4°C until evaluation after 24, 72, and 144h; the second fraction was processed by chilling for 3h at 4°C to equilibrate, then packaged in 0.25ml straws and frozen and stored in liquid nitrogen at -196°C until evaluation after 7 and 14 days. Both chilled and frozen semen samples were then thawed at 37°C and assessed for general motility using computer-assisted semen analysis (CASA), viability, acrosome integrity, and morphology (eosin-nigrosin), membrane integrity (hypo-osmotic swelling test) and lipid peroxidation (thiobarbituric acid-reactive substances (TBARS)). The results indicate treatments with 8%, 12%, 16% and 20% egg yolk with 8% VCO had greater sperm quality (Psemen. In conclusion, the use of 8% VCO combined with 20% egg yolk in a Tris-based extender enhanced the values for chilled and frozen-thawed quality variables of bull sperm. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Effect of different cryo-protectants on the viability of frozen/thawed semen from boars of the Piau breed.

    Science.gov (United States)

    Pinho, R O; Lima, D M A; Shiomi, H H; Siqueira, J B; Silva, H T; Lopes, P S; Guimarães, S E F; Guimarães, J D

    2014-05-01

    The objective of this study was to evaluate the effect of different cryo-protectants (glycerol, dimethylacetamide and dimethylformamide alone or combined and added to lactose-egg yolk extender) on the viability of frozen/thawed semen from the Piau breed as assessed by in vitro testing. Frozen semen samples (n=20) were used from five male swine. Five different freezing extenders, including 2% glycerol (Group 1 - G), 2% glycerol and 3% dimethylacetamide (Group 2 - GA), 2% glycerol and 3% dimethylformamide (Group 3 - GF), 5% dimethylacetamide (Group 4 - A) and 5% dimethylformamide (group 5 - F), were evaluated. To assess post-thawing sperm quality, sperm motility and morphology were evaluated. Sperm viability was determined using the hypoosmotic swelling test, supravital staining, and a fluorescent assay (carboxyfluorescein diacetate and propidium iodide). The mean total sperm motility of semen immediately after thawing was 46.2±1.3, 57.7±1.5, 53.2±2.1, 51.7±1.2, and 46.5±1.6% for groups 1-5, respectively. Groups 2 (GA) and 3 (GF) had greater motility values (P0.05). The other complementary tests (hypoosmotic swelling test and supra-vital staining) demonstrated that sperm in Groups 2 (GA), 3 (GF) and 4 (A) had the greatest viability and there were no significant differences among these three groups (P>0.05). The most effective cryo-protectant combinations likely minimized and controlled the deleterious processes that occur in the sperm cell during freezing/thawing, thus improving post-thawing sperm viability. In conclusion, the combination of amides (3%) and glycerol (2%) or dimethylacetamide (5%) alone were more efficient at cryo-protection than glycerol alone for semen freezing in the Piau swine breed. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Andrea D. Abranches

    2014-07-01

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

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

    Science.gov (United States)

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

    2014-01-01

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

  9. TEMPORARY STORAGE OF BOVINE SEMEN CRYOPRESERVED IN LIQUID NITROGEN ON DRY ICE AND REFREEZING OF FROZEN-THAWED SEMEN.

    Science.gov (United States)

    Abdussamad, A M; Gauly, M; Holtz, W

    2015-01-01

    Two experiments were conducted. The purpose of Experiment 1 was to investigate whether viability of bovine semen stored in liquid nitrogen (-196°C) will be adversely affected by temporary exposure to dry ice (-79°C). It was convincingly shown that post thaw-motility was not affected, regardless whether semen was thawed immediately or after being returned to liquid nitrogen. Shipping or temporary storage on dry ice, thus, is a viable option. In Experiment 2, refreezing of frozen-thawed semen was attempted. The proportion of motile spermatozoa was reduced by a factor of ten to between 6.0 % and 7.4 %, regardless whether thawing occurred directly after removal from liquid nitrogen or after an interim period on dry ice. When semen was refrozen on dry ice before being returned to liquid nitrogen, motility rates were significantly improved (13.0 % to 17.0 %, P<0.05). In both experiments sperm cells that remained motile displayed vigorous forward movement and normal morphological appearance.

  10. Meat quality of lamb frozen stored up to 21 months: instrumental analyses on thawed meat during display.

    Science.gov (United States)

    Muela, E; Monge, P; Sañudo, C; Campo, M M; Beltrán, J A

    2015-04-01

    The study analysed the effect of frozen storage duration (FSD: 0, 1, 9, 15 or 21 months) and display duration (DD: 0-24 h post-slaughter-, 3 and 6 days) in modified atmosphere packaging (MAP) on lamb quality. pH, colour, lipid oxidation, water holding capacity and instrumental texture were performed on Longissimus muscle in displayed fresh and thawed meat. FSD affected all the variables showing lower differences between fresh and 1 month storage than among them and longer FSD. Only cooking losses were not affected by DD in thawed meats. It was observed a general decrease in quality (lower redness and water holding capacity; higher yellowness and lipid oxidation) as FSD or DD increased and only texture was improved over DD being thawed meat more tender. In conclusion, lamb storage at -18°C should not exceed 1 month if thawed meat would be later displayed in MAP while meat would have an acceptable quality up to 21 months without subsequent display. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. DESCRIPTION OF ULTRASTRUCTURAL DAMAGES IN FROZEN-THAWED CANINE SPERMATOZOA DESCRIÇÃO DE DANOS ULTRAESTRUTURAIS EM ESPERMATOZOIDES CANINOS CONGELADOS-DESCONGELADOS

    Directory of Open Access Journals (Sweden)

    Viviane Helena Chirinéa

    2009-07-01

    Full Text Available In spite of the advances in transmission electronic microscopy (TEM, there are few studies presenting a systematic description of the canine spermatozoa and they are only focused on sperm cell heads. The goal of the present study was to describe ultrastructural appearance of canine fresh and frozen-thawed spermatozoa, focusing on damages induced by freezing-thawing in different sperm regions. Ten ejaculates from five proven stud dogs (two ejaculates/dog were collected, evaluated, extended in Tris-egg yolk-glycerol, frozen and stored in liquid nitrogen, and thawed six weeks later.  Samples were evaluated for progressive motility, morphology, and for ultrastructural analysis by TEM. Concerning to TEM, the most striking differences between fresh and frozen-thawed samples were observed over the mid-piece since the fresh spermatozoa showed a well preserved mid-piece. However, the frozen-thawed spermatozoa mid-piece showed signs of damage such as mitochondrial vacuolization. In conclusion, freezing-thawing processes using a Tris-egg yolk extender induce ultrastructural damages in head and mid-piece of canine sperm, affecting the mitochondrial ultrastructure besides.

    KEY WORDS: Cryopreservation, dog, semen, ultrastructure.
    Apesar dos avanços na microscopia eletrônica de transmissão (MET, existem poucos estudos apresentando uma descrição sistemática do espermatozoide canino, os quais estão focados apenas na cabeça espermática. Objetivou-se, com o presente estudo, descrever a aparência ultraestrutural do espermatozoide canino fresco e congelado-descongelado, enfocando os danos induzidos pela congelação e descongelação em diferentes regiões espermáticas. Dez ejaculados obtidos de cinco cães (dois ejaculados por cão foram coletados, avaliados e diluídos em Tris-gema-glicerol, congelados e armazenados em nitrogênio líquido, e descongelados seis semanas após. Avaliaram-se as amostras quanto à motilidade progressiva

  12. Fertility test of frozen boar semen.

    Science.gov (United States)

    Osinowa, O; Salamon, S

    1976-10-01

    The fertility results of two experiments are presented. In experiment 1, the semen was frozen in tris-fructose-EDTA or BF3 diluents at 0-25 X 10(9)/ml sperm concentration and extended after thawing with either seminal plasma (SP) or the freezing medium (FM) containing no cryoprotective agent. In the second experiment the semen was glycerolated by two methods, frozen at 1-0 X 10(9)/ml sperm concentration, and extended wtih FM before insemination. Fertility after double insemination within one oestrus with semen frozen in tris-fructose-EDTA or BF3 diluents varied depending on the medium used for extension of thawed semen. The farrowing rates for semen frozen in the former diluent with FM and SP post-thawing media were 4/8 and 1/8 respectively, and for semen frozen BF3 diluent with FM and SP post-thawing extenders 1/8 and 5/8. The mean farrowing for the 32 animals inseminasted was 34-4%. Pregnancies for semen frozen in tris-fructose-EDTA and glycerolated at 30 or 5 degrees C were 5/12 and 4/12 respectively, and for single and double inseminations 6/12 and 3/12 respectively. Of 24 animals inseminated 37-5% farrowed.

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Science.gov (United States)

    2014-01-01

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

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

    Science.gov (United States)

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

    2013-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Fatemeh Ghasemian

    2012-01-01

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

  17. Fertilisation rate obtained with frozen-thawed boar semen supplemented with rosmarinic acid using a single insemination timed according to vulvar skin temperature changes.

    Science.gov (United States)

    Luño, Victoria; Gil, Lydia; Olaciregui, Maite; Grandía, Juan; Ansó, Trinidad; De Blas, Ignacio

    2015-03-01

    Artificial insemination (AI) of sows with frozen-thawed semen usually results in lower pregnancy rates and litter sizes than the use of liquid preserved semen. The present study evaluated the effectiveness of vulvar skin temperature changes as a predictor of ovulation in sows and determined the fertility rates obtained after AI with frozen-thawed semen supplemented with rosmarinic acid (RA). Semen was collected from mature boars and cryopreserved in experimental extenders supplemented with or without 105 μM of RA. Multiparous sows were inseminated with a single dose of semen when vulvar skin temperature decreased to a value below 35 °C. Intrauterine insemination was performed using 1.5 × 109 spermatozoa. The sows were slaughtered 48 h after AI and the embryos and oocytes were recovered from the oviducts. Total and progressive motility, viability and acrosome integrity were significantly (P semen samples compared with the control. Fertilisation occurred in all sows inseminated in the study, although there were no significant differences between the experimental groups. Sows inseminated with RA-supplemented semen showed a slight increase in the number of embryos recovered as compared to sows inseminated with control semen. In conclusion, insemination according to vulvar skin temperature changes resulted in successful fertilisation in all sows, although supplementation of the freezing media with RA did not improve the fertilising ability of frozen-thawed boar sperm.

  18. Non-invasive metabolomic profiling of embryo culture media and morphology grading to predict implantation outcome in frozen-thawed embryo transfer cycles.

    Science.gov (United States)

    Li, Xiong; Xu, Yan; Fu, Jing; Zhang, Wen-Bi; Liu, Su-Ying; Sun, Xiao-Xi

    2015-11-01

    Assessment of embryo viability is a crucial component of in vitro fertilization and currently relies largely on embryo morphology and cleavage rate. Because morphological assessment remains highly subjective, it can be unreliable in predicting embryo viability. This study investigated the metabolomic profiling of embryo culture media using near-infrared (NIR) spectroscopy for predicting the implantation potential of human embryos in frozen-thawed embryo transfer (FET) cycles. Spent embryo culture media was collected on day 4 after thawed embryo transfer (n = 621) and analysed using NIR spectroscopy. Viability scores were calculated using a predictive multivariate algorithm of fresh embryos with known pregnancy outcomes. The mean viability indices of embryos resulting in clinical pregnancy following FET were significantly higher than those of non-implanted embryos and differed between the 0, 50, and 100 % implantation groups. Notably, the 0 % group index was significantly lower than the 100 % implantation group index (-0.787 ± 0.382 vs. 1.064 ± 0.331, P  0.05). NIR metabolomic profiling of thawed embryo culture media is independent of morphology and correlates with embryo implantation potential in FET cycles. The viability score alone or in conjunction with morphologic grading is a more objective marker for implantation outcome in FET cycles than morphology alone.

  19. Effect of different concentrations of soybean lecithin and virgin coconut oil in Tris-based extender on the quality of chilled and frozen-thawed bull semen

    Directory of Open Access Journals (Sweden)

    A. A. Tarig

    2017-06-01

    Full Text Available Aim: The objective of this study was to evaluate the effects of different concentrations of soybean lecithin (SL and virgin coconut oil (VCO in Tris-based extender on chilled and frozen-thawed bull semen quality parameters. Materials and Methods: A total of 24 ejaculates were collected from four bulls via an electroejaculator. Semen samples were diluted with 2% VCO in Tris-based extender which consists of various concentrations of SL (1, 1.25, 1.5, and 1.75%. A 20% egg yolk in Tris used as a positive control (C+. The diluted semen samples were divided into two fractions; one for chilling which were stored at 4°C for 24, 72, and 144 h before evaluated for semen quality parameters. The second fraction used for freezing was chilled for 3 h at 4°C, packed into 0.25 mL straws and then cryopreserved in liquid nitrogen. The samples were then evaluated after 7 and 14 days. Chilled and frozen semen samples were thawed at 37°C and assessed for general motility using computer-assisted semen analysis, viability, acrosome integrity and morphology (eosin-nigrosin stain, membrane integrity, and lipid peroxidation using thiobarbituric acid reaction test. Results: The results showed that all the quality parameters assessed were significantly (p<0.05 improved at 1.5% SL concentration in chilled semen. Treatment groups of 1, 1.25, 1.5, and 1.75% SL were higher in quality parameters than the control group (C+ in chilled semen. However, all the quality parameters in frozen-thawed semen were significantly higher in the C+ than the treated groups. Conclusion: In conclusion, supplementation of 1.5% SL in 2% VCO Tris-based extender enhanced the chilled bull semen. However, there was no marked improvement in the frozen-thawed quality parameters after treatment.

  20. Liquid nitrogen vapor is comparable to liquid nitrogen for storage of cryopreserved human sperm: evidence from the characteristics of post-thaw human sperm.

    Science.gov (United States)

    Hu, Jingmei; Zhao, Shidou; Xu, Chengyan; Zhang, Lin; Lu, Shaoming; Cui, Linlin; Ma, Jinlong; Chen, Zi-Jiang

    2015-11-01

    To compare the differences in the characteristics of post-thaw human sperm after storage in either liquid nitrogen (LN2; -196 °C) or LN2 vapor (-167 °C). Experimental study. University hospital. Thirty healthy volunteers who agreed to donate their normal semen samples for infertility or research were included in the study. Semen samples (n = 30) were divided into eight aliquots and frozen. Four aliquots of each human semen sample were stored in LN2 (-196 °C), and the other four aliquots were stored in LN2 vapor (-167 °C). After 1, 3, 6, or 12 months, samples were thawed and analyzed. The motility was evaluated by the manual counting method. The viability was estimated by eosin staining. The morphology was analyzed by Diff-Quik staining. The sperm DNA integrity was determined with acridine orange fluorescent staining, and acrosin activity was assayed by the modified Kennedy method. The characteristics of post-thaw human sperm, including motility, viability, morphology, DNA integrity, and acrosin activity, showed no significant difference between LN2 and LN2 vapor storage for the different time periods. LN2 vapor was comparable to LN2 in post-thaw sperm characteristics, suggesting that LN2 vapor may be substituted for LN2 for the long-term storage of human sperm. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    2017-11-22

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

  2. The effects of embryo culture media on the birthweight of singletons via fresh or frozen-thawed embryo transfer: a large-scale retrospective study.

    Science.gov (United States)

    Gu, Fang; Deng, Mingfen; Gao, Jun; Wang, Zilian; Ding, Chenhui; Xu, Yanwen; Zhou, Canquan

    2016-09-19

    Embryo culture media used for IVF treatment might affect fetal growth and thus birthweight of the newborns. A retrospective study was conducted in South China using data from 2370 singleton neonates born after IVF/ICSI between 2009 and 2012. Two culture media, i.e., either Vitrolife or SAGE were used as embryo culture media during the study period. Neonates' birthweights were compared between the two embryo culture media groups. Among the 2370 singletons, 1755 cases came from fresh cleavage embryo transfer while 615 were from frozen-thawed cleavage embryo transfer. Within the fresh embryo transfer newborns, no statistical difference was observed in either birthweight (mean ± SD: 3196.0 ± 468.9 versus 3168.4 ± 462.0g, p > 0.05) or adjusted birthweight controlled for gestational age and gender (z-score mean ± SD: 0.11 ± 1.02 versus 0.11 ± 0.99 g, P > 0.05) between the Vitrolife (n = 419) and the SAGE group (n = 1336). Likewise within frozen embryo transfer neotates, no statistical difference of the birthweight (3300.6 ± 441.3 vs.3256.0 ± 466.7 g, P > 0.05) and adjusted birthweight (0.30 ± 0.99 g versus 0.29 ± 0.97 g, P > 0.05) was found between the Vitrolife (n = 202) and the SAGE group (n = 413). The sex ratio [OR1.17, 95 % CI (0.94-1.46)/OR1.1, 95 % CI (0.78-1.54)], rate of small for gestational age [OR1.14, 95 % CI (0.82-1.59)/OR1.06, 95 % CI (0.56-2.02)] and large for gestational age [OR1.07, 95 % CI (0.64-1.76)/OR0.98, 95 % CI (0.47-2.02)] in fresh and frozen-thawed subgourps are all comparable respectively between the two culture media. No group differences were found in the rate of low birthweight and macosomia. Multiple linear regression analysis demonstrated that maternal weight, gestational age, frozen-thawed embryo transfer and infant gender were significantly related to neonatal birthweight (P cultured in SAGE or Vitrolife media after fresh or frozen-thawed cleavage

  3. To transfer fresh or thawed embryos?

    DEFF Research Database (Denmark)

    Pinborg, Anja

    2012-01-01

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

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

    Science.gov (United States)

    Yoo, Seon-Mi; Han, Gui-Jung

    2014-01-01

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

  5. Detection of lipid peroxidation in frozen-thawed avian spermatozoa using C(11)-BODIPY(581/591).

    Science.gov (United States)

    Partyka, Agnieszka; Lukaszewicz, Ewa; Niżański, Wojciech; Twardoń, Jan

    2011-06-01

    The aim of this study was to perform flow cytometric analysis of C11-BODIPY581/591 oxidation in fowl and geese sperm as a marker for membrane lipid peroxidation (LPO) and to establish if the cryopreservation process would make sperm membranes more susceptible to oxidative stress. The experiment was carried out on 10 meat type line Flex roosters and 10 White Koluda® geese. The semen was collected two times a week, by dorso-abdominal massage method and pooled from 10 individuals of each species. Fowl semen samples were subjected to cryopreservation using the "pellet" method and Dimethylacetamide (DMA) as a cryoprotectant. Geese semen samples were cryopreserved in plastic straws in a programmable freezing unit with Dimethyloformamide (DMF) as the cryoprotectant. A fluorescent lipid probe C11-BODIPY581/591 provided with two double bonds that are oxidized during their contact with ROS, was used for the purpose of the assessment of the LPO in freshly diluted semen samples and frozen-thawed semen samples. This probe changes its color according to its state (non peroxidized: red; peroxidized: green). Flow cytometric analysis was used to monitor these changes. The White Koluda® geese fresh semen had a higher level of LPO than the Flex fresh semen (P > 0.01). The cryopreservation of fowl semen significantly (P > 0.01) increased the percentage of live and dead spermatozoa with lipid peroxidation. In frozen-thawed semen of White Koluda® geese the percentage of live spermatozoa with LPO significantly decreased (P > 0.05) whereas significantly (P > 0.01) higher level of dead cells with LPO was observed. There were significant differences between the two studied species. After thawing, the percentage of live and dead spermatozoa with lipid peroxidation was higher in fowl semen than in geese semen (P > 0.01). In conclusion, our data clearly indicate the existence of species specific differences in susceptibility of spermatozoa to the oxidation of PUFAs in the cell membranes

  6. Digestion of frozen/thawed food waste in the hybrid anaerobic solid-liquid system

    International Nuclear Information System (INIS)

    Stabnikova, O.; Liu, X.Y.; Wang, J.Y.

    2008-01-01

    The hybrid anaerobic solid-liquid (HASL) system, which is a modified two-phase anaerobic digester, is to be used in an industrial scale operation to minimize disposal of food waste at incineration plants in Singapore. The aim of the present research was to evaluate freezing/thawing of food waste as a pre-treatment for its anaerobic digestion in the HASL system. The hydrolytic and fermentation processes in the acidogenic reactor were enhanced when food waste was frozen for 24 h at -20 deg. C and then thawed for 12 h at 25 deg. C (experiment) in comparison with fresh food waste (control). The highest dissolved COD concentrations in the leachate from the acidogenic reactors were 16.9 g/l on day 3 in the control and 18.9 g/l on day 1 in the experiment. The highest VFA concentrations in the leachate from the acidogenic reactors were 11.7 g/l on day 3 in the control and 17.0 g/l on day 1 in the experiment. The same volume of methane was produced during 12 days in the control and 7 days in the experiment. It gave the opportunity to diminish operational time of batch process by 42%. The effect of freezing/thawing of food waste as pre-treatment for its anaerobic digestion in the HASL system was comparable with that of thermal pre-treatment of food waste at 150 deg. C for 1 h. However, estimation of energy required either to heat the suspended food waste to 150 deg. C or to freeze the same quantity of food waste to -20 deg. C showed that freezing pre-treatment consumes about 3 times less energy than thermal pre-treatment

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

    Science.gov (United States)

    Fraser, L; Strzezek, J

    2007-07-15

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

  8. Assessment of the interaction between straw size and thawing rate and its impact on in vitro quality of post-thaw goat semen

    Directory of Open Access Journals (Sweden)

    Francisco Silvestre Brilhante Bezerra

    2012-03-01

    Full Text Available The objective of this study was to analyze interactions between different straw sizes and thawing rates on the post-thaw goat semen parameters. Twenty-one ejaculates (seven per animal were collected from three stud bucks by using an artificial vagina. After evaluation, the semen was extended in Tris-egg yolk-glycerol and packed in 0.25 and 0.50 mL straws, followed by storage in liquid nitrogen. Thawing was performed using two different rates: 37 ºC/1 min and 55 ºC/7 s. The interaction between the 0.5-mL straw and the thawing rate of 55 ºC/7 s promoted higher progressive motility. When the effect of straws alone was analyzed, it was verified that the use of the 0.50 mL straw promoted better conservation than the 0.25 mL one for progressive motility and acrosomal integrity, after the frozen-thawing procedures. Optimal results for progressive motility were achieved when goat semen was frozen in 0.5 mL straws and thawed in water at 55 ºC/7 s.

  9. L-Carnitine in rooster semen cryopreservation: Flow cytometric, biochemical and motion findings for frozen-thawed sperm.

    Science.gov (United States)

    Fattah, A; Sharafi, M; Masoudi, R; Shahverdi, A; Esmaeili, V; Najafi, A

    2017-02-01

    Rooster semen cryopreservation is not efficient for artificial insemination in breeder flocks. L-Carnitine (LC) has been evaluated for effectiveness in cryopreservation media on the characteristics of rooster sperm after freeze-thawing. Motility characteristics, membrane functionality, abnormal morphology, apoptotic like changes, mitochondria activity and lipid peroxidation of rooster sperms were assessed after freeze-thawing with different concentrations of LC in Beltsville medium. Semen samples were collected from 12 roosters, twice a week, and diluted in the extenders that contained different concentrations of LC. Supplementation of Beltsevile with 1 and 2 mM LC was found to result in higher total motility (68.2± 1.7% and 69.1± 1.7%, respectively), progressive motility (28.4± 1.6%, 29.8± 1.6%), membrane functionality (76.2± 1.9% and 75.9± 1.9%), viability (58.2 ± 1.1%, 59.1 ± 1.1%) and lower significant of lipid peroxidation (2.53 ± 0.08 nmol/ml, 2.49 ± 0.08 nmol/ml) compared to control group containing no LC. Lower motility, progressive motility, and viability were observed in frozen-thawed sperm in extender containing 8 mM LC (35.8± 1.7%, 9.6± 1.2% and 27.1 ± 1.2%, respectively) compared to control. Morphology and mitochondrial activity were not affected by different concentrations of LC. Our results showed that supplementation of Beltsville extender with 1 and 2 mM LC significantly improved the quality of rooster sperm quality after freeze-thawing. Copyright © 2016 Elsevier Inc. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Hyun-Wook Kim

    2017-02-01

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

  11. Effect of freezing on electrical properties and quality of thawed chicken breast meat

    Directory of Open Access Journals (Sweden)

    Ran Wei

    2017-04-01

    Full Text Available Objective The objective of this research was to study the electrical properties and quality of frozen-thawed chicken breast meat and to investigate the relationship between these parameters at different times of frozen storage. Methods Thawed samples of chicken breast muscles were evaluated after being kept in frozen storage at −18°C for different periods of time (1, 2, 3, 4, 5, 6, 7, and 8 months. Results The results showed that water-holding capacity (WHC and protein solubility decreased while thiobarbituric acid-reactive substances content increased with increasing storage time. The impedance module of samples decreased during 8-month frozen storage. Pearson correlation coefficients showed that the impedance change ratio (Q value was significantly (p<0.05 related to pH, color, WHC, lipid oxidation and protein solubility, indicating a good relationship between the electrical properties and qualities of frozen-thawed chicken breast meat. Conclusion Impedance measurement has a potential to assess the quality of frozen chicken meat combining with quality indices.

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

    Science.gov (United States)

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

    2017-08-15

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

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

    Science.gov (United States)

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

    2015-05-01

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

  14. Effect of hyaluronic acid-enriched transfer medium on frozen-thawed embryo transfer outcomes.

    Science.gov (United States)

    Fu, Wei; Yu, Min; Zhang, Xiao-Jin

    2018-04-01

    To determine if hyaluronic acid-enriched transfer medium (HETM) affects the implantation rate (IR) and clinical pregnancy rate (PR) in women undergoing frozen-thawed embryo transfer (FET). The records of women who underwent FET from May 2014 to October 2014 were retrospectively reviewed. Outcome measures were IR and PR. In all 1721 cycles of 1632 patients were included in this study. HETM was used for 347 cycles of 342 patients, and standard medium for 1374 cycles of 1290 patients. Overall, FET outcomes were similar between the groups. For patients undergoing their first FET attempt, the IR (24.3% vs 31.6%, P = 0.042) and clinical PR (34.3% vs 50.1%, P = 0.004) were lower in the HETM group. For patients undergoing their second FET attempt, pregnancy outcomes were similar between the groups. For patients undergoing their third or more FET attempt, HETM was associated with a higher IR (33.3% vs 16.4%, P Gynecology.

  15. Metagenomic analysis of permafrost microbial community response to thaw

    Energy Technology Data Exchange (ETDEWEB)

    Mackelprang, R.; Waldrop, M.P.; DeAngelis, K.M.; David, M.M.; Chavarria, K.L.; Blazewicz, S.J.; Rubin, E.M.; Jansson, J.K.

    2011-07-01

    We employed deep metagenomic sequencing to determine the impact of thaw on microbial phylogenetic and functional genes and related this data to measurements of methane emissions. Metagenomics, the direct sequencing of DNA from the environment, allows for the examination of whole biochemical pathways and associated processes, as opposed to individual pieces of the metabolic puzzle. Our metagenome analyses revealed that during transition from a frozen to a thawed state there were rapid shifts in many microbial, phylogenetic and functional gene abundances and pathways. After one week of incubation at 5°C, permafrost metagenomes converged to be more similar to each other than while they were frozen. We found that multiple genes involved in cycling of C and nitrogen shifted rapidly during thaw. We also constructed the first draft genome from a complex soil metagenome, which corresponded to a novel methanogen. Methane previously accumulated in permafrost was released during thaw and subsequently consumed by methanotrophic bacteria. Together these data point towards the importance of rapid cycling of methane and nitrogen in thawing permafrost.

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

    Directory of Open Access Journals (Sweden)

    R. Fürst

    2005-10-01

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

  17. Post-thaw sperm characteristics following long-term storage of boar semen in liquid nitrogen.

    Science.gov (United States)

    Fraser, L; Strzeżek, J; Kordan, W

    2014-06-30

    This study investigated the effect of long-term liquid nitrogen storage of semen from individual boars on post-thaw sperm characteristics. Ejaculates, collected from five Polish large white (PLW) and five Polish landrace (PLR) boars, were frozen using a standard cryopreservation protocol. Post-thaw analysis was performed within a week (Period 1) and 42-48 months (Period 2) of semen storage in liquid nitrogen. Post-thaw sperm assessments included total motility, mitochondrial function (JC-1/PI assay), plasma membrane integrity (SYBR-14/PI assay), osmotic resistance test (ORT), lipid peroxidation (LPO) status and DNA fragmentation, analysed by the neutral Comet assay. Individual boar variability within breed and cryostorage periods had significant effects on the analysed parameters of frozen-thawed spermatozoa. Prolonged semen storage in liquid nitrogen (Period 2) induced a marked reduction in post-thaw sperm motility, mitochondrial function and plasma membrane integrity in most of the boars. Post-thaw semen of eight boars exhibited a marked decrease in osmotic resistance of the sperm acrosomal membrane, whereas a significant increase in the sperm cryo-susceptibility to induced LPO and DNA fragmentation was observed only in three boars after long-term semen storage. Additionally, frozen-thawed spermatozoa of PLR boars exhibited significantly lower osmotic resistance of the acrosomal membrane than PLW boars following prolonged semen storage in liquid nitrogen. The results of this study provide evidence of ageing processes in frozen-thawed boar spermatozoa following prolonged cryostorage. It seems that, even though cryopreservation allows long-term semen storage in liquid nitrogen, spermatozoa from individual boars are more susceptible to cryo-induced damage. Copyright © 2014 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2012-03-01

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

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

    Science.gov (United States)

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

    2015-08-01

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

  20. Microparticles variability in fresh frozen plasma: preparation protocol and storage time effects.

    Science.gov (United States)

    Kriebardis, Anastasios G; Antonelou, Marianna H; Georgatzakou, Hara T; Tzounakas, Vassilis L; Stamoulis, Konstantinos E; Papassideri, Issidora S

    2016-05-01

    Extracellular vesicles or microparticles exhibiting procoagulant and thrombogenic activity may contribute to the haemostatic potential of fresh frozen plasma. Fresh frozen plasma was prepared from platelet-rich plasma at 20 °C (Group-1 donors) or directly from whole blood at 4 °C (Group-2 donors). Each unit was aseptically divided into three parts, stored frozen for specific periods of time, and analysed by flow cytometry for procoagulant activity immediately after thaw or following post-thaw storage for 24 h at 4 °C. Donors' haematologic, biochemical and life-style profiles as well as circulating microparticles were analysed in parallel. Circulating microparticles exhibited a considerable interdonor but not intergroup variation. Fresh frozen plasma units were enriched in microparticles compared to plasma in vivo. Duration of storage significantly affected platelet- and red cell-derived microparticles. Fresh frozen plasma prepared directly from whole blood contained more residual platelets and more platelet-derived microparticles compared to fresh frozen plasma prepared from platelet-rich plasma. Consequently, there was a statistically significant difference in total, platelet- and red cell-derived microparticles between the two preparation protocols over storage time in the freezer. Preservation of the thawed units for 24 h at 4 °C did not significantly alter microparticle accumulation. Microparticle accumulation and anti-oxidant capacity of fresh frozen plasma was positively or negatively correlated, respectively, with the level of circulating microparticles in individual donors. The preparation protocol and the duration of storage in the freezer, independently and in combination, influenced the accumulation of microparticles in fresh frozen plasma units. In contrast, storage of thawed units for 24 h at 4 °C had no significant effect on the concentration of microparticles.

  1. A randomized controlled, non-inferiority trial of modified natural versus artificial cycle for cryo-thawed embryo transfer

    NARCIS (Netherlands)

    Groenewoud, E. R.; Cohlen, B. J.; Al-Oraiby, A.; Brinkhuis, E. A.; Broekmans, F. J M; De Bruin, J. P.; Van Den Dool, G.; Fleisher, K.; Friederich, J.; Goddijn, M.; Hoek, A.; Hoozemans, D. A.; Kaaijk, E. M.; Koks, C. A M; Laven, J. S E; Van Der Linden, P. J Q; Manger, A. P.; Slappendel, E.; Spinder, T.; Kollen, B. J.; Macklon, N. S.

    2016-01-01

    studyquestion: Are live birth rates (LBRs) after artificial cycle frozen-thawed embryo transfer (AC-FET) non-inferior to LBRs after modified natural cycle frozen-thawed embryo transfer (mNC-FET)? summaryanswer: AC-FET is non-inferior to mNC-FET with regard to LBRs, clinical and ongoing pregnancy

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

    Directory of Open Access Journals (Sweden)

    Fagundes, B.

    2011-07-01

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

  3. A randomized controlled, non-inferiority trial of modified natural versus artificial cycle for cryo-thawed embryo transfer

    NARCIS (Netherlands)

    Groenewoud, E. R.; Cohlen, B. J.; Al-Oraiby, A.; Brinkhuis, E. A.; Broekmans, F. J. M.; de Bruin, J. P.; van den Dool, G.; Fleisher, K.; Friederich, J.; Goddijn, M.; Hoek, A.; Hoozemans, D. A.; Kaaijk, E. M.; Koks, C. A. M.; Laven, J. S. E.; van der Linden, P. J. Q.; Manger, A. P.; Slappendel, E.; Spinder, T.; Kollen, B. J.; Macklon, N. S.

    STUDY QUESTION: Are live birth rates (LBRs) after artificial cycle frozen-thawed embryo transfer (AC-FET) non-inferior to LBRs after modified natural cycle frozen-thawed embryo transfer (mNC-FET)? SUMMARY ANSWER: AC-FET is non-inferior to mNC-FET with regard to LBRs, clinical and ongoing pregnancy

  4. Human oocyte cryopreservation and the fate of cortical granules.

    Science.gov (United States)

    Ghetler, Yehudith; Skutelsky, Ehud; Ben Nun, Isaac; Ben Dor, Liah; Amihai, Dina; Shalgi, Ruth

    2006-07-01

    To examine the effect of the commonly used oocyte cryopreservation protocol on the cortical granules (CGs) of human immature germinal vesicle (GV) and mature metaphase II (MII) oocytes. Laboratory study. IVF unit. Unfertilized, intracytoplasmic sperm injected (ICSI) oocytes, and immature oocytes were cryopreserved using a slow freezing-rapid thawing program with 1,2-propanediol (PROH) as a cryoprotectant. Cortical granule exocytosis (CGE) was assessed by either confocal microscopy or transmission electron microscopy (TEM). The survival rates of frozen-thawed oocytes (mature and immature) were significantly lower compared with zygotes. Both mature and immature oocytes exhibited increased fluorescence after cryopreservation, indicating the occurrence of CGE. Mere exposure of oocytes to cryoprotectants induced CGE of 70% the value of control zygotes. The TEM revealed a drastic reduction in the amount of CGs at the cortex of frozen-thawed GV and MII oocytes, as well as appearance of vesicles in the ooplasm. The commonly used PROH freezing protocol for human oocytes resulted in extensive CGE. This finding explains why ICSI is needed to achieve fertilization of frozen-thawed human oocytes.

  5. 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.

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

    Science.gov (United States)

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

    2010-04-01

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

  7. Large baby syndrome in singletons born after frozen embryo transfer (FET)

    DEFF Research Database (Denmark)

    Pinborg, Anja; Henningsen, AA; Loft, A

    2013-01-01

    Are singletons born after frozen embryo transfer (FET) at increased risk of being born large for gestational age (LGA) and if so, is this caused by intrinsic maternal factors or related to the freezing/thawing procedures?......Are singletons born after frozen embryo transfer (FET) at increased risk of being born large for gestational age (LGA) and if so, is this caused by intrinsic maternal factors or related to the freezing/thawing procedures?...

  8. High survival of mouse embryos after rapid freezing and thawing inside plastic straws with 1-2 propanediol as cryoprotectant.

    Science.gov (United States)

    Renard, J P; Babinet, C

    1984-06-01

    A method for obtaining a high survival rate of frozen-thawed mouse embryos is presented. Eight-cell mouse embryos were frozen inside small plastic straws in the presence of 1-2 propanediol and stored at -196 C. After thawing, the embryos were diluted for only 5 min in a 1.0 M sucrose solution to remove the 1-2 propanediol from the cells. At high rate of thawing (is equivalent to 2500 C/min) more than 88% of the embryos survived in vitro to the blastocyst stage provided that the dilution of propanediol was performed rapidly during thawing. At a lower rate of thawing (is equivalent to 300 C/min), survival tended to be higher (94.7%) when dilution was done 5 min after thawing. When the frozen-thawed embryos were transferred to the oviducts of day 1 pseudopregnant recipients either directly after the dilution of 1-2 propanediol or after 24 or 48 hr of culture, a high proportion of them (65.9%) develop normally to viable fetuses.

  9. A randomized controlled, non-inferiority trial of modified natural versus artificial cycle for cryo-thawed embryo transfer

    NARCIS (Netherlands)

    Groenewoud, E. R.; Cohlen, B. J.; Al-Oraiby, A.; Brinkhuis, E. A.; Broekmans, F. J. M.; de Bruin, J. P.; van den Dool, G.; Fleisher, K.; Friederich, J.; Goddijn, M.; Hoek, A.; Hoozemans, D. A.; Kaaijk, E. M.; Koks, C. A. M.; Laven, J. S. E.; van der Linden, P. J. Q.; Manger, A. P.; Slappendel, E.; Spinder, T.; Kollen, B. J.; Macklon, N. S.

    2016-01-01

    Are live birth rates (LBRs) after artificial cycle frozen-thawed embryo transfer (AC-FET) non-inferior to LBRs after modified natural cycle frozen-thawed embryo transfer (mNC-FET)? AC-FET is non-inferior to mNC-FET with regard to LBRs, clinical and ongoing pregnancy rates (OPRs) but AC-FET does

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

    DEFF Research Database (Denmark)

    Prentø, P

    1997-01-01

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

  11. Optimizing recovery of frozen human peripheral blood mononuclear cells for flow cytometry.

    Directory of Open Access Journals (Sweden)

    Bo Langhoff Hønge

    Full Text Available Live peripheral blood mononuclear cells (PBMCs can be frozen and thawed for later analyses by adding and removing a cryoprotectant, such as dimethyl sulfoxide (DMSO. Laboratories across the world use various procedures, but published evidence of optimal thawing procedures is scarce.PBMCs were separated from blood collected from healthy Danish blood donors, and stored at -80°C after adding of DMSO. The essential steps in the thawing procedure were modified and performance was evaluated by flow cytometry with respect to the percentage and total yield of viable PMBCs.The best-performing washing medium was Roswell Park Memorial Institute (RPMI 1640 at 37°C with 20% fetal bovine serum. When using 10 mL washing medium in a 15-mL Falcon tube, samples should be centrifuged for at least 10 minutes at 500 g. We failed to detect any differences between the tested methods of mixing PBMCs with washing medium. Likewise, neither the thawing duration nor centrifugation temperature (20°C and 37°C had any effect. PBMCs could be incubated (rested for up to eight hours in a 37°C 5% CO2 incubator without affecting cell counts, but incubating PBMCs for 16 hours significantly decreased viability and recovery. In general, high viability was not necessarily associated with high recovery.Changing the thawing procedure significantly impacted PBMC viability and live cell recovery. Evaluating both viability and live PBMC recovery are necessary to evaluate method performance. Investigation of differential loss of PBMC subtypes and phenotypic changes during thawing and incubation requires further evaluation.

  12. Quality of Clotting Factor Activity in Fresh Frozen Plasma at Thaw with a Microwave System and after Storage at 4 degrees C for 48 Hours.

    Science.gov (United States)

    Kuta, Piotr; Hauck-Dlimi, Barbara; Strobel, Julian; Zimmermann, Robert; Eckstein, Reinhold

    2016-01-01

    Uncontrolled hemorrhage in polytrauma patients usually results in rapid need of blood products. Despite the shorter thawing times of microwave devices for heating fresh frozen plasma (FFP), their use has remained controversial, and just a few laboratory analyses have been published on this topic. The aim of this study was to analyse the quality of clotting factors immediately after thawing FFP with a microwave device and after 48-hour post thaw storage at 4 degrees C. 24 FFP units of all four ABO blood groups (six of each blood group) were thawed with a Transfusio-therm 2000 and later stored at 4 degrees C for 48 hours. Samples were drawn aseptically and investigated on various clotting factors and protein proteases (fibrinogen, antithrombin, FII, FV, FVII, FVIII, FIX, FX, FXI, FXIII, vWF antigen and activity, protein S, and protein C) using standard coagulation and chromogenic assays immediately after thawing and again after a 48-hour storage period at 4 degrees C. All units were tested for both anaerobic and aerobic microbial contamination using standard operating procedures immediately after thawing. After thawing, all coagulation factors and protein protease activities were within normal ranges. Blood group O individuals had approximately 25% lower plasma levels of vWF antigen and activity. After a 48-hour storage period at 4 degrees C, FVIII and FIX activities declined significantly in all blood groups, whereas the remaining clotting factors remained comparably stable. Immediately after rapid thawing using a microwave system, all FFP units contained adequate coagulation factor activities to maintain hemostatic activity at the time of product thaw. The post thaw refrigerated storage caused an anticipated decrease in factor VIII and IX activities, but retained normal coagulation factor levels of many plasma proteins. Therefore we conclude that the Transfusio-therm 2000 has no clinically significant influence on the activity of clotting factors and plasma

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

    Directory of Open Access Journals (Sweden)

    ZHUANG Hong

    2014-12-01

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

  14. Thawing of lithium in the SP-100 reactor core configuration

    International Nuclear Information System (INIS)

    Magee, P.M.; Malovrh, J.W.; REineking, W.H.

    1986-01-01

    The General Electric SP-100 Liquid Metal Reactor is designed to be launched with the lithium coolant in the reactor and primary loops frozen. Initial startup of the system in space, after a satisfactory orbit is achieved, will be accomplished by slowly increasing the power in the reactor core and using the heat generated to melt the lithium, first in the reactor, and then progressively down the primary loops. This technique significantly facilitates ground handling, reduces vibrational loads during vehicle launch and minimized the shuttle bay heat load. The challenge is to thaw the coolant and startup the system within an acceptable time without structural damage. The test results clearly demonstrate that thawing of the lithium in the SP-100 reactor core can be done rapidly without structural damage and, thus, support the selected concept of SP-100 launch with frozen lithium and thaw/startup in space

  15. Major losses of fat, carbohydrates and energy content of preterm human milk frozen at -80°C.

    Science.gov (United States)

    Lev, H M; Ovental, A; Mandel, D; Mimouni, F B; Marom, R; Lubetzky, R

    2014-05-01

    Long-term storage of human milk (HM) requires freezing at low temperatures, the consequences of which upon macronutrients are unclear. To test the null hypothesis that HM freezing and storage for a range of 1 to 10 weeks at -80 °C does not affect HM fat, protein, lactose and energy contents. Samples of HM were obtained from 20 mothers (60 samples) of preterm infants (25 to 35 weeks gestation), who routinely expressed their milk, every 3 h, using an electric pump, from the second to the seventh week after delivery. All samples were frozen at -80 °C for 8 to 83 days (43.8 days average). After thawing and homogenization, energy and macronutrient contents were measured using an HM analyzer. Fat, carbohydrates and energy contents were significantly lower in thawed HM than in fresh HM (fat, fresh vs thawed: 3.72±1.17 vs 3.36±1.19 g/100 ml, Pcarbohydrates, fresh vs thawed: 5.86±0.71 vs 4.09±0.96 g/100 ml, Pvs thawed: 64.93±12.97 vs 56.63±16.82 kcal/100 ml, Pvs thawed: 1.14±0.36 vs 1.15±0.37 g/100 ml, P=0.7). The decline in carbohydrates content but not in fat and energy correlated significantly with freezing duration. Freezing at -80 °C significantly decreases the energy content of HM, both from fat and carbohydrates. Since quantitatively the decrease in macronutrients was much higher than that published for HM storage at -20 °C, our results do not support freezing HM at -80 °C as the gold standard for long-term storage. We suggest that caloric intake calculations in preterm infants cannot be established based upon fresh HM data.

  16. Thawed cod fillets spoil less rapidly than unfrozen fillets when stored under modified atmosphere at 2 degrees C.

    Science.gov (United States)

    Guldager, H S; Bøknaes, N; Osterberg, C; Nielsen, J; Dalgaard, P

    1998-09-01

    The effect of two months of frozen storage at -20 degrees C on the spoilage characteristics and shelf life of thawed and modified atmosphere packed (MAP) cod fillets stored at 2 degrees C was studied. Thawed MAP cod fillets were compared with fresh cod fillets stored in CO2-containing modified atmospheres with and without added oxygen. The shelf life of 11 to 12 days in the fresh MAP cod was extended to more than 20 days in the thawed MAP cod at 2 degrees C. This shelf life extension was most likely due to the inactivation of the spoilage bacterium Photobacterium phosphoreum during frozen storage as reflected both in chemical analyses and sensory evaluation. In contrast to fresh MAP cod fillets no significant production of trimethylamine occurred and almost no amine odor and taste were detected during 20 days of chill storage of thawed MAP cod fillets. The use of frozen fillets as raw material not only provides a more stable product in MAP but also allows much greater flexibility for production and distribution. However, a slightly increased concentration of dimethylamine, a larger drip loss, and detection of weak frozen storage flavor were observed in the thawed MAP cod fillets.

  17. Modified hMG stimulated: an effective option in endometrial preparation for frozen-thawed embryo transfer in patients with normal menstrual cycles.

    Science.gov (United States)

    Huang, Pinxiu; Wei, Lihong; Li, Xinlin; Lin, Zhong

    2018-04-20

    To evaluate the clinical efficacy of modified human menopausal gonadotropin (hMG) stimulated, hormone replacement therapy (HRT), natural cycling and letrozole ovulation induction during endometrial preparation for frozen-thawed embryo transfer (FET) in patients with normal menstrual cycles. This retrospective analysis included a total of 5070 cycles of patients with normal menstrual patterns who underwent FET between October 2009 and September 2015. The patients were divided into four groups according to the method of endometrial preparation for FET: 1838 cycles were natural, 1666 underwent HRT, 340 underwent letrozole ovulation induction and 1226 underwent modified hMG stimulated. Reproduction-related clinical outcomes in the four groups were compared. The clinical pregnancy rates and live birth rates of patients in the modified hMG stimulated group were significantly higher than that in the other groups p .05). Modified hMG stimulated resulted in a higher pregnancy rate compared to the other treatment groups. Therefore, modified hMG stimulated may be an effective option in endometrial preparation for FET in patients with normal menstrual cycles.

  18. Conception Rate and Litter Size in Multiparous Sows after Intrauterine Insemination Using Frozen-Thawed Boar Semen in a Commercial Swine Herd in Thailand

    OpenAIRE

    CHANAPIWAT, Panida; OLANRATMANEE, Em-On; KAEOKET, Kampon; TUMMARUK, Padet

    2014-01-01

    ABSTRACT The aim of the present study was to determine the conception rate and litter size in sows after fixed time intra-uterine insemination using frozen-thawed boar semen in a commercial swine herd in Thailand. Sixty-nine Landrace multiparous sows were randomly allocated into two groups, including control (n=36) and treatment (n=33). The control sows were inseminated with extended fresh semen (3 × 109 motile sperm/dose, 100 ml) at 24, 36 and 48 hr after the onset of estrus. The treatment s...

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

    Science.gov (United States)

    Strzeżek, R; Polakiewicz, P; Kordan, W

    2015-01-01

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

  20. Limited contribution of permafrost carbon to methane release from thawing peatlands

    Science.gov (United States)

    Cooper, Mark D. A.; Estop-Aragonés, Cristian; Fisher, James P.; Thierry, Aaron; Garnett, Mark H.; Charman, Dan J.; Murton, Julian B.; Phoenix, Gareth K.; Treharne, Rachael; Kokelj, Steve V.; Wolfe, Stephen A.; Lewkowicz, Antoni G.; Williams, Mathew; Hartley, Iain P.

    2017-07-01

    Models predict that thaw of permafrost soils at northern high latitudes will release tens of billions of tonnes of carbon (C) to the atmosphere by 2100 (refs ,,). The effect on the Earth’s climate depends strongly on the proportion of this C that is released as the more powerful greenhouse gas methane (CH4), rather than carbon dioxide (CO2) (refs ,); even if CH4 emissions represent just 2% of the C release, they would contribute approximately one-quarter of the climate forcing. In northern peatlands, thaw of ice-rich permafrost causes surface subsidence (thermokarst) and water-logging, exposing substantial stores (tens of kilograms of C per square meter, ref. ) of previously frozen organic matter to anaerobic conditions, and generating ideal conditions for permafrost-derived CH4 release. Here we show that, contrary to expectations, although substantial CH4 fluxes (>20 g CH4 m-2 yr-1) were recorded from thawing peatlands in northern Canada, only a small amount was derived from previously frozen C (effect of permafrost thaw on CH4 emissions from northern peatlands.

  1. Frozen-thawed rhesus sperm retain normal morphology and highly progressive motility but exhibit sharply reduced efficiency in penetrating cervical mucus and hyualuronic acid gel

    Science.gov (United States)

    Tollner, Theodore L.; Dong, Qiaoxiang; VandeVoort, Catherine A.

    2011-01-01

    The preservation of the genetic diversity of captive populations of rhesus monkeys is critical to the future of biomedical research. Cryopreservation of rhesus macaque sperm is relatively simple to perform, yields high post-thaw motility, and theoretically, provides via artificial insemination (AI) a way to easily transfer genetics among colonies of animals. In the interest of optimizing semen cryopreservation methods for use with vaginal AI, we evaluated the ability of frozen-thawed rhesus sperm to penetrate periovulatory cervical mucus (CM). Motile sperm concentration of pre–freeze (“fresh”) and post-thawed (“thawed”) samples from 5 different males were normalized for both computer assisted sperm motion analysis and CM penetration experiments. Sperm samples were deposited into slide chambers containing CM or gel composed of hyaluronic acid (HA) as a surrogate for CM and numbers of sperm were recorded as they entered a video field a preset distance from the sperm suspension-CM (or HA) interface. Fresh and thawed sperm were dried on glass slides, “Pap”-stained, and assessed for changes in head dimensions and head and flagellar shape. While retaining better than 80% of fresh sperm progressive motility, thawed sperm from the same ejaculate retained on average only 18.6% of the CM penetration ability. Experiments using HA gel yielded similar results only with reduced experimental error and thus improved detection of treatment differences. Neither the percentage of abnormal forms nor head dimensions differed between fresh and thawed sperm. While findings suggests that sperm-CM interaction is a prominent factor in previous failures of vaginal AI with cryopreserved macaque sperm, neither sperm motility nor morphology appears to account for changes in the ability of cryopreserved sperm to penetrate CM. Our data points to a previously unidentified manifestation of cryodamage which may have implications for assessment of sperm function beyond the cervix and

  2. Monitoring Bare Soil Freeze–Thaw Process Using GPS-Interferometric Reflectometry: Simulation and Validation

    Directory of Open Access Journals (Sweden)

    Xuerui Wu

    2017-12-01

    Full Text Available Frozen soil and permafrost affect ecosystem diversity and productivity as well as global energy and water cycles. Although some space-based Radar techniques or ground-based sensors can monitor frozen soil and permafrost variations, there are some shortcomings and challenges. For the first time, we use GPS-Interferometric Reflectometry (GPS-IR to monitor and investigate the bare soil freeze–thaw process as a new remote sensing tool. The mixed-texture permittivity models are employed to calculate the frozen and thawed soil permittivities. When the soil freeze/thaw process occurs, there is an abrupt change in the soil permittivity, which will result in soil scattering variations. The corresponding theoretical simulation results from the forward GPS multipath simulator show variations of GPS multipath observables. As for the in-situ measurements, virtual bistatic radar is employed to simplify the analysis. Within the GPS-IR spatial resolution, one SNOTEL site (ID 958 and one corresponding PBO (plate boundary observatory GPS site (AB33 are used for analysis. In 2011, two representative days (frozen soil on Doy of Year (DOY 318 and thawed soil on DOY 322 show the SNR changes of phase and amplitude. The GPS site and the corresponding SNOTEL site in four different years are analyzed for comparisons. When the soil freeze/thaw process occurred and no confounding snow depth and soil moisture effects existed, it exhibited a good absolute correlation (|R| = 0.72 in 2009, |R| = 0.902 in 2012, |R| = 0.646 in 2013, and |R| = 0.7017 in 2014 with the average detrended SNR data. Our theoretical simulation and experimental results demonstrate that GPS-IR has potential for monitoring the bare soil temperature during the soil freeze–thaw process, while more test works should be done in the future. GNSS-R polarimetry is also discussed as an option for detection. More retrieval work about elevation and polarization combinations are the focus of future development.

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

    Science.gov (United States)

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

    2016-04-01

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

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

    Directory of Open Access Journals (Sweden)

    Sidney Verza Jr.

    2009-10-01

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

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

    Science.gov (United States)

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

    2010-03-01

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

  6. Culture of somatic cells isolated from frozen-thawed equine semen using fluorescence-assisted cell sorting.

    Science.gov (United States)

    Brom-de-Luna, Joao Gatto; Canesin, Heloísa Siqueira; Wright, Gus; Hinrichs, Katrin

    2018-03-01

    Nuclear transfer using somatic cells from frozen semen (FzSC) would allow cloning of animals for which no other genetic material is available. Horses are one of the few species for which cloning is commercially feasible; despite this, there is no information available on the culture of equine FzSC. After preliminary trials on equine FzSC, recovered by density-gradient centrifugation, resulted in no growth, we hypothesized that sperm in the culture system negatively affected cell proliferation. Therefore, we evaluated culture of FzSC isolated using fluorescence-assisted cell sorting. In Exp. 1, sperm were labeled using antibodies to a sperm-specific antigen, SP17, and unlabeled cells were collected. This resulted in high sperm contamination. In Exp. 2, FzSC were labeled using an anti-MHC class I antibody. This resulted in an essentially pure population of FzSC, 13-25% of which were nucleated. Culture yielded no proliferation in any of nine replicates. In Exp. 3, 5 × 10 3 viable fresh, cultured horse fibroblasts were added to the frozen-thawed, washed semen, then this suspension was labeled and sorted as for Exp. 2. The enriched population had a mean of five sperm per recovered somatic cell; culture yielded formation of monolayers. In conclusion, an essentially pure population of equine FzSC could be obtained using sorting for presence of MHC class I antigens. No equine FzSC grew in culture; however, the proliferation of fibroblasts subjected to the same processing demonstrated that the labeling and sorting methods, and the presence of few sperm in culture, were compatible with cell viability. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Effect of cyclodextrin-loaded cholesterol conjugates on plasma membrane viability of Piau swine breed frozen/thawed spermatozoa.

    Science.gov (United States)

    Pinho, R O; Lima, D M A; Shiomi, H H; Siqueira, J B; Silveira, C O; Faria, V R; Lopes, P S; Guimarães, S E F; Guimarães, J D

    2016-08-01

    The objective of this study was to investigate the effect of cyclodextrin-loaded cholesterol conjugates addition to freezing extenders on plasma membrane viability of frozen-thawed spermatozoa of the Piau swine breed. Twenty semen samples were used from five males. The freezing extender was based on lactose-egg yolk extender, added to 2% glycerol, 3% dimethylacetamide. The addition of cyclodextrin-loaded cholesterol conjugates was performed after centrifugation, when semen was diluted with the cooling extender. Four groups were subjected to the following treatment: without addition (group 1); 1.5 mg of cyclodextrin-loaded cholesterol/120 × 10(6) sperm (group 2); 1.5 mg of cyclodextrin-loaded cholestanol/120 × 10(6) sperm (group 3); 1.5 mg of cyclodextrin-loaded desmosterol/120 × 10(6) sperm (group 4). To check post-thawing sperm quality sperm motility and sperm morphology evaluation were used. Additionally, to check sperm viability the hypoosmotic swelling test, supravital staining, and fluorescent assay were used. The mean values recorded for total sperm motility of semen immediately after thawing were 54.5 ± 5.8, 55.5 ± 5.3, 53.7 ± 6.7, and 52.5 ± 6.6% respectively for groups one to four, without difference between themselves (p > 0.05). Regarding fluorescent assay the results were 28.3 ± 13.2, 26.9 ± 12.2, 22.2 ± 11.4, and 32.0 ± 15.3% respectively for groups one to four, also without difference between groups (p > 0,05). Similarly, complementary tests for evaluating the integrity and functionality of the plasma membrane showed no difference between treatments (p > 0.05). In conclusion, use of cyclodextrin-loaded cholesterol conjugates added to the plasma membrane of sperm did not demonstrate any additive effect on increasing and/or maintaining sperm motility. Copyright © 2016 Elsevier Inc. All rights reserved.

  8. Adjusting cryodiluent composition for improved post-thaw quality of rabbit spermatozoa.

    Directory of Open Access Journals (Sweden)

    Sally E Hall

    Full Text Available Improved fertility following artificial insemination with frozen-thawed spermatozoa would offer rabbit producers faster genetic improvement. Previous work investigating cryoprotectants for rabbit spermatozoa have reported inconsistent results. Semen was collected from three rabbit bucks by artificial vagina and frozen using a standard procedure with varied cryodiluent components. Post-thaw analysis encompassed motility, sperm kinematic parameters and acrosome and membrane integrity. Spermatozoa were evaluated at 0, 2 and 4 h after thawing. Experiment 1 compared diluents with 3.5% dimethyl sulfoxide (DMSO, 1.5% acetamide, 1.75% DMSO + 0.75% acetamide or 3.5% DMSO + 1.5% acetamide. The treatment that resulted in the highest post-thaw motility (P<0.001 and acrosome integrity (P<0.001 was DMSO alone. Experiment 2 compared 3.5, 7 and 10% DMSO in the cryodiluent. The best post-thaw sperm motility (P<0.001 and linearity (P = .002 was in 3.5% DMSO, while 10% DMSO afforded higher acrosome/membrane integrity at this last time point (P<0.05. Experiment 3 varied the cryodiluent to contain either 9 or 17% egg yolk or 9 or 17% low density lipoproteins extracted from whole egg yolk. The treatment with the best post-thaw result was 17% egg yolk (motility, P = 0.01; acrosome/membrane integrity, P<0.001. Experiment 4 compared different carbohydrates in the cryodiluent; 50 mM glucose (TCG, 25 mM glucose with 25 mM sucrose (TCGS low, or 50 mM glucose with 50 mM sucrose (TCGS high. When data were pooled across time points, TCG had significantly higher motility than TCGS high (P = 0.021, but was not different from TCGS low. However, TCG had fewer spermatozoa with intact acrosomes and membranes than both TCGS low and TCGS high (P = .002. Put together, these results indicate that the best cryodiluent for rabbit spermatozoa frozen under the conditions used in this paper is with 7% DMSO and 17% egg yolk in a base medium containing 25 mM glucose and 25 mM sucrose.

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

    Directory of Open Access Journals (Sweden)

    Yanlong Chen

    2017-01-01

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

  10. Revitalizing genetically-modified mouse strains using frozen-thawed sperm after up to 192 h of refrigerated epididymis transportation.

    Science.gov (United States)

    Moreno-Del Val, Gonzalo; Muñoz-Robledano, Patricia

    2017-10-01

    In the scientific interchange of genetically-modified mouse strains the transportation of refrigerated epididymis has several advantages over the transportation of live animals, especially with regard to the 3R (replacement, reduction and refinement) principles. The major limiting factor is the duration of the transportation. Previous reports have shown that sperm collected from transported epididymis maintained their fertility for around 72 h, but there are no published data with longer transportation times, and this window of time may be too short, especially for international shipments and where locations are not well connected. In this study live pups were born using frozen-thawed sperm after up to 192 h (8 days) of transportation, using a special in vitro fertilization design which resulted in a fertilization rate of 10.5%.

  11. Thawed chilled Barents Sea cod fillets in modified atmosphere packaging-application of multivariate data analysis to select key parameters in good manufacturing practice

    DEFF Research Database (Denmark)

    Bøknæs, Niels; Jensen, K.N.; Guldager, H.S.

    2002-01-01

    The purpose of the present study was to select key parameters in good manufacturing practice for production of thawed chilled modified atmosphere packed (MAP) cod (Gadus morhua) fillets. The effect of frozen storage temperature (-20 and -30 C), frozen storage period (3, 6, 9 and 12 mo) and chill...... storage periods up to 21 d at 2 C were evaluated for thawed MAP Barents Sea cod fillets. Sensory, chemical, microbiological and physical quality attributes were evaluated and multivariate data analysis (principal component analysis and partial least- squares regression) applied for identification of key...... storage was low for thawed MAP Barents Sea cod and this fish raw material seemed the more appropriate for production of thawed chilled MAP products. Frozen storage inactivation of the spoilage bacteria of Photobacterium phosphorcum was modest in Barnets Sea cod, possibly due to high trimethylamine oxide...

  12. Endometrial thickness significantly affects clinical pregnancy and live birth rates in frozen-thawed embryo transfer cycles.

    Science.gov (United States)

    Bu, Zhiqin; Wang, Keyan; Dai, Wei; Sun, Yingpu

    2016-07-01

    In order to explore the relationship between endometrial thickness on the day of embryo transfer and pregnancy outcomes in frozen-thawed embryo transfer (FET) cycles, we retrospectively analyzed data from 2997 patients undergoing their first FET cycles from January 2010 to December 2012. All patients were divided into three groups (Group A, ≤8 mm; Group B, 9-13 mm; Group C, ≥14 mm) according to the endometrial thickness on embryo transfer day. Compared with patients in the other two groups, patients with thin endometrial thickness in Group A had significantly lower clinical pregnancy rate (33.4%, 41.3% and 45.4%, p birth rate (23.8%, 32.2% and 34.0%, p confidence interval (CI): 1.10-1.77, p birth rate (aOR: 1.50; 95% CI: 1.16-1.95, p < 0.01) were significant. We conclude that for patients undergoing FET, endometrial thickness on the embryo transfer day significantly affects IVF outcomes in cleavage embryo transfer cycles independent of other factors.

  13. Are there ethnic differences in pregnancy rates in African-American versus white women undergoing frozen blastocyst transfers?

    Science.gov (United States)

    Csokmay, John M; Hill, Micah J; Maguire, Marcy; Payson, Mark D; Fujimoto, Victor Y; Armstrong, Alicia Y

    2011-01-01

    To determine whether frozen-thawed blastocyst transfer pregnancy rates (PR) are lower in African-American compared with white women. Retrospective review of frozen blastocyst cycles. University-based assisted reproductive technology (ART) program. All patients who underwent a frozen blastocyst transfer between 2003 and 2008. None. Live birth rate. One hundred sixty-nine patients underwent transfer of a frozen-thawed blastocyst. African-American women had a higher incidence of leiomyoma (40% vs. 10%) and tubal and uterine factor infertility. There was no difference in the live birth rate for African-American patients (28.0%) compared with white patients (30.2%). Of the patients who underwent a frozen-thawed blastocyst transfer, 58% (n=98) had their fresh, autologous IVF cycle, which produced the cryopreserved blastocyst, at Walter Reed Medical Center. A higher peak serum E2 level was noted in African-American patients (5,355 pg/mL) compared with white patients (4,541 pg/mL). During the fresh cycle, the live birth rates between African-American and white patients were significantly different at 16.7% versus 39.7%, respectively. Live birth rates after frozen blastocyst transfer are not different between African-American and white women despite a fourfold higher incidence of leiomyomas in African-American women. Copyright © 2011. Published by Elsevier Inc.

  14. A non-destructive method to measure the thermal properties of frozen soils during phase transition

    Directory of Open Access Journals (Sweden)

    Bin Zhang

    2015-04-01

    Full Text Available Frozen soils cover about 40% of the land surface on the earth and are responsible for the global energy balances affecting the climate. Measurement of the thermal properties of frozen soils during phase transition is important for analyzing the thermal transport process. Due to the involvement of phase transition, the thermal properties of frozen soils are rather complex. This paper introduces the uses of a multifunctional instrument that integrates time domain reflectometry (TDR sensor and thermal pulse technology (TPT to measure the thermal properties of soil during phase transition. With this method, the extent of phase transition (freezing/thawing was measured with the TDR module; and the corresponding thermal properties were measured with the TPT module. Therefore, the variation of thermal properties with the extent of freezing/thawing can be obtained. Wet soils were used to demonstrate the performance of this measurement method. The performance of individual modules was first validated with designed experiments. The new sensor was then used to monitor the properties of soils during freezing–thawing process, from which the freezing/thawing degree and thermal properties were simultaneously measured. The results are consistent with documented trends of thermal properties variations.

  15. Effects of the technique of cryopreservation and dilution/centrifugation after thawing on the motility and vitality of spermatozoa of oligoasthenozoospermic men

    Directory of Open Access Journals (Sweden)

    Esteves Sandro C.

    2003-01-01

    Full Text Available OBJECTIVE: Comparing in human semen samples with low initial quality, the effects of 2 techniques of cryopreservation and dilution/centrifugation after thawing on the spermatic motility and vitality. MATERIALS AND METHODS: Semen samples from 15 oligo and/or asthenozoospermic individuals assisted in the infertility sector of a tertiary hospital were obtained through masturbation. The samples were divided into 2 portions of equal volume, and diluted (1:1; v/v with the cryoprotector containing glycerol (Test yolk buffer. One portion was frozen through the technique of liquid nitrogen vapor with static phases (group I - GI, while the other was frozen through a programmable biological freezer with linear speed (Planer, Kryo 10, series III (group II - GII. The following parameters were assessed before freezing and after thawing: percentage of spermatozoa with progressive motility (Prog% and percentage of live spermatozoa (Vit%. After defrosting, Prog% was assessed before and after removal of cryoprotector diluent, in different time intervals (zero, 3 h, and 24 h. The statistical analysis has been accomplished by using the non-parametric tests of Wilcoxon and Friedman. RESULTS: There was significant reduction of Prog% and Vit% from before freezing to after defrosting in both groups, I and II (p < 0.001. Values of Prog% and Vit% were not statistically different between groups, after thawing. It has been observed a significant reduction in Prog% among portions frozen with the automated technique after dilution and centrifugation for removal of cryoprotector (p = 0.006. After cryoprotector removal, Prog% has been kept unaltered, in both groups, during the first 3 hours of incubation, although being superior in group I (p = 0,04. There was a significant decrease in Prog% after 24 hours of incubation, in both groups (p < 0,01. CONCLUSION: For human semen samples with low initial quality, freezing through vapor technique or through the automated technique

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

    OpenAIRE

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

    2017-01-01

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

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

    Science.gov (United States)

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

    2017-10-01

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

  18. Computer-Assisted Sperm Analysis (CASA) parameters and their evolution during preparation as predictors of pregnancy in intrauterine insemination with frozen-thawed donor semen cycles.

    Science.gov (United States)

    Fréour, Thomas; Jean, Miguel; Mirallié, Sophie; Dubourdieu, Sophie; Barrière, Paul

    2010-04-01

    To study the potential of CASA parameters in frozen-thawed donor semen before and after preparation on silica gradient as predictors of pregnancy in IUI with donor semen cycles. CASA parameters were measured in thawed donor semen before and after preparation on a silica gradient in 132 couples undergoing 168 IUI cycles with donor semen. The evolution of these parameters throughout this process was calculated. The relationship with cycle outcome was then studied. Clinical pregnancy rate was 18.4% per cycle. CASA parameters on donor semen before or after preparation were not significantly different between pregnancy and failure groups. However, amplitude of lateral head displacement (ALH) of spermatozoa improved in all cycles where pregnancy occurred, thus predicting pregnancy with a sensitivity of 100% and a specificity of 20%. Even if CASA parameters do not seem to predict pregnancy in IUI with donor semen cycles, their evolution during the preparation process should be evaluated, especially for ALH. However, the link between ALH improvement during preparation process and pregnancy remains to be explored. Copyright (c) 2009 Elsevier Ireland Ltd. All rights reserved.

  19. Pregnancy and conception rate after two intravaginal inseminations with dog semen frozen either with 5% glycerol or 5% ethylene glycol.

    Science.gov (United States)

    Rota, Ada; Milani, C; Romagnoli, S; Zucchini, P; Mollo, A

    2010-03-01

    The primary goal of this study was to compare the effects of 5% ethylene glycol (EG) and 5% glycerol (G) on fertility of frozen-thawed dog semen following intravaginal insemination. The sperm-rich fraction of the ejaculate of three male dogs was collected, pooled and divided into two aliquots, and then frozen with a Tris-glucose-egg yolk-citric acid extender containing either 5% G or 5% EG. A total of 10 bitches were inseminated twice, five with G-frozen-thawed semen and five with EG-frozen-thawed semen; intravaginal inseminations were performed the 4th and the 5th day after the estimated LH peak; four straws, thawed in a 37 degrees C water bath for 1 min and diluted in a Tris buffer, were used for insemination (200 x 10(6) spermatozoa); the insemination dose was introduced in the cranial vagina of the bitch using a sterile plastic catheter. Ovariohysterectomy was performed in all bitches between days 29 and 31 after the calculated LH surge, and pregnancy status, and the number of conceptuses and corpora lutea were recorded. All bitches were pregnant. Neither the number of conceptuses, nor the ratio of conceptuses to corpora lutea (conception rate) was significantly different between groups. In this first screening, with a limited number of bitches, EG-frozen semen did not show a higher fertility than G-frozen semen when used for two intravaginal inseminations. Irrespective of the semen used, conception rate was 0.50.

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

    Science.gov (United States)

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

    2017-09-01

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

  1. Cryopreservation of turkey semen: effect of breeding line and freezing method on post-thaw sperm quality, fertilization, and hatching

    Science.gov (United States)

    Cryopreservation methods for poultry semen are not reliable for germplasm preservation, especially for turkeys, where fertility rates from frozen/thawed semen are particularly low. The objective was to evaluate cryopreservation methods for effectiveness in promoting cryosurvival and post-thaw funct...

  2. Cytochrome oxidase as an indicator of ice storage and frozen storage

    DEFF Research Database (Denmark)

    Godiksen, Helene; Jessen, Flemming

    2001-01-01

    in different cods was 21%, and the coefficient of variation of different analyses on the same homogenate was 5%. It was shown that ice storage of muscle samples before they were frozen and thawed resulted in a major freezing-induced activation of cytochrome oxidase activity. The enzyme may therefore be used...... as an indicator of frozen fish to determine if the fish has been stored on ice before freezing. Cytochrome oxidase activity showed also potential as an indicator of frozen storage, as it was possible to distinguish between the frozen storage temperatures -9, -20, and -40 degreesC....

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

    Science.gov (United States)

    Gao, J.; Xie, Z.

    2016-12-01

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

  4. Factors Analysis of Spontaneous Abortion after Thawed-vitrified Blastocysts Transfer

    Institute of Scientific and Technical Information of China (English)

    Dong YANG; Zheng-yi SUN; Cheng-yan DENG; Qi YU; Fang-fang HE

    2008-01-01

    Objective To investigate the factors resulting in spontaneous abortion after transferring frozen-thawing blastocysts. Methods A total of 108 cases transferring vitrified blastocysts were divided into two groups: abortion group (n =20) and ongoing group (n=88). Cytogenetic analysis of apoblemas was performed in 12 cases of the abortion.Results The overall spontaneous abortion rate was 18.50%(20/108) and the early spontaneous rate was 16.67%(18/108). ,4 significant difference in maternal age was observed (abortion group: 33.3±4.0 years, ongoing group: 31.0±3.6 years, P=0.02). No difference in other parameters was found. Cytogenetic analysis of apoblemas was obtained for 12 cases, and 2 specimens were contaminated. Seven of ten patients had abnormal karyotypes. Conclusion The underlying cause of spontaneous abortion after transferring frozen thawing blastocysts appears to be abnormal karyotypes.Advancing maternal age seems to increase the risk of spontaneous abortion.

  5. Experimental studies of pipeline uplift resistance in frozen ground

    Energy Technology Data Exchange (ETDEWEB)

    Liu, B.; Crooks, J. [Golder Associates Ltd., Calgary, AB (Canada); Nixon, J.F. [Nixon Geotech Ltd., Calgary, AB (Canada); Zhou, J. [TransCanada PipeLines Ltd., Calgary, AB (Canada)

    2004-07-01

    Gas pipelines buried in permafrost are subjected to the mechanisms of frost heave and thaw settlement which induce movement in the pipe. The magnitude of loads depend on the relative displacement and the load-displacement characteristics of the surrounding soils. This paper presented the results of laboratory tests that investigated the geomechanical properties and uplift resistance of frozen Calgary silt. Compacted soil was frozen 24 hours prior to the uplift testing. A thick-wall steel pipe was laid down and more soil was subsequently placed and compacted around and above the pipe until the desired burial depth was achieved. The temperature of the frozen soil was monitored along with deformations of the soil surface in order to determine the impact of loading rate, ice content and freezing direction. The geomechanical tests included triaxial compression tests as well as tensile and creep tests. The stiffness, tensile strain limits and time dependent behaviour of the frozen soil were also defined. The issue of crack development in the frozen soil was addressed and a numerical model was developed based on measured load-displacement relationships and geomechanical properties of the frozen soil. Bilinear stress-strain relationships were consistently noted for the frozen samples under triaxial compression. The stress-strain behaviour was found to be independent of the confining stress. The measured tensile limits were found to be in good agreement with published literature. Peak uplift resistance and post-peak decreases were consistently noted in the uplift resistance. Tensile fractures were observed vertically above the crown of the pipe. Uplift resistance was highly affected by the loading rate, but the load-displacement curve prior to the peak was not affected by radial freezing or ice lenses. A new peak uplift resistance was reached following the freeze/thaw cycle, suggesting the healing of the tensile fractures. 9 refs., 1 tab., 11 figs.

  6. Thawed cod fillets spoil less rapidly than unfrozen fillets when stored under modified atmosphere at 2 C

    DEFF Research Database (Denmark)

    Guldager, H.S.; Bøknæs, Niels; Østerberg, Carsten

    1998-01-01

    The effect of two months of frozen storage at -20 degrees C on the spoilage characteristics and shelf life of thawed and modified atmosphere packed (MAP) cod fillets stored at 2 degrees C was studied. Thawed MAP cod fillets were compared with fresh cod fillets stored in CO2-containing modified at...

  7. Presence and dehydration of ikaite, calcium carbonate hexahydrate, in frozen shrimp shell.

    Science.gov (United States)

    Mikkelsen, A; Andersen, A B; Engelsen, S B; Hansen, H C; Larsen, O; Skibsted, L H

    1999-03-01

    Ikaite, calcium carbonate hexahydrate, has by means of X-ray diffraction analyses of frozen samples been identified as the mineral component of the white spots formed in the shell of frozen shrimp during storage. When the shrimp thaw and the shell material is dried and kept at room temperature, ikaite rapidly transforms into a mixture of anhydrous calcium carbonate forms. X-ray diffraction analyses and Raman spectra of synthetic ikaite as well as the dehydration product confirm the assignments, and the rate constant for dehydration is approximately 7 x 10(-)(4) s(-)(1) at ambient temperature. Differential scanning calorimetry showed that dehydration of synthetic ikaite is an entropy-driven, athermal process and confirms that a single first-order reaction is rate-determining. Ikaite is found to be stable in aqueous solution at temperatures below 5 degrees C and in the shell of frozen shrimps but decomposes on thawing to form anhydrous calcium carbonates.

  8. Effect of Adding Human Chorionic Gonadotropin to The Endometrial Preparation Protocol in Frozen Embryo Transfer Cycles

    Directory of Open Access Journals (Sweden)

    Maryam Eftekhar

    2012-01-01

    Full Text Available Background: Human chorionic gonadotropin (HCG, one of the initial embryonic signals, isprobably a major regulator of the embryo-endometrial relationship. This study aims to assess theadvantage of HCG supplementation during the secretory phase of hormonally prepared cycles forthe transfer of cryopreserved-thawed embryos.Materials and Methods: This study was a randomized clinical trial. Infertile women who werecandidates for frozen-thawed embryo transfers entered the study and were divided into two groups,HCG and control. The endometrial preparation method was similar in both groups: all women receivedestradiol valerate (6 mg po per day from the second day of the menstrual cycle and progesteronein oil (100 mg intramuscular (I.M. when the endometrial thickness reached 8 mm. Estradiol andprogesterone were continued until the tenth week of gestation. In the HCG group, patients received anHCG 5000 IU injection on the first day of progesterone administration and the day of embryo transfer.Results: In this study, 130 couples participated: 65 in the HCG group and 65 in the control group.There was no statistically significant difference between groups regarding basic characteristics.Implantation rate, chemical pregnancy, clinical pregnancy, ongoing pregnancy, and abortion rateswere similar in both groups.Conclusion: Although HCG has some advantages in assisted reproductive technology (ARTcycles, our study did not show any benefit of HCG supplementation during the secretory phase offrozen cycles (Registration Number: IRCT201107266420N4.

  9. Comparison of chemical, microbiological and histological changes in fresh, frozen and double frozen rainbow trout (Oncorhynchus mykiss

    Directory of Open Access Journals (Sweden)

    Peter Popelka

    2014-01-01

    Full Text Available The final quality of fish meat depends on the chemical and microbiological quality of fish at the time of freezing as well as on other factors including storage temperature and freezing rate. Analysis of chemical composition (water, protein and fat content, expressible drip, total volatile nitrogen levels, microbiological analyses (total viable counts, Enterobacteriaceae, psychrotrophic bacteria and histological examinations on dorsal skeletal muscles were carried out to distinguish fresh, frozen and double frozen rainbow trout (Oncorhynchus mykiss. Significantly higher expressible drip and total volatile base nitrogen concentrations (P Enterobacteriaceae and psychrotrophic bacteria were determined in double frozen trout. The light microscopy of fresh trout muscles did not show any microstructural changes, whereas deformations of muscle fibres and optically empty areas were found in frozen trout. Remarkable defects of the muscle structure in double frozen trout were demonstrated and total disruption of muscle fibres was found. The freezing of trout resulted in various structural changes in the dorsal skeletal musculature. This is a first study comparing changes in fresh, frozen and repeatedly frozen trout. Chemical, microbiological and subsequent histological examinations can be used for revealing the foul practices confusing the consumer with offering thawed fish instead of fresh cooled fish.

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

    NARCIS (Netherlands)

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

    2017-01-01

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

  11. Simulations of water, heat, and solute transport in partially frozen soils

    OpenAIRE

    Wu, Mousong; Jansson, Per-Erik; Tan, Xiao; Huang, Jiesheng; Wu, Jingwei

    2016-01-01

    Experiments for soil freezing/thawing were conducted in two seasonally frozen agricultural fields in northern China during 2011/2012 and 2012/2013 wintertime, respectively. Mass balance was checked based on measured data at various depths. Simulation work was conducted by combining CoupModel with Monte-Carlo sampling method to achieve parameter sets with equally good performance. Uncertainties existed in both measurements and model due to complexity in freezing/thawing processes as well as in...

  12. Effects of the technique of cryopreservation and dilution/centrifugation after thawing on the motility and vitality of spermatozoa of oligoasthenozoospermic men.

    Science.gov (United States)

    Esteves, Sandro C; Spaine, Deborah M; Cedenho, Agnaldo P; Srougi, Miguel

    2003-01-01

    Comparing in human semen samples with low initial quality, the effects of 2 techniques of cryopreservation and dilution/centrifugation after thawing on the spermatic motility and vitality. Semen samples from 15 oligo and/or asthenozoospermic individuals assisted in the infertility sector of a tertiary hospital were obtained through masturbation. The samples were divided into 2 portions of equal volume, and diluted (1:1; v/v) with the cryoprotector containing glycerol (Test yolk buffer). One portion was frozen through the technique of liquid nitrogen vapor with static phases (group I - GI), while the other was frozen through a programmable biological freezer with linear speed (Planer, Kryo 10, series III) (group II - GII). The following parameters were assessed before freezing and after thawing: percentage of spermatozoa with progressive motility (Prog%) and percentage of live spermatozoa (Vit%). After defrosting, Prog% was assessed before and after removal of cryoprotector diluent, in different time intervals (zero, 3 h, and 24 h). The statistical analysis has been accomplished by using the non-parametric tests of Wilcoxon and Friedman. There was significant reduction of Prog% and Vit% from before freezing to after defrosting in both groups, I and II (p live spermatozoa with progressive motility. The effects of dilution and centrifugation to remove the cryoprotector had a negative impact only in samples frozen through the automated technique. In both techniques, progressive motility is kept constant during the first 3 hours after thawing and removal of the cryoprotector, but is drastically diminished by the end of an incubation period of 24 hours.

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

    Science.gov (United States)

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

    2017-02-01

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

  14. Assessing the use of frozen pork meat in the manufacture of cooked ham

    Directory of Open Access Journals (Sweden)

    Francine Gomes Basso LOS

    2016-01-01

    Full Text Available This study aimed to evaluate the effects of slow (–18 °C and fast freezing (liquid nitrogen of pork meat, and the use of exudate released upon thawing, on the physicochemical, color, rheological, microbiological, histological, and sensory characteristics of cooked ham. The meat samples were frozen at –18 °C and thawed after 22 weeks for the production of cooked ham. No significant difference was observed regarding physicochemical, color and microbiological parameters or in sensory acceptance. The hardness and chewiness parameters showed significant differences when compared to the control sample (ham made from chilled meat. Light microscopy of cooked ham samples showed that changes in the tissues were caused by freezing and thawing the meat. The effect of exudate was significant on the sodium content and compression force parameters, but this difference was not perceived in the sensory analysis, confirming that frozen pork meat can be used to produce cooked ham without loss of quality.

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

    Science.gov (United States)

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

    2011-09-01

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

  16. Cryopreserved human aortic root allografts arterial wall: Structural changes occurring during thawing.

    Directory of Open Access Journals (Sweden)

    Robert Novotny

    Full Text Available The aim of our experimental work was to assess morphological changes of arterial wall that arise during different thawing protocols of a cryopreserved human aortic root allograft (CHARA arterial wall.The experiment was performed on CHARAs. Two thawing protocols were tested: 1, CHARAs were thawed at a room temperature at +23°C; 2, CHARAs were placed directly into a water bath at +37°C.After fixation, all samples were washed in distilled water for 5 min, and dehydrated in a graded ethanol series (70, 85, 95, and 100% for 5 min at each level. The tissue samples were then immersed in 100% hexamethyldisilazane for 10 minutes and air dried in an exhaust hood at room temperature. Processed samples were mounted on stainless steel stubs, coated with gold.Thawing protocol 1: All 6 (100% samples showed loss of the endothelium and damage to the subendothelial layers with randomly dispersed circular defects and micro-fractures without smooth muscle cells contractions in the tunica media. Thawing protocol 2: All 6 (100% samples showed loss of endothelium from the luminal surface, longitudinal corrugations in the direction of blood flow caused by smooth muscle cells contractions in the tunica media with frequent fractures in the subendothelial layer.All the samples thawed at the room temperature showed smaller structural damage to the CHARA arterial wall with no smooth muscle cell contraction in tunica media when compared to the samples thawed in a water bath.

  17. Assessment of frozen storage duration effect on quality characteristics of various horse muscles

    Directory of Open Access Journals (Sweden)

    Pil Nam Seong

    2017-12-01

    Full Text Available Objective The study aimed at assessing the effects of frozen storage duration on quality characteristics, lipid oxidation and sensory quality of various horse muscles. Methods Five representative muscles: longissimus dorsi (LD, gluteus medius (GM, semimembranosus (SM, biceps femoris (BF, and triceps brachii (TB at 24 h post-mortem obtained from 28-mo-old Jeju female breed horses (n = 8 were used in the present investigation. The muscles were vacuum-packaged and frozen at −20°C for 120, 240, and 360 days. All the samples were analyzed for thawing and cooking losses, pH, Warner–Bratzler shear forces (WBSF, color traits, total volatile basic nitrogen (TVBN, thiobarbituric acid reactive substances (TBARS and sensory traits. The muscle samples analyzed on day 0 of frozen storage (fresh, non-frozen were used for comparison. Results Results revealed that thawing and cooking losses significantly (p<0.05 increased in all the muscles after 120 days and then remained unchanged up to 360 days of frozen storage. The TBARS and TVBN contents significantly increased as increasing frozen storage time up to 360 days (p<0.05. While, significant decreases in WBSF values were observed for all the muscles with increased frozen storage time (p<0.05. Frozen storage variously affected the color traits of the muscles for instance; the redness of LD, GM, and BF muscles showed a decreasing tendency during frozen storage while it was not changed in TB and SM muscles. Furthermore, the frozen storage did not produce detrimental effects on sensory quality as it did not cause flavor and juiciness defects whereas it partially improved the tenderness of all the muscles studied. Conclusion Based on the results obtained from our work, it is concluded that frozen storage could be applied to increase the long-term shelf life of horsemeat while still retaining its sensory quality.

  18. Effect of sequence of insemination after simultaneous thawing of multiple semen straws on conception rate to timed AI in suckled multiparous Nelore cows.

    Science.gov (United States)

    Oliveira, L Z; Arruda, R P; de Andrade, A F C; Santos, R M; Beletti, M E; Peres, R F G; Martins, J P N; de Lima, V F M Hossepian

    2012-11-01

    The objective was to determine the effect of sequence of insemination after simultaneous thawing of multiple 0.5 mL semen straws on conception rate in suckled multiparous Nelore cows. The effect of this thawing procedure on in vitro sperm characteristics was also evaluated. All cows (N = 944) received the same timed AI protocol. Ten straws (0.5 mL) of frozen semen from the same batch were simultaneously thawed at 36 °C, for a minimum of 30 sec. One straw per cow was used for timed AI. Frozen semen from three Angus bulls was used. Timed AI records included sequence of insemination (first to tenth) and time of semen removal from thawing bath. For laboratory analyses, the same semen batches used in the field experiment were evaluated. Ten frozen straws from the same batch were thawed simultaneously in a thawing unit identical to that used in the field experiment. The following sperm characteristics were analyzed: sperm motility parameters, sperm thermal resistance, plasma and acrosomal membrane integrity, lipid peroxidation, chromatin structure, and sperm morphometry. Based on logistic regression, there were no significant effects of breeding group, body condition score, AI technician, and sire on conception rate, but there was an interaction between sire and straw group (P = 0.002). Semen from only one bull had decreased (P conception rates at timed AI, depending on the sire used. Nevertheless, the effects of this thawing environment on in vitro sperm characteristics, remain to be further investigated. Copyright © 2012 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    Charoenrein, Sanguansri; Preechathammawong, Nutsuda

    2012-10-01

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

  20. Improved recovery of post-thaw motility and vitality of human spermatozoa cryopreserved in the presence of dithiothreitol.

    Science.gov (United States)

    Rao, B; David, G

    1984-10-01

    Semen was collected in the laboratory from nine healthy donors. The concentrations and the percentages of live and motile spermatozoa in all semen samples were within the normal range. Each sample was diluted with citrate-egg yolk-glycerol medium with and without 5 mM dithiothreitol (DTT). Samples were frozen in liquid nitrogen vapor (-70 degrees C) for 7 min and subsequently stored in liquid nitrogen. The effect of DTT in cryopreservation of sperm was determined by comparing percentage of motile and live spermatozoa between controls and DTT-treated post-thaw samples. Percentage of motile spermatozoa was determined by two techniques, laser Doppler velocimetry (LDV) and light microscopy. The percentage of live spermatozoa was measured by microscopic evaluation after staining with eosin-nigrosin. It was shown that the addition of DTT to the freezing medium significantly improved the recovery of motile and live spermatozoa in the post-thaw samples. The mean motility recovery, as measured by LDV, was 44.9% in the controls as compared to 73.9% in the DTT-treated samples. Similarly the mean recovery of live spermatozoa in the controls and DTT-treated samples was 66.5 and 86.6%, respectively. Based on these results, a new hypothesis implicating lipid peroxidation in cryoinjury is proposed. It is also suggested that the use of DTT in the freezing medium may offer an advantage over the commonly used techniques of human sperm cryopreservation.

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

    Directory of Open Access Journals (Sweden)

    Maria - Bernadett SZMUTKU

    2013-03-01

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

  2. Supplementation of different concentrations of Orvus Es Paste (OEP) to ostrich egg yolk lipoprotein extender improves post-thaw boar semen quality.

    Science.gov (United States)

    Fraser, L; Jasiewicz, E; Kordan, W

    2014-01-01

    This study aimed to compare post-thaw quality of boar semen following freezing in an ostrich egg yolk lipoprotein (LPFo) extender supplemented with 0%, 0.25% and 0.50% Orvus Es Paste (OEP). Sperm assessments included total motility (TMOT), mitochondrial function (MF), plasma membrane integrity (PMI) and acrosome integrity (normal apical ridge, NAR). Considerable variations among boars and OEP treatments had a significant effect (P semen samples frozen in the absence of OEP. By contrast, lactose-LPFo-glycerol extender supplemented with either 0.25% OEP or 0.50% OEP markedly enhanced post-thaw sperm characteristics. In all boars, there were no marked differences in post-thaw sperm TMOT between the freezing extenders supplemented with 0.25% and 0.50% OEP. However, a decline in the percentage of post-thaw motile spermatozoa was more pronounced in the extender supplemented with 0.50% OEP following a 120-min incubation period. Furthermore, the proportions of frozen-thawed spermatozoa with MF, PMI and NAR acrosomes varied significantly among the boars in the OEP-supplemented extenders. The findings of this study indicate that different OEP concentrations, in the presence of ostrich egg yolk lipoproteins, could have varying effects on post-thaw sperm survival.

  3. SHELF LIFE OF THAWED CRUSTACEANS TREATED WITH SULPHITES

    Directory of Open Access Journals (Sweden)

    G. Smaldone

    2011-04-01

    Full Text Available The quality of fish and fish products is closely related to their freshness. Aim of this research was to evaluate the shelf life of thawed crustaceans (Aristeomorpha foliacea and Nefrops norvegicus which had been treated with sulphites and frozen on board. Organoleptic characteristics and microbiological and chemical parameters were judged favourably up to day 6 and 7 for the shrimps and Norway lobsters, respectively.

  4. Hyaluronic acid improves frozen-thawed sperm quality and fertility potential in rooster.

    Science.gov (United States)

    Lotfi, Saied; Mehri, Morteza; Sharafi, Mohsen; Masoudi, Reza

    2017-09-01

    Beneficial effects of Hyaluronic acid (HA) has not been yet assessed for cryopreservation of rooster sperm. This study was conducted to evaluate the effects of different concentrations of HA (0, 1, 2, 4 and 8mM) in Beltsville extender on the cryopreservation of rooster sperm. Semen samples were collected from six Ross broiler breeders (24-week) using abdominal massage, then divided into five equal aliquots and cryopreserved in Beltsville extender that contained different concentrations of HA. Motion characteristics, morphology, membrane functionality, viability, acrosome integrity, lipid peroxidation and fertility potential of sperm were assessed after thawing. HA at concentration of 2mM (HA2) resulted in the highest (Prooster sperm after freeze thawing. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Frozen herring as raw material for spice-salting

    NARCIS (Netherlands)

    Stefansson, G.; Nielsen, H.H.; Skara, T.; Schubring, R.; Oehlenschlager, J.; Luten, J.

    2000-01-01

    One batch of herring (Clupea harengus) was spice-salted fresh and as thawed after 32 days of frozen storage at -24 °C. After salting, samples of both groups were sent to participating laboratories in Iceland, Denmark, Norway, Germany and England. The herring was kept at 5 ± 1 °C and sampled three

  6. In search of better spermatogonial preservation by supplementation of cryopreserved human immature testicular tissue xenografts with N-acetylcysteine and testosterone

    Directory of Open Access Journals (Sweden)

    Jonathan ePoels

    2014-12-01

    Full Text Available Controlled slow-freezing is the procedure currently applied for immature testicular tissue cryobanking in clinical practice. Vitrification has been proposed as a promising alternative, with a view to better preserving spermatogonial stem cells for future fertility restoration by autografting in young boys suffering from cancer. It appears that besides the potential influence of the cryopreservation technique used, the transplantation procedure itself has a significant impact on spermatogonial loss observed in ITT xenografts. Eighteen immature testicular tissue pieces issuing from 6 patients aged 2-15 years were used. Fragments of fresh tissue (serving as ungrafted controls, frozen-thawed tissue, frozen-thawed tissue supplemented with N-acetylcysteine and frozen-thawed tissue supplemented with testosterone xenografted to nude mice for 5 days were compared. Upon graft removal, histological and immunohistochemical analyses were performed to evaluate spermatogonia, intratubular proliferation and intrinsic and extrinsic apoptosis. A significant decrease in the integrity of intact seminiferous tubules was found in all three grafted groups. Spermatogonia were observed by immunohistochemistry in all grafted groups, with recovery rates of 67%, 63% and 53% respectively for slow-frozen tissue, slow-frozen tissue supplemented with N-acetylcysteine and slow-frozen tissue supplemented with testosterone. Apoptosis evidenced by active caspase-3 and TUNEL was similar in all grafts. The study is limited by the low availability of immature testicular tissue samples of human origin, and no clear impact of graft supplementation was found. The mouse xenotransplantation model needs to be refined to investigate human spermatogenesis in human immature testicular tissue grafts.

  7. Dissolved organic carbon and nitrogen release from Holocene permafrost and seasonally frozen soils

    Science.gov (United States)

    Wickland, K.; Waldrop, M. P.; Koch, J. C.; Jorgenson, T.; Striegl, R. G.

    2017-12-01

    Permafrost (perennially frozen) soils store vast amounts of carbon (C) and nitrogen (N) that are vulnerable to mobilization to the atmosphere as greenhouse gases and to terrestrial and aquatic ecosystems as dissolved organic carbon (DOC) and total dissolved nitrogen (TDN) upon thaw. Such releases will affect the biogeochemistry of arctic and boreal regions, yet little is known about active layer (seasonally frozen) and permafrost source variability that determines DOC and TDN mobilization. We quantified DOC and TDN leachate yields from a range of active layer and permafrost soils in Alaska varying in age and C and N content to determine potential release upon thaw. Soil cores from the upper 1 meter were collected in late winter, when soils were frozen, from three locations representing a range in geographic position, landscape setting, permafrost depth, and soil types across interior Alaska. Two 15 cm-thick segments were extracted from each core: a deep active-layer horizon and a shallow permafrost horizon. Soils were thawed and leached for DOC and TDN yields, dissolved organic matter optical properties, and DOC biodegradability; soils were analyzed for C and N content, and radiocarbon content. Soils had wide-ranging C and N content (<1-44% C, <0.1-2.3% N), and varied in radiocarbon age from 450-9200 years before present - thus capturing typical ranges of boreal and arctic soils. Soil DOC and TDN yields increased linearly with soil C and N content, and decreased with increasing radiocarbon age. However, across all sites DOC and TDN yields were significantly greater from permafrost soils (0.387 ± 0.324 mg DOC g-1 soil; 0.271 ± 0.0271 mg N g-1 soil) than from active layer soils (0.210 ± 0.192 mg DOC g-1 soil; 0.00716 ± 0.00569 mg N g-1 soil). DOC biodegradability increased with increasing radiocarbon age, and was statistically similar for active layer and permafrost soils. Our findings suggest that the continuously frozen state of permafrost soils has preserved

  8. Effects of Technological Parameters and Fishing Ground on Quality Attributes of Thawed, Chilled Cod Fillets Stored in Modified Atmosphere Packaging

    DEFF Research Database (Denmark)

    Bøknæs, Niels; Østerberg, Carsten; Sørensen, Rie

    2001-01-01

    . The parameters investigated were: (1) packaging in modified atmosphere during frozen storage, (2)frozen storage period and temperature, (3),fishing ground and chill storage temperature, together with (4) the addition of trimethylamine oxide (TMAO) and sodium chloride (NaCl) to cod fillets before freezing......Effects were studied of various technological parameters and fishing ground on quality attributes of thawed, chilled cod fillets stored in modified atmosphere packaging Frozen fillets of Baltic Sea and Barents Sea cod, representing two commercial fishing grounds, were used as raw material...... of Baltic Sea cod. Therefore, addition of trimethylamine oxide and NaCl to Baltic Sea cod fillets was evaluated and shown to protect P, phosphoreum against fro::en storage inactivation and this explained the observed differences in growth of the spoilage bacteria and trimethylamine production between thawed...

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

    Directory of Open Access Journals (Sweden)

    Yahya TÜLEK

    1999-03-01

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

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

    Science.gov (United States)

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

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

  11. Factors affecting the fertility of ewes after intrauterine insemination with frozen-thawed semen during the non-breeding season.

    Science.gov (United States)

    Fukui, Yutaka; Kohno, Hirohide; Okabe, Kentaro; Katsuki, Sara; Yoshizawa, Masahiro; Togari, Tetsuro; Watanabe, Hiroyuki

    2010-08-01

    In this study, two successive field trials were conducted during the non-breeding season to investigate various factors affecting on fertility of Suffolk ewes after intrauterine insemination with frozen-thawed semen. In the first year (Experiment 1), three sperm numbers per insemination dose (0.25, 0.5 and 1 million sperm) and five sheep farms were used, and in the second year (Experiment 2), parity, age, body weight, body condition score (BCS) and postpartum days were investigated to compare pregnancy and lambing rates. High pregnancy and lambing rates (70.6 and 70.6%, respectively) were obtained with 0.25 million sperm per dose. There were no significant differences in the pregnancy and lambing rates among the five farms, but there was a tendency for one farm to have higher pregnancy (75.8%, P=0.065) and lambing (72.7%, P=0.077) rates than those (46.7-53.3% and 45.2-53.3% for the pregnancy and lambing rates, respectively) of the other farms. In Experiment 2, ewe age significantly affected both the pregnancy and lambing rates. Nulliparous ewes had a higher lambing rate (72.0%) than that (44.2%) of multiparous ewes, but a significant difference was not revealed. Regardless of body weight, BCS tended to be an important factor influencing on fertility of ewes. Body weight and the postpartum days did not affect the fertility of ewes. It was concluded from these results that the fertility of Suffolk ewes after intrauterine insemination with frozen semen was significantly influenced by sperm number per dose and ewe age. Nulliparous ewes at less than three years of age and with a BCS of more than 3.0 are expected to have higher fertility than other ewes.

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

    Directory of Open Access Journals (Sweden)

    Peng Liu

    2016-07-01

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

  13. Live births after polar body biopsy and frozen-thawed cleavage stage embryo transfer: case report.

    Science.gov (United States)

    Guimarães, Fernando; Roque, Matheus; Valle, Marcello; Kostolias, Alessandra; Azevedo, Rodrigo A de; Martinhago, Ciro D; Sampaio, Marcos; Geber, Selmo

    2016-12-01

    Pre-implantation genetic diagnosis (PGD) or screening (PGS) technology, has emerged and developed in the past few years, benefiting couples as it allows the selection and transfer of healthy embryos during IVF treatments. These techniques can be performed in oocytes (polar-body biopsy) or embryos (blastomere or trophectoderm biopsy). In this case report, we describe the first two live births to be published in Brazil after a polar-body (PB) biopsy. In case 1, a 42-year-old was submitted to PB biopsy with PGS due to advanced maternal age and poor ovarian reserve. Five MII oocytes underwent first and second polar body biopsy and four cleavage embryos were cryopreserved. The PGS analysis resulted in two euploid embryos (next generation sequence). A frozen-thawed embryo transfer (FET) was performed after endometrial priming and a healthy baby was delivered after a cesarean section (37 weeks, female, 3390g, 47.5 cm). In case 2, a 40-year old patient with balanced translocation and poor ovarian response was submitted to PB biopsy. Two MII oocytes underwent first and second polar body biopsy and two embryos were cryopreserved in cleavage stage. The analysis resulted in one euploid embryo that was transferred after endometrial priming. A preterm healthy baby (34 weeks, female, 2100g, 40 cm) was delivered via cesarean section. In conclusion, although the blastocyst biopsy is the norm when performing PGS/PGD during IVF treatments, other alternatives (as PB biopsy) should be considered in some specific situations.

  14. Frozen blood products: clinically effective and potentially ideal for remote Australia.

    Science.gov (United States)

    Holley, A; Marks, D C; Johnson, L; Reade, M C; Badloe, J F; Noorman, F

    2013-01-01

    The development of effective cryopreservation techniques for both red blood cells and platelets, which maintain ex vivo biological activity, in combination with frozen plasma, provides for a unique blood banking strategy. This technology greatly enhances the storage life of these products. The rationale and potential advantages of using cryopreservation techniques for the provision of blood products to remote and military environments have been effectively demonstrated in several conflicts over the last decade. Current haemostatic resuscitation doctrine for the exsanguinating patient supports the use of red blood cells, platelets and frozen plasma early in the resuscitation. We believe an integrated fresh-frozen blood bank inventory could facilitate provision of blood products, not only in the military setting but also in regional Australia, by overcoming many logistic and geographical challenges. The processes involved in production and point of care thawing are sufficiently well developed and achievable to make this technology a viable option. The potential limitations of cryopreservation and subsequent product thawing need to be considered if such a strategy is to be developed. A substantial body of international experience using cryopreserved products in remote settings has already been accrued. This experience provides a template for the possible creation of an Australian integrated fresh-frozen blood bank inventory that could conceivably enhance the care of patients in both regional Australia and in the military setting.

  15. Thawing of ice in porous space of building materials: Experimental monitoring and computational modelling

    Science.gov (United States)

    Kočí, Jan; Maděra, Jiří; Kočí, Václav; Hlaváčová, Zuzana; Černý, Robert

    2017-11-01

    A simple laboratory experiment for the determination of thermal response of a studied sample during thawing is described in the paper. The sample made of autoclaved aerated concrete was partially water saturated and frozen. Then, the temperature development during thawing was recorded, allowing to identify the time scale of the phase change process taking place inside the sample. The experimental data was then used in the inverse analysis, in order to find unknown parameters of the smoothed effective specific heat capacity model.

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

    International Nuclear Information System (INIS)

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

    1993-01-01

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

  17. Comparison of pregnancy rate between fresh embryo transfers and frozen-thawed embryo transfers following ICSI treatment

    Directory of Open Access Journals (Sweden)

    Zahra Basirat

    2016-01-01

    Full Text Available Background: The use of assisted reproductive technology (ART is increasing in the world. The rate, efficacy and safety of ART are very different among countries. There is an increase in the use of intra cytoplasmic sperm injection (ICSI, single fresh embryo transfer (ET and frozen-thawed embryo transfer (FET. Objective: The objective of this study was to compare pregnancy rate in fresh ET and FET. Materials and Methods: In this retrospective cross-sectional study 1014 ICSI-ET cycles (426 fresh ET and 588 FET from 753 women undergoing ICSI treatment referred to Fatemezahra Infertility and Reproductive Health Research Center in Babol, Iran from 2008 to 2013 were reviewed. Results: There were no significant differences between biochemical pregnancy rate (23% versus 18.8%, OR 1.301; 95% CI .95-1.774, gestational sac (95.6% versus 100% in FET, OR 0.60; 95% CI 0.54-0.67, and fetal heart activity (87.2% versus 93.6% OR .46; 95% CI .16-1.32 in fresh ET and FET cycles, respectively. P< 0.05 was considered statistically significant for all measures. Conclusion: Although, the result showed no significantly difference between the fresh ET and the FET cycles, however the embryos are able to be stored for subsequent ART. Therefore, we recommend FET cycles as an option alongside the fresh ET.

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

    Directory of Open Access Journals (Sweden)

    Martina Doležalová

    2015-01-01

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

  19. Biochemical effects on long-term frozen human costal cartilage

    Energy Technology Data Exchange (ETDEWEB)

    Santin, Stefany P.; Martinho Junior, Antonio C.; Yoshito, Daniele; Soares, Fernando A.N.; Mathor, Monica B., E-mail: mathor@ipen.b [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2011-07-01

    Currently, the progresses on treatment of musculoskeletal diseases with the evolving of artificial implants and the success of tissue transplantation between genetically different individuals have conducted to an increase in radiosterilization. Regarding to tissue transplantation, it is essential to have sterile tissue and many tissue banks use radiosterilization as an effective method to sterilize these tissues. However, high doses of ionizing radiation and the preservation method may induce structural modifications in the tissues, as degradation of structural scaffold, decreasing its mechanical properties. Particularly, cartilage have been preserved in high concentrations of glycerol or deep-frozen at -70 degree C for storage after radiosterilization. Therefore, it is important to study the modifications induced in cartilage by preservation methods and by radiosterilization to determine the appropriated parameters for high quality of human allografts. Costal cartilages were obtained from cadaveric donors and were frozen at -20 degree C for 2 years long in order to compare with previous studies for fresh, deep-frozen and glycerolised cartilages. The mechanical tests were carried out in a universal testing machine until sample failure. According our results, there is no significant statistical difference between stress at break of fresh, long-term - 20 degree C frozen cartilages and deep-frozen cartilage. This early result suggests, regarding to tensile property, that long-term - 20 degree C frozen cartilages corresponds to glycerolised costal cartilages irradiated with 25 kGy or deep-frozen cartilages irradiated with 25 and 50 kGy. Thus, this long-term frozen cartilages may be used for tissue banks, but more studies about effects of ionizing radiation are necessary. (author)

  20. Biochemical effects on long-term frozen human costal cartilage

    International Nuclear Information System (INIS)

    Santin, Stefany P.; Martinho Junior, Antonio C.; Yoshito, Daniele; Soares, Fernando A.N.; Mathor, Monica B.

    2011-01-01

    Currently, the progresses on treatment of musculoskeletal diseases with the evolving of artificial implants and the success of tissue transplantation between genetically different individuals have conducted to an increase in radiosterilization. Regarding to tissue transplantation, it is essential to have sterile tissue and many tissue banks use radiosterilization as an effective method to sterilize these tissues. However, high doses of ionizing radiation and the preservation method may induce structural modifications in the tissues, as degradation of structural scaffold, decreasing its mechanical properties. Particularly, cartilage have been preserved in high concentrations of glycerol or deep-frozen at -70 degree C for storage after radiosterilization. Therefore, it is important to study the modifications induced in cartilage by preservation methods and by radiosterilization to determine the appropriated parameters for high quality of human allografts. Costal cartilages were obtained from cadaveric donors and were frozen at -20 degree C for 2 years long in order to compare with previous studies for fresh, deep-frozen and glycerolised cartilages. The mechanical tests were carried out in a universal testing machine until sample failure. According our results, there is no significant statistical difference between stress at break of fresh, long-term - 20 degree C frozen cartilages and deep-frozen cartilage. This early result suggests, regarding to tensile property, that long-term - 20 degree C frozen cartilages corresponds to glycerolised costal cartilages irradiated with 25 kGy or deep-frozen cartilages irradiated with 25 and 50 kGy. Thus, this long-term frozen cartilages may be used for tissue banks, but more studies about effects of ionizing radiation are necessary. (author)

  1. The impacts of recent permafrost thaw on land-atmosphere greenhouse gas exchange

    Science.gov (United States)

    Hayes, Daniel J.; Kicklighter, David W.; McGuire, A. David; Chen, Min; Zhuang, Qianlai; Yuan, Fengming; Melillo, Jerry M.; Wullschleger, Stan D.

    2014-01-01

    Permafrost thaw and the subsequent mobilization of carbon (C) stored in previously frozen soil organic matter (SOM) have the potential to be a strong positive feedback to climate. As the northern permafrost region experiences as much as a doubling of the rate of warming as the rest of the Earth, the vast amount of C in permafrost soils is vulnerable to thaw, decomposition and release as atmospheric greenhouse gases. Diagnostic and predictive estimates of high-latitude terrestrial C fluxes vary widely among different models depending on how dynamics in permafrost, and the seasonally thawed 'active layer' above it, are represented. Here, we employ a process-based model simulation experiment to assess the net effect of active layer dynamics on this 'permafrost carbon feedback' in recent decades, from 1970 to 2006, over the circumpolar domain of continuous and discontinuous permafrost. Over this time period, the model estimates a mean increase of 6.8 cm in active layer thickness across the domain, which exposes a total of 11.6 Pg C of thawed SOM to decomposition. According to our simulation experiment, mobilization of this previously frozen C results in an estimated cumulative net source of 3.7 Pg C to the atmosphere since 1970 directly tied to active layer dynamics. Enhanced decomposition from the newly exposed SOM accounts for the release of both CO2 (4.0 Pg C) and CH4 (0.03 Pg C), but is partially compensated by CO2 uptake (0.3 Pg C) associated with enhanced net primary production of vegetation. This estimated net C transfer to the atmosphere from permafrost thaw represents a significant factor in the overall ecosystem carbon budget of the Pan-Arctic, and a non-trivial additional contribution on top of the combined fossil fuel emissions from the eight Arctic nations over this time period.

  2. Birth of puppies of predetermined sex after artificial insemination with a low number of sex-sorted, frozen-thawed spermatozoa in field conditions.

    Science.gov (United States)

    Wei, Yun-Fang; Chen, Fang-Liang; Tang, Shu-Sheng; Mao, Ai-Guo; Li, Li-Guang; Cheng, Lu-Guang; Chen, Chao; Li, Fei-Xiang; Wang, Bin; Xu, Tao; Zhang, Yue-Jun; Li, Jing; Wan, Jiu-Sheng

    2017-08-01

    The aim of this study was to evaluate fertility and sex ratios after artificial insemination in dogs under field conditions. Semen was cryopreserved as unsorted (control) or was separated into X- and Y-chromosome-bearing sperm using a cell sorter. Sixty female dogs were inseminated with frozen-thawed spermatozoa of 100 × 10 6 unsorted (a dose in practice) and 4 × 10 6 sorted (X and Y group, respectively). A total of 20 dogs became pregnant and 126 puppies were born from the three groups. The percentage of parturition was similar for the X (5/20; 25.0%) and Y (4/20; 20.0%) group (P > 0.05), but lower than controls (11/20; 55.0%) (P dog spermatozoa at a farm level, making sperm-sexing technology potentially applicable for elite breeding units. © 2017 Japanese Society of Animal Science.

  3. Modeling snowmelt infiltration in seasonally frozen ground

    Science.gov (United States)

    Budhathoki, S.; Ireson, A. M.

    2017-12-01

    In cold regions, freezing and thawing of the soil govern soil hydraulic properties that shape the surface and subsurface hydrological processes. The partitioning of snowmelt into infiltration and runoff has also important implications for integrated water resource management and flood risk. However, there is an inadequate representation of the snowmelt infiltration into frozen soils in most land-surface and hydrological models, creating the need for improved models and methods. Here we apply, the Frozen Soil Infiltration Model, FroSIn, which is a novel algorithm for infiltration in frozen soils that can be implemented in physically based models of coupled flow and heat transport. In this study, we apply the model in a simple configuration to reproduce observations from field sites in the Canadian prairies, specifically St Denis and Brightwater Creek in Saskatchewan, Canada. We demonstrate the limitations of conventional approaches to simulate infiltration, which systematically over-predict runoff and under predict infiltration. The findings show that FroSIn enables models to predict more reasonable infiltration volumes in frozen soils, and also represent how infiltration-runoff partitioning is impacted by antecedent soil moisture.

  4. THAWING PROCEDURES FOR HOSPITAL-MADE ENTERAL FEEDINGS: SURVIVAL OF COLIFORM AND MESOPHILIC AEROBIC BACTERIA

    Directory of Open Access Journals (Sweden)

    KATHIA ROSSI ROLIM LOPES

    2009-07-01

    Full Text Available

    ABSTRACT: This study had the purpose to observe the effect of thawing procedures on survival of coliform and mesophilic aerobic bacteria in hospital-made enteral feedings. The samples are represented by three different lots. The tests were realized in three moments: immediately after the sample preparation and after freezing during 1 or 2 months. The thawing procedures were denominated convencional and alternative. The first, used by hospital, utilizes water bath at 50ºC, considering the time spent from the total thawing to its distribution in the infirmaries. The second was the fast thawing made by microwaving. The results showed that the reduction of the mesophiles and coliform was related to the time the samples were frozen. The results obtained indicate an advantage of the alternative method, which presented lower total and fecal coliform counts than the conventional one. KEYWORDS: Enteral feedings; thawing; food microbiology.

  5. Assessing the use of frozen pork meat in the manufacture of cooked ham

    OpenAIRE

    LOS, Francine Gomes Basso; PRESTES, Rosa Cristina; GRANATO, Daniel; SIMÕES, Deise Rosana Silva; ROMAN, Silvane Souza; DEMIATE, Ivo Mottin

    2016-01-01

    This study aimed to evaluate the effects of slow (–18 °C) and fast freezing (liquid nitrogen) of pork meat, and the use of exudate released upon thawing, on the physicochemical, color, rheological, microbiological, histological, and sensory characteristics of cooked ham. The meat samples were frozen at –18 °C and thawed after 22 weeks for the production of cooked ham. No significant difference was observed regarding physicochemical, color and microbiological parameters or in senso...

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

    Science.gov (United States)

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

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

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

    Science.gov (United States)

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

    2007-08-11

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

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

    Science.gov (United States)

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

    2017-07-01

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

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

    Directory of Open Access Journals (Sweden)

    Alice Roffi

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Thomas Jagdhuber

    2014-03-01

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

  11. Effect of Holder pasteurization and frozen storage on macronutrients and energy content of breast milk.

    Science.gov (United States)

    García-Lara, Nadia Raquel; Vieco, Diana Escuder; De la Cruz-Bértolo, Javier; Lora-Pablos, David; Velasco, Noelia Ureta; Pallás-Alonso, Carmen Rosa

    2013-09-01

    The aim of this study was to explore the effect of Holder pasteurization and frozen storage at -20°C after pasteurization on fat, total nitrogen, lactose, and energy content of breast milk. Both procedures are routinely practiced in human milk banks. A total of 34 samples of frozen breast milk, donated by 28 women, were collected. Once thawed, an aliquot of each sample was analyzed before pasteurization; the remaining milk was pasteurized (Holder method) and split into 8 aliquots. One aliquot was analyzed after pasteurization and the remainder frozen at -20°C and analyzed 30, 60, 90, 120, and 180 days later. For every aliquot, fat, total nitrogen, lactose, and energy content were determined using the device human Milk Analyzer. We observed a significant reduction in fat (3.5%; -0.17 (-0.29; -0.04) g/dL) and energy content (2.8%; -2.03 (-3.60; -0.46) g/dL) after pasteurization. A significant decrease over time was observed for fat, lactose and energy content. No significant changes were observed for nitrogen content. Mean differences between day 0 postpasteurization and day 180 were -0.13 (-0.21; -0.06) g/dL for fat, -0.08 (-0.13; -0.03) g/dL for lactose, and -1.55 (-2.38; -0.71) kcal/dL for energy content. The relative decreases were 2.8%, 1.7%, and 2.2%, respectively. Overall (postpasteurization + frozen storage), a 6.2% and 5% decrease were observed for fat and energy, respectively. Holder pasteurization decreased fat and energy content of human milk. Frozen storage at -20°C of pasteurized milk significantly reduced fat, lactose, and energy content of human milk.

  12. The effect of kinds and concentration of cryoprotectant and thawing methods on frozen semen of Arab chicken

    Directory of Open Access Journals (Sweden)

    Sofjan Iskandar

    2006-03-01

    Full Text Available The success of freezing chicken semen is the hope for preserving Indonesian native chickens. Semen from twenty Arab roosters were collected using massage technique once in a week. Cryoprotectant DMA (dimethyl acetamide or DMF (dimethyl formalmide of 7 or 9% and thawing A at temperature of 30oC for 30 seconds or in B at 5oC for 5 minutes. The volume of fresh semen was 0.3 ± 0.072 ml/ejaculate, white colour, rather thick to thick, with 2200 ± 372 millions sperms/ml and pH 6.95 ± 0.32, 4+/3+ mass movement, 80% motility, 84 ± 4.48% and abnormality of 14.75 ± 1.28%. There were not statistically significant (P>0.05 effect of interaction of treatments (kinds and concentrations of cryoprotectant, and thawing methods on motility and live-sperms. Sperm motility preserved with DMA (34.69% significantly higher than with DMF (29.84%. Sperm motility was also significantly higher (34.53% when preserved with 7% cryoprotectant than with 9% (30%. Thawing-A significantly gave higher motility (35.31% than thawing-B did (29.22%. Live-sperms of semen preserved with DMA (46.75% was significantly higher than with DMF (41.72%. Cryoprotectant concentration of 7% gave higher live-sperms (46.98% than of 9% did (41.48%. Thawing-A left live-sperms of 47.14%, which was significantly higher than thawing-B did (41.30%.

  13. Cryo-thawed embryo transfer : natural versus artificial cycle. A non-inferiority trial. (ANTARCTICA trial)

    NARCIS (Netherlands)

    Groenewoud, Eva R.; Macklon, Nick S.; Cohlen, Ben J.

    2012-01-01

    Background: Frozen thawed embryo transfer (FET) is a cost-effective adjunct to IVF or IVF-ICSI treatment. In order to optimize treatment outcome, FET should be carried out during a period of optimal endometrial receptivity. To optimize implantation several methods for endometrium preparation have

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

    Science.gov (United States)

    Chen, Xiyu; Liu, Lin

    2017-04-01

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

  15. The transcriptional response of microbial communities in thawing Alaskan permafrost soils

    Science.gov (United States)

    Coolen, Marco J. L.; Orsi, William D.

    2015-01-01

    Thawing of permafrost soils is expected to stimulate microbial decomposition and respiration of sequestered carbon. This could, in turn, increase atmospheric concentrations of greenhouse gasses, such as carbon dioxide and methane, and create a positive feedback to climate warming. Recent metagenomic studies suggest that permafrost has a large metabolic potential for carbon processing, including pathways for fermentation and methanogenesis. Here, we performed a pilot study using ultrahigh throughput Illumina HiSeq sequencing of reverse transcribed messenger RNA to obtain a detailed overview of active metabolic pathways and responsible organisms in up to 70 cm deep permafrost soils at a moist acidic tundra location in Arctic Alaska. The transcriptional response of the permafrost microbial community was compared before and after 11 days of thaw. In general, the transcriptional profile under frozen conditions suggests a dominance of stress responses, survival strategies, and maintenance processes, whereas upon thaw a rapid enzymatic response to decomposing soil organic matter (SOM) was observed. Bacteroidetes, Firmicutes, ascomycete fungi, and methanogens were responsible for largest transcriptional response upon thaw. Transcripts indicative of heterotrophic methanogenic pathways utilizing acetate, methanol, and methylamine were found predominantly in the permafrost table after thaw. Furthermore, transcripts involved in acetogenesis were expressed exclusively after thaw suggesting that acetogenic bacteria are a potential source of acetate for acetoclastic methanogenesis in freshly thawed permafrost. Metatranscriptomics is shown here to be a useful approach for inferring the activity of permafrost microbes that has potential to improve our understanding of permafrost SOM bioavailability and biogeochemical mechanisms contributing to greenhouse gas emissions as a result of permafrost thaw. PMID:25852660

  16. The transcriptional response of microbial communities in thawing Alaskan permafrost soils

    Directory of Open Access Journals (Sweden)

    M J L Coolen

    2015-03-01

    Full Text Available Thawing of permafrost soils is expected to stimulate microbial decomposition and respiration of sequestered carbon. This could, in turn, increase atmospheric concentrations of greenhouse gases, such as carbon dioxide and methane, and create a positive feedback to climate warming. Recent metagenomic studies suggest that permafrost has a large metabolic potential for carbon processing, including pathways for fermentation and methanogenesis. Here, we performed a pilot study using ultrahigh throughput Illumina HiSeq sequencing of reverse transcribed messenger RNA to obtain a detailed overview of active metabolic pathways and responsible organisms in up to 70 cm deep permafrost soils at a moist acidic tundra location in Arctic Alaska. The transcriptional response of the permafrost microbial community was compared before and after eleven days of thaw. In general, the transcriptional profile under frozen conditions suggests a dominance of stress responses, survival strategies, and maintenance processes, whereas upon thaw a rapid enzymatic response to decomposing soil organic matter (SOM was observed. Bacteroidetes, Firmicutes, ascomycete fungi, and methanogens were responsible for largest transcriptional response upon thaw. Transcripts indicative of heterotrophic methanogenic pathways utilizing acetate, methanol, and methylamine were found predominantly in the permafrost table after thaw. Furthermore, transcripts involved in acetogenesis were expressed exclusively after thaw suggesting that acetogenic bacteria are a potential source of acetate for acetoclastic methanogenesis in freshly thawed permafrost. Metatranscriptomics is shown here to be a useful approach for inferring the activity of permafrost microbes that has potential to improve our understanding of permafrost SOM bioavailability and biogeochemical mechanisms contributing to greenhouse gas emissions as a result of permafrost thaw.

  17. Pregnancy and neonatal outcomes following letrozole use in frozen-thawed single embryo transfer cycles.

    Science.gov (United States)

    Tatsumi, T; Jwa, S C; Kuwahara, A; Irahara, M; Kubota, T; Saito, H

    2017-06-01

    Are pregnancy and neonatal outcomes following letrozole use comparable with natural and HRT cycles in patients undergoing single frozen-thawed embryo transfer (FET)? Letrozole use was significantly associated with higher rates of clinical pregnancy, clinical pregnancy with fetal heart beat and live birth, and with a lower rate of miscarriage, compared with natural and HRT cycles. Letrozole is the most commonly used aromatase inhibitor for mild ovarian stimulation in ART. However, the effect of letrozole on pregnancy and neonatal outcomes in FET are not well known. A retrospective cohort study was conducted using data from the Japanese national ART registry between 2012 and 2013. A total of 110 722 single FET cycles with letrozole (n = 2409), natural (n = 41 470) or HRT cycles (n = 66 843) were included. The main outcomes were the rates of clinical pregnancy, clinical pregnancy with fetal heart beat, miscarriage and live birth. Adjusted odds ratios and relative risks (RRs) were calculated using a generalized estimating equation adjusting for correlations within clinics. The rates of clinical pregnancy, clinical pregnancy with fetal heart beat, and live birth were significantly higher, while the rate of miscarriage was significantly lower in the letrozole group compared with the natural and HRT groups. In blastocyst stage transfers, the adjusted RRs for clinical pregnancy with fetal heart beat of letrozole compared with natural and HRT cycles were 1.48 (95% CI: 1.41-1.55) and 1.62 (95% CI: 1.54-1.70), respectively. Similarly, the adjusted RRs of letrozole for miscarriage compared with natural and HRT cycles were 0.91 (95% CI: 0.88-0.93) and 0.84 (95% CI: 0.82-0.87), respectively. Neonatal outcomes were mostly similar in letrozole, natural and HRT cycles. Important limitations of this study included the lack of information concerning the reasons for selecting the specific FET method, parity, the number of previous ART failures, embryo quality and the dose and duration

  18. Effects of single dose GnRH agonist as luteal support on pregnancy outcome in frozen-thawed embryo transfer cycles: an RCT

    Directory of Open Access Journals (Sweden)

    Robab Davar

    2015-08-01

    Full Text Available Background: There is no doubt that luteal phase support is essential to enhance the reproductive outcome in IVF cycles. In addition to progesterone and human chorionic gonadotropin, several studies have described GnRH agonists as luteal phase support to improve implantation rate, pregnancy rate and live birth rate, whereas other studies showed dissimilar conclusions. All of these studies have been done in fresh IVF cycles. Objective: To determine whether an additional GnRH agonist administered at the time of implantation for luteal phase support in frozen-thawed embryo transfer (FET improves the embryo developmental potential. Materials and Methods: This is a prospective controlled trial study in 200 FET cycles, patients were randomized on the day of embryo transfer into group 1 (n=100 to whom a single dose of GnRH agonist (0.1 mg triptorelin was administered three days after transfer and group 2 (n=100, who did not receive agonist. Both groups received daily vaginal progesterone suppositories plus estradiol valerate 6 mg daily. Primary outcome measure was clinical pregnancy rate. Secondary outcome measures were implantation rate, chemical, ongoing pregnancy rate and abortion rate. Results: A total of 200 FET cycles were analyzed. Demographic data and embryo quality were comparable between two groups. No statistically significant difference in clinical and ongoing pregnancy rates was observed between the two groups (26% versus 21%, p=0.40 and 21% versus 17%, p=0.37, respectively. Conclusion: Administration of a subcutaneous GnRH agonist at the time of implantation does not increase clinical or ongoing pregnancy.

  19. A reliable procedure for decontamination before thawing of human specimens cryostored in liquid nitrogen: three washes with sterile liquid nitrogen (SLN2).

    Science.gov (United States)

    Parmegiani, Lodovico; Accorsi, Antonio; Bernardi, Silvia; Arnone, Alessandra; Cognigni, Graciela Estela; Filicori, Marco

    2012-10-01

    To report a washing procedure, to be performed as frozen specimens are taken out of cryobanks, to minimize the risk of hypothetical culture contamination during thawing. Basic research. Private assisted reproduction center. Two batches of liquid nitrogen (LN(2)) were experimentally contaminated, one with bacteria (Pseudomonas aeruginosa, Escherichia coli, Stenotrophomonas maltophilia) and the other with fungi (Aspergillus niger). Two hundred thirty-two of the most common human gamete/embryo vitrification carriers (Cryotop, Cryoleaf, Cryopette) were immersed in the contaminated LN(2) (117 in the bacteria and 25 in the fungi-contaminated LN(2)). The carriers were tested microbiologically, one group without washing (control) and the other after three subsequent washings in certified ultraviolet sterile liquid nitrogen (SLN(2)). The carriers were randomly allocated to the "three-wash procedure" (three-wash group, 142 carriers) or "no-wash" (control group, 90 carriers) using a specific software tool. Assessment of microorganism growth. In the no-wash control group, 78.6% of the carriers were contaminated by the bacteria and 100% by the fungi. No carriers were found to be contaminated, either by bacteria or fungi, after the three-wash procedure. The three-wash procedure with SLN(2) produced an efficient decontamination of carriers in extreme experimental conditions. For this reason, this procedure could be routinely performed in IVF laboratories for safe thawing of human specimens that are cryostored in nonhermetical cryocontainers, particularly in the case of open or single-straw closed vitrification systems. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  20. Influence of immersion freezing in NaCl solutions and of frozen storage on the viscoelastic behavior of mozzarella cheese.

    Science.gov (United States)

    Ribero, G G; Rubiolo, A C; Zorrilla, S E

    2007-06-01

    The freezing of Mozzarella cheese by immersion in NaCl solutions may be an innovative procedure for the dairy industry because it combines conveniently salting and freezing processes. In this work, the influence of this type of freezing method and of the frozen storage of samples on the viscoelastic behavior of Mozzarella cheese was studied. Slabs (2 x 10 x 10 cm(3)) were immersed in 23% w/w NaCl solutions (control samples: 4 degrees C, 90 min; frozen samples: -15 degrees C, 180 min). Half of the frozen samples were immediately thawed at 4 degrees C . The other half was stored at -20 degrees C for 2 mo and then was thawed at 4 degrees C (frozen-stored samples). Samples were stored at 4 degrees C and assayed at 1, 7, 14, 20, 27, 34, and 41 d. Rheological tests were carried out in oscillatory mode (parallel-plate geometry, diameter: 20 mm, gap: 1 mm, frequency: 1 Hz). Strain sweeps were run (0.001 immersion freezing of Mozzarella cheese affects some of the studied parameters, the differences observed between frozen and frozen-stored samples with control samples were small. Therefore, it was considered that the immersion freezing might be useful for the manufacture and commercialization of Mozzarella cheese.

  1. Experimental Study and a Mathematical Model of the Processes in Frozen Soil Under a Reservoir with a Hot Heat-Transfer Agent

    Science.gov (United States)

    Kislitsyn, A. A.; Shastunova, U. Yu.; Yanbikova, Yu. F.

    2018-05-01

    On an experimental setup, the authors have measured temperature fields in frozen soil during the filling of a reservoir with hot heat-transfer agent (oil), and also the change in the shape and position of the front of ice melting (isotherms T = 0°C) with time. The approximate solution of a two-dimensional Stefan problem on thawing of frozen soil has been given; it has been shown that satisfactory agreement with experimental results can only be obtained with account taken of the convective transfer of heat due to the water motion in the region of thawed soil.

  2. Qualitative assessment of imported frozen fish fillets in Sulaimani markets

    Directory of Open Access Journals (Sweden)

    Z.K. Khidhir

    2013-06-01

    Full Text Available This study aims to determine the quality of frozen fish fillets sold in Sulaimani city markets a total number of (64 sample of frozen fish fillets belonged to 4 foreign trademarks were collected from different parts of Sulaimani markets. The samples were subjected to physical and chemical tests to determine their quality and suitability for human consumption. The proximate chemical analysis referred to presence of significant differences in moisture, fat and protein content among the four trademarks except for ash content. While the physical indices showed that White fish fillets recorded the lowest thawing and cooking loss which in return recorded the highest WHC. Chemical indices showed that the pH mean values of Myanmar and Flander mark were significantly differed (P<0.05 than Hasson and White fish fillet, Although, the results of FFA recorded no significant differences among the trademarks, and Flander mark recorded the highest PV and TBA among the other which made it significantly differed than them and White fish fillet recorded the lowest, still, they were within the international standard limits. Where, the results of TVN values recorded no significant differences ( among the inspected marks. All obtained results referred to the validity of these fish fillets for human consumption.

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

    Science.gov (United States)

    Smith, Cosmo

    2011-01-01

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

  4. Nonlinear CO2 flux response to 7 years of experimentally induced permafrost thaw.

    Science.gov (United States)

    Mauritz, Marguerite; Bracho, Rosvel; Celis, Gerardo; Hutchings, Jack; Natali, Susan M; Pegoraro, Elaine; Salmon, Verity G; Schädel, Christina; Webb, Elizabeth E; Schuur, Edward A G

    2017-09-01

    Rapid Arctic warming is expected to increase global greenhouse gas concentrations as permafrost thaw exposes immense stores of frozen carbon (C) to microbial decomposition. Permafrost thaw also stimulates plant growth, which could offset C loss. Using data from 7 years of experimental Air and Soil warming in moist acidic tundra, we show that Soil warming had a much stronger effect on CO 2 flux than Air warming. Soil warming caused rapid permafrost thaw and increased ecosystem respiration (R eco ), gross primary productivity (GPP), and net summer CO 2 storage (NEE). Over 7 years R eco , GPP, and NEE also increased in Control (i.e., ambient plots), but this change could be explained by slow thaw in Control areas. In the initial stages of thaw, R eco , GPP, and NEE increased linearly with thaw across all treatments, despite different rates of thaw. As thaw in Soil warming continued to increase linearly, ground surface subsidence created saturated microsites and suppressed R eco , GPP, and NEE. However R eco and GPP remained high in areas with large Eriophorum vaginatum biomass. In general NEE increased with thaw, but was more strongly correlated with plant biomass than thaw, indicating that higher R eco in deeply thawed areas during summer months was balanced by GPP. Summer CO 2 flux across treatments fit a single quadratic relationship that captured the functional response of CO 2 flux to thaw, water table depth, and plant biomass. These results demonstrate the importance of indirect thaw effects on CO 2 flux: plant growth and water table dynamics. Nonsummer R eco models estimated that the area was an annual CO 2 source during all years of observation. Nonsummer CO 2 loss in warmer, more deeply thawed soils exceeded the increases in summer GPP, and thawed tundra was a net annual CO 2 source. © 2017 John Wiley & Sons Ltd.

  5. The impacts of recent permafrost thaw on land–atmosphere greenhouse gas exchange

    International Nuclear Information System (INIS)

    Hayes, Daniel J; Yuan, Fengming; Wullschleger, Stan D; Kicklighter, David W; Melillo, Jerry M; McGuire, A David; Chen, Min; Zhuang, Qianlai

    2014-01-01

    Permafrost thaw and the subsequent mobilization of carbon (C) stored in previously frozen soil organic matter (SOM) have the potential to be a strong positive feedback to climate. As the northern permafrost region experiences as much as a doubling of the rate of warming as the rest of the Earth, the vast amount of C in permafrost soils is vulnerable to thaw, decomposition and release as atmospheric greenhouse gases. Diagnostic and predictive estimates of high-latitude terrestrial C fluxes vary widely among different models depending on how dynamics in permafrost, and the seasonally thawed ‘active layer’ above it, are represented. Here, we employ a process-based model simulation experiment to assess the net effect of active layer dynamics on this ‘permafrost carbon feedback’ in recent decades, from 1970 to 2006, over the circumpolar domain of continuous and discontinuous permafrost. Over this time period, the model estimates a mean increase of 6.8 cm in active layer thickness across the domain, which exposes a total of 11.6 Pg C of thawed SOM to decomposition. According to our simulation experiment, mobilization of this previously frozen C results in an estimated cumulative net source of 3.7 Pg C to the atmosphere since 1970 directly tied to active layer dynamics. Enhanced decomposition from the newly exposed SOM accounts for the release of both CO 2 (4.0 Pg C) and CH 4 (0.03 Pg C), but is partially compensated by CO 2 uptake (0.3 Pg C) associated with enhanced net primary production of vegetation. This estimated net C transfer to the atmosphere from permafrost thaw represents a significant factor in the overall ecosystem carbon budget of the Pan-Arctic, and a non-trivial additional contribution on top of the combined fossil fuel emissions from the eight Arctic nations over this time period. (paper)

  6. 21 CFR 160.110 - Frozen eggs.

    Science.gov (United States)

    2010-04-01

    ... Frozen eggs. (a) Frozen eggs, frozen whole eggs, frozen mixed eggs is the food prepared by freezing... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen eggs. 160.110 Section 160.110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...

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

    International Nuclear Information System (INIS)

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

    1994-01-01

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

  8. Obstetric outcomes after fresh versus frozen-thawed embryo transfers: A systematic review and meta-analysis.

    Science.gov (United States)

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

    2018-05-21

    To evaluate if there are differences in the risks of obstetric outcomes in IVF/ICSI singleton pregnancies when compared fresh to frozen-thawed embryo transfers (FET). This was a systematic review and meta-analysis evaluating the obstetric outcomes in singleton pregnancies after FET and fresh embryo transfer. The outcomes included in this study were pregnancy-induced hypertension (PIH), pre-eclampsia, placenta previa, and placenta accreta. The search yielded 654 papers, 6 of which met the inclusion criteria and reported on obstetric outcomes. When comparing pregnancies that arose from FET or fresh embryo transfer, there was an increase in the risk of obstetric complications in pregnancies resulting from FET when compared to those emerging from fresh embryo transfers in PIH (aOR 1.82; 95% CI 1.24-2.68), pre-eclampsia (aOR 1.32, 95% CI 1.07, 1.63), and placenta accreta (aOR 3.51, 95% CI 2.04-6.05). There were no significant differences in the risk between the FET and fresh embryo transfer groups when evaluating placenta previa (aOR 0.70; 95% CI 0.46-1.08). The obstetric outcomes observed in pregnancies arising from ART may differ among fresh and FET cycles. Thus, when evaluating to perform a fresh embryo transfer or a freeze-all cycle, these differences found in obstetric outcomes between fresh and FET should be taken into account. The adverse obstetric outcomes after FET found in this study emphasize that the freeze-all policy should not be offered to all the patients, but should be offered to those with a clear indication of the benefit of this strategy.

  9. Changes in Ultrastructure and Sensory Characteristics on Electro-magnetic and Air Blast Freezing of Beef during Frozen Storage

    Science.gov (United States)

    2015-01-01

    The ultrastructure in the beef muscle of the electro-magnetic resonance and air blast freezing during the frozen storage, and the changes in the quality characteristics after thawing were evaluated. The size of ice crystal was small and evenly formed in the initial freezing period, and it showed that the size was increased as the storage period was elapsed (pfreezing showed the size of ice crystal with a lower rate of increase than the air blast freezing during the frozen storage. The thawing loss of beef stored by the electro-magnetic resonance freezing was significantly lower than the air blast freezing during frozen storage (pfreezing was higher than the air blast on 8 month (pfreezing did not show the difference until 4 months, and it showed higher acceptability in comparison with the beef stored by the air blast freezing. Thus, it is considered that the freezing method has an effect on the change in the ultrastructure and quality characteristics of the beef. PMID:26761797

  10. QUALIY PARAMETERS AND SHELF LIFE OF GAME MEAT DURING FROZEN STORAGE

    Directory of Open Access Journals (Sweden)

    M. Spaziani

    2011-01-01

    Full Text Available This study examined the effect of the duration of frozen storage at –20°C on the game meat quality parameters, namely the pH, colour, thawing and cooking losses. The oxidative stability of game meat was evaluated by the production of thiobarbituric acid reactive substances (TBARS. Frozen storage duration did not extensively influence either the quality properties, or the oxidative stability of game meat. Therefore, it was hypothesized that the higher amount of a- tocopherol in the muscles of game compared to pellet-fed animals could be mainly responsible for the lower lipid oxidation and longer shelf life.

  11. Potential Stemness of Frozen-Thawed Testicular Biopsies without Sperm in Infertile Men Included into the In Vitro Fertilization Programme

    Directory of Open Access Journals (Sweden)

    Martin Stimpfel

    2012-01-01

    Full Text Available We describe the potential stemness of a small amount of frozen-thawed testicular tissue without sperm obtained by biopsy from six patients undergoing assisted reproductive treatment. The patients were diagnosed with Sertoli Cell-Only Syndrome alone or combined with maturation arrest. Trying to provide the natural stem cell niche for cultured stem cells, all isolated cells from enzymatically degraded biopsies where cultured together in different culture media and the presence of putative mesenchymal and putative pluripotent ES-like stem cells was indicated using different methods. High throughput real-time quantitative PCR followed by multivariate analysis revealed the formation of distinct cell clusters reflecting high degree of similarity and some of these cell clusters expressed the genes characteristic for pluripotent stem cells. In the presence of the follicular fluid, prepared as serum, putative testicular stem cells showed a certain degree of plasticity, and spontaneously differentiated into adipose-like and neuronal-like cells. Additionally, using differentiation protocols putative testicular stem cells were differentiated into neuronal- and pancreatic-like cells. This study shows that in assisted reproduction programmes, testicular tissue with no sperm might be an important source of stem cells, although it is discarded in daily medical practice; this requires further research.

  12. Cooking frozen Turkey: duration, yield and stuffing alterations with solid-state starting temperature.

    Science.gov (United States)

    Moran, E T; Bauermeister, L

    2015-08-01

    Frozen pre-stuffed turkeys from 12 week females were conductively cooked at 163°C to an 85°C internal breast end-point. Bread cubes having 50% added water filled crop and body cavity to 20% of weight the turkeys were then frozen. Each of 4 treatments employed 7 carcasses: one was thawed to +5°C with repetitions at -5, -15, and -25°C enabling a regression analysis. Time to attain breast end-point was 44 min when the starting temperature was +5°C, which increased to 63 min at -5°C, and 69 min at -25°C (P cooking was initiated at +5°C and increased to 84°C as the starting temperatures fell to -25°C (P cooking, thereafter decreasing to 59 and 60°C when at -5 and -15°C, respectively, before returning to 65°C for the -25°C case (P cooking was initiated at +5°C. This increased to 22.6% at -25°C (P cooking averaged 23.7% however, the percentage of fat in the drip increased from 51.1% when using +5°C, to 57.2% with carcasses at -25°C (P cooked after thawing, particularly for crop (59.4% using +5°C vs. 55.8% frozen, P cooking from the frozen state, while cavity stuffing consistently failed to attain a safe temperature. © 2015 Poultry Science Association Inc.

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

    Science.gov (United States)

    Sun, W.; Na, S.

    2017-12-01

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

  14. Addition of cholesterol-loaded cyclodextrins to the thawing extender: effects on boar sperm quality.

    Science.gov (United States)

    Tomás, C; Gómez-Fernández, J; Gómez-Izquierdo, E; Mocé, E; de Mercado, E

    2014-06-01

    The aim of the present study was to evaluate the effect that the addition of cholesterol-loaded cyclodextrins (CLC) to the thawing extender has on the quality of frozen-thawed boar sperm. Pooled semen (n = 5) from three boars was used for the experiments. The semen was cryopreserved with an egg-yolk-based extender, it was diluted after thawing in Beltsville thawing solution (BTS) supplemented with different concentrations of CLC (0, 12.5, 25, 50 or 100 mg/500 × 10(6) sperm), and these samples were incubated at 37°C for 150 min. The following parameters of sperm quality were evaluated 30 and 150 min after incubation: sperm with intact plasma membrane (SIPM; %), sperm with normal acrosomal ridge (NAR; %), total motile sperm (TMS; %), progressively motile sperm (PMS; %) and kinetic parameters. Both SIPM and NAR increased (p extender was supplemented with 12.5, 25 and 50 mg CLC/500 × 10(6) sperm. Nevertheless, motility decreased (p extenders with CLC improves sperm viability and reduces acrosome damage after freezing/thawing. © 2014 Blackwell Verlag GmbH.

  15. Orally administered Chrysin improves post-thawed sperm quality and fertility of rooster.

    Science.gov (United States)

    Zhandi, M; Ansari, M; Roknabadi, P; Zare Shahneh, A; Sharafi, M

    2017-12-01

    Chrysin is a bioflavonoid compound found in passion flower, chamomile, propolis and honey at high levels. Post-thawed sperm quality and fertility of Chrysin-fed roosters were assessed in this study. Twenty 40-week-old male broiler breeders were randomly divided into four groups and fed basal diet supplemented with different levels of Chrysin including 0 (Ch-0), 25 (Ch-25), 50 (Ch-50) or 75 (Ch-75) mg/day for 12 consecutive weeks. Semen samples were weekly collected from 6th to 9th week of experiment to evaluate some sperm quality parameters including total and progressive motility, plasma membrane integrity and functionality (in fresh and post-thawed samples) and mitochondrial activity (only in post-thawed samples). Also, collected semen samples from 10th, 11th and 12th week of experiment were frozen and then artificially inseminated to test fertility rate. According to the results, an improvement in both fresh and post-thawed sperm quality including total [fresh: 88.00 ± 0.58 and 87.25 ± 0.67 (p roosters could ameliorate cryopreservation-induced impairment of sperm quality and fertility rate. © 2017 Blackwell Verlag GmbH.

  16. Comparison of the Viability of Cryopreserved Fat Tissue in Accordance with the Thawing Temperature

    Directory of Open Access Journals (Sweden)

    So-Min Hwang

    2015-03-01

    Full Text Available BackgroundAdipose tissue damage of cryopreserved fat after autologous fat transfer is inevitable in several processes of re-transplantation. This study aims to compare and analyze the survivability of adipocytes after thawing fat cryopreserved at -20℃ by using thawing methods used in clinics.MethodsThe survival rates of adipocytes in the following thawing groups were measured: natural thawing at 25℃ for 15 minutes; natural thawing at 25℃ for 5 minutes, followed by rapid thawing at 37℃ in a water bath for 5 minutes; and rapid thawing at 37℃ for 10 minutes in a water bath. The survival rates of adipocytes were assessed by measuring the volume of the fat layer in the top layers separated after centrifugation, counting the number of live adipocytes after staining with trypan blue, and measuring the activity of mitochondria in the adipocytes.ResultsIn the group with rapid thawing for 10 minutes in a water bath, it was observed that the cell count of live adipocytes and the activity of the adipocyte mitochondria were significantly higher than in the other two groups (P<0.05. The volume of the fat layer separated by centrifugation was also measured to be higher, which was, however, not statistically significant.ConclusionsIt was shown that the survival rate of adipocytes was higher when the frozen fat tissue was thawed rapidly at 37℃. It can thus be concluded that if fats thawed with this method are re-transplanted, the survival rate of cryopreserved fats in transplantation will be improved, and thus, the effect of autologous fat transfer will increase.

  17. Effectiveness of sucrose during the frozen storage of arazá (Eugenia stipitata Mc. Vaugh pulp

    Directory of Open Access Journals (Sweden)

    Kelly Johanna Silva-Bustos

    2011-12-01

    Full Text Available The effects of sucrose on the quality of frozen arazá pulp were investigated. Five levels of sucrose (0, 5, 10, 20, and 30% were evaluated in combination with freezing at low speed, storage during one month, and thawing at two different speeds. Sensory quality and water loss after centrifugation (WLAC were evaluated in the samples. Before freezing sucrose addition (5-30% to the pulp did not affect the sensory quality of the samples, but a negative effect on WLAC at 5-10% sucrose was observed. Furthermore, addition of 20-30% sucrose to arazá pulp, in combination with low speed-freezing, frozen storage, and low-speed-thawing, can be a useful technique to preserve the texture of arazá pulp. The described treatment gave results comparable to those obtained for the fresh pulp and much better than those obtained when freezing was done at high speed

  18. Semen quality before cryopreservation and after thawing in 543 patients with testicular cancer

    Science.gov (United States)

    MacKenna, Antonio; Crosby, Javier; Huidobro, Cristián; Correa, Eduardo; Duque, Gonzalo

    2017-01-01

    Objective The main objective of this study was to assess semen characteristics of patients with testicular cancer before cryopreservation and after thawing, to evaluate the consequences of this technique on sperm quality in patients with testicular cancer. Methods Five hundred eighty-nine samples from 543 patients with testicular cancer were cryopreserved between 1995 and 2015, one aliquot per patient was used for a thawing test to assess the impact of cryopreservation on sperm motility; semen analysis was performed before cryo preservation and after thawing, the result interpretation was carried out using the 2010 World Health Organization (WHO) Laboratory Manual, and consent forms were signed by the patients for freezing and when sperm was used for reproductive purposes. Results Hypospermia was observed in 28.7% of samples, the median sperm concentration was 18 million/mL with 35% oligozoospermia; twenty-two patients (4.1%) had azoospermia and 12.7% had severe oligozoospermia, the median sperm count was 31.3 million and 261 semen samples (44.3%) were normal in all parameters according to the WHO; total motile sperm count before cryopreservation and after thawing was 12 (0-412.2) and 7 (0-303.9) million sperm, respectively (p < 0.00001, 95% CI 5.48-14.91), which represents a 32% reduction; concerning the utilization of cryopreserved semen samples, only twelve patients (2.2%) used their frozen sperm for reproductive purposes. Conclusions An impairment in semen quality was found in almost half of the samples from patients with testicular cancer, only few patients had azoospermia or severe oligozoospermia; sperm cryopreservation significantly reduces sperm motility and total motile sperm count and very few patients use their frozen sperm for reproductive purposes. PMID:28333030

  19. A retrospective study of artificial insemination of 251 mares using chilled and fixed time frozen-thawed semen.

    Science.gov (United States)

    Crowe, C A M; Ravenhill, P J; Hepburn, R J; Shepherd, C H

    2008-09-01

    Historically, artificial insemination (AI) using frozen semen has been perceived to have poorer success rates and be more labour intensive than using chilled semen. A retrospective study was therefore conducted to compare the conception rate achieved by AI between chilled and frozen semen, using fixed time insemination protocols over 2 breeding seasons. Artificial insemination using chilled semen produces a higher conception rate than that achieved with frozen semen. Mares (n = 251) were inseminated with either chilled (n = 112) or frozen (n = 139) semen in the 2006 and 2007 northern hemisphere breeding season. Per rectum ultrasonography of the mare's reproductive tract determined the timing of insemination, and deslorelin acetate was used to induce ovulation. Chilled semen insemination was performed using a single preovulatory dose delivered into the uterine body. Frozen semen was administered as 2 doses (pre- and post ovulation) using a deep uterine insemination technique. Pregnancy was detected ultrasonographically at 15 days post insemination. Conception rates were compared using a Chi-squared test. Insemination with frozen semen produced a significantly (P = 0.022) higher seasonal conception rate (82.0%) than that achieved with chilled semen (69.6%). Insemination with frozen semen can achieve conception rates equal to those with chilled semen, enabling the mare owner a greater selection of stallions.

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

    Science.gov (United States)

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

    2017-04-01

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

  1. Combinations of glycerol percent, glycerol equilibration time, and thawing rate upon freezability of bull spermatozoa in plastic straws.

    Science.gov (United States)

    Wiggin, H B; Almquist, J O

    1975-03-01

    Twelve ejaculates were used in a central composite experiment to test 15 combinations of glycerol (7, 9, 11, 13, or 15%), glycerol equilibration times (1, 2, 4, 8, or 16 h) and thawing rates (water at 35 C for 15 s, 50 C for 13 s, 65 C for 11 s, 80 C for 9 s, or 95 C for 7 s). Semen was diluted in heated skim milk-glycerol, packaged in .3-ml. Continental U.S. straws and frozen in liquid nitrogen vapor. Based on post-thaw progressive sperm motility after storage at -196 C for 9 to 11 days, estimated optima from multiple regression were 10.7% for glycerol, 2.0 h for glycerol equilibration time, and 76 C for thawing bath temperature. Only the linear effect for each variable was significant. Much faster thawing rates and shorter glycerol equilibration times than those for freezing bull spermatozoa in glass ampules should be used for maximum post-thaw sperm motility in straws.

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

    Science.gov (United States)

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

    2013-01-01

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

  3. Difference in birth weight of consecutive sibling singletons is not found in oocyte donation when comparing fresh versus frozen embryo replacements.

    Science.gov (United States)

    Galliano, Daniela; Garrido, Nicolás; Serra-Serra, Vicente; Pellicer, Antonio

    2015-12-01

    First, to assess if there are any differences in birth weight or gestational length in newborns from egg-donation pregnancies delivering singletons, originating from either fresh or frozen-thawed embryos when they were developed and delivered within the same mothers. Second, to determine if there are any clinical, phenotypic, or laboratory factors influencing this relationship, including the origin of the oocyte (same or different donor), the order of the children (first fresh or first frozen-thawed embryo transfer), the embryo freezing technique (vitrification or slow freezing), the in vitro embryo culture length, and the duration that embryos remained frozen. Retrospective cohorts study. University-affiliated infertility centers. A total of 360 women undergoing oocyte donation (OD), delivering (>28 weeks) at least two babies, each one from a single pregnancy, originating from at least one fresh and one frozen-thawed embryo transfer, controlling maternal and laboratory characteristics, to test the effect of embryo freezing on children size (n = 731). None. Birth weight, gestational age, weight percentile, being large for gestational age (LGA), small for gestational age (SGA), size out of normal range (ONR = LGA + SGA), and macrosomy. From fresh versus thawed embryos, respectively, mean birth weight of children was 3,183.7 g versus 3,226.4 g, gestational age was 272.1 days versus 268.8 days, and mean weight percentiles were 47.6 versus 50.1. The proportions and corresponding odds ratios (ORs) from fresh versus thawed embryos, respectively, were for LGA 13.6% versus 11.3% (OR 0.81), for SGA 9.4% versus 12.5% (OR 1.37), for ONR 23.1% versus 23.8% (OR 1.04), and for macrosomy 0.3% versus 0.8% (OR 3.1). After adjusting for clinically relevant variables, the ORs were for LGA 0.96, for SGA 1.40, for ONR 1.20, and for macrosomy not computable. None of the stated measures were significantly different. Also, independent analyses run on the origin of the oocytes

  4. Prediction of the fertility of stallion frozen-thawed semen using a combination of computer-assisted motility analysis, microscopical observation and flow cytometry.

    Science.gov (United States)

    Battut, I Barrier; Kempfer, A; Lemasson, N; Chevrier, L; Camugli, S

    2017-07-15

    Spermatozoa from some stallions do not maintain an acceptable fertility after freezing and thawing. The selection of frozen ejaculates that would be suitable for insemination is mainly based on post-thaw motility, but the prediction of fertility remains limited. A recent study in our laboratory has enabled the determination of a new protocol for the evaluation of fresh stallion semen, combining microscopical observation, computer-assisted motility analysis and flow cytometry, and providing a high level of fertility prediction. The purpose of the present experiment was to perform similar investigations on frozen semen. A panel of tests evaluating a large number of compartments or functions of the spermatozoa was applied to a population of 42 stallions, 33 of which showing widely differing fertilities (17-67% pregnancy rate per cycle [PRC]). Variability was evaluated by calculating the coefficient of variation (CV=SD/mean) and the intra-class correlation or "repeatability" for each variable. For paired variables, mean within-stallion CV% was significantly lower than between-stallion CV%, which was significantly lower than total CV%. Within-ejaculate repeatability, determined by analysing 6 straws for each of 10 ejaculates, ranged from 0.60 to 0.97. Within-stallion repeatability, determined by analysing at least 5 ejaculates for each of 38 stallions, ranged from 0.12 to 0.95. Principal component regression using a combination of 25 variables, including motility, morphology, viability, oxidation level, acrosome integrity, DNA integrity and hypoosmotic resistance, accounted for 94.5% of the variability regarding fertility, and was used to calculate a prediction of the PRC with a mean standard deviation of 2.2. The difference between the observed PRC and the calculated value ranged from -3.4 to 4.2. The 90% confidence interval (90CI) for the prediction of the PRC for the stallions of unknown fertility ranged from 8 to 30 (mean = 17). The best-fit model using only

  5. Physical-chemical basis of the protection of slowly frozen human erythrocytes by glycerol

    Energy Technology Data Exchange (ETDEWEB)

    Rall, W.F.; Mazur, P.; Souzu, H.

    1978-07-01

    One theory of freezing damage suggests that slowly cooled cells are killed by being exposed to increasing concentrations of electrolytes as the suspending medium freezes. A corollary to this view is that protective additives such as glycerol protect cells by acting colligatively to reduce the electrolyte concentration at any subzero temperature. Recently published phase-diagram data for the ternary system glycerol-NaCl-water by M.L. Shepard et al. (Cryobiology, 13: 9-23, 1976), in combination with the data on human red cell survival vs. subzero temperature presented here and in the companion study of Souzu and Mazur (Biophys. J., 23: 89-100), permit a precise test of this theory. Appropriate liquidus phase-diagram information for the solutions used in the red cell freezing experiments was obtained by interpolation of liquidus data of Shepard and his co-workers. The results of phase-diagram analysis of red cell survival indicate that the correlation between the temperature that yields 50% hemolysis (LT/sub 50/) and the electrolyte concentration attained at that temperature in various concentrations of glycerol is poor. With increasing concentrations of glycerol, the cells were killed at progressively lower concentrations of NaCl. For example, the LT/sub 50/ for cells frozen in the absence of glycerol corresponds to a NaCl concentration of 12 weight percent (2.4 molal), while for cells frozen in 1.75 M glycerol in buffered saline the LT/sub 50/ corresponds to 3.0 weight percent NaCl (1.3 molal). The data, in combination with other findings, lead to two conclusions: (a) The protection from glycerol is due to its colligative ability to reduce the concentration of sodium chloride in the external medium, but (b) the protection is less than that expected from colligative effects; apparently glycerol itself can also be a source of damage, probably because it renders the red cells susceptible to osmotic shock during thawing.

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

    Science.gov (United States)

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

    2018-05-01

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

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

    KAUST Repository

    Jadoon, Khan

    2015-09-18

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

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

    KAUST Repository

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

    2015-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Khan Zaib Jadoon

    2015-09-01

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

  10. Effect of post-thaw addition of seminal plasma on motility, viability and chromatin integrity of cryopreserved donkey jack (Equus asinus) spermatozoa.

    Science.gov (United States)

    Sabatini, C; Mari, G; Mislei, B; Love, Cc; Panzani, D; Camillo, F; Rota, A

    2014-12-01

    Pregnancy rates in donkeys after artificial insemination with cryopreserved semen are still low, compared to the horse species. Addition of autologous seminal plasma to frozen-thawed semen appeared to improve pregnancy rates. The aims of this study were to evaluate (1) sperm motility and plasma membrane integrity after thawing (T0) and after one and 2 h (T1 and T2) of post-thaw incubation in either 0% (SP0) or 70% (SP70) autologous seminal plasma and (2) sperm motility, plasma membrane integrity and DNA quality (%COMP-αt) after thawing (T0) and after 2 and 4 h (T2 and T4) of post-thaw incubation in either 0% (SP0), 5% (SP5) or 20% (SP20) homologous seminal plasma. In experiment 1, seminal plasma decreased total and progressive sperm motility and plasma membrane intact spermatozoa immediately after dilution and at all following time points (p < 0.05). In experiment 2, total and progressive motility did not differ between treatments immediately after dilution and between SP0 and SP5 at T2, while they were lower in both SP5 and SP20 than in SP0 at T4. Plasma membrane intact sperm cells did not differ between SP0 and SP5 and were lower in SP20 at all time points. DNA quality was not affected by treatment immediately after dilution and was significantly worse for SP20 after 4 h of incubation (p < 0.05). The post-thaw addition of seminal plasma at the tested concentrations did not improve donkey frozen semen characteristics in vitro over time. © 2014 Blackwell Verlag GmbH.

  11. Production of healthy cloned mice from bodies frozen at −20°C for 16 years

    OpenAIRE

    Wakayama, Sayaka; Ohta, Hiroshi; Hikichi, Takafusa; Mizutani, Eiji; Iwaki, Takamasa; Kanagawa, Osami; Wakayama, Teruhiko

    2008-01-01

    Cloning animals by nuclear transfer provides an opportunity to preserve endangered mammalian species. However, it has been suggested that the “resurrection” of frozen extinct species (such as the woolly mammoth) is impracticable, as no live cells are available, and the genomic material that remains is inevitably degraded. Here we report production of cloned mice from bodies kept frozen at −20 °C for up to 16 years without any cryoprotection. As all of the cells were ruptured after thawing, we...

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

    Directory of Open Access Journals (Sweden)

    Caldevilla M

    2016-12-01

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

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

    Science.gov (United States)

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

    2009-12-01

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

  14. Effects of permafrost thaw on CO2 and CH4 exchange in a western Alaska peatland chronosequence

    Science.gov (United States)

    Carmel E. Johnston,; Stephanie A. Ewing,; Harden, Jennifer W.; Ruth K. Varner,; Wickland, Kimberly P.; Koch, Joshua C.; Fuller, Christopher C.; Manies, Kristen L.; M. Torre Jorgenson,

    2014-01-01

    Permafrost soils store over half of global soil carbon (C), and northern frozen peatlands store about 10% of global permafrost C. With thaw, inundation of high latitude lowland peatlands typically increases the surface-atmosphere flux of methane (CH4), a potent greenhouse gas. To examine the effects of lowland permafrost thaw over millennial timescales, we measured carbon dioxide (CO2) and CH4 exchange along sites that constitute a ~1000 yr thaw chronosequence of thermokarst collapse bogs and adjacent fen locations at Innoko Flats Wildlife Refuge in western Alaska. Peak CH4exchange in July (123 ± 71 mg CH4–C m−2 d−1) was observed in features that have been thawed for 30 to 70 (peatlands, particularly those developed on poorly drained mineral substrates, are a key locus of elevated CH4 emission to the atmosphere that must be considered for a complete understanding of high latitude CH4 dynamics.

  15. Effects of adding different levels of Glutamine to modified Beltsville extender on the survival of frozen rooster semen.

    Science.gov (United States)

    Khiabani, Aytak Bakhshayesh; Moghaddam, Gholamali; Kia, Hossein Daghigh

    2017-09-01

    The aim of the present study was to investigate the effects of l-glutamine on the quality of frozen-thawed rooster semen. Semen samples were collected from eight mature roosters (Ross 308). After initial semen assessments, samples of adequate quality were mixed together and diluted with modified Beltsville extender without l-glutamine (control) and supplemented with 2.5, 5, and 7.5mM l-glutamine. Semen straws were subjected to cryopreservation and evaluated twice at 15-day intervals. After thawing, sperm viability, total and progressive sperm motilities were measured by Eosin-Nigrosine and Computer-Aided Sperm Analysis (CASA), respectively. The results showed that sperm functions decreased on day 30 compared to day 15. The extender supplemented with 5mM glutamine improved (prooster sperm to be frozen for longer. Copyright © 2017 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Muldrew Ken

    2010-10-01

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

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

    Science.gov (United States)

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

    2011-08-01

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

  18. Radio-sterilization and processing of frozen human skin

    International Nuclear Information System (INIS)

    Zarate S, Herman; Aguirre H, Paulina; Silva R, Samy; Hitschfeld G, Mario

    2006-01-01

    The Laboratory of Radio-sterilized Biological Tissues Processing (LPTR) belonging to the Chilean Commission of Nuclear Energy and the International Atomic Energy Agency have played a paramount role in our country, concerning the biological tissue processing, which can be radio-sterilized as human skin, pig skin, amniotic membrane, human bone and bovine bone. The frozen radio.-sterilized human skin processing began in 2001, by means of putting into practice the knowledge acquired in training courses through the IAEA and the experience transferred by experts who visited our laboratory. The human skin processing of dead donor can be divided into 6 stages: a) Profuse washing with physiological sterilized serum in to remove the microorganisms, chemical and pharmacological compounds; b) immersion in glycerol solution at 10% to better keep the stored tissues; c) packing, to avoid post manipulation of the sterilized tissue; d) microbiological controls which allow and guarantee a sterility assurance level of 10 6 ; e) radio-sterilization, technique that consists of exposing the grafts to electromagnetic gamma waves which eliminate the microorganisms of the tissue, f) and finally, dispatching and liberation of the frozen sterilized human skin for its clinical use in different centers that take care of burned people. The LPTR receives feedback from surgeons who have used these tissues in order to improve the processing stages based in an integral quality system ISO 9001.2000. The State Health System in our country counts on limited and scarce resources to implement synthetic substitutes that is why It is considered necessary to spread the use of these noble tissues which have sterility assurance and they are processed at low price

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

    Science.gov (United States)

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

    2009-02-01

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

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

    Science.gov (United States)

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

    2017-12-01

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

  1. 75 FR 12206 - Certain Frozen Warmwater Shrimp From the Socialist Republic of Vietnam: Preliminary Results...

    Science.gov (United States)

    2010-03-15

    ... (or thawed-from-frozen) and peeled shrimp; (2) to which a ``dusting'' layer of rice or wheat flour of... thoroughly and evenly coated with the flour; (4) with the non- shrimp content of the end product constituting...., Ltd. (``Fish One''), and Seaprodex Minh Hai (collectively ``revocation companies''). Of the revocation...

  2. Immunoreactive LH in long-term frozen human urine samples.

    Science.gov (United States)

    Singh, Gurmeet Kaur Surindar; Jimenez, Mark; Newman, Ron; Handelsman, David J

    2014-04-01

    Urine provides a convenient non-invasive alternative to blood sampling for measurement of certain hormones. Urinary luteinizing hormone (LH) measurements have been used for endocrinology research and anti-doping testing. However, the commercially available LH immunoassays are developed and validated for human blood samples but not urine so that LH assays intended for use with urine samples need thorough validation. Therefore, the present study evaluated the measurement of urinary LH immunoreactivity using previously validated immunofluorometric (IF) and immunochemiluminometric (ICL) LH assays after prolonged frozen storage. LH was measured in serial urine samples following administration of a single injection of one of two doses of recombinant human chorionic hormone (rhCG) with assays run at the end of study (2008) and again after four years of frozen (-20 °C) storage where samples were stored without adding preservatives. The ICL assay showed quantitatively reproducible LH measurements after prolonged -20 °C storage. However, the IF immunoassay gave consistently lower LH levels relative to ICL (2008) with a further proportionate reduction after four years of sample storage (2012). Yet, both the assays displayed similar patterns of the time-course of urine LH measurement both before and after four years of frozen storage. In conclusion, we found that both immunoassays are suitable for urinary LH measurements with ICL assay being more robust for quantitative urinary LH measurement such as for anti-doping purposes, whereas the IF could be applicable for research studies where urine LH levels are compared within-study but not in absolute terms. Copyright © 2013 John Wiley & Sons, Ltd.

  3. Evaluation of Container Closure System Integrity for Frozen Storage Drug Products.

    Science.gov (United States)

    Nieto, Alejandra; Roehl, Holger; Brown, Helen; Nikoloff, Jonas; Adler, Michael; Mahler, Hanns-Christian

    2016-01-01

    Sometimes, drug product for parenteral administration is stored in a frozen state (e.g., -20 °C or -80 °C), particularly during early stages of development of some biotech molecules in order to provide sufficient stability. Shipment of frozen product could potentially be performed in the frozen state, yet possibly at different temperatures, for example, using dry ice (-80 °C). Container closure systems of drug products usually consist of a glass vial, rubber stopper, and an aluminum crimped cap. In the frozen state, the glass transition temperature (Tg) of commonly used rubber stoppers is between -55 and -65 °C. Below their Tg, rubber stoppers are known to lose their elastic properties and become brittle, and thus potentially fail to maintain container closure integrity in the frozen state. Leaks during frozen temperature storage and transportation are likely to be transient, yet, can possibly risk container closure integrity and lead to microbial contamination. After thawing, the rubber stopper is supposed to re-seal the container closure system. Given the transient nature of the possible impact on container closure integrity in the frozen state, typical container closure integrity testing methods (used at room temperature conditions) are unable to evaluate and thus confirm container closure integrity in the frozen state. Here we present the development of a novel method (thermal physical container closure integrity) for direct assessment of container closure integrity by a physical method (physical container closure integrity) at frozen conditions, using a modified He leakage test. In this study, different container closure systems were evaluated with regard to physical container closure integrity in the frozen state to assess the suitability of vial/stopper combinations and were compared to a gas headspace method. In summary, the thermal physical container closure integrity He leakage method was more sensitive in detecting physical container closure

  4. Dissolved organic carbon and nitrogen release from boreal Holocene permafrost and seasonally frozen soils of Alaska

    Science.gov (United States)

    Wickland, Kimberly P.; Waldrop, Mark P.; Aiken, George R.; Koch, Joshua C.; Torre Jorgenson, M.; Striegl, Robert G.

    2018-06-01

    Permafrost (perennially frozen) soils store vast amounts of organic carbon (C) and nitrogen (N) that are vulnerable to mobilization as dissolved organic carbon (DOC) and dissolved organic and inorganic nitrogen (DON, DIN) upon thaw. Such releases will affect the biogeochemistry of permafrost regions, yet little is known about the chemical composition and source variability of active-layer (seasonally frozen) and permafrost soil DOC, DON and DIN. We quantified DOC, total dissolved N (TDN), DON, and DIN leachate yields from deep active-layer and near-surface boreal Holocene permafrost soils in interior Alaska varying in soil C and N content and radiocarbon age to determine potential release upon thaw. Soil cores were collected at three sites distributed across the Alaska boreal region in late winter, cut in 15 cm thick sections, and deep active-layer and shallow permafrost sections were thawed and leached. Leachates were analyzed for DOC, TDN, nitrate (NO3 ‑), and ammonium (NH4 +) concentrations, dissolved organic matter optical properties, and DOC biodegradability. Soils were analyzed for C, N, and radiocarbon (14C) content. Soil DOC, TDN, DON, and DIN yields increased linearly with soil C and N content, and decreased with increasing radiocarbon age. These relationships were significantly different for active-layer and permafrost soils such that for a given soil C or N content, or radiocarbon age, permafrost soils released more DOC and TDN (mostly as DON) per gram soil than active-layer soils. Permafrost soil DOC biodegradability was significantly correlated with soil Δ14C and DOM optical properties. Our results demonstrate that near-surface Holocene permafrost soils preserve greater relative potential DOC and TDN yields than overlying seasonally frozen soils that are exposed to annual leaching and decomposition. While many factors control the fate of DOC and TDN, the greater relative yields from newly thawed Holocene permafrost soils will have the largest

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

    Science.gov (United States)

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

    2008-12-01

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

  6. In vivo fertilizing capacity of deep frozen boar semen packaged in plastic bags and maxi-straws.

    Science.gov (United States)

    Bwanga, C O; Hofmo, P O; Grevle, I S; Einarsson, S; Rodriguez-Martinez

    1991-05-01

    Pooled ejaculates from six fertile boars were frozen under controlled conditions in Teflon FEP-film plastic bags (5 ml) and maxi-straws (2.5 ml) using 3% glycerol as cryoprotectant. The percentages of both post-thaw motility and normal apical ridges were significantly higher (P less than 0.001) for the bags (54.5 and 75%) than for the maxi-straws (40.1 and 59.4%) respectively. For evaluation of the in vivo fertilizing capacity of the frozen-thawed spermatozoa, 26 gilts were inseminated once 24 h after the first observation of standing reflex in their second oestrus, with 5 ml of semen (containing 5 billion spermatozoa) reconstituted in 80 ml of BTS from either bags or maxi-straws. Ova were recovered from the oviducts/uteri 2-4 days following insemination and examined for cleavage and sperm binding to the zona pellucida (ZP). Significantly higher rates (P less than 0.02) of fertilized ova were found in the bag-inseminated (75%) than in maxi-straw inseminated gilts (63%); and similarly their ova had significantly more spermatozoa in the ZP, irrespective of whether they were fertilized or nonfertilized. This study confirmed that the plastic bags are suitable and may be used for packaging single insemination doses of deep frozen boar semen for routine A.I. work.

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

    Science.gov (United States)

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

    1991-01-01

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

  8. Trehalose improves semen antioxidant enzymes activity, post-thaw quality, and fertility in Nili Ravi buffaloes (Bubalus bubalis).

    Science.gov (United States)

    Iqbal, Sajid; Andrabi, Syed Murtaza Hassan; Riaz, Amjad; Durrani, Aneela Zameer; Ahmad, Nasim

    2016-03-15

    Our objectives were to study the effect of trehalose in extender on (1) antioxidant enzymes profile during cryopreservation (after dilution, before freezing, and after thawing), (2) in vitro quality (after thawing), and (3) in vivo fertility of Nili Ravi buffalo (Bubalus bubalis) bull spermatozoa. Semen samples (n = 20) from four buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of trehalose (0.0, 15, 30, 45, and 60 mM) and frozen in French straws. At post dilution, profile of sperm catalase (U/mL) was higher (P semen doses cryopreserved in extender containing 30 mM of trehalose than the control. It is concluded that addition of 30-mM trehalose in extender improves the semen antioxidant enzymes activity, post thaw quality, and fertility in Nili Ravi buffaloes. Copyright © 2016 Elsevier Inc. All rights reserved.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2014-02-15

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

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

    International Nuclear Information System (INIS)

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

    2014-01-01

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

  11. Tundra permafrost thaw causes significant shifts in energy partitioning

    Directory of Open Access Journals (Sweden)

    Christian Stiegler

    2016-04-01

    Full Text Available Permafrost, a key component of the arctic and global climate system, is highly sensitive to climate change. Observed and ongoing permafrost degradation influences arctic hydrology, ecology and biogeochemistry, and models predict that rapid warming is expected to significantly reduce near-surface permafrost and seasonally frozen ground during the 21st century. These changes raise concern of how permafrost thaw affects the exchange of water and energy with the atmosphere. However, associated impacts of permafrost thaw on the surface energy balance and possible feedbacks on the climate system are largely unknown. In this study, we show that in northern subarctic Sweden, permafrost thaw and related degradation of peat plateaus significantly change the surface energy balance of three peatland complexes by enhancing latent heat flux and, to less degree, also ground heat flux at the cost of sensible heat flux. This effect is valid at all radiation levels but more pronounced at higher radiation levels. The observed differences in flux partitioning mainly result from the strong coupling between soil moisture availability, vegetation composition, albedo and surface structure. Our results suggest that ongoing and predicted permafrost degradation in northern subarctic Sweden ultimately result in changes in land–atmosphere coupling due to changes in the partitioning between latent and sensible heat fluxes. This in turn has crucial implications for how predictive climate models for the Arctic are further developed.

  12. Nitrogen availability increases in a tundra ecosystem during five years of experimental permafrost thaw.

    Science.gov (United States)

    Salmon, Verity G; Soucy, Patrick; Mauritz, Marguerite; Celis, Gerardo; Natali, Susan M; Mack, Michelle C; Schuur, Edward A G

    2016-05-01

    Perennially frozen soil in high latitude ecosystems (permafrost) currently stores 1330-1580 Pg of carbon (C). As these ecosystems warm, the thaw and decomposition of permafrost is expected to release large amounts of C to the atmosphere. Fortunately, losses from the permafrost C pool will be partially offset by increased plant productivity. The degree to which plants are able to sequester C, however, will be determined by changing nitrogen (N) availability in these thawing soil profiles. N availability currently limits plant productivity in tundra ecosystems but plant access to N is expected improve as decomposition increases in speed and extends to deeper soil horizons. To evaluate the relationship between permafrost thaw and N availability, we monitored N cycling during 5 years of experimentally induced permafrost thaw at the Carbon in Permafrost Experimental Heating Research (CiPEHR) project. Inorganic N availability increased significantly in response to deeper thaw and greater soil moisture induced by Soil warming. This treatment also prompted a 23% increase in aboveground biomass and a 49% increase in foliar N pools. The sedge Eriophorum vaginatum responded most strongly to warming: this species explained 91% of the change in aboveground biomass during the 5 year period. Air warming had little impact when applied alone, but when applied in combination with Soil warming, growing season soil inorganic N availability was significantly reduced. These results demonstrate that there is a strong positive relationship between the depth of permafrost thaw and N availability in tundra ecosystems but that this relationship can be diminished by interactions between increased thaw, warmer air temperatures, and higher levels of soil moisture. Within 5 years of permafrost thaw, plants actively incorporate newly available N into biomass but C storage in live vascular plant biomass is unlikely to be greater than losses from deep soil C pools. © 2015 John Wiley & Sons Ltd.

  13. The effects of permafrost thaw on soil hydrologic, thermal, and carbon dynamics in an Alaskan peatland

    Science.gov (United States)

    O'Donnell, Jonathan A.; Jorgenson, M. Torre; Harden, Jennifer W.; McGuire, A. David; Kanevskiy, Mikhail Z.; Wickland, Kimberly P.

    2012-01-01

    Recent warming at high-latitudes has accelerated permafrost thaw in northern peatlands, and thaw can have profound effects on local hydrology and ecosystem carbon balance. To assess the impact of permafrost thaw on soil organic carbon (OC) dynamics, we measured soil hydrologic and thermal dynamics and soil OC stocks across a collapse-scar bog chronosequence in interior Alaska. We observed dramatic changes in the distribution of soil water associated with thawing of ice-rich frozen peat. The impoundment of warm water in collapse-scar bogs initiated talik formation and the lateral expansion of bogs over time. On average, Permafrost Plateaus stored 137 ± 37 kg C m-2, whereas OC storage in Young Bogs and Old Bogs averaged 84 ± 13 kg C m-2. Based on our reconstructions, the accumulation of OC in near-surface bog peat continued for nearly 1,000 years following permafrost thaw, at which point accumulation rates slowed. Rapid decomposition of thawed forest peat reduced deep OC stocks by nearly half during the first 100 years following thaw. Using a simple mass-balance model, we show that accumulation rates at the bog surface were not sufficient to balance deep OC losses, resulting in a net loss of OC from the entire peat column. An uncertainty analysis also revealed that the magnitude and timing of soil OC loss from thawed forest peat depends substantially on variation in OC input rates to bog peat and variation in decay constants for shallow and deep OC stocks. These findings suggest that permafrost thaw and the subsequent release of OC from thawed peat will likely reduce the strength of northern permafrost-affected peatlands as a carbon dioxide sink, and consequently, will likely accelerate rates of atmospheric warming.

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

    Science.gov (United States)

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

    2008-06-01

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

  15. The impact of cold chain temperature abuses on the quality of frozen strawberries (Fragaria ×ananassa

    Directory of Open Access Journals (Sweden)

    Rui Cruz

    2013-04-01

    Full Text Available The quality of frozen foods can be negatively affected if improper storage and distribution temperatures are allowed. The objective of this study was to investigate the effect of freeze-thaw cycles, which may occur in the cold chain, on colour (Lab, Total Colour Differences (TCD, chroma and hue angle and vitamin C (ascorbic and dehydroascorbic acids content of frozen strawberries (Fragaria ×ananassa, Duschesne, cv. Selva. A plan of temperature abuses (TAs was established, based on a real situation, and applied to frozen strawberries during a four month frozen storage period. The results showed that the lightness (L was the only parameter that was not significantly affected by range of TAs studied. The colour showed some variation on the parameters a, b, TCD, chroma and hue angle. During TAs, ascorbic acid decreased about 75% and dehydroascorbic acid increased 73%. The non-abused strawberry samples showed better overall appearance than the abused samples. This work contributes to an understanding of the quality changes of frozen strawberries that might occur during frozen storage and cold chain distribution.

  16. Probiotic supplementation and fast freezing to improve quality attributes and oxidation stability of frozen chicken breast muscle

    Science.gov (United States)

    The objective of this study was to determine the effects of probiotic supplementation and fast freezing on quality attributes and oxidation stability of frozen/thawed chicken breast meat. Broilers were fed with a basal diet or the basal diet plus 250 ppm Sporulin (three strains of Bacillus subtilis)...

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

    Science.gov (United States)

    Ku, Jentung; Ottenstein, Laura; Krimchansky, Alexander

    2012-01-01

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

  18. Effects of soil rewetting and thawing on soil gas fluxes: a review of current literature and suggestions for future research

    Directory of Open Access Journals (Sweden)

    D.-G. Kim

    2012-07-01

    Full Text Available The rewetting of dry soils and the thawing of frozen soils are short-term, transitional phenomena in terms of hydrology and the thermodynamics of soil systems. The impact of these short-term phenomena on larger scale ecosystem fluxes is increasingly recognized, and a growing number of studies show that these events affect fluxes of soil gases such as carbon dioxide (CO2, methane (CH4, nitrous oxide (N2O, ammonia (NH3 and nitric oxide (NO. Global climate models predict that future climatic change is likely to alter the frequency and intensity of drying-rewetting events and thawing of frozen soils. These future scenarios highlight the importance of understanding how rewetting and thawing will influence dynamics of these soil gases. This study summarizes findings using a new database containing 338 studies conducted from 1956 to 2011, and highlights open research questions. The database revealed conflicting results following rewetting and thawing in various terrestrial ecosystems and among soil gases, ranging from large increases in fluxes to non-significant changes. Studies reporting lower gas fluxes before rewetting tended to find higher post-rewetting fluxes for CO2, N2O and NO; in addition, increases in N2O flux following thawing were greater in warmer climate regions. We discuss possible mechanisms and controls that regulate flux responses, and recommend that a high temporal resolution of flux measurements is critical to capture rapid changes in gas fluxes after these soil perturbations. Finally, we propose that future studies should investigate the interactions between biological (i.e., microbial community and gas production and physical (i.e., porosity, diffusivity, dissolution changes in soil gas fluxes, apply techniques to capture rapid changes (i.e., automated measurements, and explore synergistic experimental and modelling approaches.

  19. High serum IGF-1 levels are associated with pregnancy loss following frozen-thawed euploid embryo transfer cycles.

    Science.gov (United States)

    Irani, Mohamad; Nasioudis, Dimitrios; Witkin, Steven S; Gunnala, Vinay; Spandorfer, Steven D

    2018-06-01

    An elevated level of insulin growth factor (IGF-1) in rat uterine fluid has been shown to exert detrimental effects of embryo development possibly leading to an increase in pregnancy loss. Interestingly, the administration of somatostatin to rats undergoing superovulation reduced IGF-1 levels in uterine luminal fluid and thus reversed its deleterious effects on embryo development and increased the number of normal embryos. Therefore, we investigated whether serum levels of IGF-1 correlate with the incidence of pregnancy loss following IVF. To account for aneuploidy and the effect of hormonal supplementation on serum IGF levels, we only included natural frozen-thawed euploid embryo transfer (N-FET) cycles. Sera collected in the follicular phase (cycle day 10) were tested for levels of IGF-1, IGF-2, and IGF-binding protein 1 (IGFBP-1) using quantitative ELISA. A total of 156 N-FET cycles were included: 120 resulted in a live birth whereas 36 led to a first trimester pregnancy loss. Women with a pregnancy loss had significantly higher serum IGF-1 levels compared to those who achieved a live birth (18.0 ± 1.1 vs. 14.6 ± 0.7 ng/mL, respectively). The two groups had comparable serum IGF-2 and IGFBP-1 levels. There was no significant difference in maternal age, body mass index, gravidity, parity, number of prior miscarriages, peak endometrial thickness, or infertility diagnosis between the two groups. In conclusion, women undergoing euploid blastocyst transfer with elevated serum IGF-1 concentrations may be at increased risk of pregnancy loss. This may constitute a novel molecular explanation of pregnancy loss of euploid conceptus. Copyright © 2018. Published by Elsevier B.V.

  20. An estimated cost of lost climate regulation services caused by thawing of the Arctic cryosphere.

    Science.gov (United States)

    Euskirchen, Eugénie S; Goodstein, Eban S; Huntington, Henry P

    2013-12-01

    Recent and expected changes in Arctic sea ice cover, snow cover, and methane emissions from permafrost thaw are likely to result in large positive feedbacks to climate warming. There is little recognition of the significant loss in economic value that the disappearance of Arctic sea ice, snow, and permafrost will impose on humans. Here, we examine how sea ice and snow cover, as well as methane emissions due to changes in permafrost, may potentially change in the future, to year 2100, and how these changes may feed back to influence the climate. Between 2010 and 2100, the annual costs from the extra warming due to a decline in albedo related to losses of sea ice and snow, plus each year's methane emissions, cumulate to a present value cost to society ranging from US$7.5 trillion to US$91.3 trillion. The estimated range reflects uncertainty associated with (1) the extent of warming-driven positive climate feedbacks from the thawing cryosphere and (2) the expected economic damages per metric ton of CO2 equivalents that will be imposed by added warming, which depend, especially, on the choice of discount rate. The economic uncertainty is much larger than the uncertainty in possible future feedback effects. Nonetheless, the frozen Arctic provides immense services to all nations by cooling the earth's temperature: the cryosphere is an air conditioner for the planet. As the Arctic thaws, this critical, climate-stabilizing ecosystem service is being lost. This paper provides a first attempt to monetize the cost of some of those lost services.

  1. Production of bovine cloned embryos with donor cells frozen at a slow cooling rate in a conventional freezer (20 C)

    Science.gov (United States)

    Chacon, L.; Gomez, M.C.; Jenkins, J.A.; Leibo, S.P.; Wirtu, G.; Dresser, B.L.; Pope, C.E.

    2009-01-01

    Summary Usually, fibroblasts are frozen in dimethyl sulphoxide (DMSO, 10% v/v) at a cooling rate of 1 C/min in a low-temperature (80 C) freezer (LTF) before storage in liquid nitrogen (LN2); however, a LTF is not always available. The purpose of the present study was to evaluate apoptosis and viability of bovine fibroblasts frozen in a LTF or conventional freezer (CF; 20 C) and their subsequent ability for development to blastocyst stage after fusion with enucleated bovine oocytes. Percentages of live cells frozen in LTF (49.5%) and CF (50.6%) were similar, but significantly less than non-frozen control (88%). In both CF and LTF, percentages of live apoptotic cells exposed to LN2 after freezing were lower (4% and 5%, respectively) as compared with unexposed cells (10% and 18%, respectively). Cells frozen in a CF had fewer cell doublings/24 h (0.45) and required more days (9.1) to reach 100% confluence at the first passage (P) after thawing and plating as compared with cells frozen in a LTF (0.96 and 4.0 days, respectively). Hypoploidy at P12 was higher than at P4 in cells frozen in either a CF (37.5% vs. 19.2%) or in a LTF (30.0% vs. 15.4%). A second-generation cryo-solution reduced the incidence of necrosis (29.4%) at 0 h after thawing as compared with that of a first generation cryo-solution (DMEM + DMSO, 60.2%). The percentage of apoptosis in live cells was affected by cooling rate (CF = 1.9% vs. LFT = 0.7%). Development of bovine cloned embryos to the blastocyst stage was not affected by cooling rate or freezer type. ?? 2009 Cambridge University Press.

  2. Separation of Contaminants in The Freeze/Thaw Process

    Directory of Open Access Journals (Sweden)

    Szpaczyński Janusz A.

    2017-06-01

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

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

    DEFF Research Database (Denmark)

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

    2003-01-01

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

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

    Science.gov (United States)

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

    2017-09-01

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

  5. Effects of frozen storage on survival of Staphylococcus aureus and enterotoxin production in precooked tuna meat.

    Science.gov (United States)

    Wu, Xulei; Su, Yi-Cheng

    2014-08-01

    This study investigated the survival of Staphylococcus aureus in precooked tuna meat for producing canned products during frozen storage (-20 ± 2 °C) as well as its growth and enterotoxin production at 35 to 37 °C after the storage. Samples (50 ± 5 g) of precooked albacore (loin, chunk, and flake) and skipjack (chunk and flake) tuna were inoculated with 5 enterotoxin-producing strains of S. aureus at a level of approximately 3.5 log CFU/g and individually packed in a vacuum bag after 3 h incubation at 35 to 37 °C. Vacuum-packed samples were stored in a freezer (-20 ± 2 °C) for 4 wk. The frozen samples were then thawed in 37 °C circulating water for 2 h and incubated at 35 to 37 °C for 22 h. Populations of S. aureus in all precooked tuna samples decreased slightly (canned tuna within 6 to 8 h of thawing to avoid product spoilage and potential enterotoxin production by S. aureus in contaminated precooked tuna meat. © 2014 Institute of Food Technologists®

  6. A disease of frozen feelings: ethically working on emotional worlds in a Russian Orthodox Church drug rehabilitation program

    NARCIS (Netherlands)

    Zigon, J.

    2010-01-01

    In a Russian Orthodox Church drug rehabilitation program in St. Petersburg, drug addiction was often described as a disease of frozen feelings. This image suggests that rehabilitation is a process of thawing emotional worlds and, thus, allows the emotions to flow once again. In this article I argue

  7. Project to develop and demonstrate methods to eliminate frozen coal handling problems. Status report I

    Energy Technology Data Exchange (ETDEWEB)

    None

    1980-09-15

    For too many years, problems associated with frozen coal have plagued the companies who mine it, the companies who handle it in transit and the utilities and other industrial concerns that finally burn it. But never before has the magnitude of the frozen coal problem been as great as it is today because of two primary factors, i.e. (1) the majority of coal currently transported and used has been ground to a very fine mesh that absorbs water readily, thus providing more surface area for freezing, and (2) the substantially increased importance of coal, indeed, the now critical necessity for more coal to be used in displacing dangerously uncertain foreign oil supplies that currently account for 50 percent of our daily domestic oil consumption. Frozen coal problems can and do have a devastating effect upon the ability to provide energy from coal during harsh winter months when it is not needed. The majority of these problems have been involved with removing frozen coal from rail cars. To allay the problem, numerous techniques have been tried, all with some measure of success. As an example, certain chemicals have been sprayed on the coal; another common treatment has been widespread use of thaw sheds, which, whether electrically or gas-fired, are all energy intensive, time consuming, hard on rail equipment and expensive to operate over long periods of time. From sledge hammers and crow bars to gas-fired jets and electric thaw sheds, available mechanical de-icing methods often damage coal handling equipment, are time consuming and, therefore, very expensive when demurrage losses must be added to significant investment and/or operating costs.

  8. Rapid runoff via shallow throughflow and deeper preferential flow in a boreal catchment underlain by frozen silt (Alaska, USA)

    Science.gov (United States)

    Koch, Joshua C.; Ewing, Stephanie A.; Striegl, Robert G.; McKnight, Diane M.

    2013-01-01

    In high-latitude catchments where permafrost is present, runoff dynamics are complicated by seasonal active-layer thaw, which may cause a change in the dominant flowpaths as water increasingly contacts mineral soils of low hydraulic conductivity. A 2-year study, conducted in an upland catchment in Alaska (USA) underlain by frozen, well-sorted eolian silt, examined changes in infiltration and runoff with thaw. It was hypothesized that rapid runoff would be maintained by flow through shallow soils during the early summer and deeper preferential flow later in the summer. Seasonal changes in soil moisture, infiltration, and runoff magnitude, location, and chemistry suggest that transport is rapid, even when soils are thawed to their maximum extent. Between June and September, a shift occurred in the location of runoff, consistent with subsurface preferential flow in steep and wet areas. Uranium isotopes suggest that late summer runoff erodes permafrost, indicating that substantial rapid flow may occur along the frozen boundary. Together, throughflow and deep preferential flow may limit upland boreal catchment water and solute storage, and subsequently biogeochemical cycling on seasonal to annual timescales. Deep preferential flow may be important for stream incision, network drainage development, and the release of ancient carbon to ecosystems

  9. Survival of pathogenic enterohemorrhagic Escherichia coli (EHEC) and control with calcium oxide in frozen meat products.

    Science.gov (United States)

    Ro, Eun Young; Ko, Young Mi; Yoon, Ki Sun

    2015-08-01

    This study investigated both the level of microbial contamination and the presence of enterohemorrhagic Escherichia coli (EHEC) in frozen meat products, followed by the evaluation of its survival over 180 days under frozen temperature. We also examined the effect of calcium oxide on the populations of EHEC, E. coli O157:H7 and EPEC under both 10 °C and -18 °C storage conditions. Afterward, the morphological changes occurring in EHEC cells in response to freezer storage temperature and calcium oxide (CaO) treatments were examined using transmission electron microscopy. Among the frozen meat products tested, the highest contamination levels of total aerobic counts, coliforms and E. coli were observed in pork cutlets. Examination showed that 20% of the frozen meat products contained virulence genes, including verotoxin (VT) 1 and 2. Over 180 days of frozen storage and after 3 freeze-thaw cycles, the population of EHEC did not change regardless of the type of products or initial inoculated concentration, indicating the strong survival ability of EHEC. Subsequent testing revealed that the growth of three pathogenic E. coli strains was completely inhibited in meat patties prepared with 1% CaO, stored at 10 °C. However, the addition of 2% CaO was necessary to control the survival of EHEC, E. coli O157:H7 and EPEC in meat patties stored at -18 °C. CaO reduced the population of E. coli O157:H7 more effectively than the other EHEC and EPEC strains at both 10 °C and -18 °C. Transmission electron microscopy analysis revealed that exposed EHEC cells were resistant to the freezer storage temperature, although some cells incurred injury and death after several freeze-thaw cycles. Most of the cells exposed to CaO were found to have died or lost their cellular integrity and membranes, indicating that CaO has the potential to be used as a powerful antimicrobial agent for manufacturing frozen meat products. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Zona pellucida damage to human embryos after cryopreservation and the consequences for their blastomere survival and in-vitro viability.

    Science.gov (United States)

    Van Den Abbeel, E; Van Steirteghem, A

    2000-02-01

    The study objective was to quantify zona pellucida (ZP) damage in cryopreserved human embryos. The influence of two different freezing containers was investigated, and the influence of freezing damage on the survival and viability of the embryos evaluated. ZP damage did not differ according to whether embryos originated from in-vitro fertilization (IVF) cycles or from IVF cycles in association with intracytoplasmic sperm injection (ICSI). The freezing container, however, significantly influenced the occurrence of ZP damage after cryopreservation. More damage was observed when the embryos were frozen-thawed using plastic cryovials than using plastic mini-straws (16.6% versus 2.3%; P plastic mini-straws. The further cleavage of frozen-thawed embryos suitable for transfer was not different whether there was ZP damage or not; however, it was higher when there was 100% blastomere survival as compared with when some blastomeres were damaged (79.0% versus 43.7%; P plastic mini-straws. In conclusion, the aim of a cryopreservation programme should be to have as many fully intact embryos as possible after thawing. Increased ZP damage might indicate a suboptimal cryopreservation procedure.

  11. Frozen-Ground Cartoons: An international collaboration between artists and permafrost scientists

    OpenAIRE

    Fritz, Michael; Bouchard, Fréderic; Deshpande, Bethany; Malenfant-Lepage, Julie; Nieuwendam, Alexandre; Paquette, Michel; Rudy, Ashley; Siewert, Matthias; Veillete, Audrey; Weege, Stefanie; Harbor, Jonathan M.; Habeck, J. O.; Sjöberg, Ylva

    2018-01-01

    Communicating science about a phenomenon found under ground and defined by its thermal properties in an easy, funny, and engaging way, can be a challenge. Two years ago, a group of young researchers from Canada and Europe united to tackle this problem by combining arts and science to produce a series of outreach comic strips about permafrost (frozen ground). Because this concerns us all. As the climate warms, permafrost thaws and becomes unstable for houses, roads and airports. The thawi...

  12. [Soil hydrolase characteristics in late soil-thawing period in subalpine/alpine forests of west Sichuan].

    Science.gov (United States)

    Tan, Bo; Wu, Fu-Zhong; Yang, Wan-Qin; Yu, Sheng; Yang, Yu-Lian; Wang, Ao

    2011-05-01

    Late soil-thawing period is a critical stage connecting winter and growth season. The significant temperature fluctuation at this stage might have strong effects on soil ecological processes. In order to understand the soil biochemical processes at this stage in the subalpine/alpine forests of west Sichuan, soil samples were collected from the representative forests including primary fir forest, fir and birch mixed forest, and secondary fir forest in March 5-April 25, 2009, with the activities of soil invertase, urease, and phosphatase (neutral, acid and alkaline phosphatases) measured. In soil frozen period, the activities of the three enzymes in test forests still kept relatively higher. With the increase of soil temperature, the activities of hydrolases at the early stage of soil-thawing decreased rapidly after a sharp increase, except for neutral phosphatease. Thereafter, there was an increase in the activities of urease and phosphatase. Relative to soil mineral layer, soil organic layer had higher hydrolase activity in late soil-thawing period, and showed more obvious responses to the variation of soil temperature.

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

    Directory of Open Access Journals (Sweden)

    Qiang Fu

    2017-05-01

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

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

    Science.gov (United States)

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

    2014-12-01

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

  15. 21 CFR 160.150 - Frozen egg whites.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen egg whites. 160.150 Section 160.150 Food... HUMAN CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.150 Frozen egg whites. (a) Frozen egg whites, frozen egg albumen is the food prepared by freezing...

  16. Effects of Bamboo Shoot Dietary Fiber on Mechanical Properties, Moisture Distribution, and Microstructure of Frozen Dough

    Directory of Open Access Journals (Sweden)

    Hua Zhang

    2017-01-01

    Full Text Available In this paper, the effects of Bamboo shoot dietary fiber (BSDF on the mechanical properties, moisture distribution, and microstructure of frozen dough were investigated. The state and distribution of water in frozen dough was determined by differential scanning calorimetry (DSC and low-field nuclear magnetic resonance (LNMR spectroscopy. The microstructure of frozen dough was studied. The structure of the gluten protein network found in wheat flour dough was studied by scanning electron microscopy (SEM. The result showed that the BSDF could significantly improve the viscoelasticity and extensibility of frozen dough after thawing in a dose-dependent manner. It was significantly improved with the increase in the addition amount of BSDF (P<0.05. DSC analysis showed that the freezable water content and thermal stability of frozen dough were increased after the addition of BSDF. LNMR analysis showed that the appropriate (<0.1% addition amount of BSDF could significantly (P<0.05 decline the contents of bound water. Meanwhile, the loose bound water and free water were raised significantly (P<0.05 after the addition of BSDF. Moreover, the addition of BSDF induces arrangement of starch granule and gluten network in frozen dough. BSDF can be used as a novel quality improver of frozen dough.

  17. Frozen-thawed embryo transfer in a natural or mildly hormonally stimulated cycle in women with regular ovulatory cycles: a RCT.

    Science.gov (United States)

    Peeraer, Karen; Couck, Isabelle; Debrock, Sophie; De Neubourg, Diane; De Loecker, Peter; Tomassetti, Carla; Laenen, Annouschka; Welkenhuysen, Myriam; Meeuwis, Luc; Pelckmans, Sofie; Meuleman, Christel; D'Hooghe, Thomas

    2015-11-01

    Can ovarian stimulation with low dose hMG improve the implantation rate (IR) per frozen-thawed embryo transferred (FET) when compared with natural cycle in an FET programme in women with a regular ovulatory cycle? Both IR and live birth rate (LBR) per FET were similar in the group with mild ovarian stimulation and the natural cycle group. Different cycle regimens for endometrial preparation are used prior to FET: spontaneous ovulatory cycles, cycles with artificial endometrial preparation using estrogen and progesterone hormones, and cycles stimulated with gonadotrophins or clomiphene citrate. At present, it is not clear which regimen results in the highest IR or LBR. More specifically, there are no RCTs in ovulatory women comparing reproductive outcome after FET during a natural cycle and during a hormonally stimulated cycle. A total of 410 women scheduled for FET during 579 cycles (December 2003-September 2013) were enrolled in an open-label RCT to natural cycle (NC FET group, n = 291) or to a cycle hormonally stimulated with s.c. gonadotrophins (hMG FET group, 37.5-75 IU per day, n = 288). A total of 672 embryos were transferred during 434 cycles (332 embryos and 213 cycles in the NC FET group; 340 embryos and 221 cycles in the hMG FET group). Assuming a = 0.05 and 80% power, it was calculated that 219 frozen-thawed embryos were required for transfer in each group to demonstrate a difference of 10% in IR. Women were eligible according to the following inclusion criteria: regular ovulatory cycle, female age ≥21 years and ≤45 years, informed consent. FET cycles with preimplantation genetic screening were excluded. The primary outcome was IR per embryo transferred. Secondary outcomes included IR with fetal heart beat (FHB), LBR per embryo transferred and endometrial thickness on the day of hCG administration. Statistical analysis was by intention to treat and controlled for the presence of multiple measures, as eligible women could be randomized in more than

  18. A rapid and efficient DNA extraction protocol from fresh and frozen human blood samples.

    Science.gov (United States)

    Guha, Pokhraj; Das, Avishek; Dutta, Somit; Chaudhuri, Tapas Kumar

    2018-01-01

    Different methods available for extraction of human genomic DNA suffer from one or more drawbacks including low yield, compromised quality, cost, time consumption, use of toxic organic solvents, and many more. Herein, we aimed to develop a method to extract DNA from 500 μL of fresh or frozen human blood. Five hundred microliters of fresh and frozen human blood samples were used for standardization of the extraction procedure. Absorbance at 260 and 280 nm, respectively, (A 260 /A 280 ) were estimated to check the quality and quantity of the extracted DNA sample. Qualitative assessment of the extracted DNA was checked by Polymerase Chain reaction and double digestion of the DNA sample. Our protocol resulted in average yield of 22±2.97 μg and 20.5±3.97 μg from 500 μL of fresh and frozen blood, respectively, which were comparable to many reference protocols and kits. Besides yielding bulk amount of DNA, our protocol is rapid, economical, and avoids toxic organic solvents such as Phenol. Due to unaffected quality, the DNA is suitable for downstream applications. The protocol may also be useful for pursuing basic molecular researches in laboratories having limited funds. © 2017 Wiley Periodicals, Inc.

  19. Modelling and validation of robust partial thawing of frozen convenience foods during distribution in the cold chain

    DEFF Research Database (Denmark)

    Adler-Nissen, Jens; Zammit, Gine Ørnholt

    2011-01-01

    with small blocks of a frozen model food (23 pct. Tylose® gel) and quipped with temperature loggers were distributed by trucks operating in the cold chain. In addition, controlled storage and temperature abuse experiments were conducted. To predict the product temperature–time relationship we developed a new...... frozen even after two days or more of distribution at +5oC, and that the temperatures inside the product and in the middle of the box were quite stable against the normal oscillations of the ambient temperature in the cold chain. The product temperature was also robust against temperature abuse......In collaboration with two commercial distributors we have tested a new concept for distribution, where convenience products for the food service industry are prepared, frozen and packed in cardboard boxes, but distributed in the chill chain at +5°C instead of in the frost chain. This will lead...

  20. Inducing Heat Shock Proteins Enhances the Stemness of Frozen-Thawed Adipose Tissue-Derived Stem Cells.

    Science.gov (United States)

    Shaik, Shahensha; Hayes, Daniel; Gimble, Jeffrey; Devireddy, Ram

    2017-04-15

    Extensive research has been performed to determine the effect of freezing protocol and cryopreservation agents on the viability of adipose tissue-derived stromal/stem cells (ASCs) as well as other cells. Unfortunately, the conclusion one may draw after decades of research utilizing fundamentally similar cryopreservation techniques is that a barrier exists, which precludes full recovery. We hypothesize that agents capable of inducing a subset of heat shock proteins (HSPs) and chaperones will reduce the intrinsic barriers to the post-thaw recovery of ASCs. ASCs were exposed to 43°C for 1 h to upregulate HSPs, and the temporal HSP expression profile postheat shock was determined by performing quantitative polymerase chain reaction (PCR) and western blotting assays. The expression levels of HSP70 and HSP32 were found to be maximum at 3 h after the heat shock, whereas HSP90 and HSP27 remain unchanged. The heat shocked ASCs cryopreserved during maximal HSPs expression exhibited increased post-thaw viability than the nonheat shocked samples. Histochemical staining and quantitative reverse transcription-PCR indicated that the ASC differentiation potential was retained. Thus, suggesting that the upregulation of HSPs before a freezing insult is beneficial to ASCs and a potential alternative to the use of harmful cryoprotective agents.

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

    Science.gov (United States)

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

    1999-01-01

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

  2. Is frozen embryo transfer better for mothers and babies? Can cumulative meta-analysis provide a definitive answer?

    Science.gov (United States)

    Maheshwari, Abha; Pandey, Shilpi; Amalraj Raja, Edwin; Shetty, Ashalatha; Hamilton, Mark; Bhattacharya, Siladitya

    2018-01-01

    Initial observational studies and a systematic review published 5 years ago have suggested that obstetric and perinatal outcomes are better in offspring conceived following frozen rather than fresh embryo transfers, with reduced risks of preterm birth, small for gestational age, low birth weight and pre-eclampsia. More recent primary studies are beginning to challenge some of these findings. We therefore conducted an updated systematic review and cumulative meta-analysis to examine if these results have remained consistent over time. The aim of this study was to perform a systematic review and cumulative meta-analysis (trend with time) of obstetric and perinatal complications in singleton pregnancies following the transfer of frozen thawed and fresh embryos generated through in-vitro fertilisation. Data Sources from Medline, EMBASE, Cochrane Central Register of Clinical Trials DARE and CINAHL (1984-2016) were searched using appropriate key words. Observational and randomised studies comparing obstetric and perinatal outcomes in singleton pregnancies conceived through IVF using either fresh or frozen thawed embryos. Two independent reviewers extracted data in 2 × 2 tables and assessed the methodological quality of the relevant studies using CASP scoring. Both aggregated as well as cumulative meta-analysis was done using STATA. Twenty-six studies met the inclusion criteria. Singleton babies conceived from frozen thawed embryos were at lower relative risk (RR) of preterm delivery (0.90; 95% CI 0.84-0.97) low birth weight (0.72; 95% CI 0.67-0.77) and small for gestational age (0.61; 95% CI 0.56-0.67) compared to those conceived from fresh embryo transfers, but faced an increased risk (RR) of hypertensive disorders of pregnancy (1.29; 95% CI 1.07-1.56) large for gestational age (1.54; 95% CI 1.48-1.61) and high birth weight (1.85; 95% CI 1.46-2.33). There was no difference in the risk of congenital anomalies and perinatal mortality between the two groups. The direction

  3. Proteome stability analysis of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human colon mucosal biopsies

    DEFF Research Database (Denmark)

    Bjerg Bennike, Tue; Kastaniegaard, Kenneth; Padurariu, Simona

    2016-01-01

    Large repositories of well characterized RNAlater preserved samples and formalin-fixed, paraffin-embedded samples have been generated worldwide. However, the impact on the proteome of the preservation methods remain poorly described. Therefore, we analyzed the impact on the proteome of preserving...... samples in RNAlater, and by formalin-fixation, paraffin-embedding on human soft tissue, using directly frozen samples as a control ("Comparing the proteome of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human tissue samples" [1]). We here report the data from the analysis...

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

    Science.gov (United States)

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

    2009-08-01

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

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

    Science.gov (United States)

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

    2017-07-01

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

  6. Production of healthy cloned mice from bodies frozen at -20 degrees C for 16 years.

    Science.gov (United States)

    Wakayama, Sayaka; Ohta, Hiroshi; Hikichi, Takafusa; Mizutani, Eiji; Iwaki, Takamasa; Kanagawa, Osami; Wakayama, Teruhiko

    2008-11-11

    Cloning animals by nuclear transfer provides an opportunity to preserve endangered mammalian species. However, it has been suggested that the "resurrection" of frozen extinct species (such as the woolly mammoth) is impracticable, as no live cells are available, and the genomic material that remains is inevitably degraded. Here we report production of cloned mice from bodies kept frozen at -20 degrees C for up to 16 years without any cryoprotection. As all of the cells were ruptured after thawing, we used a modified cloning method and examined nuclei from several organs for use in nuclear transfer attempts. Using brain nuclei as nuclear donors, we established embryonic stem cell lines from the cloned embryos. Healthy cloned mice were then produced from these nuclear transferred embryonic stem cells by serial nuclear transfer. Thus, nuclear transfer techniques could be used to "resurrect" animals or maintain valuable genomic stocks from tissues frozen for prolonged periods without any cryopreservation.

  7. Evaluation of ebselen supplementation on cryopreservation medium in human semen.

    Science.gov (United States)

    Khodayari Naeini, Zohreh; Hassani Bafrani, Hassan; Nikzad, Hossein

    2014-04-01

    An effect of cryopreservation on human sperm is sublethal cryodamage, in which cell viability post-thaw is lost more rapidly at later times than in fresh cells. This study examined whether the addition of an antioxidant to cryopreservation medium could improve the post-thaw parameters and evaluation of sperm chromatin quality of cryopreserved human spermatozoa from men with normal semen parameters. Semen samples (n=35) were collected by masturbation and assessed following WHO standards. Individual samples were classified as two portions. One portion (n=10) was for elucidate the concentration of ebselen.Then the samples(n=25) were divided in to 5groups.The first aliquot remained fresh.The second aliquots was mixed with cryopreservation medium.The third aliquots were mixed with cryopreservation medium containing solvent of ebselen.The forth and fifth aliquots were mixed with cryopreservation medium containing 1.25 and 2.5 µm of ebselen.Samples were frozen and thawed samples were assessed for sperm parameters.Three-way ANOVA Multivariate measures were used to assess. According to this assesment the differences are observed in existent groups in post-thaw count, motility index, vitality staining, and morphology and DNA fragmentation. After freezing the media containing of ebselen, DNA fragmentation is significantly different in comparison with control group. ebselen with 1.25 µm dose was significantly associated with post-thaw DNA fragmentation (p=0.047). Similarly ebselen with 2.5 µm dose was significantly associated with post-thaw DNA fragmentation (p=0.038). But other parameters were not altered. These results suggest that the addition of ebselen to cryopreservation medium doesnot improve post-thaw parameters and DNA fragmentation of sperm.

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

    Science.gov (United States)

    Fraser, L; Strzezek, J

    2007-06-01

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

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

    Science.gov (United States)

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

    2016-03-01

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

  10. Effect of permafrost thaw on CO2 and CH4 exchange in a western Alaska peatland chronosequence

    International Nuclear Information System (INIS)

    Johnston, Carmel E; Ewing, Stephanie A; Harden, Jennifer W; Fuller, Christopher C; Manies, Kristen; Varner, Ruth K; Wickland, Kimberly P; Koch, Joshua C; Jorgenson, M Torre

    2014-01-01

    Permafrost soils store over half of global soil carbon (C), and northern frozen peatlands store about 10% of global permafrost C. With thaw, inundation of high latitude lowland peatlands typically increases the surface-atmosphere flux of methane (CH 4 ), a potent greenhouse gas. To examine the effects of lowland permafrost thaw over millennial timescales, we measured carbon dioxide (CO 2 ) and CH 4 exchange along sites that constitute a ∼1000 yr thaw chronosequence of thermokarst collapse bogs and adjacent fen locations at Innoko Flats Wildlife Refuge in western Alaska. Peak CH 4 exchange in July (123 ± 71 mg CH 4 –C m −2 d −1 ) was observed in features that have been thawed for 30 to 70 (<100) yr, where soils were warmer than at more recently thawed sites (14 to 21 yr; emitting 1.37 ± 0.67 mg CH 4 –C m −2 d −1 in July) and had shallower water tables than at older sites (200 to 1400 yr; emitting 6.55 ± 2.23 mg CH 4 –C m −2 d −1 in July). Carbon lost via CH 4 efflux during the growing season at these intermediate age sites was 8% of uptake by net ecosystem exchange. Our results provide evidence that CH 4 emissions following lowland permafrost thaw are enhanced over decadal time scales, but limited over millennia. Over larger spatial scales, adjacent fen systems may contribute sustained CH 4 emission, CO 2 uptake, and DOC export. We argue that over timescales of decades to centuries, thaw features in high-latitude lowland peatlands, particularly those developed on poorly drained mineral substrates, are a key locus of elevated CH 4 emission to the atmosphere that must be considered for a complete understanding of high latitude CH 4 dynamics. (paper)

  11. Potential carbon emissions dominated by carbon dioxide from thawed permafrost soils

    Science.gov (United States)

    Schädel, Christina; Bader, Martin K.-F.; Schuur, Edward A.G.; Biasi, Christina; Bracho, Rosvel; Čapek, Petr; De Baets, Sarah; Diáková, Kateřina; Ernakovich, Jessica; Estop-Aragones, Cristian; Graham, David E.; Hartley, Iain P.; Iversen, Colleen M.; Kane, Evan S.; Knoblauch, Christian; Lupascu, Massimo; Martikainen, Pertti J.; Natali, Susan M.; Norby, Richard J.; O'Donnell, Jonathan A.; Roy Chowdhury, Taniya; Šantrůčková, Hana; Shaver, Gaius; Sloan, Victoria L.; Treat, Claire C.; Turetsky, Merritt R.; Waldrop, Mark P.; Wickland, Kimberly P.

    2016-01-01

    Increasing temperatures in northern high latitudes are causing permafrost to thaw, making large amounts of previously frozen organic matter vulnerable to microbial decomposition. Permafrost thaw also creates a fragmented landscape of drier and wetter soil conditions that determine the amount and form (carbon dioxide (CO2), or methane (CH4)) of carbon (C) released to the atmosphere. The rate and form of C release control the magnitude of the permafrost C feedback, so their relative contribution with a warming climate remains unclear. We quantified the effect of increasing temperature and changes from aerobic to anaerobic soil conditions using 25 soil incubation studies from the permafrost zone. Here we show, using two separate meta-analyses, that a 10 °C increase in incubation temperature increased C release by a factor of 2.0 (95% confidence interval (CI), 1.8 to 2.2). Under aerobic incubation conditions, soils released 3.4 (95% CI, 2.2 to 5.2) times more C than under anaerobic conditions. Even when accounting for the higher heat trapping capacity of CH4, soils released 2.3 (95% CI, 1.5 to 3.4) times more C under aerobic conditions. These results imply that permafrost ecosystems thawing under aerobic conditions and releasing CO2 will strengthen the permafrost C feedback more than waterlogged systems releasing CO2 and CH4 for a given amount of C.

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

    Science.gov (United States)

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

    2015-01-01

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

  13. Derivation of the human embryonic stem cell line RCe010-A (RC-6

    Directory of Open Access Journals (Sweden)

    P.A. De Sousa

    2016-03-01

    Full Text Available The human embryonic stem cell line RCe010-A (RC-6 was derived from a frozen and thawed blastocyst voluntarily donated as unsuitable and surplus to fertility requirements following ethics committee approved informed consent under licence from the UK Human Fertilisation and Embryology Authority. The cell line shows normal pluripotency marker expression and differentiation to the three germ layers in vitro. It has a normal 46XY male karyotype and microsatellite PCR identity, HLA and blood group typing data are available.

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

    Science.gov (United States)

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

    1990-04-01

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

  15. Effect of lactose, skim milk and Tris diluents on frozen buffalo spermatozoa

    Directory of Open Access Journals (Sweden)

    Ali Rastegarnia

    2007-08-01

    Full Text Available The composition of the extender in which semen is diluted before freezing plays a major role in successful cryopreservation of spermatozoa. This study was carried out to identify the suitable buffer for cryopreservation of buffalo semen. Sixteen split pooled ejaculates from two buffalo bulls possessing more than 70% visual sperm motility, were diluted at 370c either in lactose, skim milk or Tris extenders. The diluted semen was cooled to 40c within 2 hours, equilibrated at 40c for 4-6 hours following the addition of glycerol, filled in 0.5 ml French straws and frozen in a programmable cell freezer before plunging into liquid nitrogen. Semen was thawed at 370c for 30 seconds after 48 hours of storage inside liquid nitrogen. Post thaw visual sperm motility, plasma membrane integrity and acrosome morphology of each semen sample were assessed by warm plate microscopy at 370c, hypo-osmotic swelling test (HOST and giemsa staining, respectively. Analysis of variance revelated that percentage of post thaw visual sperm motility (Mean± standard deviation tended to be higher in Tris (50±3.6 than skim milk (44.5±2.5 and lower in lactose (24.4±10.5 extenders (P

  16. HPTAM, a two-dimensional Heat Pipe Transient Analysis Model, including the startup from a frozen state

    Science.gov (United States)

    Tournier, Jean-Michel; El-Genk, Mohamed S.

    1995-01-01

    A two-dimensional Heat Pipe Transient Analysis Model, 'HPTAM,' was developed to simulate the transient operation of fully-thawed heat pipes and the startup of heat pipes from a frozen state. The model incorporates: (a) sublimation and resolidification of working fluid; (b) melting and freezing of the working fluid in the porous wick; (c) evaporation of thawed working fluid and condensation as a thin liquid film on a frozen substrate; (d) free-molecule, transition, and continuum vapor flow regimes, using the Dusty Gas Model; (e) liquid flow and heat transfer in the porous wick; and (f) thermal and hydrodynamic couplings of phases at their respective interfaces. HPTAM predicts the radius of curvature of the liquid meniscus at the liquid-vapor interface and the radial location of the working fluid level (liquid or solid) in the wick. It also includes the transverse momentum jump condition (capillary relationship of Pascal) at the liquid-vapor interface and geometrically relates the radius of curvature of the liquid meniscus to the volume fraction of vapor in the wick. The present model predicts the capillary limit and partial liquid recess (dryout) in the evaporator wick, and incorporates a liquid pooling submodel, which simulates accumulation of the excess liquid in the vapor core at the condenser end.

  17. Production of healthy cloned mice from bodies frozen at −20°C for 16 years

    Science.gov (United States)

    Wakayama, Sayaka; Ohta, Hiroshi; Hikichi, Takafusa; Mizutani, Eiji; Iwaki, Takamasa; Kanagawa, Osami; Wakayama, Teruhiko

    2008-01-01

    Cloning animals by nuclear transfer provides an opportunity to preserve endangered mammalian species. However, it has been suggested that the “resurrection” of frozen extinct species (such as the woolly mammoth) is impracticable, as no live cells are available, and the genomic material that remains is inevitably degraded. Here we report production of cloned mice from bodies kept frozen at −20 °C for up to 16 years without any cryoprotection. As all of the cells were ruptured after thawing, we used a modified cloning method and examined nuclei from several organs for use in nuclear transfer attempts. Using brain nuclei as nuclear donors, we established embryonic stem cell lines from the cloned embryos. Healthy cloned mice were then produced from these nuclear transferred embryonic stem cells by serial nuclear transfer. Thus, nuclear transfer techniques could be used to “resurrect” animals or maintain valuable genomic stocks from tissues frozen for prolonged periods without any cryopreservation. PMID:18981419

  18. Coleopterofauna found on fresh and frozen rabbit carcasses in Curitiba, Paraná, Brazil

    Directory of Open Access Journals (Sweden)

    KM. Mise

    Full Text Available Many arthropod species are associated with carrion and some of them can be used as forensic indicators in murder investigations to estimate the time of death. Different physical conditions of cadavers may influence the fauna and the importance of freezing and thawing is unknown. The present paper sought to survey the Coleoptera species encountered in frozen and fresh rabbit carcasses, at a forest in Curitiba, Brazil. Four rabbit carcasses, two of them fresh and the other frozen were used. The coleopterofauna was sampled daily, and analyzed using non-parametric tests. A total of 666 beetles were sampled, belonging to 28 species in 10 families. Most of the beetles captured were larvae of Oxelytrum spp. (433 which are known to be necrophagous. The two frozen carcasses accounted for most of the beetles (338 and 180 in comparison with the two fresh carcasses (103 and 45. The tests were based on the most abundant species. Oxelytrum spp. median differed significantly between carcasses (H = 12.47844; p = 0.0059. The two fresh carcasses differed significantly (U = 190.0; p = 0.00019, but there was no significant difference between the frozen carcasses (U = 336.0; p = 0.29755. The data indicate that the freezing process prevents certain species to colonize carcasses, in this case with the dominance of species of Oxelytrum. These data also indicate that careful attention is necessary before using frozen carcasses in forensic entomology studies.

  19. Frozen dispersions of nanomaterials are a useful operational procedure in nanotoxicology.

    Science.gov (United States)

    Vila, Laura; Rubio, Laura; Annangi, Balasubramanyam; García-Rodríguez, Alba; Marcos, Ricard; Hernández, Alba

    2017-02-01

    The variability observed in nanoparticle (NP) dispersions can affect the reliability of the results obtained in short-term tests, and mainly in long-term experiments. In addition, obtaining a good dispersion is time-consuming and acts as a bottleneck in the development of high-throughput screening methodologies. The freezing of different aliquots from a stock dispersion would overcome such limitations, but no studies have explored the impact of freezing thawing the samples on the physico-chemical and biological properties of the nanomaterial (NM). This work aims to compare fresh-prepared and frozen MWCNT, ZnO-, Ag-, TiO 2- and CeO 2 -NP dispersions, used as models. NP characterization (size and morphology by TEM), hydrodynamic size and zeta potential were performed. Viability comparisons were determined in BEAS-2B cells. Cellular NP uptake and induced ROS production was assessed by TEM and flow cytometry, respectively. The obtained results show no important differences between frozen and fresh NP in their physico-chemical characteristics or their biological effects. This study is the first to demonstrate that there is no scientific evidence to dismiss the use of frozen NP, opening the door to the development of short- and long-term experiments with higher consistency, accuracy and reproducibility in a much shorter time and using a simplified procedure.

  20. Frozen Soil Barrier. Subsurface Contaminants Focus Area. OST Reference No. 51

    International Nuclear Information System (INIS)

    1999-01-01

    Problem: Hazardous and radioactive materials have historically been disposed of at the surface during operations at Department of Energy facilities. These contaminants have entered the subsurface, contaminating soils and groundwater resources. Remediation of these groundwater plumes using the baseline technology of pump and treat is expensive and takes a long time to complete. Containment of these groundwater plumes can be alternative or an addition to the remediation activities. Standard containment technologies include slurry walls, sheet piling, and grouting. These are permanent structures that once installed are difficult to remove. How It Works: Frozen Soil Barrier technology provides a containment alternative, with the key difference being that the barrier can be easily removed after a period of time, such as after the remediation or removal of the source is completed. Frozen Soil Barrier technology can be used to isolate and control the migration of underground radioactive or other hazardous contaminants subject to transport by groundwater flow. Frozen Soil Barrier technology consists of a series of subsurface heat transfer devices, known as thermoprobes, which are installed around a contaminant source and function to freeze the soil pore water. The barrier can easily be maintained in place until remediation or removal of the contaminants is complete, at which time the barrier is allowed to thaw.

  1. Proteome stability analysis of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human colon mucosal biopsies

    Directory of Open Access Journals (Sweden)

    Tue Bjerg Bennike

    2016-03-01

    Full Text Available Large repositories of well characterized RNAlater preserved samples and formalin-fixed, paraffin-embedded samples have been generated worldwide. However, the impact on the proteome of the preservation methods remain poorly described. Therefore, we analyzed the impact on the proteome of preserving samples in RNAlater, and by formalin-fixation, paraffin-embedding on human soft tissue, using directly frozen samples as a control (“Comparing the proteome of snap frozen, RNAlater preserved, and formalin-fixed paraffin-embedded human tissue samples” [1]. We here report the data from the analysis. The comparative analysis was performed on 24 colon mucosa biopsies, extracted from the sigmoideum of two gastroenterologically healthy participants for the purpose of this study. A set of biopsies were additionally stored for 30 min at room temperature prior to formalin-fixation. The samples were analyzed by high throughput gel free quantitative proteomics. The MS proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PRIDE: http://www.ebi.ac.uk/pride/archive/projects/PXD002029. Keywords: Human, Colon, Mucosa, RNAlater, FFPE, Snap-frozen, Stability, LC–MS, Proteomics

  2. [In vitro activity of human bone marrow cells after cryopreservation in liquid nitrogen for 21 - 25 years].

    Science.gov (United States)

    Huang, You-Zhang; Shen, Jian-Liang; Gong, Li-Zhong; Zheng, Pei-Hao; Liu, Yi; Yin, Wen-Jie; Cen, Jian; Wang, Ning; Zhao, De-Feng

    2010-02-01

    The aim of this study was to investigate the best method to preserve human bone marrow cells and the effectiveness of long term cryopreservation at -80 degrees C. The human bone marrow cells in 20 samples were firstly frozen by a programmed freezer or -80 degrees C refrigerator, and then were preserved in liquid nitrogen with DMSO-AuP (10% dimethylsulfonamide, 10% autologous plasma) or DMSO-HES-HuA (5% dimethylsulfonamide, 6% hydroxyethyl starch, 4% human serum albumin) as cryoprotectant for 21 to 25 years. They were thawed in 38 degrees C. The cell sample frozen in -80 degrees C refrigerator was frozen at a low frozen speed of 1 degrees C/min which was the same as the programmed freezer before -30 degrees C. Before detection the bone marrow cells were taken from liquid nitrogen and were thawed in 38 degrees C, then the suspension of bone marrow cells was prepared for detection. The cell morphology and recovery rate of erythrocytes, nucleocytes and platelets; the recovery rate of hematopoietic stem progenitors cells, as well as mesenchymal stem cells were determined. The results showed that the protective effectiveness of DMSO-HES-HuA was better than DMSO-AuP. The mature erythrocytes were destroyed lightly [(3.5 +/- 1.5)% versus (12.6 +/- 4.8)%], the hemolysis rate was lower [(3.3 +/- 1.6)% versus (23.1 +/- 5.1)%]. Osmotic fragility of erythrocytes in the former was not changed, but was dropped in the latter. The recovery rates of red cell, platelet, granulocyte-macrophage colony forming units and long term culture-initiating cells were higher in the former than that in the latter [(96.1 +/- 1.8)%, (70.0 +/- 9.5)%, (49.2 +/- 10.9)%, (54.2 +/- 13.8)% versus (76.3 +/- 5.6)%, (52.7 +/- 8.1)%, (43.5 +/- 12.3)%, (47.2 +/- 13.6)% respectively]. With each kind of cryoprotectant or frozen method, the frozen MSC could keep the original growth properties. With the same cryoprotectant and different frozen method, the cryopreservative effectiveness was not different. The

  3. Dissolved organic carbon loss from Yedoma permafrost amplified by ice wedge thaw

    International Nuclear Information System (INIS)

    Vonk, J E; Mann, P J; Spencer, R G M; Bulygina, E B; Holmes, R M; Dowdy, K L; Davydova, A; Davydov, S P; Zimov, N; Eglinton, T I

    2013-01-01

    Pleistocene Yedoma permafrost contains nearly a third of all organic matter (OM) stored in circum-arctic permafrost and is characterized by the presence of massive ice wedges. Due to its rapid formation by sediment accumulation and subsequent frozen storage, Yedoma OM is relatively well preserved and highly biologically available (biolabile) upon thaw. A better understanding of the processes regulating Yedoma degradation is important to improve estimates of the response and magnitude of permafrost carbon feedbacks to climate warming. In this study, we examine the composition of ice wedges and the influence of ice wedge thaw on the biolability of Yedoma OM. Incubation assays were used to assess OM biolability, fluorescence spectroscopy to characterize the OM composition, and potential enzyme activity rates to examine the controls and regulation of OM degradation. We show that increasing amounts of ice wedge melt water in Yedoma-leached incubations enhanced the loss of dissolved OM over time. This may be attributed to the presence of low-molecular weight compounds and low initial phenolic content in the OM of ice wedges, providing a readily available substrate that promotes the degradation of Yedoma OC. The physical vulnerability of ice wedges upon thaw (causing irreversible collapse), combined with the composition of ice wedge-engrained OM (co-metabolizing old OM), underlines the particularly strong potential of Yedoma to generate a positive feedback to climate warming relative to other forms of non-ice wedge permafrost. (letter)

  4. Methane production as key to the greenhouse gas budget of thawing permafrost

    Science.gov (United States)

    Knoblauch, Christian; Beer, Christian; Liebner, Susanne; Grigoriev, Mikhail N.; Pfeiffer, Eva-Maria

    2018-04-01

    Permafrost thaw liberates frozen organic carbon, which is decomposed into carbon dioxide (CO2) and methane (CH4). The release of these greenhouse gases (GHGs) forms a positive feedback to atmospheric CO2 and CH4 concentrations and accelerates climate change1,2. Current studies report a minor importance of CH4 production in water-saturated (anoxic) permafrost soils3-6 and a stronger permafrost carbon-climate feedback from drained (oxic) soils1,7. Here we show through seven-year laboratory incubations that equal amounts of CO2 and CH4 are formed in thawing permafrost under anoxic conditions after stable CH4-producing microbial communities have established. Less permafrost carbon was mineralized under anoxic conditions but more CO2-carbon equivalents (CO2-Ce) were formed than under oxic conditions when the higher global warming potential (GWP) of CH4 is taken into account8. A model of organic carbon decomposition, calibrated with the observed decomposition data, predicts a higher loss of permafrost carbon under oxic conditions (113 ± 58 g CO2-C kgC-1 (kgC, kilograms of carbon)) by 2100, but a twice as high production of CO2-Ce (241 ± 138 g CO2-Ce kgC-1) under anoxic conditions. These findings challenge the view of a stronger permafrost carbon-climate feedback from drained soils1,7 and emphasize the importance of CH4 production in thawing permafrost on climate-relevant timescales.

  5. Ancient low-molecular-weight organic acids in permafrost fuel rapid carbon dioxide production upon thaw.

    Science.gov (United States)

    Drake, Travis W; Wickland, Kimberly P; Spencer, Robert G M; McKnight, Diane M; Striegl, Robert G

    2015-11-10

    Northern permafrost soils store a vast reservoir of carbon, nearly twice that of the present atmosphere. Current and projected climate warming threatens widespread thaw of these frozen, organic carbon (OC)-rich soils. Upon thaw, mobilized permafrost OC in dissolved and particulate forms can enter streams and rivers, which are important processors of OC and conduits for carbon dioxide (CO2) to the atmosphere. Here, we demonstrate that ancient dissolved organic carbon (DOC) leached from 35,800 y B.P. permafrost soils is rapidly mineralized to CO2. During 200-h experiments in a novel high-temporal-resolution bioreactor, DOC concentration decreased by an average of 53%, fueling a more than sevenfold increase in dissolved inorganic carbon (DIC) concentration. Eighty-seven percent of the DOC loss to microbial uptake was derived from the low-molecular-weight (LMW) organic acids acetate and butyrate. To our knowledge, our study is the first to directly quantify high CO2 production rates from permafrost-derived LMW DOC mineralization. The observed DOC loss rates are among the highest reported for permafrost carbon and demonstrate the potential importance of LMW DOC in driving the rapid metabolism of Pleistocene-age permafrost carbon upon thaw and the outgassing of CO2 to the atmosphere by soils and nearby inland waters.

  6. Ancient low–molecular-weight organic acids in permafrost fuel rapid carbon dioxide production upon thaw

    Science.gov (United States)

    Drake, Travis W.; Wickland, Kimberly P.; Spencer, Robert G. M.; McKnight, Diane M.; Striegl, Robert G.

    2015-01-01

    Northern permafrost soils store a vast reservoir of carbon, nearly twice that of the present atmosphere. Current and projected climate warming threatens widespread thaw of these frozen, organic carbon (OC)-rich soils. Upon thaw, mobilized permafrost OC in dissolved and particulate forms can enter streams and rivers, which are important processors of OC and conduits for carbon dioxide (CO2) to the atmosphere. Here, we demonstrate that ancient dissolved organic carbon (DOC) leached from 35,800 y B.P. permafrost soils is rapidly mineralized to CO2. During 200-h experiments in a novel high–temporal-resolution bioreactor, DOC concentration decreased by an average of 53%, fueling a more than sevenfold increase in dissolved inorganic carbon (DIC) concentration. Eighty-seven percent of the DOC loss to microbial uptake was derived from the low–molecular-weight (LMW) organic acids acetate and butyrate. To our knowledge, our study is the first to directly quantify high CO2 production rates from permafrost-derived LMW DOC mineralization. The observed DOC loss rates are among the highest reported for permafrost carbon and demonstrate the potential importance of LMW DOC in driving the rapid metabolism of Pleistocene-age permafrost carbon upon thaw and the outgassing of CO2 to the atmosphere by soils and nearby inland waters.

  7. Derivation of the human embryonic stem cell line RCe006-A (RC-2

    Directory of Open Access Journals (Sweden)

    P.A. De Sousa

    2016-03-01

    Full Text Available The human embryonic stem cell line RCe006-A (RC-2 was derived from a frozen and thawed blastocyst voluntarily donated as surplus to fertility requirements following ethics committee approved informed consent under licence from the UK Human Fertilisation and Embryology Authority. The cell line exhibits expression of expected pluripotency markers and in vitro differentiation potential to three germinal lineage representative cell populations. It has a male trisomy 12 karyotype (47XY, +12. Microsatellite DNA marker identity and HLA and blood group typing data are available.

  8. Derivation of the human embryonic stem cell line RCe012-A (RC-8

    Directory of Open Access Journals (Sweden)

    P.A. De Sousa

    2016-03-01

    Full Text Available The human embryonic stem cell line RCe012-A (RC-8 was derived from a frozen and thawed day 5 embryo cultivated to the blastocyst stage. The embryo was voluntarily donated as unsuitable and surplus to fertility requirements following ethics committee approved informed consent under licence from the UK Human Fertilisation and Embryology Authority. The cell line shows normal pluripotency marker expression and differentiation to the three germ layers in vitro. It has a normal 46XX female karyotype and microsatellite PCR identity, HLA and blood group typing data is available.

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

    Science.gov (United States)

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

    2013-12-01

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

  10. Hydrolysis of Marine Cod (Gadus Morhua) Head - Utilization of rest raw material from cod for production of ingredients for human consumption

    OpenAIRE

    Meldstad, Fataneh

    2015-01-01

    The main objective of this study was to determine the possibility of utilizing frozen marine cod head for further processing into high-quality and high-yield fish protein hydrolysates. The project included determination of the effect of mincing versus whole head, frozen storage and thawing methods on the composition and yield of FPH. The results show that freezing and thawing of cod head lead to small changes in the composition of raw material and consequently the resulting ...

  11. Cryo-scanning electron microscopy discloses differences in dehydration of frozen boar semen stored in large containers.

    Science.gov (United States)

    Ekwall, H

    2009-02-01

    In general, freezing in flat plastic polyethylene terephthalate (PET) bags (FlatPacks) at 50 degrees C/min gives better post-thaw viability, in terms of sperm motility and membrane integrity, than does freezing in plastic maxi-straws, probably owing to differences in cryobiology. To test the hypothesis that this better survival post-thaw relates to the degree of sperm dehydration during freezing, the present study investigated the structure of boar semen in a frozen state using cryo-scanning electron microscopy (cryo-SEM) to compare two different packages (FlatPacks and maxi-straws) for single artificial insemination (AI) doses, and three different freezing rates. The semen was split-sample frozen in maxi-straws or FlatPacks (both holding 5 ml) using 3% glycerol as cryoprotectant. Three freezing rates were applied from -5 degrees C to -100 degrees C, namely 2 degrees C/min, 50 degrees C/min and 1200 degrees C/min, the lattermost by plunging the samples into liquid nitrogen (LN(2)). The samples were thereafter fractured into LN(2) and larger areas of extra-cellular, unbound frozen water ('ice lakes') were measured to determine the degree of dehydration of the spermatozoa. These areas decreased in size with an increase in cooling rate, the differences in size being more dramatic for maxi-straws than for FlatPacks. Size of ice lakes was also influenced by location within package in relation to cooling rate, the central values being always smaller in maxi-straws than in Flatpacks (p < 0.05 at 2 degrees C/min and 50 degrees C/min) but not at 1200 degrees C/min, which suggested the FlatPack allows for more homogenous freezing of boar semen.

  12. Oxygenated thawing and rewarming alleviate rewarming injury of cryopreserved pancreatic islets.

    Science.gov (United States)

    Komatsu, Hirotake; Barriga, Alyssa; Medrano, Leonard; Omori, Keiko; Kandeel, Fouad; Mullen, Yoko

    2017-05-06

    Pancreatic islet transplantation is an effective treatment for Type 1 diabetic patients to eliminate insulin injections; however, a shortage of donor organs hinders the widespread use. Although long-term islet storage, such as cryopreservation, is considered one of the key solutions, transplantation of cryopreserved islets is still not practical due to the extensive loss during the cryopreservation-rewarming process. We have previously reported that culturing islets in a hyperoxic environment is an effective treatment to prevent islet death from the hypoxic injury during culture. In this study, we explored the effectiveness of thawing and rewarming cryopreserved islets in a hyperoxic environment. Following cryopreservation of isolated human islets, the thawing solution and culture media were prepared with or without pre-equilibration to 50% oxygen. Thawing/rewarming and the pursuant two-day culture were performed with or without oxygenation. Short-term recovery rate, defined as the volume change during cryopreservation and thawing/rewarming, was assessed. Ischemia-associated and inflammation-associated gene expressions were examined using qPCR after the initial rewarming period. Long-term recovery rate, defined as the volume change during the two-day culture after the thawing/rewarming, was also examined. Islet metabolism and function were assessed by basal oxygen consumption rate and glucose stimulated insulin secretion after long-term recovery. Oxygenated thawing/rewarming did not alter the short-term recovery rate. Inflammation-associated gene expressions were elevated by the conventional thawing/rewarming method and suppressed by the oxygenated thawing/rewarming, whereas ischemia-associated gene expressions did not change between the thawing/rewarming methods. Long-term recovery rate experiments revealed that only the combination therapy of oxygenated thawing/rewarming and oxygenated culture alleviated islet volume loss. These islets showed higher metabolism

  13. Mercury speciation in thawed out and refrozen fish samples by gas chromatography coupled to inductively coupled plasma mass spectrometry and atomic fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Krystek, Petra; Ritsema, Rob [National Institute for Public Health and the Environment (RIVM), Laboratory for Analytical Chemistry, Bilthoven (Netherlands)

    2005-01-01

    Different sub-sampling procedures were applied for the determination of mercury species (as total mercury Hg, methylmercury MeHg{sup +} and inorganic mercury Hg{sup 2+}) in frozen fish meat. Analyses were carried out by two different techniques. After the sample material was pre-treated by microwave digestion, atomic fluorescence spectroscopy (AFS) was used for the determination of total Hg. Speciation analysis was performed according to the following procedure: dissolution of sample material in tetramethylammonium hydroxide (TMAH), derivatisation with sodium tetraethylborate (NaBEt{sub 4}), extraction into isooctane and measurement with gas chromatography inductively coupled plasma mass spectrometry (GC-ICPMS) for the identification and quantification of methylmercury (MeHg{sup +}) and inorganic mercury (Hg{sup 2+}). The concentration range of total Hg measured in the shark fillets is between 0.9 and 3.6 {mu}g g{sup -1} thawed out shark fillet. Speciation analysis leads to {>=}94% Hg present as MeHg{sup +}. Homogeneity, storage conditions and stability of analytical species and sample materials have great influence on analytical results. Sub-sampling of half-frozen/partly thawed out fish and analysis lead to significantly different concentrations, which are on average a factor of two lower. (orig.)

  14. Type of homogenization and fat loss during continuous infusion of human milk.

    Science.gov (United States)

    García-Lara, Nadia Raquel; Escuder-Vieco, Diana; Alonso Díaz, Clara; Vázquez Román, Sara; De la Cruz-Bértolo, Javier; Pallás-Alonso, Carmen Rosa

    2014-11-01

    Substantial fat loss may occur during continuous feeding of human milk (HM). A decrease of fat loss has been described following homogenization. Well-established methods of homogenization of HM for routine use in the neonatal intensive care unit (NICU) would be desirable. We compared the loss of fat based on the use of 3 different methods for homogenizing thawed HM during continuous feeding. Sixteen frozen donor HM samples were thawed, homogenized with ultrasound and separated into 3 aliquots ("baseline agitation," "hourly agitation," and "ultrasound"), and then frozen for 48 hours. Aliquots were thawed again and a baseline agitation was applied. Subsequently, aliquots baseline agitation and hourly agitation were drawn into a syringe, while ultrasound was applied to aliquot ultrasound before it was drawn into a syringe. The syringes were loaded into a pump (2 mL/h; 4 hours). At hourly intervals the hourly agitation infusion was stopped, the syringe was disconnected and gently shaken. During infusion, samples from the 3 groups were collected hourly for analysis of fat and caloric content. The 3 groups of homogenization showed similar fat content at the beginning of the infusion. For fat, mean (SD) hourly changes of -0.03 (0.01), -0.09 (0.01), and -0.09 (0.01) g/dL were observed for the hourly agitation, baseline agitation, and ultrasound groups, respectively. The decrease was smaller for the hourly agitation group (P homogenization is used. © The Author(s) 2014.

  15. Frozen and fresh ovarian tissue require different culture media to promote in vitro development of bovine preantral follicles.

    Science.gov (United States)

    Castro, Simone Vieira; Carvalho, Adeline Andrade; Silva, Cleidson Manoel Gomes; Santos, Francielli Weber; Campello, Cláudio Cabral; de Figueiredo, José Ricardo; Rodrigues, Ana Paula Ribeiro

    2014-10-01

    The aim of this study was to evaluate the efficiency of different media in the in vitro culture of bovine preantral follicles that were used either fresh or following slow freezing treatment. Frozen and fresh noncultured or cultured ovarian fragments were processed for histological, viability, and cell proliferation analyses. For cryopreservation, a solution containing 1.5 M ethylene glycol was frozen in a programmable biological freezer. After thawing, a portion of the samples was destined for frozen controls. The remainder were cultured in vitro for 5 days in three media: α-MEM, McCoy, or M199. Samples from these culture media were collected on days 1 and 5 for quantification of reactive oxygen species (ROS) and for hormonal assays. In fresh-cultured tissues, the percentage of morphologically normal follicles was significantly higher when cultured in M199 compared to that in the other media. In frozen-cultured tissues, McCoy medium was significantly superior to the other media, and was the only treatment that helped in maintaining the viability similar to fresh and frozen controls. Upon quantification of the nucleolus organizer region, we observed greater proliferation of granulosa cells in the frozen-cultured tissues with McCoy medium, and lesser proliferation in fresh-cultured tissues only with α-MEM. In frozen-cultured tissues, ROS levels were highest at day 1 and progressively reduced during culture, independent of the media used. In conclusion, under the conditions used in this study, the M199 and McCoy media are recommended for the culture of follicles derived from fresh and frozen ovarian tissues, respectively.

  16. Effect of different concentration of fish oil in skim milk-egg yolk extenders on post- thawed semen qualities of Kalang swamp buffalo bull

    Directory of Open Access Journals (Sweden)

    Abdul Malik

    2018-01-01

    Full Text Available Objective: To explore the effect of fish oil at different concentrations on post-thawed semen of Kalang swamp buffalo. Methods: A total of 4 Kalang swamp buffalo bulls with 3-5 years of age and weighed about 340-360 kg were slected. Semen was regularly collected from these buffalo bulls once a week by an artificial vagina. Fish oil was supplementary at the dosages of 0 mg (control, 50 mg, 100 mg, 150 mg, and 200 mg to the extender (skim milk-egg yolk. Fresh, pre-freezing and frozen semen were thawed at 37 °C and evaluated for motility, viability, morphology, and plasma integrity of membrane. Results: The study results indicated that before freezing, supplementation of fish oil at the dose of 150 mg in the extender had significantly motility. And a significant (P<0.05 increase was observed in viability and motility of post-thawed semen at the dose of 150 mg fish oil, which was in difference with other treatment groups. Conclusions: Addition of 150 mg fish oil in the extender could be positive for the enhancement of the quality of post-thawed semen of Kalang swamp buffaloes.

  17. The protective effect of rapid cold-hardening develops more quickly in frozen versus supercooled larvae of the Antarctic midge, Belgica antarctica.

    Science.gov (United States)

    Kawarasaki, Yuta; Teets, Nicholas M; Denlinger, David L; Lee, Richard E

    2013-10-15

    During the austral summer, larvae of the terrestrial midge Belgica antarctica (Diptera: Chironomidae) experience highly variable and often unpredictable thermal conditions. In addition to remaining freeze tolerant year-round, larvae are capable of swiftly increasing their cold tolerance through the rapid cold-hardening (RCH) response. The present study compared the induction of RCH in frozen versus supercooled larvae. At the same induction temperature, RCH occurred more rapidly and conferred a greater level of cryoprotection in frozen versus supercooled larvae. Furthermore, RCH in frozen larvae could be induced at temperatures as low as -12°C, which is the lowest temperature reported to induce RCH. Remarkably, as little as 15 min at -5°C significantly enhanced larval cold tolerance. Not only is protection from RCH acquired swiftly, but it is also quickly lost after thawing for 2 h at 2°C. Because the primary difference between frozen and supercooled larvae is cellular dehydration caused by freeze concentration of body fluids, we also compared the effects of acclimation in dehydrated versus frozen larvae. Because slow dehydration without chilling significantly increased larval survival to a subsequent cold exposure, we hypothesize that cellular dehydration caused by freeze concentration promotes the rapid acquisition of cold tolerance in frozen larvae.

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

    International Nuclear Information System (INIS)

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

    2009-01-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Duarte, R.C. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Av. Professor Lineu Prestes 2242 Zip code, 05508-000 Cidade Universitaria Sao Paulo, SP-Brazil (Brazil)], E-mail: renatocduarte@yahoo.com.br; Araujo, M.M.; Salum, D.C. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Av. Professor Lineu Prestes 2242 Zip code, 05508-000 Cidade Universitaria Sao Paulo, SP-Brazil (Brazil); Marchioni, E. [IPHC-UMR 7178-Faculte de Pharmacie, 74 route du Rhin, 67400 Illkirch (France); Villavicencio, A.L.C.H. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Av. Professor Lineu Prestes 2242 Zip code, 05508-000 Cidade Universitaria Sao Paulo, SP-Brazil (Brazil)], E-mail: villavic@ipen.br

    2009-07-15

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

  20. Frozen-Ground Cartoons: An international collaboration between artists and permafrost scientists

    Science.gov (United States)

    Sjoberg, Y.; Bouchard, F.; Deshpande, B.; Fritz, M.; Malenfant-Lepage, J.; Nieuwendam, A.; Paquette, M.; Rudy, A.; Siewert, M. B.; Veillette, A.; Weege, S.; Habeck, J. O.; Harbor, J.

    2017-12-01

    Communicating science about a phenomenon found under ground and defined by its thermal properties in an easy, fun, and engaging way, can be a challenge. Two years ago, a group of young researchers from Canada and Europe united to tackle this problem by combining arts and science to produce a series of outreach comic strips about permafrost (frozen ground). Because this concerns us all. As the climate warms, permafrost thaws and becomes unstable for houses, roads and airports.The thawing also disrupts ecosystems, impacts water quality, and releases greenhouse gases into the atmosphere, making climate change even stronger. The Frozen Ground Cartoon project aims to present and explain permafrost research, placing emphasis on field work and the rapidly changing northern environment. The target audience is kids, youth, parents and teachers, with the general goal of making permafrost science more fun and accessible to the public. The project has so far produced 22 pages of comics through an iterative process of exchanging ideas between two artists and thirteen scientists. The project artists were selected through an application call that received 49 applications from artists in 16 countries. With input from scientists, artists Noémie Ross (Canada) and Heta Nääs (Finland) have created a set of beautiful, artistic, humoristic, and pedagogic comics.. The comics are available for free download through the project web page (in English and Swedish), and printed copies have so far been handed out to school kids and general public in Europe. Prints in North America are planned for the fall of 2017. The next steps of the project are (1) to distribute the comics as wide as possible, (2) work towards translations into more languages, and (3) to evaluate the effectiveness of the science communication through the comics, in collaboration with schools and pedagogic experts.

  1. 14-Day thawed plasma retains clot enhancing properties and inhibits tPA-induced fibrinolysis.

    Science.gov (United States)

    Huebner, Benjamin R; Moore, Ernest E; Moore, Hunter B; Shepherd-Singh, Raymond; Sauaia, Angela; Stettler, Gregory R; Nunns, Geoffrey R; Silliman, Christopher C

    2017-11-01

    Plasma-first resuscitation attenuates trauma-induced coagulopathy (TIC); however, the logistics of plasma-first resuscitation require thawed plasma (TP) be readily available due to the obligatory thawing time of fresh frozen plasma (FFP). The current standard is storage of TP for up to 5 days at 4°C, based on factor levels at outdate, for use in patients at risk for TIC, but there remains a 2.2% outdated wastage rate. However, the multitude of plasma proteins in attenuating TIC remains unknown. We hypothesize that TP retains the ability to enhance clotting and reduce tPA-induced fibrinolysis at 14-day storage. FFP was thawed and stored at 4°C at the following intervals: 14, 10, 7, 5, 3, and 1-day prior to the experiment. Healthy volunteers underwent blood draws followed by 50% dilution with TP stored at previously mentioned intervals as well as FFP, normal saline (NS), albumin, and whole blood (WB) control. Samples underwent tPA-modified (75 ng/mL) thrombelastography (TEG) with analysis of R-time, angle, maximum amplitude (MA), and LY30. TEG properties did not change significantly over the thawed storage. 14-day TP retained the ability to inhibit tPA-induced hyperfibrinolysis (median LY30% 9.6%) similar to FFP (5.6%), WB (14.6%), and superior to albumin (59.3%) and NS (58.1%). 14-day TP also retained faster clot formation (median angle, 66.2°) and superior clot strength (MA, 61.5 mm) to albumin (34.8°, 21.6 mm) and NS (41.6°, 32.2 mm). TP plasma stored for 14 days retains clot-enhancing ability and resistance to clot degradation similar to FFP. A clinical trial is needed to validate these in vitro results. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    1997-10-01

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

  3. Soy lecithin interferes with mitochondrial function in frozen-thawed ram spermatozoa.

    Science.gov (United States)

    Del Valle, I; Gómez-Durán, A; Holt, W V; Muiño-Blanco, T; Cebrián-Pérez, J A

    2012-01-01

    Egg yolk and milk are the 2 major membrane cryoprotectants commonly used in freezing media for the long-term preservation of semen (alone or in combination with others). However, in recent years, there have been increasing arguments against the use of egg yolk or milk because of the risk of introducing diseases through the use of cryopreserved semen. In this study, we analyzed the protective effect of lecithin as an alternative to egg yolk for the cryopreservation of ram semen, using a range of functional markers for sperm viability, motility, apoptosis, and mitochondrial functionality analyses (mitochondrial inner membrane surface [MIMS], mitochondrial inner membrane potential [MIMP], and cell membrane potential) as methods of assessment in samples diluted in 3 different media: Tris-citrate-glucose as control and 2 media supplemented with soy lecithin or egg yolk. The results showed that lecithin was able to effectively protect certain sperm quality characteristics against freezing-induced damage. However, lecithin induced loss of mitochondrial membrane potential or mitochondrial loss that was not reflected by modifications in sperm motility in fresh semen. MIMS and MIMP values decreased in thawed lecithin-treated samples, concomitant with a lower (P lecithin may have affected the inner mitochondrial membrane in frozenthawed spermatozoa and confirmed that sublethal damages that seriously affect sperm functionality, not detected by classic sperm quality analyses, can be evidenced by changes in the inner mitochondrial membrane surface. These findings strengthen the relationship between mitochondrial membrane potential and motility and show that the mitochondrial alterations induced by the cryopreservation process could be specific targets for the improvement of semen cryopreservation protocols.

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

    Directory of Open Access Journals (Sweden)

    Katarzyna Kozłowicz

    2013-06-01

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

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

    Science.gov (United States)

    Arora, Rajeev; Palta, Jiwan P.

    1988-01-01

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

  6. Petroleum hydrocarbon remediation in frozen soil using a meat and bonemeal biochar plus fertilizer.

    Science.gov (United States)

    Karppinen, Erin M; Stewart, Katherine J; Farrell, Richard E; Siciliano, Steven D

    2017-04-01

    Petroleum hydrocarbon (PHC) degradation slows significantly during the winter which substantially increases the time it takes to remediate soil in Arctic landfarms. The aim of this laboratory trial was to assess the potential of a meat and bonemeal (MBM) biochar to stimulate PHC degradation in contaminated soil collected from Iqaluit, Canada. Over 90 days, 3% (w/w) MBM biochar significantly increased F3- (equivalent nC 16 -C 34 ) PHC degradation rate constants (k) in frozen soils when compared to the fertilizer (urea and monoammonium phosphate) control. Taking into consideration extensive variability within treatments and negative k values, this difference may not reflect significant remediation. Decreasing C 17 /Pr and C 18 /Ph ratios in the frozen soil suggest that this reduction is a result of microbial degradation rather than volatilization. Amendment type and application rate affected the immediate abiotic losses of F2 and F3-PHC in sterile soils, with the greatest losses occurring in compost-amended treatments in the first 24 h. In frozen soils, MBM biochar was found to increase liquid water content (θ liquid ) but not nutrient supply rates. Under frozen but not thawed conditions, genes for aromatic (C2,3O and nahAc) but not aliphatic (alkB) PHC degradation increased over time in both biochar-amended and control treatments but total viable PHC-degrading populations only increased in biochar-amended soils. Based on these results, it is possible that PHC degradation in biochar-amended soils is active and even enhanced under frozen conditions, but further investigation is required. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Improvement of post-thawed sperm quality and fertility of Arian rooster by oral administration of d-aspartic acid.

    Science.gov (United States)

    Ansari, Mahdi; Zhandi, Mahdi; Kohram, Hamid; Zaghari, Mojtaba; Sadeghi, Mostafa; Sharafi, Mohsen

    2017-04-01

    This study was conducted to investigate the effect of d-Aspartic acid (D-Asp) on post-thawed sperm quality, fertility and hatchability outcomes in male broiler breeders. Twenty 55-week-old roosters were selected and equally split into four groups (n = 5 rooster/group). Different daily D-Asp doses including 0 (D-0), 100 (D-100), 200 (D-200) or 300 (D-300) mg/kg BW were capsulated and individually administered for 12 weeks to roosters in each group. Semen samples were weekly collected from 7th to 12th week of experiment. Sperm quality from 7th to 11th week was evaluated in both fresh (total and forward motility and plasma membrane functionality) and post-thawed (total and forward motility, plasma membrane functionality, apoptosis status and mitochondrial activity) conditions. Also, collected semen samples on the 12th week were frozen and artificially inseminated to evaluate fertility and hatchability. The results from fresh condition showed that total and forward motility and plasma membrane functionality were significantly higher in D-200 compared to other groups. Also, interaction effect of time and treatment was not significant for all assessed parameters in fresh condition. In post-thawed condition, D-200 showed significantly higher total and forward motility, fertility and hatchability compared to other groups. The higher value for plasma membrane functionality and mitochondrial activity was observed in D-200 compared to D-0 and D300 groups. However, the percentage of live, early apoptotic and dead spermatozoa were not significantly affected by applied treatment in the current study. No significant difference for time and treat interaction effect was observed for all assessed parameters except forward motility. In conclusion, it seems that D-Asp administration could improve fresh and post-thawed sperm quality and post-thawed sperm fertility in male broiler breeders. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    Pietrasik, Z; Janz, J A M

    2009-03-01

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

  9. l-Cysteine improves antioxidant enzyme activity, post-thaw quality and fertility of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa.

    Science.gov (United States)

    Iqbal, S; Riaz, A; Andrabi, S M H; Shahzad, Q; Durrani, A Z; Ahmad, N

    2016-11-01

    The effects of l-cysteine in extender on antioxidant enzymes profile during cryopreservation, post-thaw quality parameters and in vivo fertility of Nili-Ravi buffalo bull spermatozoa were studied. Semen samples from 4 buffalo bulls were diluted in Tris-citric acid-based extender having different concentrations of l-cysteine (0.0, 0.5, 1.0, 2.0 and 3.0 mm) and frozen in 0.5-ml French straws. The antioxidative enzymes [catalase, super oxide dismutase and total glutathione (peroxidase and reductase)] were significantly higher (P l-cysteine as compared to other groups. Post-thaw total motility (%), progressive motility (%), rapid velocity (%), average path velocity (μm s -1 ), straight line velocity (μm s -1 ), curvilinear velocity (μm s -1 ), beat cross frequency (Hz), viable spermatozoa with intact plasmalemma (%), acrosome and DNA integrity (%) were higher with the addition of 2.0 mm l-cysteine as compared to other groups (P l-cysteine than in the control. In conclusion, the addition of 2.0 mm l-cysteine in extender improved the antioxidant enzymes profile, post-thaw quality and in vivo fertility of Nili-Ravi buffalo bull spermatozoa. © 2016 Blackwell Verlag GmbH.

  10. Fill and spill drives runoff connectivity over frozen ground

    Science.gov (United States)

    Coles, A. E.; McDonnell, J. J.

    2018-03-01

    Snowmelt-runoff processes on frozen ground are poorly understood at the hillslope scale. This is especially true for hillslopes on the northern Great Plains of North America where long periods of snow-covered frozen ground with very shallow slopes mask any spatial patterns and process controls on connectivity and hillslope runoff generation. This study examines a 4.66 ha (46,600 m2) hillslope on the northern Great Plains during the 2014 spring snowmelt season to explore hillslope runoff processes. Specifically, we explore the spatial patterns of runoff production source areas and examine how surface topography and patterns of snow cover, snow water equivalent, soil water content, and thawed layer depth - which we measured on a 10 m grid across our 46,600 m2 hillslope - affect melt water partitioning and runoff connectivity. A key question was whether or not the controls on connectivity are consistent with the fill and spill mechanism found in rain-dominated and unfrozen soil domains. The contrast between the slow infiltration rates into frozen soil and the relatively fast rates of snowmelt delivery to the soil surface resulted in water accumulation in small depressions under the snowpack. Consequently, infiltration was minimal over the 12 day melt period. Instead, nested filling of micro- and meso-depressions was followed by macro-scale, whole-slope spilling. This spilling occurred when large patches of ponded water exceeded the storage capacity behind downslope micro barriers in the surface topography, and flows from them coalesced to drive a rapid increase in runoff at the hillslope outlet. These observations of ponded water and flowpaths followed mapable fill and spill locations based on 2 m resolution digital topographic analysis. Interestingly, while surface topography is relatively unimportant under unfrozen conditions at our site because of low relief and high infiltrability, surface topography shows episodically critical importance for connectivity and

  11. Effect of gamma irrigation followed by moderate temperature abuse on the psychrotrophic and mesophilic microbial association of frozen chicken

    International Nuclear Information System (INIS)

    Osman, M.E.

    2001-01-01

    Commercially scalded, plucked and eviscerated broilers, aerobically packed into polyethylene bags were irradiated at - 18 deg C. A dose of 2.5 kilogay was effective in the elimination of salmonella spp. and staphylococcus aureus. Psychrotrophic and mesophilic aerobic colony counts of the non-irradiated chicken were of the order 10**4 and 10**5 gE-1, respectively. Radiation resulted in approximately two long cycles reduction in the count. The surviving microflora in frozen chicken after irradiation consisted mainly of lactobacillus spp. and micrococcus spp. which dominated the mesophilic flora. Moraxella spp. amounted to 70% of the total psychrotrophic flora. The microflora of frozen chicken after temperature abuse at 12 deg C were maily of the moraxella-acinetobacter group and to a lesser extent of Pseudomonas spp. The results demonstrated that the microflora after temperature abuse of irradiated chicken was similar to the microflora of non-irradiated chicken. This supports the view that irradiation of chicken does not entail a hazard, resulting from a shift in the micrflora in case frozen chicken are thawed and stored at increased temperatures

  12. Effects of chilled-then-frozen storage (up to 52weeks) on lamb M. longissimus lumborum quality and safety parameters.

    Science.gov (United States)

    Coombs, Cassius E O; Holman, Benjamin W B; Collins, Damian; Friend, Michael A; Hopkins, David L

    2017-12-01

    This study evaluated the effect of chilled followed by frozen storage on lamb quality and safety parameters. Experimental (n=360) M. longissimus lumborum (LL) were randomly sampled from the boning room of a commercial Australian abattoir, at 24 h post-mortem, and assigned to five chilled storage periods (0, 2, 4, 6 and 8 weeks) and six subsequent frozen storage periods (0, 4, 8, 12, 24 and 52 weeks). Upon completion of each storage treatment combination, corresponding LL were sub-sectioned and analysed for colour stability (0, 1, 2 and 3 days), shear force, fluid losses (purge, thaw and cooking losses), intramuscular fat content, sarcomere length, water activity and microbial load (lactic acid bacteria, Enterobacteriaceae sp., Brochothrix thermosphacta, Clostridium perfringens and Escherichia coli). LL stored chilled for 2-4 weeks prior to freezing presented superior results for shear force, display colour and low levels of spoilage microbes, correlating with good eating quality and safety following more than one year of frozen storage. Crown Copyright © 2017. Published by Elsevier Ltd. All rights reserved.

  13. The Role of Frozen Soil in Groundwater Discharge Predictions for Warming Alpine Watersheds

    Science.gov (United States)

    Evans, Sarah G.; Ge, Shemin; Voss, Clifford I.; Molotch, Noah P.

    2018-03-01

    Climate warming may alter the quantity and timing of groundwater discharge to streams in high alpine watersheds due to changes in the timing of the duration of seasonal freezing in the subsurface and snowmelt recharge. It is imperative to understand the effects of seasonal freezing and recharge on groundwater discharge to streams in warming alpine watersheds as streamflow originating from these watersheds is a critical water resource for downstream users. This study evaluates how climate warming may alter groundwater discharge due to changes in seasonally frozen ground and snowmelt using a 2-D coupled flow and heat transport model with freeze and thaw capabilities for variably saturated media. The model is applied to a representative snowmelt-dominated watershed in the Rocky Mountains of central Colorado, USA, with snowmelt time series reconstructed from a 12 year data set of hydrometeorological records and satellite-derived snow covered area. Model analyses indicate that the duration of seasonal freezing in the subsurface controls groundwater discharge to streams, while snowmelt timing controls groundwater discharge to hillslope faces. Climate warming causes changes to subsurface ice content and duration, rerouting groundwater flow paths but not altering the total magnitude of future groundwater discharge outside of the bounds of hydrologic parameter uncertainties. These findings suggest that frozen soil routines play an important role for predicting the future location of groundwater discharge in watersheds underlain by seasonally frozen ground.

  14. The role of frozen soil in groundwater discharge predictions for warming alpine watersheds

    Science.gov (United States)

    Evans, Sarah G.; Ge, Shemin; Voss, Clifford I.; Molotch, Noah P.

    2018-01-01

    Climate warming may alter the quantity and timing of groundwater discharge to streams in high alpine watersheds due to changes in the timing of the duration of seasonal freezing in the subsurface and snowmelt recharge. It is imperative to understand the effects of seasonal freezing and recharge on groundwater discharge to streams in warming alpine watersheds as streamflow originating from these watersheds is a critical water resource for downstream users. This study evaluates how climate warming may alter groundwater discharge due to changes in seasonally frozen ground and snowmelt using a 2‐D coupled flow and heat transport model with freeze and thaw capabilities for variably saturated media. The model is applied to a representative snowmelt‐dominated watershed in the Rocky Mountains of central Colorado, USA, with snowmelt time series reconstructed from a 12 year data set of hydrometeorological records and satellite‐derived snow covered area. Model analyses indicate that the duration of seasonal freezing in the subsurface controls groundwater discharge to streams, while snowmelt timing controls groundwater discharge to hillslope faces. Climate warming causes changes to subsurface ice content and duration, rerouting groundwater flow paths but not altering the total magnitude of future groundwater discharge outside of the bounds of hydrologic parameter uncertainties. These findings suggest that frozen soil routines play an important role for predicting the future location of groundwater discharge in watersheds underlain by seasonally frozen ground.

  15. What is the optimal means of preparing the endometrium in frozen-thawed embryo transfer cycles? A systematic review and meta-analysis.

    Science.gov (United States)

    Groenewoud, Eva R; Cantineau, Astrid E P; Kollen, Boudewijn J; Macklon, Nick S; Cohlen, Ben J

    2013-01-01

    BACKGROUND Frozen-thawed embryo transfer (FET) enables surplus embryos derived from IVF or IVF-ICSI treatment to be stored and transferred at a later date. In recent years the number of FET cycles performed has increased due to transferring fewer embryos per transfer and improved laboratory techniques. Currently, there is little consensus on the most effective method of endometrium preparation prior to FET. METHODS Using both MEDLINE and EMBASE database a systematic review and meta-analysis of literature was performed. Case-series, case-control studies and articles in languages other than English, Dutch or Spanish were excluded. Those studies comparing clinical and ongoing pregnancy rates as well as live birth rates in (i) true natural cycle FET (NC-FET) versus modified NC-FET, (ii) NC-FET versus artificial cycle FET (AC-FET), (iii) AC-FET versus artificial with GnRH agonist cycle FET and (iv) NC-FET versus artificial with GnRH agonist cycle FET were included. Forest plots were constructed and relative risks or odds ratios were calculated. RESULTS A total of 43 publications were selected for critical appraisal and 20 articles were included in the final review. For all comparisons, no differences in the clinical pregnancy rate, ongoing pregnancy rate or live birth rate could be found. Based on information provided in the articles no conclusions could be drawn with regard to cancellation rates. CONCLUSIONS Based on the current literature it is not possible to identify one method of endometrium preparation in FET as being more effective than another. Therefore, all of the current methods of endometrial preparation appear to be equally successful in terms of ongoing pregnancy rate. However, in some comparisons predominantly retrospective studies were included leaving these comparisons subject to selection and publication bias. Also patients' preferences as well as cost-efficiency were not addressed in any of the included studies. Therefore, prospective randomized

  16. Optimal Thawing of Cryopreserved Peripheral Blood Mononuclear Cells for Use in High-Throughput Human Immune Monitoring Studies

    Directory of Open Access Journals (Sweden)

    Ramu A. Subbramanian

    2012-07-01

    Full Text Available Cryopreserved peripheral blood mononuclear cells (PBMC constitute an important component of immune monitoring studies as they allow for efficient batch- testing of samples as well as for the validation and extension of original studies in the future. In this study, we systematically test the permutations of PBMC thawing practices commonly employed in the field and identify conditions that are high and low risk for the viability of PBMC and their functionality in downstream ELISPOT assays. The study identifies the addition of ice-chilled washing media to thawed cells at the same temperature as being a high risk practice, as it yields significantly lower viability and functionality of recovered PBMC when compared to warming the cryovials to 37 °C and adding a warm washing medium. We found thawed PBMC in cryovials could be kept up to 30 minutes at 37 °C in the presence of DMSO before commencement of washing, which surprisingly identifies exposure to DMSO as a low risk step during the thawing process. This latter finding is of considerable practical relevance since it permits batch-thawing of PBMC in high-throughput immune monitoring environments.

  17. Cryopreservation of human sperm: efficacy and use of a new nitrogen-free controlled rate freezer versus liquid nitrogen vapour freezing.

    Science.gov (United States)

    Creemers, E; Nijs, M; Vanheusden, E; Ombelet, W

    2011-12-01

    Preservation of spermatozoa is an important aspect of assisted reproductive medicine. The aim of this study was to investigate the efficacy and use of a recently developed liquid nitrogen and cryogen-free controlled rate freezer and this compared with the classical liquid nitrogen vapour freezing method for the cryopreservation of human spermatozoa. Ten patients entering the IVF programme donated semen samples for the study. Samples were analysed according to the World Health Organization guidelines. No significant difference in total sperm motility after freeze-thawing between the new technique and classical technique was demonstrated. The advantage of the new freezing technique is that it uses no liquid nitrogen during the freezing process, hence being safer to use and clean room compatible. Investment costs are higher for the apparatus but running costs are only 1% in comparison with classical liquid nitrogen freezing. In conclusion, post-thaw motility of samples frozen with the classical liquid nitrogen vapour technique was comparable with samples frozen with the new nitrogen-free freezing technique. This latter technique can thus be a very useful asset to the sperm cryopreservation laboratory. © 2011 Blackwell Verlag GmbH.

  18. Sensory Properties of Frozen Herring (Clupea harengus) from Different Catch Seasons and Locations

    DEFF Research Database (Denmark)

    Hyldig, Grethe; Jørgensen, Bo Munk; Undeland, Ingrid

    2012-01-01

    Freezing of herring (Clupea harengus) for human consumption is increasing in the Nordic herring industry, either onboard the fishing vessels or right after landing. The quality of frozen herring as a raw material does not only depend on the frozen storage conditions applied, but also...... on compositional features, something which in turn can vary with season and catching ground. To unravel the link between biological variations, basic muscle composition, and sensory properties of frozen herring, a unique herring raw material was caught by commercial fishing vessels at three locations: around...... the utilization of herring for frozen storage for human consumption....

  19. Effect of gamma irradiation followed by moderate temperature a buse on the psychrotrophic and mesophilic microbial association of frozen chicken

    International Nuclear Information System (INIS)

    Osman, M. E.

    2001-01-01

    commercially scalded, plucked and eviscerated broilers, aerobically packed into polyethylene bags were irradiated at -18 C. A dose of 2.5 kilo gay was effective in the elimination of salmonella spp and staphylococcus aureus. Psychrotrophic and mesophilic aerobic colony counts of the non-irradiated chicken were of the order 10 4 and 10 5 g -1 , respectively. Radiation resulted in approximately tow long cycles reduction in the count. The surviving microflora in frozen chicken after irradiation consisted mainly of lactobacillus spp. and micrococcus spp. which dominated the mesophilic flora. moraxella spp.amounted to 70% of the total Psychrotrophic flora. The microflora of frozen chicken after temperature abuse at 12 C were mainly of the moraxella-acinetobacter group and to a lesser extent of pseudomonas spp. The results demonstrated that the microflora after temperature abuse irradiated chicken was similar to the microflora of non-abuse irradiated chicken. This supports the view that irradiation of chicken dose not entail a hazard, resulting from a shift in the microflora in case frozen chicken are thawed and stored at increased temperatures.(Author)

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

    Science.gov (United States)

    Shang, Huai-Shuai; Yi, Ting-Hua

    2013-01-01

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

  1. Non-climatic factors and long-term, continental-scale changes in seasonally frozen ground

    Science.gov (United States)

    Shiklomanov, Nikolay I.

    2012-03-01

    Numerous studies indicate that the northern high latitudes are experiencing an unprecedented rate of environmental change, including an increase in air temperatures (e.g. Serreze and Francis 2006), reduction of snow cover (e.g. Brown and Robinson 2011), ecosystem transformations and land cover changes (e.g. Callaghan et al 2011). Many of the potential environmental impacts of global warming in the high latitudes are associated with frozen ground, which occupies about 55% of the unglaciated land area in the northern hemisphere and consists of both permafrost and seasonally frozen ground. Frozen soils have a tremendous impact on hydrologic, climatic and biologic systems. Periodic freezing and thawing promote changes in soil structure, affect the surface and subsurface water cycle, and regulate the availability of nutrients in the soil for plants and biota that depend upon them. Freezing and thawing cycles can affect the decomposition of organic substances in the soil and greenhouse gas exchange between the atmosphere and land surface. Significant efforts have been devoted to permafrost-related studies, including the establishment of standardized observations (e.g. Romanovsky et al 2010, Shiklomanov et al 2008), modeling (e.g. Riseborough et al 2008), and climate-related feedback processes (e.g. Schuur et al 2008). Despite its vast extent and importance, seasonally frozen ground has received much less attention. One of the major obstacles in assessing changes in seasonally frozen ground is the lack of long-term data. In general, observations on soil temperature and freeze propagation are available for a limited area and involve a relatively short time period, precluding assessment of long-term, climate-driven change. A few known exceptions include shallow soil temperature and freeze/thaw depth observations conducted as part of the standard hydrometeorological monitoring system in China (e.g. Zhao et al 2004) and the Soviet Union/Russia (e.g. Gilichinsky et al 2000

  2. Effects of Oxidized Glutathione, Bovine Serum Albumin, Cysteine and Lycopene on the Quality of Frozen-Thawed Ram Semen

    Directory of Open Access Journals (Sweden)

    O. Uysal

    2007-01-01

    Full Text Available Free radicals are known to be involved in lipid peroxidation as well as DNA and sperm membrane damages that may lead to decreased sperm motility or cell death. The balance between free radical production and their detoxification may be an important factor in sperm survival and function before, during and after cryopreservation. The aim of this study was to determine the effects of the addition of the antioxidants of oxidized glutathione (GSSG, bovine serum albumin (BSA, cysteine and lycopene to freezing media on the post-thawing sperm characteristics, including motility, morphology, acrosome integrity, viability and membrane integrity. A total number of 42 ejaculates were collected using the artifi cial vagina from 4 Akkaraman rams and 10 replicates of the ejaculates were diluted with a Tris-based extender containing additives and no additives as control. GSSG (5 mM, BSA (20 mg/ml, cysteine (10 mM and lycopene (800 μg showed more positive effects than other concentrations of the supplements and controls in protecting sperm characteristics after the freezing-thawing process (P < 0.001. Many aspects of sperm protection, e.g. sperm motility, viability and membrane stabilisation of the sperm cells during relative cryopreservation, are the key factors in determining the preservation of sperm function. The results of this study provide a new approach to the cryopreservation of sperm from rams and related breeds, and thereby contribute to the improvement of these breeds for the world sheep industry.

  3. Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples.

    Science.gov (United States)

    Roque, M; Valle, M; Marques, F; Sampaio, M; Geber, S

    2016-04-01

    Intracytoplasmic sperm injection (ICSI) may be performed with testicular frozen-thawed spermatozoa in patients with nonobstructive azoospermia (NOA). Sperm retrieval can be performed in advance of oocyte aspiration, as it may avoid the possibility of no recovery of spermatozoa on the day of oocyte pickup. There are few studies available in the literature concerning the use of frozen-thawed spermatozoa obtained from testicular sperm aspiration (TESA). To evaluate the effects and the outcomes of ICSI with frozen-thawed spermatozoa obtained by TESA, we performed a retrospective analysis of 43 ICSI cycles using frozen-thawed TESA. We obtained acceptable results with a fertilisation rate of 67.9%, an implantation rate (IR) of 17.1%, and clinical and ongoing pregnancy rates of 41.9% and 37.2% respectively. The results of this study suggest that performing ICSI using cryopreserved frozen-thawed testicular spermatozoa with TESA as a first option is a viable, safe, economic and effective method for patients with NOA. © 2015 Blackwell Verlag GmbH.

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

    Science.gov (United States)

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

    2004-08-01

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

  5. Construct Validity of Fresh Frozen Human Cadaver as a Training Model in Minimal Access Surgery

    Science.gov (United States)

    Macafee, David; Pranesh, Nagarajan; Horgan, Alan F.

    2012-01-01

    Background: The construct validity of fresh human cadaver as a training tool has not been established previously. The aims of this study were to investigate the construct validity of fresh frozen human cadaver as a method of training in minimal access surgery and determine if novices can be rapidly trained using this model to a safe level of performance. Methods: Junior surgical trainees, novices (cadavers. Expert laparoscopists (>100 laparoscopic procedures) performed 3 repetitions of identical tasks. Performances were scored using a validated, objective Global Operative Assessment of Laparoscopic Skills scale. Scores for 3 consecutive repetitions were compared between experts and novices to determine construct validity. Furthermore, to determine if the novices reached a safe level, a trimmed mean of the experts score was used to define a benchmark. Mann-Whitney U test was used for construct validity analysis and 1-sample t test to compare performances of the novice group with the benchmark safe score. Results: Ten novices and 2 experts were recruited. Four out of 5 tasks (nondominant to dominant hand transfer; simulated appendicectomy; intracorporeal and extracorporeal knot tying) showed construct validity. Novices’ scores became comparable to benchmark scores between the eighth and tenth repetition. Conclusion: Minimal access surgical training using fresh frozen human cadavers appears to have construct validity. The laparoscopic skills of novices can be accelerated through to a safe level within 8 to 10 repetitions. PMID:23318058

  6. Soil data from fire and permafrost-thaw chronosequences in upland Picea mariana stands near Hess Creek and Tok, interior Alaska

    Science.gov (United States)

    O'Donnell, Jonathan A.; Harden, Jennifer W.; Manies, Kristen L.; Jorgenson, M. Torre; Kanevskiy, Mikhail; Xu, Xiaomei

    2013-01-01

    Soils of the Northern Circumpolar Permafrost region harbor 1,672 petagrams (Pg) (1 Pg = 1,000,000,000 kilograms) of organic carbon (OC), nearly 50 percent of the global belowground OC pool (Tarnocai and others, 2009). Of that soil OC, nearly 88 percent is presently stored in perennially frozen ground. Recent climate warming at northern latitudes has resulted in warming and thawing of permafrost in many regions (Osterkamp, 2007), which might mobilize OC stocks from associated soil reservoirs via decomposition, leaching, or erosion. Warming also has increased the magnitude and severity of wildfires in the boreal region (Turetsky and others, 2011), which might exacerbate rates of permafrost degradation relative to warming alone. Given the size and vulnerability of the soil OC pool in permafrost soils, permafrost thaw will likely function as a strong positive feedback to the climate system (Koven and others, 2011; Schaefer and others, 2011). In this report, we report soil OC inventories from two upland fire chronosequences located near Hess Creek and Tok in Interior Alaska. We sampled organic and mineral soils in the top 2 meters (m) across a range of stand ages to evaluate the effects of wildfire and permafrost thaw on soil C dynamics. These data were used to parameterize a simple process-based fire-permafrost-carbon model, which is described in detail by O’Donnell and others (2011a, b). Model simulations examine long-term changes in soil OC storage in response to fire, permafrost thaw, and climate change. These data also have been used in other papers, including Harden and others (2012), which examines C recovery post-fire, and Johnson and others (2011), which synthesizes data within the Alaska Soil Carbon Database. Findings from these studies highlight the importance of climate and disturbance (wildfire, permafrost thaw) on soil C storage, and loss of soil C from high-latitude ecosystems.

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

    Science.gov (United States)

    Mayr, Stefan; Sperry, John S

    2010-03-01

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

  8. Freeze-Thaw Durability of Air-Entrained Concrete

    Directory of Open Access Journals (Sweden)

    Huai-Shuai Shang

    2013-01-01

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

  9. 21 CFR 135.110 - Ice cream and frozen custard.

    Science.gov (United States)

    2010-04-01

    ...) of this section, the name of the food is “frozen custard” or “french ice cream” or “french custard... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Ice cream and frozen custard. 135.110 Section 135.110 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...

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

    Science.gov (United States)

    Zheng, G.; Yang, D.

    2017-12-01

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

  11. Post-thaw non-cultured and post-thaw cultured equine cord blood mesenchymal stromal cells equally suppress lymphocyte proliferation in vitro.

    Directory of Open Access Journals (Sweden)

    Lynn B Williams

    Full Text Available Multipotent mesenchymal stromal cells (MSC are receiving increased attention for their non-progenitor immunomodulatory potential. Cryopreservation is commonly used for long-term storage of MSC. Post-thaw MSC proliferation is associated with a lag-phase in vitro. How this lag-phase affect MSC immunomodulatory properties is unknown. We hypothesized that in vitro there is no difference in lymphocyte suppression potential between quick-thawed cryopreserved equine cord blood (CB MSC immediately included in mixed lymphocyte reaction (MLR and same MSC allowed post-thaw culture time prior to inclusion in MLR. Cryopreserved CB-MSC from five unrelated foals were compared using two-way MLR. For each of the five unrelated MSC cultures, paired MLR assays of MSC allowed five days of post-thaw culture and MSC included in MLR assay immediately post-thawing were evaluated. We report no difference in the suppression of lymphocyte proliferation by CB-MSC that had undergone post-thaw culture and MSC not cultured post-thaw (p<0.0001. Also, there was no inter-donor variability between the lymphocyte suppressive properties of MSC harvested from the five different donors (p = 0.13. These findings suggest that cryopreserved CB-MSC may have clinical utility immediately upon thawing. One implication hereof is the possibility of using cryopreserved CB-MSC at third party locations without the need for cell culture equipment or competencies.

  12. Cryopreservation of boar semen. II: Effect of cooling rate and duration of freezing point plateau on boar semen frozen in mini- and maxi-straws and plastic bags.

    Science.gov (United States)

    Bwanga, C O; Einarsson, S; Rodriguez-Martinez, H

    1991-01-01

    The post-thaw motility and the acrosome integrity of semen from 4 boars frozen with a programmable freezing machine, in mini (0.25 ml) and maxi (5 ml) plastic straws and in 10 x 5 cm Teflon FEP-plastic bags (0.12 mm thick, 5 ml), were compared. The freezing of the semen was monitored by way of thermo-couples placed in the straws and the bags. Three freezing programmes were used, namely A: from +5 degrees C, at a rate of 3 degrees C/min, to -6 degrees C, held for 1 min at -6 degrees C, and followed by a cooling rate of 20 degrees C/min to -100 degrees C; B: a similar curve except that there was no holding time at -6 degrees C and that the cooling rate was 30 degrees C/min, and C: from +5 degrees C to -100 degrees C, with a cooling rate of 35 degrees C/min, followed by storage in liquid N2. Despite the freezing curve assayed, both the mini-straws and the bags depicted much shorter freezing point plateaus as compared to the maxi-straws. Post-thaw sperm motility as well as the amount of normal apical ridges were equally significantly higher when semen was frozen in mini-straws or in bags than in maxi-straws. Significant differences in these post-thawing parameters were obtained between the freezing curves used. The stepwise freezing procedure A appeared as the best alternative for boar semen, considering this in vitro evaluation.

  13. Baker’s yeasts for use in frozen-dough technology : sugar utilization in freeze tolerant Torulaspora delbrueckii strains and elucidation of cryo-resistance mechanisms

    OpenAIRE

    Pacheco, Andreia

    2009-01-01

    Tese de doutoramento em Ciências (ramo de conhecimento em Biologia) Bread is a central dietary item in most countries of the world. Currently, frozen dough technology is extensively used in the baking industry to supply oven-fresh bakery products to consumers and to improve labor conditions for bakers. Since freeze–thaw stress affects the viability and activity of yeast cells, one serious disadvantage of this technology is a significant reduction in leavening activity during fr...

  14. Factors affecting storage of Slovak native rabbit semen in the gene bank.

    Science.gov (United States)

    Kulíková, Barbora; Oravcová, Marta; Baláži, Andrej; Supuka, Peter; Chrenek, Peter

    2017-10-01

    In this study, fresh and frozen-thawed semen of Nitra and Zobor rabbit breeds were evaluated for potential inter-breed or inter-male differences in sperm quality traits. Individual male semen from four rabbits of each breed were diluted (v:v; 1:1) in a freezing medium composed of a commercial diluent, 16% of dimethyl sulphoxide (DMSO), 4% of Ficoll 70 and 2% of sucrose and frozen in liquid nitrogen vapours before being plunged into liquid nitrogen. Different motility traits, viability and plasma membrane integrity of fresh and frozen-thawed semen were evaluated in vitro using computer-assisted sperm analysis and flow cytometry. To evaluate the sperm fertilization ability, artificial insemination of fresh and frozen-thawed sperm was performed. Our results showed the effect of breed (P ≤ 0.05) on frozen-thawed sperm viability and plasma membrane integrity. Moreover, individual variability in semen quality among the rabbits was revealed (0.31 to 0.71 among quality traits). Our results thereby confirmed that the cryopreservation procedure could not ensure comparable sperm post-thaw survival for different breeds or males. Nevertheless, correlations between numbers of fresh total motile and progressively moving sperm and several quality parameters measured post thawing were revealed. Therefore, we suggest that the objective assessment of fresh rabbit sperm motility may be an effective indicator of frozen-thawed semen quality. Consequently, regular semen assessment is required in order to preserve good-quality insemination doses from native breeds.

  15. Effects of ionizing radiation on proteins in demineralized, lyophilized or frozen human bone

    Energy Technology Data Exchange (ETDEWEB)

    Antebi, Uri; Mathor, Monica B., E-mail: uri@usp.br, E-mail: mathor@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil); Guimaraes, Rodrigo P., E-mail: clinicaguimaraes@gmail.com [Santa Casa de Sao Paulo (FCM/SCSP), Sao Paulo, SP (Brazil). Faculdade de Ciencias Medicas

    2015-07-01

    The aim is the study of the application of ionizing radiation (gamma and electron) as sterilizing agents at doses of 15 kGy, 25 kGy and 50 kGy, the demineralized bone tissue frozen and freeze-dried for use in transplants. Five human femoral diaphysis of different donors demineralized bone tissues were preserved as lyophilized or frozen at - 80 deg C. The samples were divided into non-irradiated groups (control) and irradiated by gamma rays or electron beam. The bone proteins were extracted and used to determine the concentrations of total protein, BMP 2 and 7. It was observed a decrease in total protein concentrations, and BMP 2 and 7. The decrease in total protein concentrations, as compared to respective control groups was significant in the lyophilized and frozen samples irradiated at a dose of 50 kGy gamma radiation and beam electrons with greater than 30% reduction. The significant decrease in the levels of BMP 2 and 7 were also observed in higher doses and especially by electron beam. The reductions in the concentrations of total protein and osteoinductive proteins (BMP 2 and 7), were related to the radiation dose, i.e., increase with higher doses of ionizing radiation type and the type of preservation of the bones. The largest reductions in concentrations were observed in bone irradiated by electron beam and at a dose of 50 kGy. But this type of radiation and this high dose are not usual practice for the sterilization of bone tissue. Keywords: demineralized bone tissue, ionizing radiation, Tissue Bank, BMP 2, BMP 7, bone proteins. (author)

  16. Effects of ionizing radiation on proteins in demineralized, lyophilized or frozen human bone

    International Nuclear Information System (INIS)

    Antebi, Uri; Mathor, Monica B.; Guimaraes, Rodrigo P.

    2015-01-01

    The aim is the study of the application of ionizing radiation (gamma and electron) as sterilizing agents at doses of 15 kGy, 25 kGy and 50 kGy, the demineralized bone tissue frozen and freeze-dried for use in transplants. Five human femoral diaphysis of different donors demineralized bone tissues were preserved as lyophilized or frozen at - 80 deg C. The samples were divided into non-irradiated groups (control) and irradiated by gamma rays or electron beam. The bone proteins were extracted and used to determine the concentrations of total protein, BMP 2 and 7. It was observed a decrease in total protein concentrations, and BMP 2 and 7. The decrease in total protein concentrations, as compared to respective control groups was significant in the lyophilized and frozen samples irradiated at a dose of 50 kGy gamma radiation and beam electrons with greater than 30% reduction. The significant decrease in the levels of BMP 2 and 7 were also observed in higher doses and especially by electron beam. The reductions in the concentrations of total protein and osteoinductive proteins (BMP 2 and 7), were related to the radiation dose, i.e., increase with higher doses of ionizing radiation type and the type of preservation of the bones. The largest reductions in concentrations were observed in bone irradiated by electron beam and at a dose of 50 kGy. But this type of radiation and this high dose are not usual practice for the sterilization of bone tissue. Keywords: demineralized bone tissue, ionizing radiation, Tissue Bank, BMP 2, BMP 7, bone proteins. (author)

  17. Cryopreservation of ovarian tissue for a decade in Denmark: a view of the technique

    DEFF Research Database (Denmark)

    Rosendahl, Mikkel; Schmidt, Kirsten Louise Tryde; Ernst, Erik

    2011-01-01

    evaluation of mouse and human ovarian tissue after freezing with four different combinations of cryoprotectants. Viability was confirmed by transplantation of frozen-thawed human ovarian tissue (n = 49) to oophorectomized Nude mice. Viability after transport of fresh tissue 4-5 h prior to freezing had...... previously been validated. Overnight transport of fresh ovarian tissue prior to cryopreservation was evaluated when human ovarian tissue was kept on ice for 20 h and then cryopreserved. The thawed ovarian tissue was transplanted to an oophorectomized Nude mouse and histology confirmed viability. In Denmark...

  18. Spring thaw ionic pulses boost nutrient availability and microbial growth in entombed Antarctic Dry Valley cryoconite holes.

    Science.gov (United States)

    Telling, Jon; Anesio, Alexandre M; Tranter, Martyn; Fountain, Andrew G; Nylen, Thomas; Hawkings, Jon; Singh, Virendra B; Kaur, Preeti; Musilova, Michaela; Wadham, Jemma L

    2014-01-01

    The seasonal melting of ice entombed cryoconite holes on McMurdo Dry Valley glaciers provides oases for life in the harsh environmental conditions of the polar desert where surface air temperatures only occasionally exceed 0°C during the Austral summer. Here we follow temporal changes in cryoconite hole biogeochemistry on Canada Glacier from fully frozen conditions through the initial stages of spring thaw toward fully melted holes. The cryoconite holes had a mean isolation age from the glacial drainage system of 3.4 years, with an increasing mass of aqueous nutrients (dissolved organic carbon, total nitrogen, total phosphorus) with longer isolation age. During the initial melt there was a mean nine times enrichment in dissolved chloride relative to mean concentrations of the initial frozen holes indicative of an ionic pulse, with similar mean nine times enrichments in nitrite, ammonium, and dissolved organic matter. Nitrate was enriched twelve times and dissolved organic nitrogen six times, suggesting net nitrification, while lower enrichments for dissolved organic phosphorus and phosphate were consistent with net microbial phosphorus uptake. Rates of bacterial production were significantly elevated during the ionic pulse, likely due to the increased nutrient availability. There was no concomitant increase in photosynthesis rates, with a net depletion of dissolved inorganic carbon suggesting inorganic carbon limitation. Potential nitrogen fixation was detected in fully melted holes where it could be an important source of nitrogen to support microbial growth, but not during the ionic pulse where nitrogen availability was higher. This study demonstrates that ionic pulses significantly alter the timing and magnitude of microbial activity within entombed cryoconite holes, and adds credence to hypotheses that ionic enrichments during freeze-thaw can elevate rates of microbial growth and activity in other icy habitats, such as ice veins and subglacial regelation zones.

  19. Spring thaw ionic pulses boost nutrient availability and microbial growth in entombed Antarctic Dry Valley cryoconite holes

    Directory of Open Access Journals (Sweden)

    Jon eTelling

    2014-12-01

    Full Text Available The seasonal melting of ice entombed cryoconite holes on McMurdo Dry Valley glaciers provides oases for life in the harsh environmental conditions of the polar desert where surface air temperatures only occasionally exceed 0°C during the Austral summer. Here we follow temporal changes in cryoconite hole biogeochemistry on Canada Glacier from fully frozen conditions through the initial stages of spring thaw towards fully melted holes. The cryoconite holes had a mean isolation age from the glacial drainage system of 3.4 years, with an increasing mass of aqueous nutrients (dissolved organic carbon, total nitrogen, total phosphorus with longer isolation age. During the initial melt there was a mean nine times enrichment in dissolved chloride relative to mean concentrations of the initial frozen holes indicative of an ionic pulse, with similar mean nine times enrichments in nitrite, ammonium, and dissolved organic matter. Nitrate was enriched twelve times and dissolved organic nitrogen six times, suggesting net nitrification, while lower enrichments for dissolved organic phosphorus and phosphate were consistent with net microbial phosphorus uptake. Rates of bacterial production were significantly elevated during the ionic pulse, likely due to the increased nutrient availability. There was no concomitant increase in photosynthesis rates, with a net depletion of dissolved inorganic carbon suggesting inorganic carbon limitation. Potential nitrogen fixation was detected in fully melted holes where it could be an important source of nitrogen to support microbial growth, but not during the ionic pulse where nitrogen availability was higher. This study demonstrates that ionic pulses significantly alter the timing and magnitude of microbial activity within entombed cryoconite holes, and adds credence to hypotheses that ionic enrichments during freeze-thaw can elevate rates of microbial growth and activity in other icy habitats, such as ice veins and

  20. Modeling Aspect Controlled Formation of Seasonally Frozen Ground on Montane Hillslopes: a Case Study from Gordon Gulch, Colorado

    Science.gov (United States)

    Rush, M.; Rajaram, H.; Anderson, R. S.; Anderson, S. P.

    2017-12-01

    The Intergovernmental Panel on Climate Change (2013) warns that high-elevation ecosystems are extremely vulnerable to climate change due to short growing seasons, thin soils, sparse vegetation, melting glaciers, and thawing permafrost. Many permafrost-free regions experience seasonally frozen ground. The spatial distribution of frozen soil exerts a strong control on subsurface flow and transport processes by reducing soil permeability and impeding infiltration. Accordingly, evolution of the extent and duration of frozen ground may alter streamflow seasonality, groundwater flow paths, and subsurface storage, presenting a need for coupled thermal-hydrologic models to project hydrologic responses to climate warming in high-elevation regions. To be useful as predictive tools, such models should incorporate the heterogeneity of solar insolation, vegetation, and snowpack dynamics. We present a coupled thermal-hydrologic modeling study against the backdrop of field observations from Gordon Gulch, a seasonally snow-covered montane catchment in the Colorado Front Range in the Boulder Creek Critical Zone Observatory. The field site features two instrumented hillslopes with opposing aspects: the snowpack on the north-facing slope persists throughout much of the winter season, while the snowpack on the south-facing slope is highly ephemeral. We implemented a surface energy balance and snowpack accumulation and ablation model that is coupled to the subsurface flow and transport code PFLOTRAN-ICE to predict the hydrologic consequences of aspect-controlled frozen soil formation during water years 2013-2016. Preliminary model results demonstrate the occurrence of seasonally-frozen ground on the north-facing slope that directs snowmelt to the stream by way of shallow subsurface flow paths. The absence of persistently frozen ground on the south-facing slope allows deeper infiltration of snowmelt recharge. The differences in subsurface flow paths also suggest strong aspect

  1. Short communication: The effects of frozen storage on the survival of probiotic microorganisms found in traditionally and commercially manufactured kefir.

    Science.gov (United States)

    O'Brien, K V; Aryana, K J; Prinyawiwatkul, W; Ordonez, K M Carabante; Boeneke, C A

    2016-09-01

    Kefir is a fermented milk traditionally made from a unique starter culture, which consists of numerous bacteria and yeast species bound together in an exopolysaccharide matrix produced by certain lactic acid bacteria. Many health benefits are associated with traditionally produced kefir; however, bulging and leaking packaging, caused by secondary yeast fermentation during storage, has limited large-scale manufacture. Commercial kefir products have been designed to reduce these effects by using a pure starter culture consisting of a mixture of bacteria and yeast species that give a flavor similar to traditional kefir, but some health benefits may be lost in commercial production due to reduced microbial diversity and lack of beneficial exopolysaccharides. In this study, traditional and commercial kefir was frozen to study the effects of frozen storage on the viability of probiotic bacteria over time. Traditional kefir was prepared by inoculating 1L of pasteurized whole goat milk with approximately 30g of kefir grains. Commercial kefir was prepared by inoculating 1L of full-fat, pasteurized goat milk with a commercial kefir starter. The milk was allowed to ferment at room temperature (24-28°C) until pH 4.6 was reached. Samples were frozen (-8 to -14°C) immediately following the completion of fermentation and were thawed and plated for lactobacilli, lactococci, and yeasts on d 0, 7, 14, and 30 of frozen storage. Lactobacilli, lactococci, and yeasts were significantly reduced in number during frozen storage; however, the traditionally produced kefir was shown to have significantly higher counts of bacteria and yeast at each sampling. We concluded that frozen storage and the development of frozen kefir products could eliminate most packaging concerns associated with the large-scale manufacture of traditionally produced kefir, resulting in increased production and marketability of this healthful product. Copyright © 2016 American Dairy Science Association. Published

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

    Science.gov (United States)

    Zschocke, Anne; Schönborn, Maike; Eschenbach, Annette

    2010-05-01

    the frozen part of the column. However these results were not statistically significant they could proof results from Chuvilin et al. (2001). Who found similar results in sandy and clayey material and presumed the expulsion of petroleum hydrocarbons. Barnes et al. (2004) specified that the exclusion of petroleum hydrocarbons due to freezing is caused by displacement from the pore spaces due to expansion of the ice and the forming of crystalline ice structure. Further experimental approaches to investigate the effect of freezing and thawing of permafrost influenced soils on PAH migration will be discussed. References: Barnes, D. L.; Wolfe, S. M. & Filler, D. M. (2004): Equilibrium distribution of petroleum hydrocarbons in freezing ground, Polar Record, 40, 245-251. Börresen M.H., Barnes D.L., Rike A.G. (2007): Repeated freeze-thaw cycles and their effects on mineralization of hexadecane and phenanthrene in cold climate soils. Cold Regions Science and Technology 49, 215-225. Chuvilin, E.; Naletova, N.; Miklyaeva, E.; Kozlova, E. & Instanes, A. (2001): Factors affecting spreadability and transportation of oil in regions of frozen ground. Polar Record 37, 229-238. Curtosi, A.; Pelletier, E.; Vodopivez, C.L.; Mac Cormack, W.P. (2007): Polycyclic aromatic hydrocarbons in soil and surface marine sediment near Jubany Station (Antarctica). Role of permafrost as a low-permeability barrier. Science of The Total Environment 383, 1-3, 193-204. Eschenbach, A.; Wienberg, R.; Mahro, B. (2000): Formation, long-term stability and fate of non-extractable 14C-PAH-residues in contaminated soils. In: Wise, D.L.; Trantolo, D.J.; Cichon, E.J.; Inyang, H.J.; Stottmeister, U. (eds): Remediation Engineering of Contaminated Soils, 2nd Edition; Marcel-Dekker, New York, p. 427-446.

  3. Conservation science in a terrorist age: the impact of airport security screening on the viability and DNA integrity of frozen felid spermatozoa.

    Science.gov (United States)

    Gloor, Kayleen T; Winget, Doug; Swanson, William F

    2006-09-01

    In response to growing terrorism concerns, the Transportation Security Administration now requires that all checked baggage at U.S. airports be scanned through a cabinet x-ray system, which may increase risk of radiation damage to transported biologic samples and other sensitive genetic material. The objective of this study was to investigate the effect of these new airport security regulations on the viability and DNA integrity of frozen felid spermatozoa. Semen was collected from two domestic cats (Felis silvestris catus) and one fishing cat (Prionailurus viverrinus), cryopreserved in plastic freezing straws, and transferred into liquid nitrogen dry shippers for security screening. Treatment groups included frozen samples from each male scanned once or three times using a Transportation Security Administration-operated cabinet x-ray system, in addition to non-scanned samples (i.e., negative control) and samples previously scanned three times and exposed to five additional high-intensity x-ray bursts (i.e., positive control). Dosimeters placed in empty dry shippers were used to quantify radiation exposure. Following treatment, straws were thawed and spermatozoa analyzed for post-thaw motility (percentage motile and rate of progressive movement), acrosome status, and DNA integrity using single-cell gel electrophoresis (i.e., the comet assay). Dosimeter measurements determined that each airport screening procedure produced approximately 16 mrem of radiation exposure. Our results indicated that all levels of radiation exposure adversely affected (P 0.05) among treatment groups. Results also showed that the amount of double-stranded DNA damage was greater (P cat species scanned three times compared to samples scanned once or negative controls. Findings suggest that new airport security measures may cause radiation-induced damage to frozen spermatozoa and other valuable biologic samples transported on passenger aircraft and that alternative modes of sample

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

    Science.gov (United States)

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

    2014-01-01

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

  5. Large scale CMB anomalies from thawing cosmic strings

    Energy Technology Data Exchange (ETDEWEB)

    Ringeval, Christophe [Centre for Cosmology, Particle Physics and Phenomenology, Institute of Mathematics and Physics, Louvain University, 2 Chemin du Cyclotron, 1348 Louvain-la-Neuve (Belgium); Yamauchi, Daisuke; Yokoyama, Jun' ichi [Research Center for the Early Universe (RESCEU), Graduate School of Science, The University of Tokyo, Tokyo 113-0033 (Japan); Bouchet, François R., E-mail: christophe.ringeval@uclouvain.be, E-mail: yamauchi@resceu.s.u-tokyo.ac.jp, E-mail: yokoyama@resceu.s.u-tokyo.ac.jp, E-mail: bouchet@iap.fr [Institut d' Astrophysique de Paris, UMR 7095-CNRS, Université Pierre et Marie Curie, 98bis boulevard Arago, 75014 Paris (France)

    2016-02-01

    Cosmic strings formed during inflation are expected to be either diluted over super-Hubble distances, i.e., invisible today, or to have crossed our past light cone very recently. We discuss the latter situation in which a few strings imprint their signature in the Cosmic Microwave Background (CMB) Anisotropies after recombination. Being almost frozen in the Hubble flow, these strings are quasi static and evade almost all of the previously derived constraints on their tension while being able to source large scale anisotropies in the CMB sky. Using a local variance estimator on thousand of numerically simulated Nambu-Goto all sky maps, we compute the expected signal and show that it can mimic a dipole modulation at large angular scales while being negligible at small angles. Interestingly, such a scenario generically produces one cold spot from the thawing of a cosmic string loop. Mixed with anisotropies of inflationary origin, we find that a few strings of tension GU = O(1) × 10{sup −6} match the amplitude of the dipole modulation reported in the Planck satellite measurements and could be at the origin of other large scale anomalies.

  6. Carbon dioxide gas hydrates accumulation in freezing and frozen sediments

    Energy Technology Data Exchange (ETDEWEB)

    Chuvilin, E.; Guryeva, O. [Moscow State Univ., Moscow (Russian Federation). Dept. of Geology

    2008-07-01

    Carbon dioxide (CO{sub 2}) hydrates and methane hydrates can be formed, and exist under natural conditions. The permafrost area has been considered as an environment for the potential disposal of CO{sub 2}. The favorable factors for preserving CO{sub 2} in liquid and gas hydrate states in frozen sediments and under permafrost horizons are great thickness of frozen sediments; low permeability in comparison with thawed sediments; and favourable conditions for hydrates formation. Therefore, research on the formation and existence conditions of CO{sub 2} gas hydrates in permafrost and under permafrost sediments are of great importance for estimation of CO{sub 2} disposal conditions in permafrost, and for working out specific sequestration schemes. This paper presented the results of an experimental study on the process of carbon dioxide (CO{sub 2}) gas hydrates formation in the porous media of sediments under positive and negative temperatures. Sediment samples of various compositions including those selected in the permafrost area were used. The research was conducted in a special pressure chamber, which allowed to monitor pressure and temperature. The study used the monitoring results in order to make quantitative estimation of the kinetics of CO{sub 2} hydrates accumulation in the model sediments. Results were presented in terms of kinetics of CO{sub 2} hydrates accumulation in the porous media at positive and negative temperatures; kinetics of CO{sub 2} hydrates accumulation in various porous media; gas hydrate-former influence on kinetics of hydrates accumulation in frozen sediments; and influence of freezing on CO{sub 2} hydrates accumulation in porous media. It was concluded that hydrate accumulation took an active place in porous media not only under positive, but also under high negative temperatures, when the water was mainly in the form of ice in porous media. 27 refs., 3 tabs., 5 figs.

  7. Cost-effectiveness of single versus double embryo transfer in IVF in relation to female age.

    Science.gov (United States)

    van Loendersloot, Laura L; Moolenaar, Lobke M; van Wely, Madelon; Repping, Sjoerd; Bossuyt, Patrick M; Hompes, Peter G A; van der Veen, Fulco; Mol, Ben Willem J

    2017-07-01

    To evaluate the cost-effectiveness of single embryo transfer followed by an additional frozen-thawed single embryo transfer, if more embryos are available, as compared to double embryo transfer in relation to female age. We used a decision tree model to evaluate the costs from a healthcare provider perspective and the pregnancy rates of two embryo transfer policies: one fresh single embryo transfer followed by an additional frozen-thawed single embryo transfer, if more embryos are available (strategy I), and double embryo transfer (strategy II). The analysis was performed on an intention-to-treat basis. Sensitivity analyses were carried out to evaluate the robustness of our model and to identify which model parameters had the strongest impact on the results. SET followed by an additional frozen-thawed single embryo transfer if available was dominant, less costly and more effective, over DET in women under 32 years. In women aged 32 or older DET was more effective than SET followed by an additional frozen-thawed single embryo transfer if available but also more costly. SET followed by an additional frozen-thawed single embryo transfer should be the preferred strategy in women under 32 undergoing IVF. The choice for SET followed by an additional frozen-thawed single embryo transfer or DET in women aged 32 or older depends on individual patient preferences and on how much society is willing to pay for an extra child. There is a strong need for a randomized clinical trial comparing the cost and effects of SET followed by an additional frozen-thawed single embryo transfer and DET in the latter category of women. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Particle size distribution of wheat starch granules in relation to baking properties of frozen dough.

    Science.gov (United States)

    Tao, Han; Wang, Pei; Wu, Fengfeng; Jin, Zhengyu; Xu, Xueming

    2016-02-10

    The impact of freezing on the wheat starches with different particle size was studied using a range of characterization methods including X-ray diffraction, differential scanning calorimetry, the Rapid Visco Analyser and a reconstitution dough system. Wheat starches were fractionated into A- and B-type granules, and then subjected to freezing/thawing treatment for 3 cycles. The freezing treatment did not cause apparent damage on A-type granular surface but induced cracked structure on B-type granules. It facilitated materials such as amylose, proteins, and lipids leaching from starch granule and an increase in gelatinization temperatures, melting enthalpy, and pasting viscosities. A smaller bread specific volume was obtained from freezing-treated B-granules while the crumb firmness significantly increased (p>0.05). No marked differences were observed in the counterparts of A-granules after freezing treatment. It seemed that the B-type granules were more sensitive to the freezing/thawing treatment, thus facilitating structural transformations from dough to bread. Results indicated that the deterioration in frozen bread quality derived from starch could be minimized by increasing the A-granules content. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Constraining Parameter Uncertainty in Simulations of Water and Heat Dynamics in Seasonally Frozen Soil Using Limited Observed Data

    Directory of Open Access Journals (Sweden)

    Mousong Wu

    2016-02-01

    Full Text Available Water and energy processes in frozen soils are important for better understanding hydrologic processes and water resources management in cold regions. To investigate the water and energy balance in seasonally frozen soils, CoupModel combined with the generalized likelihood uncertainty estimation (GLUE method was used. Simulation work on water and heat processes in frozen soil in northern China during the 2012/2013 winter was conducted. Ensemble simulations through the Monte Carlo sampling method were generated for uncertainty analysis. Behavioral simulations were selected based on combinations of multiple model performance index criteria with respect to simulated soil water and temperature at four depths (5 cm, 15 cm, 25 cm, and 35 cm. Posterior distributions for parameters related to soil hydraulic, radiation processes, and heat transport indicated that uncertainties in both input and model structures could influence model performance in modeling water and heat processes in seasonally frozen soils. Seasonal courses in water and energy partitioning were obvious during the winter. Within the day-cycle, soil evaporation/condensation and energy distributions were well captured and clarified as an important phenomenon in the dynamics of the energy balance system. The combination of the CoupModel simulations with the uncertainty-based calibration method provides a way of understanding the seasonal courses of hydrology and energy processes in cold regions with limited data. Additional measurements may be used to further reduce the uncertainty of regulating factors during the different stages of freezing–thawing.

  10. Dissolved organic carbon biodegradability from thawing permafrost stimulated by sunlight rather than inorganic nitrogen

    Science.gov (United States)

    Liu, F.; Chen, L.; Zhang, B.; Wang, G.; Qin, S.; Yang, Y.

    2017-12-01

    Permafrost thaw could result in a large portion of frozen carbon being laterally transferred to aquatic ecosystems as dissolved organic carbon (DOC). During this delivery process, the size of biodegradable DOC (BDOC) determines the proportion of DOC mineralized by microorganisms and associated carbon loss to the atmosphere, which may further trigger positive carbon-climate feedback. Thermokarst is an abrupt permafrost thaw process that can enhance DOC export and also impact DOC processing through increased inorganic nitrogen (N) and sunlight exposure. However, it remains unclear how thermokarst-impacted BDOC responds to inorganic N addition and ultraviolet (UV) light irradiation. Here we explored the responses of DOC concentration, composition and its biodegradability to inorganic N and UV light in a typical thermokarst on the Tibetan Plateau, by combining field observation and laboratory incubation with spectra analyses (UV-visible absorption and three-dimensional fluorescence spectra) and parallel factor analyses. Our results showed that BDOC in thermokarst feature outflows was significantly higher than in reference water. Furthermore, inorganic N addition had no influence on thermokarst-impacted BDOC, whereas exposure to UV light significantly increased BDOC by as much as 2.3 times higher than the dark-control. Moreover, N addition and UV irradiation did not generate additive effects on BDOC. These results imply that sunlight rather than inorganic N can increase thermokarst-derived BDOC, potentially strengthening the positive permafrost carbon-climate feedback.

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

    KAUST Repository

    Jadoon, Khan

    2013-07-01

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

  12. Magnetic resonance of field-frozen and zero-field-frozen magnetic fluids

    International Nuclear Information System (INIS)

    Pereira, A.R.; Pelegrini, F.; Neto, K. Skeff; Buske, N.; Morais, P.C.

    2004-01-01

    In this study magnetic resonance was used to investigate magnetic fluid samples frozen under zero and non-zero (15 kG) external fields. The magnetite-based sample containing 2x10 17 particle/cm 3 was investigated from 100 to 400 K. Analysis of the temperature dependence of the resonance field revealed bigger magnetic structures in the frozen state than in the liquid phase. Also, differences in the mesoscopic organization in the frozen state may explain the data obtained from samples frozen under zero and non-zero fields

  13. Case report

    DEFF Research Database (Denmark)

    Ernst, Erik; Kjærsgaard, Mimi; Birkebæk, Niels

    2013-01-01

    To induce puberty by transplantation of frozen/thawed ovarian tissue collected prior to gonadotoxic treatment for a cancer.......To induce puberty by transplantation of frozen/thawed ovarian tissue collected prior to gonadotoxic treatment for a cancer....

  14. Microsatellite analysis of cryopreserved stallion semen stored on FTA(R paper : research communication

    Directory of Open Access Journals (Sweden)

    M.L. Schulman

    2002-07-01

    Full Text Available The aim of this study was to establish and validate a method to permit microsatellite analysis of DNA profiles obtained from frozen-thawed stallion sperm cells. This would provide reliable and accurate verification of the identification of a semen donor. Ejaculates from 5 pony stallions were collected, processed and frozen in 0.5 m plastic straws. Aliquots of 100 m of the frozen-thawed semen thus obtained were either placed directly, or diluted (1 : 10 ; 1 : 100 ; and 1 : 1000 and placed on slides of FTA(R paper. Similarly, blood samples obtained from each of the stallions were placed onto slides of FTA(R paper. A punch was removed from each sample after drying. Each sample was mixed with FTA(R purification reagent, Dithiothreitol and Proteinase K before incubation and processing. All samples were processed with a set of 13 microsatellite markers. Further analysis permitted a comparison of the DNA profiles of the frozen-thawed semen and the blood samples. A full profile of markers was obtained from the 1 : 10 and 1 : 100 dilutions of the frozen-thawed semen samples as well as from the blood samples. The DNA profiles from the frozen-thawed semen and blood samples obtained from the stallions matched in all cases.

  15. Frozen Mummies from Andean Mountaintop Shrines: Bioarchaeology and Ethnohistory of Inca Human Sacrifice.

    Science.gov (United States)

    Ceruti, Maria Constanza

    2015-01-01

    This study will focus on frozen mummies of sacrificial victims from mounts Llullaillaco (6739 m), Quehuar (6130 m), El Toro (6160 m), and the Aconcagua massif. These finds provide bioarchaeological data from mountaintop sites that has been recovered in scientifically controlled excavations in the northwest of Argentina, which was once part of the southern province of the Inca Empire. Numerous interdisciplinary studies have been conducted on the Llullaillaco mummies, including radiological evaluations by conventional X-rays and CT scans, which provided information about condition and pathology of the bones and internal organ, as well as dental studies oriented to the estimation of the ages of the three children at the time of death. Ancient DNA studies and hair analysis were also performed in cooperation with the George Mason University, the University of Bradford, and the Laboratory of Biological Anthropology at the University of Copenhagen. Ethnohistorical sources reveal interesting aspects related to the commemorative, expiatory, propitiatory, and dedicatory aspects of human sacrifice performed under Inca rule. The selection of the victims along with the procedures followed during the performance of the capacocha ceremony will be discussed, based on the bioarchaeological evidences from frozen mummies and the accounts recorded by the Spanish chroniclers.

  16. Frozen Mummies from Andean Mountaintop Shrines: Bioarchaeology and Ethnohistory of Inca Human Sacrifice

    Directory of Open Access Journals (Sweden)

    Maria Constanza Ceruti

    2015-01-01

    Full Text Available This study will focus on frozen mummies of sacrificial victims from mounts Llullaillaco (6739 m, Quehuar (6130 m, El Toro (6160 m, and the Aconcagua massif. These finds provide bioarchaeological data from mountaintop sites that has been recovered in scientifically controlled excavations in the northwest of Argentina, which was once part of the southern province of the Inca Empire. Numerous interdisciplinary studies have been conducted on the Llullaillaco mummies, including radiological evaluations by conventional X-rays and CT scans, which provided information about condition and pathology of the bones and internal organ, as well as dental studies oriented to the estimation of the ages of the three children at the time of death. Ancient DNA studies and hair analysis were also performed in cooperation with the George Mason University, the University of Bradford, and the Laboratory of Biological Anthropology at the University of Copenhagen. Ethnohistorical sources reveal interesting aspects related to the commemorative, expiatory, propitiatory, and dedicatory aspects of human sacrifice performed under Inca rule. The selection of the victims along with the procedures followed during the performance of the capacocha ceremony will be discussed, based on the bioarchaeological evidences from frozen mummies and the accounts recorded by the Spanish chroniclers.

  17. Cryopreservation of human insulin expressing cells macro-encapsulated in a durable therapeutic immunoisolating device theracyte.

    Science.gov (United States)

    Yakhnenko, Ilya; Wong, Wallace K; Katkov, Igor I; Itkin-Ansari, Pamela

    2012-01-01

    Encapsulating insulin producing cells (INPCs) in an immunoisolation device have been shown to cure diabetes in rodents without the need for immunosuppression. However, micro-encapsulation in semi-solid gels raises longevity and safety concerns for future use of stem cell derived INPCs. We have focused on a durable and retrievable macro-encapsulation (> 10(6) cells) device (TheraCyte). Cryopreservation (CP) of cells preloaded into the device is highly desirable but may require prolonged exposure to cryoprotectants during loading and post-thaw manipulations. Here, we are reporting survival and function of a human islet cell line frozen as single cells or as islet-like cell clusters. The non-clusterized cells exhibited high cryosurvival after prolonged pre-freeze or post-thaw exposure to 10 percent DMSO. However, both clusterization and especially loading INPCs into the device reduced viable yield even without CP. The survived cryopreserved macro-encapsulated INPCs remained fully functional suggesting that CP of macro-encapsulated cells is a promising tool for cell based therapies.

  18. Large for gestational age and macrosomia in singletons born after frozen/thawed embryo transfer (FET) in assisted reproductive technology (ART)

    DEFF Research Database (Denmark)

    Berntsen, Sine; Pinborg, Anja

    2018-01-01

    Increase in success rates with frozen embryo transfer (FET) and reduced risk of ovarian hyperstimulation syndrome (OHSS) and multiple pregnancies has lead to a steady rise in FET. Further, FET is associated with lower risk of prematurity and low birth weight in singletons, when compared with fres...

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

    Science.gov (United States)

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

    2017-06-01

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

  20. 21 CFR 101.95 - “Fresh,” “freshly frozen,” “fresh frozen,” “frozen fresh.”

    Science.gov (United States)

    2010-04-01

    ... freezing will not preclude use of the term “fresh frozen” to describe the food. “Quickly frozen” means... 21 Food and Drugs 2 2010-04-01 2010-04-01 false âFresh,â âfreshly frozen,â âfresh frozen,â âfrozen fresh.â 101.95 Section 101.95 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND...

  1. Effects of alginate on frozen-thawed boar spermatozoa quality, lipid peroxidation and antioxidant enzymes activities.

    Science.gov (United States)

    Hu, Jinghua; Geng, Guoxia; Li, Qingwang; Sun, Xiuzhu; Cao, Hualin; Liu, Yawei

    2014-06-30

    Although alginate was reported to play an important role as free radical scavengers in vitro and could be used as sources of natural antioxidants, there was no study about the cryoprotective effects of alginate on boar spermatozoa freezing. The objective of this research was to evaluate the effects of different concentrations of alginate added to the freezing extenders on boar spermatozoa motility, plasma membrane integrity, acrosomal integrity, mitochondrial activities, lipid peroxidation and antioxidative enzymes activities (SOD and GSH-Px) after thawing. Alginate was added to the TCG extender to yield six different final concentrations: 0, 0.2, 0.4, 0.6, 0.8, and 1.0mg/mL. The semen extender supplemented with various doses of alginate increased (Pboar spermatozoa acrosomal integrity at concentrations of 0.6, 0.8, 1.0mg/mL, compared with that of the control (Pextenders with the presence of alginate led to higher SOD and GSH-Px activities and lower MDA levels, in comparison to the control (Pboar spermatozoa motility, functional integrity and antioxidative capacity at appropriate concentrations. Therefore alginate could be employed as an effective cryoprotectant in boar spermatozoa cryopreservation. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Climate Change and Thawing Permafrost in Two Iñupiaq Communities of Alaska's Arctic: Observations, Implications, and Resilience

    Science.gov (United States)

    Woodward, A.; Kofinas, G.

    2013-12-01

    For thousands of years the Iñupiat of northern Alaska have relied on ecosystems underlain by permafrost for material and cultural resources. As permafrost thaws across the Arctic, these social-ecological systems are changing rapidly. Community-based research and extensive local knowledge of Iñupiaq villagers offer unique and valuable contributions to understanding permafrost change and its implications for humans. We partnered with two Iñupiaq communities in Alaska's Arctic to investigate current and potential effects of thawing permafrost on social-ecological systems. Anaktuvuk Pass is situated on thaw-stable consolidated gravel in the Brooks Range, while Selawik rests on ice-rich permafrost in Beringia lowland tundra. Using the transdisciplinary approach of resilience theory and mixed geophysical and ethnographic methods, we measured active layer thaw depths and documented local knowledge about climate and permafrost change. Thaw depths were greater overall in Selawik. Residents of both communities reported a variety of changes in surface features, hydrology, weather, flora, and fauna that they attribute to thawing permafrost and / or climate change. Overall, Selawik residents described more numerous and extreme examples of such changes, expressed higher degrees of certainty that change is occurring, and anticipated more significant and negative implications for their way of life than did residents of Anaktuvuk Pass. Of the two villages, Selawik faces greater and more immediate challenges to the resilience of its social-ecological system as permafrost thaws.

  3. Male gender explains increased birthweight in children born after transfer of blastocysts.

    Science.gov (United States)

    Kaartinen, N M; Kananen, K M; Rodriguez-Wallberg, K A; Tomás, C M; Huhtala, H Sa; Tinkanen, H I

    2015-10-01

    = 0.478). The study material was large enough to detect differences between birthweights as a whole, but a larger study group would confirm these new findings. To avoid selection bias, the next possible control candidate, fulfilling the selection criteria, was included for matching cases and controls. We have matched the cases and controls by gender and gestational week at birth, with an aim to reduce their impact as confounding factors. Our findings of a similar weight at birth of newborns after frozen-thawed blastocysts and frozen-thawed cleavage stage embryos, when matching for age and duration of pregnancy, are novel. The gender of the newborn has an impact on the birthweight, and the extended embryo culture increases the weight difference between the genders, which is a new finding as well. The study was funded by the Fertility Society of Finland. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm

    NARCIS (Netherlands)

    Guasti, P. N.; Monteiro, G. A.; Maziero, R. R.D.; Carmo, M. T.; Dell'Aqua, J. A.; Crespilho, A. M.; Rifai, E. A.; Papa, F. O.

    2017-01-01

    The aims of this study were to determinate whether pentoxifylline (PTX) increases the motion parameters of fresh and frozen-thawed equine epididymal spermatozoa, to evaluate the tyrosine phosphorylation of frozen-thawed epididymal sperm in the presence of PTX and to determine whether the

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

    Science.gov (United States)

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

    2014-01-01

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

  6. Thaw flow control for liquid heat transport systems

    Science.gov (United States)

    Kirpich, Aaron S.

    1989-01-01

    In a liquid metal heat transport system including a source of thaw heat for use in a space reactor power system, the thaw flow throttle or control comprises a fluid passage having forward and reverse flow sections and a partition having a plurality of bleed holes therein to enable fluid flow between the forward and reverse sections. The flow throttle is positioned in the system relatively far from the source of thaw heat.

  7. Thermokarst and thaw-related landscape dynamics -- an annotated bibliography with an emphasis on potential effects on habitat and wildlife

    Science.gov (United States)

    Jones, Benjamin M.; Amundson, Courtney L.; Koch, Joshua C.; Grosse, Guido

    2013-01-01

    , and heat and mass transfer processes. Typical Arctic thermokarst landforms include thermokarst lakes, collapsed pingos, sinkholes, and pits. Thermokarst is differentiated from thermal erosion, which refers to the erosion of the land surface by thermal and mechanical processes (Mackay, 1970; van Everdingen, 1998). Typical thermal erosional features include thermo-erosional gullies. Thermal abrasion is further differentiated from thermokarst and thermal erosion by association with the reworking of ocean, river, and lake bluffs (Are, 1988). Typical thermo-abrasion features include erosional niches at the base of bluffs. Thermal denudation is another distinct term that refers to the effect of incoming solar energy on the thaw of frozen slopes and permafrost bodies that subsequently become transported downhill by gravity (Shur and Osterkamp, 2007). Active layer detachment slides and thaw slumps are typical thermal denudation features. Shur and Osterkamp (2007) noted that these various transport processes may occur together with thermokarst or in instances that would not be considered thermokarst. This compilation of references regarding thermokarst and other thaw-related features is focused on the Arctic and the Subarctic. References were drawn from North America as well as Siberia. English-language literature mostly was targeted, with 167 references annotated in version 1.0; however, an additional 28 Russian-language references were taken from Shur and Osterkamp (2007) and are provided at the end of this document. This compilation may be missing key references and inevitably will become outdated soon after publication. We hope that this document, version 1.0, will serve as the foundation for a comprehensive compilation of thermokarst and permafrost-terrain stability references, and that it will be updated continually over the coming years.

  8. IN VITRO CULTURE OF FROZEN AND THAWED MOUSE OVA

    OpenAIRE

    KANAGAWA, Hiroshi

    1980-01-01

    Seventy mouse ova at the 4--8-cell stages were collected at room temperature from mice treated with pregnant mare's serum gonadotrophin and human chorionic gonadotrophin. Seventy ova were divided into 5 groups, and each group was placed in a 0.5 ml plastic straw with 0.2 ml of Brinster's medium. Then, the straws were immersed into an ice bath (0℃) for 5 minutes. Next, an equal volume of 2 M dimethyl sulfoxide was added to the sample straw. The medium with ova was then seeded at -6℃ and cooled...

  9. The Effects of Permafrost Thaw on Organic Matter Quality and Availability Along a Hill Slope in Northeastern Siberia

    Science.gov (United States)

    Connolly, C. T.; Spawn, S.; Ludwig, S.; Schade, J. D.; Natali, S.

    2014-12-01

    Climate warming and permafrost thaw in northeastern Siberia are expected to change the quantity and quality of organic matter (OM) transported through watersheds, releasing previously frozen carbon (C) to biologically available pool. Hill slopes have shown to influence the distribution of OM, resulting in a downhill accumulation of available C and nutrients relative to uphill. Here we examine how future permafrost thaw will change OM quality and availability along a hill slope in a larch-dominated watershed. We collected soils from the thawed organic and mineral layers, and 1m deep permafrost cores for dissolved organic C (DOC) and total dissolved N (TDN), C composition from measures of colored dissolved organic matter (CDOM), DOC lability from biodegradable DOC (BDOC) incubations, C and nutrient availability from extracellular-enzyme assays (EEA's), and microbial respiration from aerobic soil incubations. Here we show that organic soils (O), in comparison to mineral soils (M) and permafrost (P) are the most abundant source of C (avg O DOC: 51.6mg/L), exhibiting low molecular complexity (avg O SUVA254: 4.05) and high quality. Evidence suggests permafrost OM may be an equally abundant, and more labile source of C than mineral soils (highest P DOC: 16.1 mg/L, lowest P SUVA254: 6.32; median M DOC: 18.5 mg/L, median M SUVA254: 24.0). Furthermore, we demonstrate that there may be a positive relationship in the rate of C mineralization and distance downhill, showing 15-30% greater CO2 production/gC downhill relative to uphill. Evidence also supports a similar relationship in permafrost DOC content and molecular complexity, showing more DOC of a lower complexity further downhill. This indicates DOC transport may have been occurring through the active layer and downhill during ice-rich permafrost formation, and may supply a labile source of carbon to lowland areas and adjacent stream networks upon thaw.

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

    Science.gov (United States)

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

    2011-06-01

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

  11. Improved Performance of Unpaved Roads During Spring Thaw

    National Research Council Canada - National Science Library

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

    2005-01-01

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

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

    Science.gov (United States)

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

    2014-02-19

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

  13. Reviews and syntheses: Effects of permafrost thaw on Arctic aquatic ecosystems

    Science.gov (United States)

    Vonk, J.E.; Tank, S.E.; Bowden, W.B.; Laurion, I.; Vincent, W.F.; Alekseychik, P.; Amyot, Y.; Billet, M.F.; Canario, J.; Cory, R.M.; Deshpande, B.N.; Helbig, M.; Jammet, M.; Karlsson, J.; Larouche, J.; MacMillan, G.; Rautio, Milla; Walter Anthony, K.M.; Wickland, Kimberly P.

    2015-01-01

    The Arctic is a water-rich region, with freshwater systems covering about 16 % of the northern permafrost landscape. Permafrost thaw creates new freshwater ecosystems, while at the same time modifying the existing lakes, streams, and rivers that are impacted by thaw. Here, we describe the current state of knowledge regarding how permafrost thaw affects lentic (still) and lotic (moving) systems, exploring the effects of both thermokarst (thawing and collapse of ice-rich permafrost) and deepening of the active layer (the surface soil layer that thaws and refreezes each year). Within thermokarst, we further differentiate between the effects of thermokarst in lowland areas vs. that on hillslopes. For almost all of the processes that we explore, the effects of thaw vary regionally, and between lake and stream systems. Much of this regional variation is caused by differences in ground ice content, topography, soil type, and permafrost coverage. Together, these modifying factors determine (i) the degree to which permafrost thaw manifests as thermokarst, (ii) whether thermokarst leads to slumping or the formation of thermokarst lakes, and (iii) the manner in which constituent delivery to freshwater systems is altered by thaw. Differences in thaw-enabled constituent delivery can be considerable, with these modifying factors determining, for example, the balance between delivery of particulate vs. dissolved constituents, and inorganic vs. organic materials. Changes in the composition of thaw-impacted waters, coupled with changes in lake morphology, can strongly affect the physical and optical properties of thermokarst lakes. The ecology of thaw-impacted lakes and streams is also likely to change; these systems have unique microbiological communities, and show differences in respiration, primary production, and food web structure that are largely driven by differences in sediment, dissolved organic matter, and nutrient delivery. The degree to which thaw enables the delivery

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

    Science.gov (United States)

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

    2017-12-01

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

  15. Production of Molecular Iodine and Tri-iodide in the Frozen Solution of Iodide: Implication for Polar Atmosphere.

    Science.gov (United States)

    Kim, Kitae; Yabushita, Akihiro; Okumura, Masanori; Saiz-Lopez, Alfonso; Cuevas, Carlos A; Blaszczak-Boxe, Christopher S; Min, Dae Wi; Yoon, Ho-Il; Choi, Wonyong

    2016-02-02

    The chemistry of reactive halogens in the polar atmosphere plays important roles in ozone and mercury depletion events, oxidizing capacity, and dimethylsulfide oxidation to form cloud-condensation nuclei. Among halogen species, the sources and emission mechanisms of inorganic iodine compounds in the polar boundary layer remain unknown. Here, we demonstrate that the production of tri-iodide (I3(-)) via iodide oxidation, which is negligible in aqueous solution, is significantly accelerated in frozen solution, both in the presence and the absence of solar irradiation. Field experiments carried out in the Antarctic region (King George Island, 62°13'S, 58°47'W) also showed that the generation of tri-iodide via solar photo-oxidation was enhanced when iodide was added to various ice media. The emission of gaseous I2 from the irradiated frozen solution of iodide to the gas phase was detected by using cavity ring-down spectroscopy, which was observed both in the frozen state at 253 K and after thawing the ice at 298 K. The accelerated (photo-)oxidation of iodide and the subsequent formation of tri-iodide and I2 in ice appear to be related with the freeze concentration of iodide and dissolved O2 trapped in the ice crystal grain boundaries. We propose that an accelerated abiotic transformation of iodide to gaseous I2 in ice media provides a previously unrecognized formation pathway of active iodine species in the polar atmosphere.

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

    Science.gov (United States)

    Xiao, qianhui; Cao, zhiyuan; Li, qiang

    2018-03-01

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

  17. The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF.

    Science.gov (United States)

    Vergouw, Carlijn G; Kostelijk, E Hanna; Doejaaren, Els; Hompes, Peter G A; Lambalk, Cornelis B; Schats, Roel

    2012-09-01

    Does the type of medium used to culture fresh and frozen-thawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean birthweight adjusted for gestational age, gender and parity (z-scores) of singletons born after a fresh or frozen-thawed SET. Furthermore, we show that embryo freezing and thawing cycles may lead to a significantly higher mean birthweight. Animal studies have shown that culture media constituents are responsible for changes in birthweight of offspring. In human IVF, there is still little knowledge of the effect of medium type on birthweight. Until now, only a small number of commercially available culture media have been investigated (Vitrolife, Cook(®) Medical and IVF online medium). Our study adds new information: it has a larger population of singleton births compared with the previously published studies, it includes outcomes of other media types (HTF and Sage(®)), not previously analysed, and it includes data on frozen-thawed SETs. This study was a retrospective analysis of birthweights of singleton newborns after fresh (Day 3) or frozen-thawed (Day 5) SET cycles, using embryos cultured in either of two different types of commercially available culture media, between 2008 and 2011. Before January 2009, a single-step culture medium was used: human tubal fluid (HTF) with 4 mg/ml human serum albumin. From January 2009 onwards, a commercially available sequential medium was introduced: Sage(®), Quinn's advantage protein plus medium. Singletons born after a fresh SET (99 embryos cultured in HTF and 259 in Sage(®)) and singletons born after a frozen-thawed SET (32 embryos cultured in HTF only, 41 in HTF and Sage(®) and 86 in Sage(®) only) were analysed. Only patients using autologous gametes without the use of a gestational carrier were considered. Also excluded were (vanishing) twins, triplets

  18. Discovery of a novel methanogen prevalent in thawing permafrost.

    Science.gov (United States)

    Mondav, Rhiannon; Woodcroft, Ben J; Kim, Eun-Hae; McCalley, Carmody K; Hodgkins, Suzanne B; Crill, Patrick M; Chanton, Jeffrey; Hurst, Gregory B; VerBerkmoes, Nathan C; Saleska, Scott R; Hugenholtz, Philip; Rich, Virginia I; Tyson, Gene W

    2014-01-01

    Thawing permafrost promotes microbial degradation of cryo-sequestered and new carbon leading to the biogenic production of methane, creating a positive feedback to climate change. Here we determine microbial community composition along a permafrost thaw gradient in northern Sweden. Partially thawed sites were frequently dominated by a single archaeal phylotype, Candidatus 'Methanoflorens stordalenmirensis' gen. nov. sp. nov., belonging to the uncultivated lineage 'Rice Cluster II' (Candidatus 'Methanoflorentaceae' fam. nov.). Metagenomic sequencing led to the recovery of its near-complete genome, revealing the genes necessary for hydrogenotrophic methanogenesis. These genes are highly expressed and methane carbon isotope data are consistent with hydrogenotrophic production of methane in the partially thawed site. In addition to permafrost wetlands, 'Methanoflorentaceae' are widespread in high methane-flux habitats suggesting that this lineage is both prevalent and a major contributor to global methane production. In thawing permafrost, Candidatus 'M. stordalenmirensis' appears to be a key mediator of methane-based positive feedback to climate warming.

  19. Permafrost Thaw increases Emissions of Nitrous Oxide from Subarctic Peatlands

    Science.gov (United States)

    Voigt, C.; Marushchak, M. E.; Lamprecht, R. E.; Jackowicz-Korczynski, M.; Lindgren, A.; Mastepanov, M.; Christensen, T. R.; Granlund, L.; Tahvanainen, T.; Martikainen, P. J.; Biasi, C.

    2017-12-01

    Permafrost soils in the Arctic are thawing, exposing not only carbon but also large nitrogen stocks. The decomposition of this vast pool of long-term immobile C and N stocks results in the release of greenhouse gases to the atmosphere. Among these, carbon dioxide (CO2) and methane (CH4) are being studied extensively, and gaseous C release from thawing permafrost is known to be substantial. Most recent studies, however, show that Arctic soils may further be a relevant source of the strong greenhouse gas nitrous oxide (N2O). As N2O is almost 300 times more powerful in warming the climate than CO2 based on a 100-yr time horizon, the release of N2O from thawing permafrost could create a significant non-carbon permafrost-climate feedback. To study the effect of permafrost thaw on N2O fluxes, we collected peat mesocosms from a Subarctic permafrost peatland, and subjected these intact soil-plant systems to sequential thawing from the top of the active layer down to the upper permafrost layer. Measurements of N2O fluxes were coupled with detailed soil analyses and process studies. Since N2O fluxes are highly dependent on moisture conditions and vegetation cover, we applied two distinct moisture treatments (dry vs. wet) and simulated permafrost thaw in vegetated as well as in naturally bare mesocosms. Under dry conditions, permafrost thaw clearly increased N2O emissions. We observed the largest post-thaw emissions from bare peat surfaces, a typical landform in subarctic peatlands previously identified as hot spots for Arctic N2O emissions. There, permafrost thaw caused a five-fold increase in emissions (0.56 vs. 2.81 mg N2O m-2 d-1). While water-logged conditions suppressed N2O emissions, the presence of vegetation lowered, but did not prevent post-thaw N2O release. Based on these findings, we show that one fourth of the Arctic land area could be vulnerable for N2O emissions when permafrost thaws. Our results demonstrate that Arctic N2O emissions may be larger than

  20. Future active layer dynamics and carbon dioxide production from thawing permafrost layers in Northeast Greenland

    DEFF Research Database (Denmark)

    Hollesen, Jørgen; Elberling, Bo; Jansson, P.E.

    2011-01-01

    Thawing permafrost and the resulting mineralization of previously frozen organic carbon (C) is considered an important future feedback from terrestrial ecosystems to the atmosphere. Here, we use a dynamic process oriented permafrost model, the CoupModel, to link surface and subsurface temperatures....... The model is successfully adjusted and applied for the study area and shown to be able to simulate active layer dynamics. Subsequently, the model is used to predict the active layer thickness under future warming scenarios. The model predicts an increase of maximum active layer thickness from today 70 to 80......–105 cm as a result of a 2–6 °C warming. An additional increase in the maximum active layer thickness of a few centimetres may be expected due to heat production from decomposition of organic matter. Simulated future soil temperatures and water contents are subsequently used with measured basal soil...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1982-09-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Suzuki, E.; Nagashima, N.

    1982-01-01

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

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

    International Nuclear Information System (INIS)

    Suzuki, Eiichiro; Nagashima, Nobuya

    1982-01-01

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

  4. Human milk donation is an alternative to human milk bank.

    Science.gov (United States)

    Hsu, Ho-Torng; Fong, Tze-Vun; Hassan, Nurulhuda Mat; Wong, Hoi-Ling; Rai, Jasminder Kaur; Khalid, Zorina

    2012-04-01

    Human milk bank is a source of human milk supply in many neonatal intensive care units. However, there are some hospitals without this facility because of financial or religious impediments, such as the Muslim community. We introduced human milk donation as an alternative to human milk banking based on Islamic principles. The suitable donor is a healthy rooming-in mother whose expressed breastmilk is in excess of her baby's demand. The milk is used after 72 hours of freezing at -20°C. The donor must fulfill the criteria for selection of donors and be nonreactive to human immunodeficiency virus and syphilis. Once the recipient's family and the donor state their desire for the human milk donation, a meeting with both parties is made. Unpasteurized frozen-thawed donor's milk will be provided to the recipient after written consents are obtained from both parties. This study was carried out in the Duchess of Kent Hospital (Sandakan, Sabah, Malaysia) between January 2009 and December 2010. A total of 48 babies received donated breastmilk. Forty-two infants were from the special care nursery, and the remaining six were from the pediatric ward. Eighty-eight percent of the donors and 77% of the recipients were Muslims. Sixty percent of the infants who received donated human milk were premature. Two infants died because of the underlying nature of their disease. Human milk donation is an option for hospitals without a human milk bank or in the Muslim community.

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

    Science.gov (United States)

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

    2008-01-01

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

  6. Proteolysis, lipolysis, volatile compounds and sensory characteristics of Hispánico cheeses made using frozen curd from raw and pasteurized ewe milk.

    Science.gov (United States)

    Alonso, Rocío; Picon, Antonia; Gaya, Pilar; Nuñez, Manuel

    2013-02-01

    Hispánico cheese, manufactured from a mixture of cow and ewe milk, is representative of cheese varieties made using milk from more than one animal species in Mediterranean countries. The shortage of ewe milk production in autumn hinders the uniformity of Hispánico cheese composition throughout the year. To surmount this inconvenience of ewe milk seasonality, curds made in spring from raw and pasteurized ewe milk were stored frozen and used four months later for the manufacture of Hispánico cheese. Experimental cheeses were made by mixing fresh curd from pasteurized cow milk with thawed curd from raw or pasteurized ewe milk, and control cheese from a mixture of pasteurized cow and ewe milk in the same proportion. Characteristics of experimental and control cheeses throughout a 60-d ripening period were investigated. On the one hand, the experimental cheese containing frozen curd from raw ewe milk showed the highest counts of staphylococci, Gram-negative bacteria and coliforms, the highest levels of aminopeptidase and esterase activity, and the highest concentrations of free amino acids, free fatty acids, alcohols and esters. On the other, the experimental cheese containing frozen curd from pasteurized ewe milk had concentrations of free amino acids, free fatty acids and volatile compounds similar to those of control cheese, with the only exception being a higher level of ketones. Flavour intensity reached the highest scores in the experimental cheese containing frozen curd from raw ewe milk, followed by the experimental cheese containing frozen curd from pasteurized ewe milk. Flavour quality scores of both experimental cheeses were similar, and lower than those of control cheese.

  7. Autotransplantation of cryopreserved ovarian tissue in 12 women with chemotherapy-induced premature ovarian failure: the Danish experience

    DEFF Research Database (Denmark)

    Schmidt, Kirsten Tryde; Rosendahl, Mikkel; Ernst, Erik

    2011-01-01

    To describe a cohort of 12 Danish women who received autotransplantation of frozen-thawed cryopreserved ovarian tissue because of premature ovarian failure after cancer treatment.......To describe a cohort of 12 Danish women who received autotransplantation of frozen-thawed cryopreserved ovarian tissue because of premature ovarian failure after cancer treatment....

  8. Dimethylformamide as a cryoprotectant for canine semen diluted and frozen in ACP-106C.

    Science.gov (United States)

    Mota Filho, A C; Teles, C H A; Jucá, R P; Cardoso, J F S; Uchoa, D C; Campello, C C; Silva, A R; Silva, L D M

    2011-10-15

    The objective was to assess the effect of adding various concentrations of dimethylformamide on characteristics of canine semen diluted in powdered coconut water (ACP-106C; ACP Biotecnologia, Fortaleza, CE, Brazil) and frozen at -196°C. Fifteen ejaculates were collected by manual stimulation from five adult Boxer dogs. The sperm-rich fraction was diluted in ACP-106C (ACP Biotecnologia) containing 10% egg yolk and divided into four aliquots. The cryoprotectants used for each aliquot were 6% glycerol (control group; CG) or 2%, 4%, or 6% dimethylformamide (DF2, DF4, and DF6, respectively). After thawing, total motility (mean ± SEM) for CG (58.4 ± 24.6) was higher (P Biotecnologia) and 10% egg yolk as a diluent, yielded unsatisfactory in vitro results for freezing canine semen. Copyright © 2011 Elsevier Inc. All rights reserved.

  9. Seasonal thaw settlement at drained thermokarst lake basins, Arctic Alaska

    Science.gov (United States)

    Liu, Lin; Schaefer, Kevin; Gusmeroli, Alessio; Grosse, Guido; Jones, Benjamin M.; Zhang, Tinjun; Parsekian, Andrew; Zebker, Howard

    2014-01-01

    Drained thermokarst lake basins (DTLBs) are ubiquitous landforms on Arctic tundra lowland. Their dynamic states are seldom investigated, despite their importance for landscape stability, hydrology, nutrient fluxes, and carbon cycling. Here we report results based on high-resolution Interferometric Synthetic Aperture Radar (InSAR) measurements using space-borne data for a study area located on the North Slope of Alaska near Prudhoe Bay, where we focus on the seasonal thaw settlement within DTLBs, averaged between 2006 and 2010. The majority (14) of the 18 DTLBs in the study area exhibited seasonal thaw settlement of 3–4 cm. However, four of the DTLBs examined exceeded 4 cm of thaw settlement, with one basin experiencing up to 12 cm. Combining the InSAR observations with the in situ active layer thickness measured using ground penetrating radar and mechanical probing, we calculated thaw strain, an index of thaw settlement strength along a transect across the basin that underwent large thaw settlement. We found thaw strains of 10–35% at the basin center, suggesting the seasonal melting of ground ice as a possible mechanism for the large settlement. These findings emphasize the dynamic nature of permafrost landforms, demonstrate the capability of the InSAR technique to remotely monitor surface deformation of individual DTLBs, and illustrate the combination of ground-based and remote sensing observations to estimate thaw strain. Our study highlights the need for better description of the spatial heterogeneity of landscape-scale processes for regional assessment of surface dynamics on Arctic coastal lowlands.

  10. Use of fresh versus frozen or blast-frozen grapes for small-scale fermentation

    Directory of Open Access Journals (Sweden)

    Schmid F

    2011-10-01

    Full Text Available Frank Schmid, Vladimir Jiranek School of Agriculture, Food and Wine, The University of Adelaide; and Wine Innovation Cluster, The Waite Campus, Glen Osmond, South Australia, Australia Background: This paper firstly examines the validity of using laboratory-scale fermentations as a means of correlating winemaking outcomes with larger industrial scale fermentations. Secondly, conventional and blast-freezing of whole bunches were investigated for their relative suitability as methods of preservation as determined by the nature of the resulting wines. Methods: Red must fermentations were compared at the laboratory 80 kg scale, and the more industrially representative 500 kg pilot scale. Fermentation profiles and duration for both scales were found to be very similar. Whole bunches were either slow/conventionally frozen (−20°C, or quickly/blast-frozen (−25°C. Results: Wines made from frozen grapes compared well with the wine made from the fresh must. Color and chemical analyses of the wines revealed few differences. A duo-trio sensory evaluation showed that wine from blast-frozen grapes was more similar to the fresh wines than wines from conventional frozen grapes. Conclusion: The findings of this research suggest that whole-bunch blast-freezing of grapes is preferable to conventional freezing. Keywords: wine color, research winemaking, frozen grapes

  11. Long-Term Survival of Enteric Microorganisms in Frozen Wastewater

    Science.gov (United States)

    2002-10-01

    sewage.............31 Table 12. Percentage of cysts of Giardia muris recovered after one and three cycles of freezing and thawing...of Giardia muris and Giardia lamblia that had been subjected to one to three cycles of freezing at –16°C and thawing at 20°C. Examination Long-Term... Giardia muris recovered after one and three cycles of freezing and thawing. (From Erlandsen et al. 1990.)a Number of freeze–thaw cycles Initial

  12. Fresh frozen cadaver workshops for advanced vascular surgical training.

    Science.gov (United States)

    Jansen, Shirley; Cowie, Margaret; Linehan, John; Hamdorf, Jeffery M

    2014-11-01

    Reduction in working hours, streamlined training schemes and increasing use of endovascular techniques has meant a reduction in operative experience for newer vascular surgical trainees, especially those exposures which are not routinely performed such as thoracoabdominal, thoracotomy and retroperitoneal aortic, for example. This paper describes an Advanced Anatomy of Exposure course which was designed and convened at the Clinical Training & Evaluation Centre in Western Australia and uses fresh frozen cadavers. Feedback was obtained from the participants who attended over three courses by questionnaire. Feedback was strongly positive for the course meeting both its learning outcomes and personal learning objectives, and in addition, making a significant contribution to specialty skills. Most participants thought the fresh frozen cadaveric model significantly improved the learning objectives for training. The fresh frozen cadaver is an excellent teaching model highly representative of the living open surgical scenario where advanced trainees and newly qualified consultants can improve their operative confidence and consequently patient safety in vascular surgery. An efficient fresh frozen cadaver teaching programme can benefit many health professionals simultaneously maximizing the use of donated human tissue. © 2013 Royal Australasian College of Surgeons.

  13. Experimental research of joint influence of salinization and petroleum pollution on thermal capacity of frozen ground

    International Nuclear Information System (INIS)

    Motenko, R.G.

    2010-01-01

    Most gas and petroleum fields are located in permafrost zones, with some being on saline territories. Oil pollution of soils can occur in different ways and at different points such as during the extraction, processing and storage, and during transportation of oil and petroleum products. Oil producing pollution and salinization of soil often happen together. In this case, the sources of salts are the formation fluid, commercial waste water, the contents of the granaries and other geochemically active substances used for the extraction and desalting of crude oil. Joint salinization and contamination can also happen during the rupture of oil pipelines in saline areas. Although there is research available on the properties of saline soils and on properties of soils polluted with petroleum, there are no studies that describe changes of ground properties with joint pollution of salt and petroleum. This paper presented a study that examined the joint influence of salinization and petroleum pollution on the thermal characteristics of thawed and frozen grounds, particularly on thermal capacity. The paper outlined the purpose of the research and described the experimental methods. It was concluded that an increase of salinization increases the heat capacity of frozen soil because the amount of unfrozen water increases with increasing salinization. 10 refs., 5 figs.

  14. Experimental research of joint influence of salinization and petroleum pollution on thermal capacity of frozen ground

    Energy Technology Data Exchange (ETDEWEB)

    Motenko, R.G. [Moscow State Univ., Moscow (Russian Federation). Dept. of Geocryology; Grechishcheva, E.S. [Fundamentproek, Moscow (Russian Federation)

    2010-07-01

    Most gas and petroleum fields are located in permafrost zones, with some being on saline territories. Oil pollution of soils can occur in different ways and at different points such as during the extraction, processing and storage, and during transportation of oil and petroleum products. Oil producing pollution and salinization of soil often happen together. In this case, the sources of salts are the formation fluid, commercial waste water, the contents of the granaries and other geochemically active substances used for the extraction and desalting of crude oil. Joint salinization and contamination can also happen during the rupture of oil pipelines in saline areas. Although there is research available on the properties of saline soils and on properties of soils polluted with petroleum, there are no studies that describe changes of ground properties with joint pollution of salt and petroleum. This paper presented a study that examined the joint influence of salinization and petroleum pollution on the thermal characteristics of thawed and frozen grounds, particularly on thermal capacity. The paper outlined the purpose of the research and described the experimental methods. It was concluded that an increase of salinization increases the heat capacity of frozen soil because the amount of unfrozen water increases with increasing salinization. 10 refs., 5 figs.

  15. Net haemoglobin increase from reinfusion of refrigerated vs. frozen red blood cells after autologous blood transfusions

    DEFF Research Database (Denmark)

    Ashenden, M; Mørkeberg, Jakob Sehested

    2011-01-01

    BACKGROUND AND OBJECTIVES  Two main blood storage procedures can be used for storing red blood cells: refrigeration and freezing. Nevertheless, the efficiency of these procedures measured as the increase in haemoglobin after reinfusion compared with baseline has never been examined. The main...... objective was to examine which storage procedure yielded the largest increase in circulating haemoglobin after reinfusion compared to baseline. MATERIALS AND METHODS  Equal volumes of blood from 15 men were withdrawn and stored either frozen or refrigerated as packed red blood cells. Serial measures...... of circulating haemoglobin by carbon monoxide rebreathing provided an opportunity to monitor recovery from anaemia, as well as the net increase in circulating haemoglobin after transfusion. RESULTS  The post-thaw yield of haemoglobin in the bags was 72% after refrigerated storage compared with only 52% after...

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

    Directory of Open Access Journals (Sweden)

    Yanjun Zhang

    2014-02-01

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

  17. Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

    Science.gov (United States)

    Rungsiwiwut, Ruttachuk; Numchaisrika, Pranee; Ahnonkitpanit, Vichuda; Isarasena, Nipan; Virutamasen, Pramuan

    2012-01-01

    Abstract Human embryonic stem (hES) cells are considered to be a potential source for the therapy of human diseases, drug screening, and the study of developmental biology. In the present study, we successfully derived hES cell lines from blastocysts developed from frozen and fresh embryos. Seventeen- to eighteen-year-old frozen embryos were thawed, cultured to the blastocyst stage, and induced to form hES cells using human foreskin fibroblasts. The Chula2.hES cell line and the Chula4.hES and Chula5.hES cell lines were derived from blastocysts developed from frozen and fresh embryos, respectively. The cell lines expressed pluripotent markers, including alkaline phosphatase (AP), Oct3/4, stage-specific embryonic antigen (SSEA)-4, and tumor recognition antigen (TRA)-1-60 and TRA-1-81 as detected with immunocytochemistry. The real-time polymerase chain reaction (RT-PCR) results showed that the cell lines expressed pluripotent genes, including OCT3/4, SOX2, NANOG, UTF, LIN28, REX1, NODAL, and E-Cadherin. In addition, the telomerase activities of the cell lines were higher than in the fibroblast cells. Moreover, the cell lines differentiated into all three germ layers both in vitro and in vivo. The cell lines had distinct identities, as revealed with DNA fingerprinting, and maintained their normal karyotype after a long-term culture. This study is the first to report the successful derivation of hES cell lines in Thailand and that frozen embryos maintained their pluripotency similar to fresh embryos, as shown by the success of hES cell derivation, even after years of cryopreservation. Therefore, embryos from prolonged cryopreservation could be an alternative source for embryonic stem cell research. PMID:23514952

  18. Characterization of cryopreserved primary human corneal endothelial cells cultured in human serum-supplemented media

    Directory of Open Access Journals (Sweden)

    Lucas Monferrari Monteiro Vianna

    2016-02-01

    Full Text Available ABSTRACT Purpose: To compare cryopreserved human corneal endothelial cells (HCECs grown in human serum-supplemented media (HS-SM with cryopreserved HCECs grown in fetal bovine serum-supplemented media (FBS-SM. Methods: Three pairs of human corneas from donors aged 8, 28, and 31 years were obtained from the eye bank. From each pair, one cornea was used to start a HCEC culture using HS-SM; the other cornea was grown in FBS-SM. On reaching confluence, the six cell populations were frozen using 10% dimethyl sulfoxidecontaining medium. Thawed cells grown in HS-SM were compared with those grown in FBS-SM with respect to morphology, growth curves, immunohistochemistry, real time-reverse transcriptase polymerase chain reaction (RT-PCR for endothelial cell markers, and detachment time. Results: No difference in morphology was observed for cells grown in the two media before or after cryopreservation. By growth curves, cell counts after thawing were similar in both media, with a slight trend toward higher cell counts in FBS-SM. Cells grown in both the media demonstrated a similar expression of endothelial cell markers when assessed by immunohistochemistry, although HCEC marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM as assessed by RT-PCR. With FBS-SM, there was a tendency of longer detachment time and lower cell passages. Conclusions: HS-SM was similar to FBS-SM for cryopreservation of cultured HCECs as assessed by analysis of cell morphology, proliferation, and protein expression, although marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM. Detachment time was longer with FBS-SM and in lower passages.

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

    Science.gov (United States)

    Cheng, Yanqiu; Shang, Xiaoyu; Zhang, Youjia

    2017-07-01

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

  20. Fertility and flow cytometry study of frozen-thawed sperm in cryopreservation medium supplemented with soybean lecithin.

    Science.gov (United States)

    Masoudi, R; Sharafi, M; Zareh Shahneh, A; Towhidi, A; Kohram, H; Esmaeili, V; Shahverdi, A; Davachi, N Dadashpour

    2016-08-01

    Semen cryopreservation can provide genetic resources for a large number of females from a small number of superior males. Optimization of cryopreservation media to achieve the highest quality of post-thaw semen is crucial. Soybean lecithin has evaluated as a plant-based cryoprotectant for substitution of egg yolk in ram semen extender. Flow cytometric and fertility assessments were applied following cryopreservation procedure in two experimental groups (SL group: extender containing 1% w/v soybean lecithin and EY group: extender containing 20% v/v egg yolk). The higher percentage of live sperm and the lower percentage of dead sperm were obtained in SL (47.66 ± 1.38, 52.33 ± 1.69, respectively) extender compared to EY (41.16 ± 1.38, 58.83 ± 1.69). For motion characteristics, plasma membrane integrity, acrosome integrity and mitochondria activity, no significant difference was observed between SL and EY extenders. In artificial insemination experiment, there was no significant difference in pregnancy rate, lambing rate and twining rate between SL and EY extenders. It can be concluded that SL extender can be an efficient alternative extender to preserve ram sperm during cryopreservation procedure without adverse effects. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Safe and efficient method for cryopreservation of human induced pluripotent stem cell-derived neural stem and progenitor cells by a programmed freezer with a magnetic field.

    Science.gov (United States)

    Nishiyama, Yuichiro; Iwanami, Akio; Kohyama, Jun; Itakura, Go; Kawabata, Soya; Sugai, Keiko; Nishimura, Soraya; Kashiwagi, Rei; Yasutake, Kaori; Isoda, Miho; Matsumoto, Morio; Nakamura, Masaya; Okano, Hideyuki

    2016-06-01

    Stem cells represent a potential cellular resource in the development of regenerative medicine approaches to the treatment of pathologies in which specific cells are degenerated or damaged by genetic abnormality, disease, or injury. Securing sufficient supplies of cells suited to the demands of cell transplantation, however, remains challenging, and the establishment of safe and efficient cell banking procedures is an important goal. Cryopreservation allows the storage of stem cells for prolonged time periods while maintaining them in adequate condition for use in clinical settings. Conventional cryopreservation systems include slow-freezing and vitrification both have advantages and disadvantages in terms of cell viability and/or scalability. In the present study, we developed an advanced slow-freezing technique using a programmed freezer with a magnetic field called Cells Alive System (CAS) and examined its effectiveness on human induced pluripotent stem cell-derived neural stem/progenitor cells (hiPSC-NS/PCs). This system significantly increased cell viability after thawing and had less impact on cellular proliferation and differentiation. We further found that frozen-thawed hiPSC-NS/PCs were comparable with non-frozen ones at the transcriptome level. Given these findings, we suggest that the CAS is useful for hiPSC-NS/PCs banking for clinical uses involving neural disorders and may open new avenues for future regenerative medicine. Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

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

    Science.gov (United States)

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

    2011-11-01

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

  3. Effective melanin depigmentation of human and murine ocular tissues: an improved method for paraffin and frozen sections.

    Directory of Open Access Journals (Sweden)

    Caroline Manicam

    Full Text Available PURPOSE: The removal of excessive melanin pigments that obscure ocular tissue morphology is important to address scientific questions and for differential diagnosis of ocular tumours based on histology. Thus, the goal of the present study was to establish an effective and fast melanin bleaching method for paraffin and frozen mouse and human ocular tissues. METHODS: Paraffin-embedded and frozen ocular specimens from mice and human donors were subjected to bleaching employing two methods. The first employed potassium permanganate (KMnO4 with oxalic acid, and the second 10% hydrogen peroxide (H2O2. To determine optimal bleaching conditions, depigmentation was carried out at various incubation times. The effect of diluents used for 10% H2O2 was assessed using phosphate-buffered saline (PBS, and deionized water. Three different slide types and two fixatives, which were ice-cold acetone with 80% methanol, and 4% paraformaldehyde (PFA were used to determine the optimal conditions for better tissue adherence during bleaching. All tissues were stained in hematoxylin and eosin for histological evaluation. RESULTS: Optimal bleaching was achieved using warm 10% H2O2 diluted in PBS at 65°C for 120 minutes. Chromium-gelatin-coated slides prevented tissue detachment. Adherence of cryosections was also improved with post-fixation using 4% PFA and overnight air-drying at RT after cryosectioning. Tissue morphology was preserved under these conditions. Conversely, tissues bleached in KMnO4/oxalic acid demonstrated poor depigmentation with extensive tissue damage. CONCLUSIONS: Warm dilute H2O2 at 65°C for 120 minutes rapidly and effectively bleached both cryo- and paraffin sections of murine and human ocular tissues.

  4. Effective melanin depigmentation of human and murine ocular tissues: an improved method for paraffin and frozen sections.

    Science.gov (United States)

    Manicam, Caroline; Pitz, Susanne; Brochhausen, Christoph; Grus, Franz H; Pfeiffer, Norbert; Gericke, Adrian

    2014-01-01

    The removal of excessive melanin pigments that obscure ocular tissue morphology is important to address scientific questions and for differential diagnosis of ocular tumours based on histology. Thus, the goal of the present study was to establish an effective and fast melanin bleaching method for paraffin and frozen mouse and human ocular tissues. Paraffin-embedded and frozen ocular specimens from mice and human donors were subjected to bleaching employing two methods. The first employed potassium permanganate (KMnO4) with oxalic acid, and the second 10% hydrogen peroxide (H2O2). To determine optimal bleaching conditions, depigmentation was carried out at various incubation times. The effect of diluents used for 10% H2O2 was assessed using phosphate-buffered saline (PBS), and deionized water. Three different slide types and two fixatives, which were ice-cold acetone with 80% methanol, and 4% paraformaldehyde (PFA) were used to determine the optimal conditions for better tissue adherence during bleaching. All tissues were stained in hematoxylin and eosin for histological evaluation. Optimal bleaching was achieved using warm 10% H2O2 diluted in PBS at 65°C for 120 minutes. Chromium-gelatin-coated slides prevented tissue detachment. Adherence of cryosections was also improved with post-fixation using 4% PFA and overnight air-drying at RT after cryosectioning. Tissue morphology was preserved under these conditions. Conversely, tissues bleached in KMnO4/oxalic acid demonstrated poor depigmentation with extensive tissue damage. Warm dilute H2O2 at 65°C for 120 minutes rapidly and effectively bleached both cryo- and paraffin sections of murine and human ocular tissues.

  5. Poorly known microbial taxa dominate the microbiome of permafrost thaw ponds.

    Science.gov (United States)

    Wurzbacher, Christian; Nilsson, R Henrik; Rautio, Milla; Peura, Sari

    2017-08-01

    In the transition zone of the shifting permafrost border, thaw ponds emerge as hotspots of microbial activity, processing the ancient carbon freed from the permafrost. We analyzed the microbial succession across a gradient of recently emerged to older ponds using three molecular markers: one universal, one bacterial and one fungal. Age was a major modulator of the microbial community of the thaw ponds. Surprisingly, typical freshwater taxa comprised only a small fraction of the community. Instead, thaw ponds of all age classes were dominated by enigmatic bacterial and fungal phyla. Our results on permafrost thaw ponds lead to a revised perception of the thaw pond ecosystem and their microbes, with potential implications for carbon and nutrient cycling in this increasingly important class of freshwaters.

  6. Monitoring recharge in areas of seasonally frozen ground in the Columbia Plateau and Snake River Plain, Idaho, Oregon, and Washington

    Science.gov (United States)

    Mastin, Mark; Josberger, Edward

    2014-01-01

    Seasonally frozen ground occurs over approximately one‑third of the contiguous United States, causing increased winter runoff. Frozen ground generally rejects potential groundwater recharge. Nearly all recharge from precipitation in semi-arid regions such as the Columbia Plateau and the Snake River Plain in Idaho, Oregon, and Washington, occurs between October and March, when precipitation is most abundant and seasonally frozen ground is commonplace. The temporal and spatial distribution of frozen ground is expected to change as the climate warms. It is difficult to predict the distribution of frozen ground, however, because of the complex ways ground freezes and the way that snow cover thermally insulates soil, by keeping it frozen longer than it would be if it was not snow covered or, more commonly, keeping the soil thawed during freezing weather. A combination of satellite remote sensing and ground truth measurements was used with some success to investigate seasonally frozen ground at local to regional scales. The frozen-ground/snow-cover algorithm from the National Snow and Ice Data Center, combined with the 21-year record of passive microwave observations from the Special Sensor Microwave Imager onboard a Defense Meteorological Satellite Program satellite, provided a unique time series of frozen ground. Periodically repeating this methodology and analyzing for trends can be a means to monitor possible regional changes to frozen ground that could occur with a warming climate. The Precipitation-Runoff Modeling System watershed model constructed for the upper Crab Creek Basin in the Columbia Plateau and Reynolds Creek basin on the eastern side of the Snake River Plain simulated recharge and frozen ground for several future climate scenarios. Frozen ground was simulated with the Continuous Frozen Ground Index, which is influenced by air temperature and snow cover. Model simulation results showed a decreased occurrence of frozen ground that coincided with

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

    Science.gov (United States)

    2013-10-01

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

  8. 21 CFR 146.137 - Frozen orange juice.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen orange juice. 146.137 Section 146.137 Food... Beverages § 146.137 Frozen orange juice. (a) Frozen orange juice is orange juice as defined in § 146.135, except that it is frozen. (b) The name of the food is “Frozen orange juice”. Such name may be preceded on...

  9. Toxigenic penicillia spoiling frozen chicken nuggets

    DEFF Research Database (Denmark)

    Wigmann, Evelin Francine; Saccomori, Fernanda; Bernardi, Angelica Olivier

    2015-01-01

    Frozen chicken nuggets are classified as pre-prepared frozen meals. These products are convenient to consumers as they are easy to prepare and allow for long storage by freezing. Over the years, spoilage of frozen food products caused by fungi has been a continual problem for the food industry...... of filamentous fungi involved in the spoilage of frozen chicken nuggets and determine their ability to produce mycotoxins under laboratorial conditions. A total of 7 samples of frozen chicken nuggets were analyzed by dilution plating in potato dextrose agar (PDA). These products had been returned by customers...

  10. The Eccentric Behavior of Nearly Frozen Orbits

    Science.gov (United States)

    Sweetser, Theodore H.; Vincent, Mark A.

    2013-01-01

    Frozen orbits are orbits which have only short-period changes in their mean eccentricity and argument of periapse, so that they basically keep a fixed orientation within their plane of motion. Nearly frozen orbits are those whose eccentricity and argument of periapse have values close to those of a frozen orbit. We call them "nearly" frozen because their eccentricity vector (a vector whose polar coordinates are eccentricity and argument of periapse) will stay within a bounded distance from the frozen orbit eccentricity vector, circulating around it over time. For highly inclined orbits around the Earth, this distance is effectively constant over time. Furthermore, frozen orbit eccentricity values are low enough that these orbits are essentially eccentric (i.e., off center) circles, so that nearly frozen orbits around Earth are bounded above and below by frozen orbits.

  11. Toxicity potential of residual ethylene oxide on fresh or frozen embryos maintained in plastic straws.

    Science.gov (United States)

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

    1988-01-01

    The toxic effects of residual ethylene oxide (EtO), a frequently used gas-sterilant, on embryos either frozen for long-term purposes or stored acutely for 30 min to 9 hr in a fresh condition in 0.25-ml straw containers were evaluated. In Experiment 1, fresh embryos were frozen (using conventional technology) in straws previously aerated for 0 hr to 8 mo after EtO sterilization. With the exception of the 8-mo group in which survival and quality ratings were depressed, embryo viability was not affected significantly by short-term prefreeze and post-thaw exposure to EtO residues. Experiment 2 was conducted to analyze the influence of prefreeze exposure to EtO residues on embryo development in vitro for embryos temporarily stored in previously sterilized straws aerated for different intervals. Compared to non-EtO-sterilized control straws, the development, quality, and viability of embryos exposed to EtO-treated straws were compromised (p less than 0.05) as the aeration interval decreased and the exposure interval increased. The combined results of both experiments indicate that EtO-treated straws can be used to cryopreserve gametes efficiently, but only if the aeration interval is greater than or equal to 72 hr and the prefreeze duration of exposure is less than or equal to 3 hr.

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

    Science.gov (United States)

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

    2009-11-01

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

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

    Science.gov (United States)

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

    1989-01-01

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

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

    Science.gov (United States)

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

    2015-04-01

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

  15. The effect of quercetin on fertility of frozen-thawed ram epididymal ...

    African Journals Online (AJOL)

    mz

    2017-03-13

    Mar 13, 2017 ... The mean number of zygote, morula, and blastocyst stage embryos ... rabbits, bulls and human beings (Silva et al., 2013; Martínez-Páramo et al., 2012; Akalin et ..... pathway in buffalo (Bubalus bubalis) embryonic stem cells.

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

    Directory of Open Access Journals (Sweden)

    S Ali

    2016-03-01

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

  17. [Chapter 7. The frozen embryo in the light of a jurist : beyond qualification].

    Science.gov (United States)

    Neirinck, Claire

    2018-03-07

    The legal qualification of the embryo does not pose any particular difficulties : this human being is a bodily thing of human nature, devoid of legal personality.However the freezing affects its humanity : it is no more than a thing made in laboratory, out of time. Stored in liquid nitrogen, it does not die, so storage must be ended.As long as they respond to a specific parental project, the one for which they were made and kept, the frozen embryos are identified by this given project.They are unique and not interchangeable. On the other hand, without a parental project, frozen embryos that can be accommodated by any infertile couple or those given to research, become interchangeable gender things.Although human beings, they are treated as the elements and products of the human body, human things.

  18. Frozen shoulder and risk of cancer

    DEFF Research Database (Denmark)

    Pedersen, Alma B; Horváth-Puhó, Erzsébet; Ehrenstein, Vera

    2017-01-01

    BACKGROUND: Frozen shoulder might be a complication or a presenting symptom of cancer. We examined the risk of a cancer diagnosis after an incident diagnosis of frozen shoulder. METHODS: We used prospectively collected data from Danish registries to identify patients with frozen shoulder during 1...

  19. Cumulative live birth rates after one ART cycle including all subsequent frozen-thaw cycles in 1050 women

    DEFF Research Database (Denmark)

    Toftager, M; Bogstad, J; Løssl, K

    2017-01-01

    birth increases. There are no previous randomized controlled trials (RCTs) comparing CLBRs in GnRH-antagonist versus GnRH-agonist protocols. Previous studies on CLBR are either retrospective cohort studies including multiple fresh cycles or RCTs comparing single embryo transfer (SET) with double embryo...... transfer (DET). STUDY DESIGN, SIZE, DURATION: CLBR was a secondary outcome in a Phase IV, dual-center, open-label, RCT including 1050 women allocated to a short GnRH-antagonist or a long GnRH-agonist protocol in a 1:1 ratio over a 5-year period using a web-based concealed randomization code. The minimum...... follow-up time from the first IVF cycle was 2 years. The aim was to compare CLBR between the two groups following utilization of all fresh and frozen embryos from the first ART cycle. PARTICIPANTS/MATERIALS, SETTING, METHODS: All women referred for their first ART cycle at two public fertility clinics...

  20. The water regime of the long-seasonally-frozen peat soils of the Northern Trans-Ural

    Directory of Open Access Journals (Sweden)

    Motorin Alexandr

    2017-01-01

    Full Text Available The results of many-years research of the water regime of the long-seasonally-frozen peat soils of the Northern Trans-Uralare described. It is shown that the fluctuation of groundwater in the Tarmanskoe swamp before drying is characterized by a sharp increase in levels during the spring snowmelt, then – a minimal level in summer, increase in levels in autumn, and winter minimum. The intensity of the decline in groundwater depends on precipitation (r=0.83 and evaporation. The change of groundwater level in winter is significantly affected by the progress of freezing of the upper layer and thaw. After drying at atmospheric-alluvial type of water supply of the swamp, the groundwater level during the vegetation period is determined mainly by the amount of rainfall (r=0.76. The deepest groundwater table (1.97 m on average during the growing season set in 2012, when 56.7% of the average annual norm of precipitation fell. On the dried potter’s drainage (To=24 m, H=1.5 m land there is no increase of the groundwater in the autumn. The lowest possible (2.5 m and more level of the groundwater table reaches in the beginning of snowmelt in late March - early April. The magnitude of the spring rise is 1-1.5 m and depends on winter moisture (r=0.65, the snow cover and the intensity of the melting of solid precipitation. The humidity of the root layer (0.3 m medium peat soil with a deep groundwater table (1.3 to 1.9 m under perennial grasses is in the range of 0.5-0.6 LMC (the least moisture capacity. In the formation of the first mowing of perennial grasses, soil moisture is in the optimum range (0.6-0.85 LMC; in the high-draught years for a full second mowing has a deficit. On the boundary of the thawed and frozen layers, soil moisture is always at the upper limit of the optimum (0.85-0.95 LMC. During the winter period, the moisture reserves in the upper layer 0.5 m up to 20% due to the underlying horizons.

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

    Directory of Open Access Journals (Sweden)

    MR Oliveira

    2015-06-01

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

  2. Frozen in Time? Microbial strategies for survival and carbon metabolism over geologic time in a Pleistocene permafrost chronosequence

    Science.gov (United States)

    Mackelprang, R.; Douglas, T. A.; Waldrop, M. P.

    2014-12-01

    Permafrost soils have received tremendous interest due to their importance as a global carbon store with the potential to be thawed over the coming centuries. Instead of being 'frozen in time,' permafrost contains active microbes. Most metagenomic studies have focused on Holocene aged permafrost. Here, we target Pleistocene aged ice and carbon rich permafrost (Yedoma), which can differ in carbon content and stage of decay. Our aim was to understand how microbes in the permafrost transform organic matter over geologic time and to identify physiological and biochemical adaptations that enable long-term survival. We used next-generation sequencing to characterize microbial communities along a permafrost age gradient. Samples were collected from the Cold Regions Research and Engineering Laboratory (CRREL) Permafrost Tunnel near Fox, AK, which penetrates a hillside providing access to permafrost ranging in age from 12 to 40 kyr. DNA was extracted directly from unthawed samples. 16S rRNA amplicon (16S) and shotgun metagenome sequencing revealed significant age-driven differences. First, microbial diversity declines with permafrost age, likely due to long-term exposure to environmental stresses and a reduction in metabolic resources. Second, we observed taxonomic differences among ages, with an increasing abundance of Firmicutes (endospore-formers) in older samples, suggesting that dormancy is a common survival strategy in older permafrost. Ordination of 16S and metagenome data revealed age-based clustering. Genes differing significantly between age categories included those involved in lipopolysaccharide assembly, cold-response, and carbon processing. These data point to the physiological adaptations to long-term frozen conditions and to the metabolic processes utilized in ancient permafrost. In fact, a gene common in older samples is involved in cadaverine production, which could potentially explain the putrefied smell of Pleistocene aged permafrost. Coupled with soil

  3. Thaw pond dynamics and carbon emissions in a Siberian lowland tundra landscape

    Science.gov (United States)

    van Huissteden, Ko; Heijmans, Monique; Dean, Josh; Meisel, Ove; Goovaerts, Arne; Parmentier, Frans-Jan; Schaepman-Strub, Gabriela; Belelli Marchesini, Luca; Kononov, Alexander; Maximov, Trofim; Borges, Alberto; Bouillon, Steven

    2017-04-01

    Arctic climate change induces drastic changes in permafrost surface wetness. As a result of thawing ground ice bodies, ice wedge troughs and thaw ponds are formed. Alternatively, ongoing thaw may enhance drainage as a result of increased interconnectedness of thawing ice wedge troughs, as inferred from a model study (Liljedahl et al., 2016, Nature Geoscience, DOI: 10.1038/NGEO2674). However, a recent review highlighted the limited predictability of consequences of thawing permafrost on hydrology (Walvoord and Kurylyk, 2016, Vadose Zone J., DOI:10.2136/vzj2016.01.0010). Overall, these changes in tundra wetness modify carbon cycling in the Arctic and in particular the emissions of CO2 and CH4 to the atmosphere, providing a possibly positive feedback on climate change. Here we present the results of a combined remote sensing, geomorphological, vegetation and biogechemical study of thaw ponds in Arctic Siberian tundra, at Kytalyk research station near Chokurdakh, Indigirka lowlands. The station is located in an area dominated by Pleistocene ice-rich 'yedoma' sediments and drained thaw lake bottoms of Holocene age. The development of three types of ponds in the Kytalyk area (polygon centre ponds, ice wedge troughs and thaw ponds) has been traced with high resolution satellite and aerial imagery. The remote sensing data show net areal expansion of all types of ponds. Next to formation of new ponds, local vegetation change from dry vegetation types to wet, sedge-dominated vegetation is common. Thawing ice wedges and thaw ponds show an increase in area and number at most studied locations. In particular the area of polygon centre ponds increased strongly between 2010 and 2015, but this is highly sensitive to antecedent precipitation conditions. Despite a nearly 60% increase of the area of thawing ice wedge troughs, there is no evidence of decreasing water surfaces by increasing drainage through connected ice wedge troughs. The number of thaw ponds shows an equilibrium

  4. Using Distributed Fiber Optic Sensing to Monitor Large Scale Permafrost Transitions: Preliminary Results from a Controlled Thaw Experiment

    Science.gov (United States)

    Ajo Franklin, J. B.; Wagner, A. M.; Lindsey, N.; Dou, S.; Bjella, K.; Daley, T. M.; Freifeld, B. M.; Ulrich, C.; Gelvin, A.; Morales, A.; James, S. R.; Saari, S.; Ekblaw, I.; Wood, T.; Robertson, M.; Martin, E. R.

    2016-12-01

    In a warming world, permafrost landscapes are being rapidly transformed by thaw, yielding surface subsidence and groundwater flow alteration. The same transformations pose a threat to arctic infrastructure and can induce catastrophic failure of the roads, runways, and pipelines on which human habitation depends. Scalable solutions to monitoring permafrost thaw dynamics are required to both quantitatively understand biogeochemical feedbacks as well as to protect built infrastructure from damage. Unfortunately, permafrost alteration happens over the time scale of climate change, years to decades, a decided challenge for testing new sensing technologies in a limited context. One solution is to engineer systems capable of rapidly thawing large permafrost units to allow short duration experiments targeting next-generation sensing approaches. We present preliminary results from a large-scale controlled permafrost thaw experiment designed to evaluate the utility of different geophysical approaches for tracking the cause, precursors, and early phases of thaw subsidence. We focus on the use of distributed fiber optic sensing for this challenge and deployed distributed temperature (DTS), strain (DSS), and acoustic (DAS) sensing systems in a 2D array to detect thaw signatures. A 10 x 15 x 1 m section of subsurface permafrost was heated using an array of 120 downhole heaters (60 w) at an experimental site near Fairbanks, AK. Ambient noise analysis of DAS datasets collected at the plot, coupled to shear wave inversion, was utilized to evaluate changes in shear wave velocity associated with heating and thaw. These measurements were confirmed by seismic surveys collected using a semi-permanent orbital seismic source activated on a daily basis. Fiber optic measurements were complemented by subsurface thermistor and thermocouple arrays, timelapse total station surveys, LIDAR, secondary seismic measurements (geophone and broadband recordings), timelapse ERT, borehole NMR, soil

  5. 21 CFR 158.170 - Frozen peas.

    Science.gov (United States)

    2010-04-01

    ... CONSUMPTION FROZEN VEGETABLES Requirements for Specific Standardized Frozen Vegetables § 158.170 Frozen peas... two or more, of the following safe and suitable optional ingredients: (i) Natural and artificial... “early June” shall precede or follow the name in the case of smooth-skin or substantially smooth-skin...

  6. Cryopreservation of canine ovarian and testicular fibroblasts.

    Science.gov (United States)

    Yu, Il-Jeoung; Leibo, S P; Songsasen, Nucharin; Dresser, Betsy L; Kim, In-Shik

    2009-01-01

    To derive a practical procedure to store canine somatic cells, fibroblasts isolated from testicular or ovarian tissues were cryopreserved in 1.2 M ethylene glycol or in 1.2 M dimethylsulfoxide prepared in Dulbecco's Modified Eagle Medium as cryoprotectants, and were frozen either in plastic straws or vials. Thawed cells were cultured for 24 hr at 38.5 degree C in a humidified atmosphere of 5 percent CO2 95 percent air, and then their membrane integrity was assayed with a double fluorescent stain, Fertilight. In addition, frozen-thawed fibroblasts were cultured for 4 days, and then their functional survival was measured after staining small colonies with trypan blue. After freezing and thawing, membrane integrity of testicular fibroblasts was 55-70 percent and functional survival ranged from 20-40 percent. With frozen-thawed ovarian cells, the average membrane integrity was 55-75 percent and the average functional survival was 35-40 percent. When frozen in ethylene glycol, functional survival of ovarian fibroblasts was significantly higher than that of testicular cells (P less than 0.05). These methods should prove useful to preserve cells collected from canids in the wild.

  7. Antimüllerian hormone as a predictor of live birth following assisted reproduction: an analysis of 85,062 fresh and thawed cycles from the Society for Assisted Reproductive Technology Clinic Outcome Reporting System database for 2012-2013.

    Science.gov (United States)

    Tal, Reshef; Seifer, David B; Wantman, Ethan; Baker, Valerie; Tal, Oded

    2018-02-01

    To determine if serum antimüllerian hormone (AMH) is associated with and/or predictive of live birth assisted reproductive technology (ART) outcomes. Retrospective analysis of Society for Assisted Reproductive Technology Clinic Outcome Reporting System database from 2012 to 2013. Not applicable. A total of 69,336 (81.8%) fresh and 15,458 (18.2%) frozen embryo transfer (FET) cycles with AMH values. None. Live birth. A total of 85,062 out of 259,499 (32.7%) fresh and frozen-thawed autologous non-preimplantation genetic diagnosis cycles had AMH reported for cycles over this 2-year period. Of those, 70,565 cycles which had embryo transfers were included in the analysis. Serum AMH was significantly associated with live birth outcome per transfer in both fresh and FET cycles. Multiple logistic regression demonstrated that AMH is an independent predictor of live birth in fresh transfer cycles and FET cycles when controlling for age, body mass index, race, day of transfer, and number of embryos transferred. Receiver operating characteristic (ROC) curves demonstrated that the areas under the curve (AUC) for AMH as predictors of live birth in fresh cycles and thawed cycles were 0.631 and 0.540, respectively, suggesting that AMH alone is a weak independent predictor of live birth after ART. Similar ROC curves were obtained also when elective single-embryo transfer (eSET) cycles were analyzed separately in either fresh (AUC 0.655) or FET (AUC 0.533) cycles, although AMH was not found to be an independent predictor in eSET cycles. AMH is a poor independent predictor of live birth outcome in either fresh or frozen embryo transfer for both eSET and non-SET transfers. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. 21 CFR 160.190 - Frozen egg yolks.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen egg yolks. 160.190 Section 160.190 Food and... CONSUMPTION EGGS AND EGG PRODUCTS Requirements for Specific Standardized Eggs and Egg Products § 160.190 Frozen egg yolks. (a) Frozen egg yolks, frozen yolks is the food prepared by freezing egg yolks that...

  9. 21 CFR 161.175 - Frozen raw breaded shrimp.

    Science.gov (United States)

    2010-04-01

    ... additives as defined in section 201(s) of the Federal Food, Drug, and Cosmetic Act; or if they are food... 21 Food and Drugs 2 2010-04-01 2010-04-01 false Frozen raw breaded shrimp. 161.175 Section 161.175 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD...

  10. Frozen shoulder and the Big Five personality traits.

    Science.gov (United States)

    Debeer, Philippe; Franssens, Fien; Roosen, Isabelle; Dankaerts, Wim; Claes, Laurence

    2014-02-01

    In the past, several studies have suggested the existence of a "periarthritic personality" in patients with frozen shoulder. We conducted a study to determine differences in personality traits in patients with primary and secondary frozen shoulders. We prospectively evaluated 118 patients (84 women and 34 men; mean age, 53.8 years; SD 7.56) with a frozen shoulder. Of these patients, 48 had an idiopathic frozen shoulder and 70 had a secondary frozen shoulder. Personality traits were determined by the NEO Five-Factor Inventory (NEO-FFI) scale. This questionnaire measures the 5 major personality traits and is based on the norms determined in a neutral test situation for 2415 controls. Compared with healthy controls, no differences in personality traits were found in patients with primary and secondary frozen shoulder, except for Conscientiousness and Extraversion, for which patients with secondary frozen shoulder scored significantly higher than healthy controls. Patients with primary frozen shoulder scored significantly higher on Openness to Experience than did patients with secondary frozen shoulder; on the other 4 Big Five personality traits, no significant differences were found between patients with primary and secondary frozen shoulder. More specifically, patients with idiopathic frozen shoulder did not score higher on the trait Neuroticism as would be expected from previous publications. Our study results do not indicate that patients with an idiopathic frozen shoulder have a specific personality compared with healthy controls. Only a few differences were found in personality traits when the entire frozen shoulder group was compared with healthy controls and between patients with primary and secondary frozen shoulders. The results of this study suggest that these differences are not sufficient to speak about a specific "frozen shoulder personality." Copyright © 2014 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Mosby, Inc. All rights

  11. 21 CFR 864.9145 - Processing system for frozen blood.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Processing system for frozen blood. 864.9145 Section 864.9145 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Products Used In Establishments That Manufacture...

  12. Long-term frozen storage of urine samples: a trouble to get PCR results in Schistosoma spp. DNA detection?

    Science.gov (United States)

    Fernández-Soto, Pedro; Velasco Tirado, Virginia; Carranza Rodríguez, Cristina; Pérez-Arellano, José Luis; Muro, Antonio

    2013-01-01

    Human schistosomiasis remains a serious worldwide public health problem. At present, a sensitive and specific assay for routine diagnosis of schistosome infection is not yet available. The potential for detecting schistosome-derived DNA by PCR-based methods in human clinical samples is currently being investigated as a diagnostic tool with potential application in routine schistosomiasis diagnosis. Collection of diagnostic samples such as stool or blood is usually difficult in some populations. However, urine is a biological sample that can be collected in a non-invasive method, easy to get from people of all ages and easy in management, but as a sample for PCR diagnosis is still not widely used. This could be due to the high variability in the reported efficiency of detection as a result of the high variation in urine samples' storage or conditions for handling and DNA preservation and extraction methods. We evaluate different commercial DNA extraction methods from a series of long-term frozen storage human urine samples from patients with parasitological confirmed schistosomiasis in order to assess the PCR effectiveness for Schistosoma spp. detection. Patients urine samples were frozen for 18 months up to 7 years until use. Results were compared with those obtained in PCR assays using fresh healthy human urine artificially contaminated with Schistosoma mansoni DNA and urine samples from mice experimentally infected with S. mansoni cercariae stored frozen for at least 12 months before use. PCR results in fresh human artificial urine samples using different DNA based extraction methods were much more effective than those obtained when long-term frozen human urine samples were used as the source of DNA template. Long-term frozen human urine samples are probably not a good source for DNA extraction for use as a template in PCR detection of Schistosoma spp., regardless of the DNA method of extraction used.

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

    Science.gov (United States)

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

    2016-12-01

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

  14. Impacts of post-mortem ageing prior to freezing on technological and oxidative properties of coarse ground lamb sausage in a model system

    OpenAIRE

    Juhui Choe; Hyun-Wook Kim; Mustafa M. Farouk; Yuan H. Brad Kim

    2017-01-01

    Objective The objective of this study was to evaluate the effects of ageing time of lamb loins prior to freezing on technological characteristics and oxidation stability of coarse ground lamb loin sausage using in a model system. Methods Lamb loins (M. longissimus lumborum, n = 25) were aged at ?1.5?C for 0, 1, 2, 3, and 8 wk and then frozen for the remaining days (a total of 30 wk). The aged/frozen/thawed lamb loins were ground, and model sausages were formulated with 75% aged/frozen/thawed ...

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

    Directory of Open Access Journals (Sweden)

    Xinbao Yu

    2010-01-01

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

  16. Hydrologic impacts of thawing permafrost—A review

    Science.gov (United States)

    Walvoord, Michelle Ann; Kurylyk, Barret L.

    2016-01-01

    Where present, permafrost exerts a primary control on water fluxes, flowpaths, and distribution. Climate warming and related drivers of soil thermal change are expected to modify the distribution of permafrost, leading to changing hydrologic conditions, including alterations in soil moisture, connectivity of inland waters, streamflow seasonality, and the partitioning of water stored above and below ground. The field of permafrost hydrology is undergoing rapid advancement with respect to multiscale observations, subsurface characterization, modeling, and integration with other disciplines. However, gaining predictive capability of the many interrelated consequences of climate change is a persistent challenge due to several factors. Observations of hydrologic change have been causally linked to permafrost thaw, but applications of process-based models needed to support and enhance the transferability of empirical linkages have often been restricted to generalized representations. Limitations stem from inadequate baseline permafrost and unfrozen hydrogeologic characterization, lack of historical data, and simplifications in structure and process representation needed to counter the high computational demands of cryohydrogeologic simulations. Further, due in part to the large degree of subsurface heterogeneity of permafrost landscapes and the nonuniformity in thaw patterns and rates, associations between various modes of permafrost thaw and hydrologic change are not readily scalable; even trajectories of change can differ. This review highlights promising advances in characterization and modeling of permafrost regions and presents ongoing research challenges toward projecting hydrologic and ecologic consequences of permafrost thaw at time and spatial scales that are useful to managers and researchers.

  17. The effects of commercial fibres on frozen bread dough

    Directory of Open Access Journals (Sweden)

    JELENA FILIPOVIĆ

    2010-02-01

    Full Text Available The daily intake of dietary fibres in highly industrialized countries is at a low level and, therefore, adversely affecting human health. The objective of this research was to analyze the influence of different commercial fibres (originating from sugar beet pulp fibrex, and Jerusalem artichoke inulin HPX and GR in yeast dough at a level of 5 %, on the rheological properties of dough and the quality of bread during frozen storage. Frozen dough characteristics were determined using a Brabender maturograph and test baking was followed according the AACC procedure. The dough was frozen at –18 °C and stored over a period of 60 days. The results concerning the dough (proving time and stability and bread quality (volume and crumb quality were statistically analyzed by multivariance Manova and discriminative analysis, which indicated that there was a significant difference between dough without fibres and dough with different fibres (fibrex, inulin HPX and GR. The discrimination coefficient points that the greatest influence of fibres on the final proof and proving stability is after 30 days (6.250 and after 0 days (6.158, respectively, but the greatest influence of fibres on bread volume and bread crumb quality (15.488 and 3.638, respectively can be expected on non frozen dough, due to above mention their adverse the effect on gluten network.

  18. Life in Ice: Microbial Growth Dynamics and Greenhouse Gas Production During Winter in a Thermokarst Bog Revealed by Stable Isotope Probing Targeted Metagenomics

    Science.gov (United States)

    Blazewicz, S.; White, R. A., III; Tas, N.; Euskirchen, E. S.; Mcfarland, J. W.; Jansson, J.; Waldrop, M. P.

    2016-12-01

    Permafrost contains a reservoir of frozen C estimated to be twice the size of the current atmospheric C pool. In response to changing climate, permafrost is rapidly warming which could result in widespread seasonal thawing. When permafrost thaws, soils that are rich in ice and C often transform into thermokarst wetlands with anaerobic conditions and significant production of atmospheric CH4. While most C flux research in recently thawed permafrost concentrates on the few summer months when seasonal thaw has occurred, there is mounting evidence that sizeable portions of annual CO2 and CH4 efflux occurs over winter or during a rapid burst of emissions associated with seasonal thaw. A potential mechanism for such efflux patterns is microbial activity in frozen soils over winter where gasses produced are partially trapped within ice until spring thaw. In order to better understand microbial transformation of soil C to greenhouse gas over winter, we applied stable isotope probing (SIP) targeted metagenomics combined with process measurements and field flux data to reveal activities of microbial communities in `frozen' soil from an Alaskan thermokarst bog. Field studies revealed build-up of CO2 and CH4 in frozen soils suggesting that microbial activity persisted throughout the winter in soils poised just below the freezing point. Laboratory incubations designed to simulate in-situ winter conditions (-1.5 °C and anaerobic) revealed continuous CH4 and CO2 production. Strikingly, the quantity of CH4 produced in 6 months in frozen soil was equivalent to approximately 80% of CH4 emitted during the 3 month summer `active' season. Heavy water SIP targeted iTag sequencing revealed growing bacteria and archaea in the frozen anaerobic soil. Growth was primarily observed in two bacterial phyla, Firmicutes and Bacteroidetes, suggesting that fermentation was likely the major C mineralization pathway. SIP targeted metagenomics facilitated characterization of the primary metabolic

  19. A comparative study of vascular injection fluids in fresh-frozen and embalmed human cadaver forearms.

    Science.gov (United States)

    Doomernik, D E; Kruse, R R; Reijnen, M M P J; Kozicz, T L; Kooloos, J G M

    2016-10-01

    Over the years, various vascular injection products have been developed to facilitate anatomical dissections. This study aimed to compare the most commonly used vascular injection products in fresh-frozen and formalin-embalmed cadaver specimens. An overview of the properties, advantages and limitations of each substance was given, and a comparison of vascular infusion procedures in both preservation methods was made. A literature search was performed in order to identify the most commonly used vascular injection products. Acrylic paint, latex, gelatin, silicone, Araldite F and Batson's No. 17 were selected for the study. One fresh-frozen and one embalmed cadaver forearm were infused with each injection product according to a uniform protocol. The curing time, skin- and subcutaneous tissue penetration, degree of filling of the arterial tree, extravasations, consistency of the injected vessels during dissection, and the costs of each injection fluid were noted. There was a large variation between the injection fluids in processing- and curing time, colour intensity, flexibility, fragility, elasticity, strength, toxicity and costs. All fluids were suitable for infusion. The penetration of injection fluid into the skin and subcutaneous tissue was significantly better in fresh-frozen specimens (P = 0.002 and P = 0.009, respectively), with significantly smaller branches casted (P = 0.004). Vascular infusion of fresh-frozen cadaver specimens results in a significantly better filled coloured arterial tree, enabling more detail to be achieved and smaller branches casted. The biomechanical properties of fresh-frozen soft tissues are less affected compared with formalin fixation. All the injection fluids studied are suitable for vascular infusion, but their different properties ensure that certain products and procedures are more suitable for specific study purposes. © 2016 Anatomical Society.