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

PubMed

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.

Thawing as a critical pre-analytical step in the lipidomic profiling of plasma samples: New standardized protocol.

PubMed

Pizarro, Consuelo; Arenzana-Rámila, Irene; Pérez-del-Notario, Nuria; Pérez-Matute, Patricia; González-Sáiz, José María

2016-03-17

Lipid profiling is a promising tool for the discovery and subsequent identification of biomarkers associated with various diseases. However, data quality is quite dependent on the pre-analytical methods employed. To date, potential confounding factors that may affect lipid metabolite levels after the thawing of plasma for biomarker exploration studies have not been thoroughly evaluated. In this study, by means of experimental design methodology, we performed the first in-depth examination of the ways in which thawing conditions affect lipid metabolite levels. After the optimization stage, we concluded that temperature, sample volume and the thawing method were the determining factors that had to be exhaustively controlled in the thawing process to ensure the quality of biomarker discovery. Best thawing conditions were found to be: 4 °C, with 0.25 mL of human plasma and ultrasound (US) thawing. The new US proposed thawing method was quicker than the other methods we studied, allowed more features to be identified and increased the signal of the lipids. In view of its speed, efficiency and detectability, the US thawing method appears to be a simple, economical method for the thawing of plasma samples, which could easily be applied in clinical laboratories before lipid profiling studies. Copyright © 2016 Elsevier B.V. All rights reserved.

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

PubMed Central

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

Numerical Simulation of Thawing Process of Biological Tissue

NASA Astrophysics Data System (ADS)

Momose, Noboru; Tada, Yukio; Hayashi, Yujiro

Heat transfer and simplified physicochemical model for thawing of the frozen biological cell element consisting of cell and extracellular region was proposed. The melting of intra-and extra-cellular ice, the water transport through cell membrane and other microscale behavior during thawing process were discussed as a function of temperature. Recovery of the cell volume and change of osmotic pressure difference during thawing were clarified theortically in connection with heating velocity, initial cell volume and membrane permeability. Extending this model, the thawing of cellular tissue consisted of numerous cell elements was also simulated. There was a position where osmotic pressure difference became maximum during thawing. Summarizing these results, the thawing damage due to osmotic stress was discussed in relation with the heating operation and the size effect of tissue.

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

NASA Astrophysics Data System (ADS)

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.

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

NASA Astrophysics Data System (ADS)

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.

Stability of hemostatic proteins in canine fresh-frozen plasma thawed with a modified commercial microwave warmer or warm water bath.

PubMed

Pashmakova, Medora B; Barr, James W; Bishop, Micah A

2015-05-01

To compare stability of hemostatic proteins in canine fresh-frozen plasma (FFP) thawed with a modified commercial microwave warmer (MCM) or warm water bath (37°C; WWB) or at room temperature (22°C). Fresh-frozen plasma obtained from 8 canine donors of a commercial blood bank. A commercial microwave warmer was modified with a thermocouple to measure surface temperature of bags containing plasma. The MCM and a WWB were each used to concurrently thaw a 60-mL bag of plasma obtained from the same donor. Two 3-mL control aliquots of FFP from each donor were thawed to room temperature without use of a heating device. Concentrations of hemostatic proteins, albumin, and D-dimers; prothrombin time (PT); and activated partial thromboplastin time (aPTT) were determined for all samples. Significant decreases in concentrations of factors II, IX, X, XI, fibrinogen, von Willebrand factor, antithrombin, protein C, and albumin and significant increases in PT and aPTT were detected for plasma thawed with the MCM, compared with results for samples thawed with the WWB. Concentrations of factors VII, VIII, and XII were not significantly different between plasma thawed with the MCM and WWB. Concentrations of D-dimers were above the reference range for all thawed samples regardless of thawing method. No significant differences in factor concentrations were detected between control and WWB-thawed samples. Significant differences in hemostatic protein concentrations and coagulation times were detected for plasma thawed with an MCM but not between control and WWB-thawed samples. Clinical importance of these changes should be investigated.

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

PubMed

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.

Prediction of Quality Change During Thawing of Frozen Tuna Meat by Numerical Calculation I

NASA Astrophysics Data System (ADS)

Murakami, Natsumi; Watanabe, Manabu; Suzuki, Toru

A numerical calculation method has been developed to determine the optimum thawing method for minimizing the increase of metmyoglobin content (metMb%) as an indicator of color changes in frozen tuna meat during thawing. The calculation method is configured the following two steps: a) calculation of temperature history in each part of frozen tuna meat during thawing by control volume method under the assumption of one-dimensional heat transfer, and b) calculation of metMb% based on the combination of calculated temperature history, Arrenius equation and the first-order reaction equation for the increase rate of metMb%. Thawing experiments for measuring temperature history of frozen tuna meat were carried out under the conditions of rapid thawing and slow thawing to compare the experimental data with calculated temperature history as well as the increase of metMb%. The results were coincident with the experimental data. The proposed simulation method would be useful for predicting the optimum thawing conditions in terms of metMb%.

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

PubMed

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.

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

PubMed

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

2017-07-01

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

Experimentally increased nutrient availability at the permafrost thaw front selectively enhances biomass production of deep-rooting subarctic peatland species.

PubMed

Keuper, Frida; Dorrepaal, Ellen; van Bodegom, Peter M; van Logtestijn, Richard; Venhuizen, Gemma; van Hal, Jurgen; Aerts, Rien

2017-10-01

Climate warming increases nitrogen (N) mineralization in superficial soil layers (the dominant rooting zone) of subarctic peatlands. Thawing and subsequent mineralization of permafrost increases plant-available N around the thaw-front. Because plant production in these peatlands is N-limited, such changes may substantially affect net primary production and species composition. We aimed to identify the potential impact of increased N-availability due to permafrost thawing on subarctic peatland plant production and species performance, relative to the impact of increased N-availability in superficial organic layers. Therefore, we investigated whether plant roots are present at the thaw-front (45 cm depth) and whether N-uptake ( 15 N-tracer) at the thaw-front occurs during maximum thaw-depth, coinciding with the end of the growing season. Moreover, we performed a unique 3-year belowground fertilization experiment with fully factorial combinations of deep- (thaw-front) and shallow-fertilization (10 cm depth) and controls. We found that certain species are present with roots at the thaw-front (Rubus chamaemorus) and have the capacity (R. chamaemorus, Eriophorum vaginatum) for N-uptake from the thaw-front between autumn and spring when aboveground tissue is largely senescent. In response to 3-year shallow-belowground fertilization (S) both shallow- (Empetrum hermaphroditum) and deep-rooting species increased aboveground biomass and N-content, but only deep-rooting species responded positively to enhanced nutrient supply at the thaw-front (D). Moreover, the effects of shallow-fertilization and thaw-front fertilization on aboveground biomass production of the deep-rooting species were similar in magnitude (S: 71%; D: 111% increase compared to control) and additive (S + D: 181% increase). Our results show that plant-available N released from thawing permafrost can form a thus far overlooked additional N-source for deep-rooting subarctic plant species and increase their biomass production beyond the already established impact of warming-driven enhanced shallow N-mineralization. This may result in shifts in plant community composition and may partially counteract the increased carbon losses from thawing permafrost. © 2017 John Wiley & Sons Ltd.

Repeated freeze-thaw cycles induce embolism in drought stressed conifers (Norway spruce, stone pine).

PubMed

Mayr, Stefan; Gruber, Andreas; Bauer, Helmut

2003-07-01

Freezing and thawing lead to xylem embolism when gas bubbles caused by ice formation expand during the thaw process. However, previous experimental studies indicated that conifers are resistant to freezing-induced embolism, unless xylem pressure becomes very negative during the freezing. In this study, we show that conifers experienced freezing-induced embolism when exposed to repeated freeze-thaw cycles and simultaneously to drought. Simulating conditions at the alpine timberline (128 days with freeze-thaw events and thawing rates of up to 9.5 K h(-1) in the xylem of exposed twigs during winter), young trees of Norway spruce [Picea abies (L.) Karst.] and stone pine (Pinus cembra L.) were exposed to 50 and 100 freeze-thaw cycles. This treatment caused a significant increase in embolism rates in drought-stressed samples. Upon 100 freeze-thaw cycles, vulnerability thresholds (50% loss of conductivity) were shifted 1.8 MPa (Norway spruce) and 0.8 MPa (stone pine) towards less negative water potentials. The results demonstrate that freeze-thaw cycles are a possible reason for winter-embolism in conifers observed in several field studies. Freezing-induced embolism may contribute to the altitudinal limits of conifers.

Reviews and syntheses: Effects of permafrost thaw on Arctic aquatic ecosystems

NASA Astrophysics Data System (ADS)

Vonk, J. E.; Tank, S. E.; Bowden, W. B.; Laurion, I.; Vincent, W. F.; Alekseychik, P.; Amyot, M.; Billet, M. F.; Canário, J.; Cory, R. M.; Deshpande, B. N.; Helbig, M.; Jammet, M.; Karlsson, J.; Larouche, J.; MacMillan, G.; Rautio, M.; Anthony, K. M. Walter; Wickland, K. P.

2015-12-01

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 of dissolved vs. particulate organic matter, coupled with the composition of that organic matter and the morphology and stratification characteristics of recipient systems will play an important role in determining the balance between the release of organic matter as greenhouse gases (CO2 and CH4), its burial in sediments, and its loss downstream. The magnitude of thaw impacts on northern aquatic ecosystems is increasing, as is the prevalence of thaw-impacted lakes and streams. There is therefore an urgent need to quantify how permafrost thaw is affecting aquatic ecosystems across diverse Arctic landscapes, and the implications of this change for further climate warming.

Evaluation of Physicochemical Deterioration and Lipid Oxidation of Beef Muscle Affected by Freeze-thaw Cycles

PubMed Central

Rahman, M. H.; Hossain, M. M.; Rahman, S. M. E.; Amin, M. R.; Oh, Deog-Hwan

2015-01-01

This study was performed to explore the deterioration of physicochemical quality of beef hind limb during frozen storage at −20℃, affected by repeated freeze-thaw cycles. The effects of three successive freeze-thaw cycles on beef hind limb were investigated comparing with unfrozen beef muscle for 80 d by keeping at −20±1℃. The freeze-thaw cycles were subjected to three thawing methods and carried out to select the best one on the basis of deterioration of physicochemical properties of beef. As the number of repeated freeze-thaw cycles increased, drip loss decreased and water holding capacity (WHC) increased (p<0.05) till two cycles and then decreased. Cooking loss increased in cycle one and three but decreased in cycle two. Moreover, drip loss, WHC and cooking loss affected (p<0.05) by thawing methods within the cycles. However, pH value decreased (p<0.05), but peroxide value (p<0.05), free fatty acids value (p<0.05) and TBARS value increased (p<0.05) significantly as the number of repeated freeze-thaw cycles increased. Moreover, significant (p<0.05) interactive effects were found among the thawing methods and repeated cycles. As a result, freeze-thaw cycles affected the physicochemical quality of beef muscle, causing the degradation of its quality. PMID:26877637

A simple, field-friendly technique for cryopreserving semen from Asian elephants (Elephas maximus).

PubMed

Arnold, Danielle M; Gray, Charlie; Roth, Terri L; Mitchell, Sebastian; Graham, Laura H

2017-07-01

The specific objectives of the present study were to investigate the effects of manual seeding, differing freeze and thaw rates as well as storage for 24h at 4°C prior to cryopreservation on post-thaw sperm quality in Asian elephants. Extended semen was cooled in an equitainer to 4°C, frozen in liquid nitrogen vapour at various rates with and without manual seeding or in a dry shipper and thawed at 37, 50 and 75°C. There was a significant effect of freeze rate on post-thaw motility (P<0.0001) and acrosomal integrity (P<0.005). The faster freeze rates in the dry shipper and at 1cm or 2cm above liquid nitrogen consistently provided better cryopreservation than slower freezing rates. Thaw temperature had no effect on post-thaw semen quality but there was an interaction between freeze and thaw rates with higher thaw rates resulting in superior post-thaw semen quality in straws frozen at fast rates. Storage of samples prior to freezing had a detrimental effect on post-thaw semen quality. In summary, our results indicate cooling extended semen in an equitainer and cryopreserving it by placing straws directly in a dry shipper is a simple technique for effectively cryopreserving Asian elephant semen in the field or zoo. Copyright © 2017 Elsevier B.V. All rights reserved.

Quality Evaluation of Pork with Various Freezing and Thawing Methods

PubMed Central

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

Influence of seminal plasma on fresh and post-thaw parameters of stallion epididymal spermatozoa.

PubMed

Heise, A; Thompson, P N; Gerber, D

2011-02-01

Fresh and post-thaw parameters (motility, morphology and viability) of stallion epididymal spermatozoa that have been and have not been exposed to seminal plasma were evaluated, and directly compared to fresh and post-thaw parameters of ejaculated spermatozoa. Six sperm categories of each stallion (n=4) were evaluated for motility, morphology and viability. These categories were fresh ejaculated spermatozoa (Fr-E), fresh epididymal spermatozoa that had been exposed to seminal plasma (Fr-SP+), fresh epididymal spermatozoa that had never been exposed to seminal plasma (Fr-SP-), frozen-thawed ejaculated spermatozoa (Cr-E), frozen-thawed epididymal spermatozoa that had been exposed to seminal plasma prior to freezing (Cr-SP+) and frozen-thawed epididymal spermatozoa that had never been exposed to seminal plasma (Cr-SP-). Results show that seminal plasma stimulates initial motility of fresh epididymal stallion spermatozoa while this difference in progressive motility is no longer present post-thaw; and that progressive motility of fresh or frozen-thawed ejaculated stallion spermatozoa is not always a good indicator for post-thaw progressive motility of epididymal spermatozoa. This study shows that seminal plasma has a positive influence on the incidence of overall sperm defects, midpiece reflexes and distal cytoplasmic droplets in frozen-thawed stallion epididymal spermatozoa while the occurance of midpiece reflexes is likely to be linked to distal cytoplasmic droplets. Furthermore, seminal plasma does not have an influence on viability of fresh and frozen-thawed morphologically normal epididymal spermatozoa. We recommend the retrograde flushing technique using seminal plasma as flushing medium to harvest and freeze stallion epididymal spermatozoa. Copyright © 2010 Elsevier B.V. All rights reserved.

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

PubMed

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.

BERG2 Micro-computer Estimation of Freeze and Thaw Depths and Thaw Consolidation (PDF file)

DOT National Transportation Integrated Search

1989-06-01

The BERG2 microcomputer program uses a methology similar to the Modified Berggren method (Aldrich and Paynter, 1953) to estimate the freeze and thaw depths in layered soil systems. The program also provides an estimate of the thaw consolidation in ic...

SP-100 system thaw and start-up strategies

NASA Astrophysics Data System (ADS)

Kirpich, A.; Choe, H.

The authors review several strategies that have been considered during calendar year 1990 for SP-100 system start-up and thaw, and provide results of screening analyses. Screening studies have identified three concepts which are capable of thawing the SP-100 system: (1) a heat pipe approach in which reactor-heated heat pipes are routed to equipment enclosed within a thaw cavity; (2) a bleed-tube approach in which perforated tubes routed through ducting and components achieve thaw progressively as the result of successively thawing bleed-holes; and (3) a NaK traceline approach in which reactor-heated NaK is routed along ducting and components and achieves lithium thaw by both radiative and conductive coupling methods. Key issues have been identified in connection with pump start-up, reactor and power converter transient behavior, and hydraulic circuit stability. Pump start-up stability is accomplished by a linked-pump hydraulic arrangement.

Freezing and thawing or freezing, thawing, and aging effects on beef tenderness

USDA-ARS?s Scientific Manuscript database

The objective of this study was to determine the effect of freezing and thawing or freezing and thawing with an additional aging period after frozen storage on the tenderness of longissimus lumborum (LL) and semitendinosus (ST) steaks relative to aged, fresh steaks. Left-side LL and ST (n=35 each) ...

Moisture controls decomposition rate in thawing tundra

Treesearch

C.E. Hicks-Pries; E.A.G. Schuur; S.M. Natali; J.G. Vogel

2013-01-01

Permafrost thaw can affect decomposition rates by changing environmental conditions and litter quality. As permafrost thaws, soils warm and thermokarst (ground subsidence) features form, causing some areas to become wetter while other areas become drier. We used a common substrate to measure how permafrost thaw affects decomposition rates in the surface soil in a...

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

PubMed

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

2009-11-01

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

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

PubMed

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

2007-03-01

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

Artificial insemination with frozen-thawed boar sperm.

PubMed

Yeste, Marc; Rodríguez-Gil, Joan E; Bonet, Sergi

2017-09-01

Artificial insemination with frozen-thawed semen in pigs is not a routine technique; its use is restricted to specific cases, such as preservation of valuable genetic material (germplasm banks), safety strategies in case of natural disasters, long-distance transport of sperm, and in combination with sex-sorting. Cryoinjuries resulting from freeze-thawing protocols are a major concern with regard to the fertilization capacity of the treated sperm, which is lower than that of liquid-stored semen. Here, we provide an overview of artificial insemination using cryopreserved sperm, and summarize the factors that influence cryopreservation success before, during, and after freeze-thaw (i.e., sperm selection before starting the cryopreservation process, holding time, use of cryoprotectants, and rates of freezing and thawing) and that are driving the identification of biomarkers to predict sensitivity to cryodamage. Three different artificial insemination techniques (conventional or intracervical; intrauterine; and deep intrauterine) are also discussed with regards to their relevance when using frozen-thawed semen. Finally, we review the use of additives to freezing and thawing media, given reports that they may maintain and improve the quality and fertilizing capacity of frozen-thawed sperm. In sum, artificial insemination with frozen-thawed boar sperm can provide reasonable fertility outcomes, if freezable ejaculates, specific additives, and appropriate insemination techniques are used. © 2017 Wiley Periodicals, Inc.

Creep characteristics and process analyses of a thaw slump in the permafrost region of the Qinghai-Tibet Plateau, China

NASA Astrophysics Data System (ADS)

Sun, Zhe; Wang, Yibo; Sun, Yan; Niu, Fujun; Li, Guoyu; Gao, Zeyong

2017-09-01

A thaw slump in the permafrost region of the Qinghai-Tibet Plateau was monitored to investigate typical characteristics of creep positions and processes in combination with soil property analyses. The results show that the thaw settlement exhibits a contraction effect in the horizontal direction because of uneven thaw settlement. Slope displacement of creep occurs only in the top 50 cm of the soil. The gravimetric water content, soil porosity, and soil temperature are higher near the thaw slump in thaw seasons compared with the undisturbed soil; however, the shear strength is lower. Melting ground ice releases thaw water that converges along the slope and forms an overland flow at the front part of the gentle slope area and a ponding depression at the slope bottom. The analyses of slope stability using the infinite slope model shows that the headwall of the slope is inevitably unstable and slides under saturated conditions, whereas the gentle slope area and slope bottom with slight creep displacement are relatively stable. The small retrogressive thaw slump is in an early development stage. With increasing degree of thaw settlement and rate of erosion, the headwall will become steeper and a thermokarst lake will form at the slope bottom.

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

NASA Astrophysics Data System (ADS)

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.

Effects of freezing and thawing on texture, microstructure and cell wall composition changes in papaya tissues.

PubMed

Phothiset, Suphatta; Charoenrein, Sanguansri

2014-01-30

During storage, frozen fruit may be thawed and refrozen many times before consumption, which may be extremely damaging to the texture of the frozen fruit and reverse the advantage of fast freezing. The effects of freezing and thawing on texture, microstructure and cell wall composition changes in papaya tissues were investigated. The frozen-thawed papayas had an increase in drip loss and a decrease in firmness with increasing number of freeze-thaw cycles. Light microscopy showed irregular shapes and cell damage in parenchyma cells of frozen-thawed papayas, whereas transmission electron microscopy showed loss of cell wall materials in middle lamella. Moreover, destruction of cell wall was observed after being subjected to five freeze-thaw cycles. These changes related with a significant decrease in alcohol-insoluble solids, Na₂CO₃- and 24% KOH-soluble fractions and an increase in the water-, EDTA- and 4% KOH-soluble fractions. This was due to a decrease in the molecular mass of pectic and hemicellulosic polymers in frozen-thawed papayas using high-performance size-exclusion chromatography. The freezing and thawing processes caused fine structural damage and cell wall composition changes which contributed to a loss of drip volume and firmness of papaya tissues. © 2013 Society of Chemical Industry.

Moisture drives surface decomposition in thawing tundra

NASA Astrophysics Data System (ADS)

Hicks Pries, Caitlin E.; Schuur, E. A. G.; Vogel, Jason G.; Natali, Susan M.

2013-07-01

Permafrost thaw can affect decomposition rates by changing environmental conditions and litter quality. As permafrost thaws, soils warm and thermokarst (ground subsidence) features form, causing some areas to become wetter while other areas become drier. We used a common substrate to measure how permafrost thaw affects decomposition rates in the surface soil in a natural permafrost thaw gradient and a warming experiment in Healy, Alaska. Permafrost thaw also changes plant community composition. We decomposed 12 plant litters in a common garden to test how changing plant litter inputs would affect decomposition. We combined species' tissue-specific decomposition rates with species and tissue-level estimates of aboveground net primary productivity to calculate community-weighted decomposition constants at both the thaw gradient and warming experiment. Moisture, specifically growing season precipitation and water table depth, was the most significant driver of decomposition. At the gradient, an increase in growing season precipitation from 200 to 300 mm increased mass loss of the common substrate by 100%. At the warming experiment, a decrease in the depth to the water table from 30 to 15 cm increased mass loss by 100%. At the gradient, community-weighted decomposition was 21% faster in extensive than in minimal thaw, but was similar when moss production was included. Overall, the effect of climate change and permafrost thaw on surface soil decomposition are driven more by precipitation and soil environment than by changes to plant communities. Increasing soil moisture is thereby another mechanism by which permafrost thaw can become a positive feedback to climate change.

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

PubMed

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 < .001). Frozen-thawed embryo transfer is associated with a lower incidence of ectopic pregnancy per clinical pregnancy, compared with fresh embryo transfers (odds ratio = 0.31; 95% confidence interval = 0.24-0.39). Female age and body mass index have no influence on ectopic pregnancy. In the frozen-thawed embryo transfer 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.

Rapid carbon loss and slow recovery following permafrost thaw in boreal peatlands

USGS Publications Warehouse

Jones, Miriam C.; Harden, Jennifer W.; O'Donnell, Jonathan A.; Manies, Kristen L.; Jorgenson, Torre; Treat, Claire C.; Ewing, Stephanie

2017-01-01

Permafrost peatlands store one-third of the total carbon (C) in the atmosphere and are increasingly vulnerable to thaw as high-latitude temperatures warm. Large uncertainties remain about C dynamics following permafrost thaw in boreal peatlands. We used a chronosequence approach to measure C stocks in forested permafrost plateaus (forest) and thawed permafrost bogs, ranging in thaw age from young (<10 years) to old (>100 years) from two interior Alaska chronosequences. Permafrost originally aggraded simultaneously with peat accumulation (syngenetic permafrost) at both sites. We found that upon thaw, C loss of the forest peat C is equivalent to ~30% of the initial forest C stock and is directly proportional to the prethaw C stocks. Our model results indicate that permafrost thaw turned these peatlands into net C sources to the atmosphere for a decade following thaw, after which post-thaw bog peat accumulation returned sites to net C sinks. It can take multiple centuries to millennia for a site to recover its prethaw C stocks; the amount of time needed for them to regain their prethaw C stocks is governed by the amount of C that accumulated prior to thaw. Consequently, these findings show that older peatlands will take longer to recover prethaw C stocks, whereas younger peatlands will exceed prethaw stocks in a matter of centuries. We conclude that the loss of sporadic and discontinuous permafrost by 2100 could result in a loss of up to 24 Pg of deep C from permafrost peatlands.

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

USGS Publications Warehouse

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.

The carbon cycle implications of chemical weathering in retrogressive thaw slump-impacted streams

NASA Astrophysics Data System (ADS)

Zolkos, S.; Tank, S. E.; Kokelj, S. V.

2016-12-01

Permafrost thaw is "unlocking" and exposing significant amounts of sediment, solutes and organic carbon previously maintained in frozen soils to biochemical processing and fluvial transport. While microbial respiration of permafrost organic carbon contributes significantly to CO2 in Arctic headwater streams, chemical weathering of minerals unearthed by thawing permafrost may fix CO2 as bicarbonate (HCO3), thus removing it from the active carbon cycle. However, the degree to which mineral weathering acts to temper CO2 generated during permafrost thaw is largely unknown. During summer 2015, we investigated these dynamics in eight streams (orders 1-3) impacted by retrogressive thaw slumps across the Peel Plateau (NT, Canada), where thaw slumps expose permafrost that is comprised of abundant glacial tills, and glaciofluvial and glaciolacustrine sediments. Thaw slump activity had a discernible signature in all streams: conductivity, pH, dissolved inorgnaic carbon (DIC), and solute concentrations (Ca, Mg, Na, K, SO4, Cl) increased in the downstream (thaw slump-impacted) reach, relative to upstream, while CO2 decreased. This corresponded with an isotopically-enriched DIC pool in impacted streams (mean δ13CDIC = -9.80‰), perhaps indicating the dissolution of carbonate minerals following exposure by thaw slump activity. Despite a general decrease downstream of thaw slumps, CO2 remained supersaturated in impacted streams (mean pCO2 = 915 µatm). However, the highest partial pressures of CO2 were found in thaw slump runoff (mean pCO2 = 4,600 µatm), above the point where runoff entered downstream systems. High pCO2 levels in slump runoff may be derived from microbial respiration of slump-released dissolved organic carbon or, for some slumps, carbonate dissolution (range δ13CDIC = 0.67 - -23.37‰). While this work suggests thaw slumps in the Western Canadian Arctic may act to partially temper CO2 in headwater streams, these stream networks will likely persist as significant sources of CO2 to the atmosphere.

Remote sensing of freeze-thaw transitions in Arctic soils using the complex resistivity method

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu, Yuxin; Hubbard, Susan S; Ulrich, Craig

2013-01-01

Our ability to monitor freeze - thaw transitions is critical to developing a predictive understanding of biogeochemical transitions and carbon dynamics in high latitude environments. In this study, we conducted laboratory column experiments to explore the potential of the complex resistivity method for monitoring the freeze - thaw transitions of the arctic permafrost soils. Samples for the experiment were collected from the upper active layer of Gelisol soils at the Barrow Environmental Observatory, Barrow Alaska. Freeze - thaw transitions were induced through exposing the soil column to controlled temperature environments at 4 C and -20 C. Complex resistivity and temperaturemore » measurements were collected regularly during the freeze - thaw transitions using electrodes and temperature sensors installed along the column. During the experiments, over two orders of magnitude of resistivity variations were observed when the temperature was increased or decreased between -20 C and 0 C. Smaller resistivity variations were also observed during the isothermal thawing or freezing processes that occurred near 0 C. Single frequency electrical phase response and imaginary conductivity at 1 Hz were found to be exclusively related to the unfrozen water in the soil matrix, suggesting that these geophysical 24 attributes can be used as a proxy for the monitoring of the onset and progression of the freeze - thaw transitions. Spectral electrical responses and fitted Cole Cole parameters contained additional information about the freeze - thaw transition affected by the soil grain size distribution. Specifically, a shift of the observed spectral response to lower frequency was observed during isothermal thawing process, which we interpret to be due to sequential thawing, first from fine then to coarse particles within the soil matrix. Our study demonstrates the potential of the complex resistivity method for remote monitoring of freeze - thaw transitions in arctic soils. Although conducted at the laboratory scale, this study provides the foundation for exploring the potential of the complex resistivity signals for monitoring spatiotemporal variations of freeze - thaw transitions over field-relevant scales.« less

Breed differences of bull frozen-thawed semen.

PubMed

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.

Oxygenated thawing and rewarming alleviate rewarming injury of cryopreserved pancreatic islets.

PubMed

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 and better function among the conditions examined. Oxygenated thawing/rewarming alleviated islet volume loss, with the help of oxygenated culture. Copyright © 2017. Published by Elsevier Inc.

Cryopreservation of American kestrel semen with dimethylsulfoxide

USGS Publications Warehouse

Gee, G.F.; Morrell, C.A.; Franson, J. Christian; Pattee, Oliver H.

1993-01-01

Semen samples from 15 male American Kestrels (Falco sparverius) were frozen in dimethyl sulfoxide (DMSO). The semen was thawed 1-14 mo later and used to inseminate six females during three breeding seasons. Kestrels inseminated with thawed semen containing 4% DMSO produced only infertile eggs (N = 14). Kestrels inseminated with thawed semen containing 6%, 8%, or 10% DMSO produced fertile eggs (N = 14) and live chicks (N = 6). Progressive motility of spermatozoa in thawed semen containing 10% DMSO was less (44 ? 6%) than in thawed semen containing 6% (62 ? 10%) or 8% (61 ? 1%) DMSO.

Poorly known microbial taxa dominate the microbiome of permafrost thaw ponds.

PubMed

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.

Rapid carbon loss and slow recovery following permafrost thaw in boreal peatlands.

PubMed

Jones, Miriam C; Harden, Jennifer; O'Donnell, Jonathan; Manies, Kristen; Jorgenson, Torre; Treat, Claire; Ewing, Stephanie

2017-03-01

Permafrost peatlands store one-third of the total carbon (C) in the atmosphere and are increasingly vulnerable to thaw as high-latitude temperatures warm. Large uncertainties remain about C dynamics following permafrost thaw in boreal peatlands. We used a chronosequence approach to measure C stocks in forested permafrost plateaus (forest) and thawed permafrost bogs, ranging in thaw age from young (<10 years) to old (>100 years) from two interior Alaska chronosequences. Permafrost originally aggraded simultaneously with peat accumulation (syngenetic permafrost) at both sites. We found that upon thaw, C loss of the forest peat C is equivalent to ~30% of the initial forest C stock and is directly proportional to the prethaw C stocks. Our model results indicate that permafrost thaw turned these peatlands into net C sources to the atmosphere for a decade following thaw, after which post-thaw bog peat accumulation returned sites to net C sinks. It can take multiple centuries to millennia for a site to recover its prethaw C stocks; the amount of time needed for them to regain their prethaw C stocks is governed by the amount of C that accumulated prior to thaw. Consequently, these findings show that older peatlands will take longer to recover prethaw C stocks, whereas younger peatlands will exceed prethaw stocks in a matter of centuries. We conclude that the loss of sporadic and discontinuous permafrost by 2100 could result in a loss of up to 24 Pg of deep C from permafrost peatlands. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

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

NASA Technical Reports Server (NTRS)

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

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

PubMed

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 < 0.001) on post-thaw sperm characteristics. It was observed that post-thaw sperm characteristics were significantly compromised in 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.

Reduced glutathione and procaine hydrochloride protect the nucleoprotein structure of boar spermatozoa during freeze-thawing by stabilising disulfide bonds.

PubMed

Yeste, Marc; Flores, Eva; Estrada, Efrén; Bonet, Sergi; Rigau, Teresa; Rodríguez-Gil, Joan E

2013-01-01

One important change the head of boar spermatozoa during freeze-thawing is the destabilisation of its nucleoprotein structure due to a disruption of disulfide bonds. With the aim of better understanding these changes in frozen-thawed spermatozoa, two agents, namely reduced glutathione (GSH) and procaine hydrochloride (ProHCl), were added at different concentrations to the freezing media at different concentrations and combinations over the range 1-2mM. Then, 30 and 240 min after thawing, cysteine-free residue levels of boar sperm nucleoproteins, DNA fragmentation and other sperm functional parameters were evaluated. Both GSH and ProHCl, at final concentrations of 2mM, induced a significant (P<0.05) increase in the number of non-disrupted sperm head disulfide bonds 30 and 240 min after thawing compared with the frozen-thawed control. This effect was accompanied by a significant (P<0.05) decrease in DNA fragmentation 240 min after thawing. Concomitantly, 1 and 2mM GSH, but not ProHCl at any of the concentrations tested, partially counteracted the detrimental effects caused by freeze-thawing on sperm peroxide levels, motility patterns and plasma membrane integrity. In conclusion, the results show that both GSH and ProHCl have a stabilising effect on the nucleoprotein structure of frozen-thawed spermatozoa, although only GSH exerts an appreciable effect on sperm viability.

Microwave thawing apparatus and method

DOEpatents

Fathi, Zakaryae; Lauf, Robert J.; McMillan, April D.

2004-06-01

An apparatus for thawing a frozen material includes: a microwave energy source; a microwave applicator which defines a cavity for applying microwave energy from the microwave source to a material to be thawed; and a shielded region which is shielded from the microwave source, the shielded region in fluid communication with the cavity so that thawed material may flow from the cavity into the shielded region.

Effects of type of freeze straw and thaw temperature on the fertilizing capacity of frozen chicken semen.

PubMed

Duplaix, M; Sexton, T J

1984-04-01

Two experiments were conducted to determine the relationship between thawing temperature and the type of straw in which chicken semen was frozen. In Experiment 1, semen was frozen in three different types of plastic straws: US (.5-ml capacity), French (.5-ml capacity), and French mini (.25-ml capacity). Experiment 1 was divided into two trials to compare semen packaged in the different straws and thawed at 15 (Trial 1) or .5 C (Trial 2). Although there were distinguishable features of the freeze and thaw curves between samples frozen in the different straws, the type of freeze straw had no effect on the fertilizing capacity of frozen semen when thaw temperature was held constant: fertility, Days 2 to 4 after artificial insemination, ranged from 16 to 27% for Trial 1 and 45 to 47% for Trial 2. In Experiment 2, semen was frozen in US straws and thawed at either .5 or 15 C to assess the effect of the thaw temperature. Fertility of frozen semen, Days 2 to 4 after artificial insemination, was significantly higher when semen was thawed at .5 than at 15 C (62 vs. 20%).

Fat loss in thawed breast milk: comparison between refrigerator and warm water.

PubMed

Thatrimontrichai, A; Janjindamai, W; Puwanant, M

2012-11-01

To compare the fat loss between refrigerator and warm water thawed breast milk. Experimental. Tertiary-care pediatric university hospital. Ninety samples of expressed breast milk were collected from mothers with singleton babies of a gestational age 32-42 weeks. Fat content in fresh breast milk (FM); thawed breast milk by refrigeration (RM); and thawed breast milk by warm water (WM). The mean (SD) total fat content in FM, RM and WM were 2.98 (0.97), 2.76 (0.99) and 2.66 (0.88) g/100 mL, respectively. The mean difference (SD) of the total fat in FM declined significantly after the frozen milk was thawed by refrigeration or warm water at -0.22 (0.50) g/100 mL (P=0.0001) and -0.32 (0.45) g/100 mL (P<0.0001), respectively. The mean (SD) total fat loss of frozen breast milk thawed by refrigeration was less than thawing in warm water at 0.094 (0.38) g/100 mL (P=0.02). The fat loss of thawed breast milk by refrigeration was significantly less than placing it in warm water.

Freeze-Thaw Durability of Air-Entrained Concrete

PubMed Central

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. PMID:23576906

Freeze-thaw durability of air-entrained concrete.

PubMed

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.

Degradation potentials of dissolved organic carbon (DOC) from thawed permafrost peat

PubMed Central

Panneer Selvam, Balathandayuthabani; Lapierre, Jean-François; Guillemette, Francois; Voigt, Carolina; Lamprecht, Richard E.; Biasi, Christina; Christensen, Torben R.; Martikainen, Pertti J.; Berggren, Martin

2017-01-01

Global warming can substantially affect the export of dissolved organic carbon (DOC) from peat-permafrost to aquatic systems. The direct degradability of such peat-derived DOC, however, is poorly constrained because previous permafrost thaw studies have mainly addressed mineral soil catchments or DOC pools that have already been processed in surface waters. We incubated peat cores from a palsa mire to compare an active layer and an experimentally thawed permafrost layer with regard to DOC composition and degradation potentials of pore water DOC. Our results show that DOC from the thawed permafrost layer had high initial degradation potentials compared with DOC from the active layer. In fact, the DOC that showed the highest bio- and photo-degradability, respectively, originated in the thawed permafrost layer. Our study sheds new light on the DOC composition of peat-permafrost directly upon thaw and suggests that past estimates of carbon-dioxide emissions from thawed peat permafrost may be biased as they have overlooked the initial mineralization potential of the exported DOC. PMID:28378792

Thaw pond dynamics and carbon emissions in a Siberian lowland tundra landscape

NASA Astrophysics Data System (ADS)

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 between newly formed and disappearing ponds, although their net area increased by 16%. The disappearing of ponds was mostly the result of vegetation succession, rather than drainage. This vegetation succession results from an invasion by sedges, followed by establishment of Sphagnum and seedlings of dwarf shrubs. The formation of thaw ponds and troughs resulting from small-scale permafrost collapse results in a drastic change of CH4 and CO2 emissions, from near-zero emission or uptake to high emission. New water surfaces with drowned dry tundra vegetation show the highest emission. However, rapid vegetation succession may mitigate these emissions over time, in particular in the relatively shallow thaw ponds. In contrast, the polygon centre ponds with a stable, oligotrophic vegetation show modest and constant CH4 emission and CO2 uptake.

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

PubMed

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 (p<0.05) higher for samples frozen in FlatPacks than in Maxi-straws, 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 boar spermatozoa for artificial insemination.

Permafrost Thaw increases Emissions of Nitrous Oxide from Subarctic Peatlands

NASA Astrophysics Data System (ADS)

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 previously thought, and that the source strength will be crucially governed by moisture conditions at times of thaw, as well as on future changes in vegetation coverage.

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

Treesearch

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

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...

Directions, Objectives, and Author's Concepts of Audiovisual Media Interpretations of School and University Theme in the Soviet Cinema of the "Thaw" Period (1956-1968)

ERIC Educational Resources Information Center

Fedorov, Alexander; Levitskaya, Anastasia; Gorbatkova, Olga; Mamadaliev, Anvar M.

2017-01-01

The "thaw period" films (1956-1968) on the school/university topic can be conditionally divided into two stages: early (1956-1963) and late "thaw" (1964-1968), although, naturally, there was a somewhat diffusion between these periods. The "thaw" audiovisual texts about school and university life, according to the…

Midwinter dehardening of montane red spruce during a natural thaw

Treesearch

G.R. Strimbeck; P.G. Schaberg; D.H. DeHayes; J.B. Shane; G.J. Hawley

1995-01-01

We documented 3 to 14°C of dehardening in current-year foliage of 10 mature, montane red spruce (Picea rubens Sarg.) trees during a natural thaw from 12 to 21 January 1995. Mean cold tolerance was about -47°C before the onset of thaw conditions, and individuals ranged from -38 to -52°C. After 3 days of thaw, mean cold...

Microwave thawing package and method

DOEpatents

Fathi, Zakaryae; Lauf, Robert J.

2004-03-16

A package for containing frozen liquids during an electromagnetic thawing process includes: a first section adapted for containing a frozen material and exposing the frozen material to electromagnetic energy; a second section adapted for receiving thawed liquid material and shielding the thawed liquid material from further exposure to electromagnetic energy; and a fluid communication means for allowing fluid flow between the first section and the second section.

Mapping Microbial Carbon Substrate Utilization Across Permafrost Thaw

NASA Astrophysics Data System (ADS)

Anderson, D.; Rich, V. I.; Hodgkins, S. B.; Tfaily, M.; Chanton, J.

2014-12-01

Permafrost thaw is likely to create a substantial positive feedback to climate warming, as previously frozen carbon becomes bioavailable and is released to the atmosphere. Microbes mediate this release, while also consuming "new" carbon from plant inputs and middle-aged soil carbon pools in the seasonally-thawed active layer overlying permafrost. This carbon consumption releases carbon dioxide (CO2) and methane (CH4), both potent greenhouse gases. To investigate microbial carbon cycling in this changing habitat, we examined how microbial communities' carbon substrate degradation changes along a natural permafrost thaw gradient in Stordalen Mire (68.35°N, 19.05°E), northern Sweden. At this location, intermediate thaw creates Sphagnum moss-dominated bogs, while complete thaw results in Eriophorum sedge-dominated fens. The progression of thaw results in increasing organic matter lability (Hodgkins et al, 2014), shifting microbial community composition (Mondav & Woodcroft et al 2014), and changing carbon gas emissions (McCalley et al, in review). However, the inter-relationship of the first two in producing the third remains unclear. We analyzed microbial carbon substrate utilization in the intermediate-thaw and full-thaw sites by two incubation-based methods. We used Biolog EcoPlates, which contain 31 ecologically relevant carbon substrates and a colorimetric marker of their consumption, and into which we added a soil liquid suspension. In addition, we performed mason-jar incubations of peat with carbon substrate amendments and measured CH4 and CO2 emissions. Preliminary Biolog Ecoplate incubations showed that intermediate-thaw features responded faster and more strongly overall to a wide range of substrates relative to the full-thaw features. Preliminary mason jar incubations showed that acetate amendment elicited the greatest response increase in CH4 production and the second greatest increase in CO2 production relative to the controls, in samples from both habitats. In addition, the lowest CH4 and CO2 production was seen in amendments of sphagnum acid. It is important to understand the carbon substrate utilization occurring at these initial and advanced thaw features to speculate the degree to which various carbon inputs are being metabolized to produce the observed gas emissions.

Antimicrobial efficiency of essential oil and freeze-thaw treatments against Escherichia coli O157:H7 and Salmonella enterica Ser. Enteritidis in strawberry juice.

PubMed

Duan, J; Zhao, Y

2009-04-01

This study investigated the antimicrobial efficiency of 3 essential oils (EOs), lemongrass, cinnamon leaf, and basil, and freeze-thaw treatment, alone or in combination, against Escherichia coli O157:H7 and Salmonella enterica Ser. Enteritidis inoculated in strawberry juice stored at 7 degrees C. EO of lemongrass or cinnamon leaf at 0.1 to 2 microL/mL and freezing at -23 degrees C for 24 or 48 h followed by thawing at 7 degrees C for 4 h all showed significant antimicrobial activities (P < 0.05) against E. coli O157:H7 and S. Enteritidis in strawberry juice. The antimicrobial activity increased with increasing EO concentration and storage time, but extending freezing time from 24 to 48 h did not enhance the antimicrobial activity of freeze-thaw treatment (P > 0.05). EO of lemongrass or cinnamon leaf at 0.1 microL/mL and freeze-thaw treatment alone obtained a 5 log(10) reduction in the population of S. Enteritidis, while EOs at 0.1 to 0.3 microL/mL or freeze-thaw alone could not achieve a satisfactory protection against E. coli O157:H7 in strawberry juice. Combined EO and freeze-thaw treatment enhanced the overall antimicrobial effect against E. coli O157:H7, with adding EO before the freeze-thaw treatment showed a faster decontamination rate than when added EO after the freeze-thaw. EOs of lemongrass and cinnamon leaf at 0.1 or 0.3 microL/mL followed by the freeze-thawing resulted in a 5 log(10) reduction in E. coli O157:H7 on the 5th and 2nd day of storage, respectively. This study suggested that combined EO and freeze-thaw treatment may be a suitable and inexpensive method to eliminate microorganisms that can be a hazard for the consumers of unpasteurized berry juices.

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.

PubMed

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 proteases in FFP units.

A preliminary study on using autologous and heterologous boar sperm supernatant from freezing processes as post-thawing solution: its effect on sperm motility.

PubMed

Kaeoket, Kampon; Chanapiwat, Panida; Tummaruk, Padet; Techakumphu, Mongkol; Kunavongkrit, Annop

2011-06-01

The aim of this study was to evaluate the effect of post-thawing dilution with autologous and heterologous sperm supernatant on motility of frozen-thawed boar spermatozoa. During the cryopreservation, sperm supernatant (a combination of seminal plasma and semen extender, 50% v/v) or seminal plasma from nine boars (Duroc, Large White, and Landrace; three in each) was collected by centrifugation and stored frozen until use as post-thawing solution. Sperm pellets were further processed and cryopreserved using control-rate freezer and was thawed at 50°C for 12 s. After thawing, frozen thawed semen samples were diluted with seminal plasma (group A), supernatant from Landrace (group B), supernatant from Large White (group C), supernatant from Duroc (group D), and Modena™ semen extender (group E). Post-thawing motility was evaluated using a phase-contrast microscope after thawing at 1, 10, 20, and 30 min. The present results show that at 1 min, a significantly higher percentage (P ≤ 0.001) of progressive motility was found in groups B (53.3%) and C (53.9%) than the other groups. At 10 min, the highest (P ≤ 0.001) progressive motility was found in groups B (65%) and C (61%). At 20 and 30 min, a significantly higher percentage (P ≤ 0.001) of progressive motility was found in groups B (58.9%), C (53.5%), and D (45.6%) than groups A (3.9%) and E (20.6%). It can be stated that supernatant from the freezing processes (consisting of seminal plasma and Modena™, 50% v/v) had a beneficial effect on post-thawing progressive motility of frozen boar semen.

Permafrost soil characteristics and microbial community structure across a boreal forest watershed vary over short spatial scales and dictate community responses to thaw.

NASA Astrophysics Data System (ADS)

Stegen, J.; Bottos, E. M.; Kennedy, D.; Romero, E. B.; Fansler, S.; Chu, R. K.; Tfaily, M.; Jansson, J.; Bernstein, H. C.; Brown, J. M.; Markillie, L. M.

2017-12-01

Understanding drivers of permafrost microbial community structure and function is critical for understanding permafrost microbiology and predicting ecosystem responses to thaw; however, studies describing ecological controls on these communities are lacking. We hypothesize that permafrost communities are uniquely shaped by constraints imposed by prolonged freezing, and decoupled from the selective factors that influence non-permafrost soil communities, but that pre-thaw environmental and community characteristics will be strong determinants of community structure and function post-thaw. We characterized patterns of environmental variation and microbial community composition in sixty permafrost samples spanning landscape gradients in a boreal forest watershed, and monitored community responses to thaw. Consistent with our hypothesis, we found that, proportionally, the strongest process influencing permafrost community composition was dispersal limitation (0.36), exceeding the influence of homogenous selection (0.21) and variable selection (0.16), and that deterministic selection arose primarily from energetic constraints of the permafrost environment. Our data supported a structural equation model in which organic carbon thermodynamics and organic acid content, influenced redox conditions and total selection. Post-thaw community composition was found to be driven primarily by pre-thaw community composition, indicating a strong influence of historical conditions. Together, these results suggest that community responses to thaw may be highly varied over short distances and that changes in community structure and function are likely to be drastic, as changes to system hydrology mobilize organisms and nutrients, thereby relieving the primary constraints on the system. These findings are being integrated with metabolomic and metatranscriptomic analyses to improve understanding of how pre-thaw conditions can be used to predict microbial activity post-thaw.

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

NASA Technical Reports Server (NTRS)

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.

Combining reduced glutathione and ascorbic acid has supplementary beneficial effects on boar sperm cryotolerance.

PubMed

Giaretta, Elisa; Estrada, Efrén; Bucci, Diego; Spinaci, Marcella; Rodríguez-Gil, Joan E; Yeste, Marc

2015-02-01

The main aim of this work was to evaluate how supplementing freezing and thawing media with reduced glutathione (GSH) and l-ascorbic acid (AA) affected the quality parameters of frozen-thawed boar spermatozoa. With this purpose, semen samples of 12 ejaculates coming from 12 boars were used. Each ejaculate was split into seven aliquots to which 5 mM of GSH and 100 μM of AA were added separately or together at two different steps of freeze-thawing. Various sperm parameters (levels of free cysteine residues in sperm nucleoproteins, sperm viability, acrosome membrane integrity, intracellular peroxide and superoxide levels [ROS], and total and progressive motility) were evaluated before freezing and at 30 and 240 minutes after thawing. Both GSH and AA significantly improved boar sperm cryotolerance when they were separately added to freezing and thawing media. However, the highest improvement was recorded when both freezing and thawing media were supplemented with 5 mM of GSH plus 100 μM of AA. This improvement was observed in sperm viability and acrosome integrity, sperm motility, and nucleoprotein structure. Although ROS levels were not much increased by freeze-thawing procedures, the addition of GSH and AA to both freezing and thawing extenders significantly decreased intracellular peroxide levels and had no impact on superoxide levels. According to our results, we can conclude that supplementation of freezing and thawing media with both GSH and AA has a combined, beneficial effect on frozen-thawed boar sperm, which is greater than that obtained with the separate addition of either GSH or AA. Copyright © 2015 Elsevier Inc. All rights reserved.

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

PubMed

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 (<3) freeze/thaw 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.

Determination of glutation peroxidase and superoxide dismutase activities in canine seminal plasma and its relation with sperm quality and lipid peroxidation post thaw.

PubMed

Neagu, V R; García, B Macías; Rodríguez, A Morillo; Ferrusola, C Ortega; Bolaños, J M Gallardo; Fernández, L González; Tapia, J A; Peña, F J

2011-01-01

Lipid peroxidation (LPO) of dog spermatozoa was assessed in fresh semen and in samples of the same ejaculates after freezing and thawing. Particular attention was paid to individual differences in the susceptibility to LPO and its possible relationship with freezeability. Innate levels of LPO were low in fresh spermatozoa but increased after thawing in one of the dogs included in our study. The level of lipid peroxidation in fresh spermatozoa was not correlated with that of thawed spermatozoa. Negative correlations were detected between the activity in seminal plasma of GPx and sperm velocities post thaw (P < 0.01), however SOD activity was positively correlated with the percentage of linear motile sperm post thaw (P < 0.05). Copyright © 2011 Elsevier Inc. All rights reserved.

Interactive effects of wildfire and permafrost thaw on peatland carbon cycling

NASA Astrophysics Data System (ADS)

Olefeldt, David; Heffernan, William; Gibson, Carolyn; Burd, Katheryn; Estop-Aragones, Cristian

2017-04-01

Boreal peatland complexes in western Canada are fine-scale mosaics of permafrost affected peat plateaus interspersed with Sphagnum dominated thermokarst bogs where permafrost is absent. Wildfire further affects landscape patterning of peatland complexes, where virtually all peat plateaus are in a stage of secondary succession following wildfire. With climate change we expect both permafrost thaw and wildfire activity to increase in these landscapes, and to have important impacts on carbon cycling. In a number of studies, we have used soil chamber techniques to assess the influence of both permafrost thaw and wildfire on soil respiration, net ecosystem exchange and methane emissions. We used chronosequences to assess the influence of time since both permafrost thaw (3 - 15 years) and wildfire (20 - 150 years). Radiocarbon signatures of soil respiration in both burned and thawed locations was used to determine the contribution of aged soil carbon to soil respiration. We furthermore characterized individual and interactive effects of fire and thaw on microbial and photochemical lability of dissolved organic matter. At many field sites it was clear that recent wildfire had accelerated permafrost thaw, and we combined field observations of soil thermal regimes with remote sensing approaches to assess the role of wildfire for accelerating permafrost thaw over the last 50 years at a regional scale. Overall, our results highlight the need to consider both individual and interacting effects of thaw and fire for projections of the future carbon cycling at the regional level.

Seasonal thaw settlement at drained thermokarst lake basins, Arctic Alaska

USGS Publications Warehouse

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.

The Classification and Geomorphic Implications of Thaw Lakes on the Arctic Coastal Plain, Alaska

DTIC Science & Technology

1975-12-01

Plain is underlain by ice-rich marine sediments , the product of several marine transgressions and regressions. Numerous thaw lake basins of...variable morphology and distribution have developed on the perennially frozen sediments (permafrost) of this low-lying plain. Most notable are the large...mechanism of thaw lake formation was recognized whereby sediment laden ice rafts initiated thawing of the permafrost and formation of lake basins

Characterization of Propylene Glycol-Mitigated Freeze/Thaw Agglomeration of a Frozen Liquid nOMV Vaccine Formulation by Static Light Scattering and Micro-Flow Imaging.

PubMed

Mensch, Christopher D; Davis, Harrison B; Blue, Jeffrey T

2015-01-01

The purpose of this work was to investigate the susceptibility of an aluminum adjuvant and an aluminum-adjuvanted native outer membrane vesicle (nOMV) vaccine formulation to freeze/thaw-induced agglomeration using static light scattering and micro-flow Imaging analysis; and to evaluate the use of propylene glycol as a vaccine formulation excipient by which freeze/thaw-induced agglomeration of a nOMV vaccine formulation could be mitigated. Our results indicate that including 7% v/v propylene glycol in an nOMV containing aluminum adjuvanted vaccine formulation, mitigates freeze/thaw-induced agglomeration. We evaluated the effect of freeze-thawing on an aluminum adjuvant and an aluminum adjuvanted native outer membrane vesicle (nOMV) vaccine formulation. Specifically, we characterized the freeze/thaw-induced agglomeration through the use of static light scattering, micro-flow imaging, and cryo-electron microscopy analysis. Further, we evaluated the use of 0-9% v/v propylene glycol as an excipient which could be included in the formulation for the purpose of mitigating the agglomeration induced by freeze/thaw. The results indicate that using 7% v/v propylene glycol as a formulation excipient is effective at mitigating agglomeration of the nOMV vaccine formulation, otherwise induced by freeze-thawing. © PDA, Inc. 2015.

Human oocyte cryopreservation: 5-year experience with a sodium-depleted slow freezing method.

PubMed

Boldt, Jeffrey; Tidswell, Non; Sayers, Amy; Kilani, Rami; Cline, Donald

2006-07-01

A slow freezing/rapid thawing method for the cryopreservation of human oocytes has been employed using a sodium-depleted culture media. In 53 frozen egg-embryo transfer (FEET) cycles, a 60.4% survival rate post-thaw was obtained and a 62.0% fertilization rate following intracytoplasmic sperm injection. Overall pregnancy rates were 26.4% per thaw attempt, 30.4% per patient, and 32.6% per embryo transfer. Pregnancy rates using sodium-depleted phosphate-buffered saline (PBS) as the base medium were 20.0% per thaw, 21.7% per patient, and 26.3% per transfer. With sodium-depleted modified human tubal fluid (mHTF) as the base for the cryopreservation medium, rates were 32.1% per thaw attempt, 39.1% per patient, 37.5% per transfer. The overall implantation rates were 4.2% per thawed oocyte and 13.6% per embryo, (PBS: 3.0% per egg, 10.6% per embryo; mHTF:5.3% per oocyte; 15.9% per embryo). These data indicate that the use of a sodium-depleted media with slow freezing and rapid thawing can yield acceptable pregnancy rates after FEET.

Shifts of methanogenic communities in response to permafrost thaw results in rising methane emissions and soil property changes.

PubMed

Wei, Shiping; Cui, Hongpeng; Zhu, Youhai; Lu, Zhenquan; Pang, Shouji; Zhang, Shuai; Dong, Hailiang; Su, Xin

2018-05-01

Permafrost thaw can bring negative consequences in terms of ecosystems, resulting in permafrost collapse, waterlogging, thermokarst lake development, and species composition changes. Little is known about how permafrost thaw influences microbial community shifts and their activities. Here, we show that the dominant archaeal community shifts from Methanomicrobiales to Methanosarcinales in response to the permafrost thaw, and the increase in methane emission is found to be associated with the methanogenic archaea, which rapidly bloom with nearly tenfold increase in total number. The mcrA gene clone libraries analyses indicate that Methanocellales/Rice Cluster I was predominant both in the original permafrost and in the thawed permafrost. However, only species belonging to Methanosarcinales showed higher transcriptional activities in the thawed permafrost, indicating a shift of methanogens from hydrogenotrophic to partly acetoclastic methane-generating metabolic processes. In addition, data also show the soil texture and features change as a result of microbial reproduction and activity induced by this permafrost thaw. Those data indicate that microbial ecology under warming permafrost has potential impacts on ecosystem and methane emissions.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Allen, M.F.

A freeze-thaw power cycle is described that includes a piston driven by the expansion power of a fluid such as water in a cyclinder on freezing and the thawing thereof with alternate, rapid freezing and thawing of the fluid by low and high temperature means with heat transfer rates facilitated by the use of heat pipes or tubes or other conductor means to produce a continuous single or multi cyclinder quick, polution free and noiseless freeze/thaw power cycle.

Thaw flow control for liquid heat transport systems

DOEpatents

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.

Permafrost thaw and wildfire: Equally important drivers of boreal tree cover changes in the Taiga Plains, Canada

NASA Astrophysics Data System (ADS)

Helbig, M.; Pappas, C.; Sonnentag, O.

2016-02-01

Boreal forests cover vast areas of the permafrost zones of North America, and changes in their composition and structure can lead to pronounced impacts on the regional and global climate. We partition the variation in regional boreal tree cover changes between 2000 and 2014 across the Taiga Plains, Canada, into its main causes: permafrost thaw, wildfire disturbance, and postfire regrowth. Moderate Resolution Imaging Spectroradiometer Percent Tree Cover (PTC) data are used in combination with maps of historic fires, and permafrost and drainage characteristics. We find that permafrost thaw is equally important as fire history to explain PTC changes. At the southern margin of the permafrost zone, PTC loss due to permafrost thaw outweighs PTC gain from postfire regrowth. These findings emphasize the importance of permafrost thaw in controlling regional boreal forest changes over the last decade, which may become more pronounced with rising air temperatures and accelerated permafrost thaw.

Discovery of a novel methanogen prevalent in thawing permafrost.

PubMed

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.

A practical algorithm to estimate soil thawing onset with the soil moisture active passive (SMAP) data

NASA Astrophysics Data System (ADS)

Chen, X.; Liu, L.

2016-12-01

The Soil Moisture Active Passive (SMAP) satellite simultaneously collected active and passive microwave data at L-band from April to July, 2015. The L-band radiometer brightness temperature (TB) data are strongly sensitive to the change of soil moisture, therefore, can be used to estimate freeze/thaw state of soil. We applied an edge detection method to detect the onset of thawing based on the SMAP level-1C TB data. This method convolves the first derivative of the Gaussian function as a kernel with the TB time series. When thawing occurs, soil moisture increases abruptly and leads to a decrease in TB. Therefore, a primary thaw event can be identified when the convolved signal reaches a local minimum. Considering the noise of the radiometer data, not all local minimums correspond to a thaw event. Therefore, we further applied a filter based on a priori or in situ soil temperature observation to eliminate false events. We compared the TB-based estimates with in situ measurements of soil temperature, moisture, and snow depth from April to June from 5 SNOTEL sites in Alaska. Our results show that at 4 out of the 5 sites the estimated thawing onsets and in-situ data agree within 5 to 10 days. However, we found a distinct inconsistency of 41 days at the fifth site. One possible reason is the mismatch in spatial coverage: one pixel of SMAP radiometer data has a size of 36 km, within which different areas may have different freeze/thaw states. The SMAP radar backscatter coefficient (σ0) data are also very sensitive to soil moisture, and has finer spatial resolution of 1 km, making it more directly comparable with the in situ measurements. We applied a seasonal threshold method to estimate thawing onset based on this data. Firstly, we set a thaw onset based on the in situ soil temperature and moisture measurements at 5 cm depth. Then we averaged σ0 observations from April 14th to 7 days before the thaw onset to represent the frozen soil, and used the mean value from 7 days after the thawing onset to June 1st as thawed reference. Next, the σ0-based freeze/thaw distribution within radiometer pixel can be obtained. Assuming TB and have a linear relationship in 36 km scale during a short time, SMAP provide a down scaling method to obtain 9 km resolution TB data. For further work, we plan to apply the edge detection method on this TB data to estimate the soil state in 9 km.

The effects of freezing and thawing rates on tenderness, sensory quality, and retail display of beef subprimals.

PubMed

Hergenreder, J E; Hosch, J J; Varnold, K A; Haack, A L; Senaratne, L S; Pokharel, S; Beauchamp, C; Lobaugh, B; Calkins, C R

2013-01-01

The objective of this study was to evaluate processing methods for frozen beef subprimals; the effects of freezing and thawing rates on tenderness, sensory properties, and retail display were evaluated. There were 6 treatments: fresh, never frozen 14 d wet aged (14D); fresh, never frozen 21 d wet aged (21D); blast frozen-fast thawed (BF); blast frozen-slow thawed (BS); conventionally frozen-fast thawed (CF); and conventionally frozen-slow thawed (CS). All frozen beef subprimals were aged for 14 d before freezing. Three beef subprimal cuts, rib eye roll (n=90), strip loin (n=90), and top sirloin butt (n=90), were used with 3 replications of 5 samples per treatment per week (total of 9 wk, n=270). Blast freezing occurred by placing spacers between the boxes of meat on pallets at -28°C with high air velocity for 3 to 5 d. Conventional freezing occurred with boxes of meat stacked on pallets and placed in a -28°C freezer with minimal air movement for at least 10 d. Fast thawing of subprimals (to an internal temperature of -1°C to 1°C) occurred by immersion in a circulating water bath (<12°C) for 21 h, and slow thawing of subprimals occurred over a 2-wk period by placing individual subprimals on tables at 0°C. Steaks (2.5 cm thick) were cut from the longissimus thoracis (LT), longissimus lumborum (LL), and gluteus medius (GM) for Warner-Bratzler shear force (WBS), trained sensory evaluation, and retail display. For LL and GM beef steaks, frozen treatments were equal or lower in WBS values to 14D and 21D beef steaks. No differences were detected in WBS among the treatments applied to GM beef steaks (P=0.08). There were no differences in sensory tenderness among the LL, LT, and GM (P>0.05). All LL and LT beef steaks had approximately 4 d to 40% discoloration, and all GM steaks had over 3 d to 40% discoloration. Steaks from the LL and LT began to discolor at about 3 d, and the GM began to discolor after 1 d. For all beef subprimals, purge loss during storage and thawing was significantly greater for the slow-thawed subprimals (P<0.01), and all fast-thawed subprimals were equal or superior to 14D and 21D (P<0.01) in storage and thawing purge. During retail display, the greatest purge loss occurred in fast-thawed treatments (P<0.01). Overall, freezing rate did not affect purge loss, and neither freezing nor thawing rates had significant meaningful effects on WBS, and sensory properties were comparable with fresh, never-frozen subprimals.

Studies on the intracellular Ca2+ concentration of thawed dog spermatozoa: influence of Equex from different sources, two thawing diluents and post-thaw incubation in capacitating conditions.

PubMed

Peña, A I; López-Lugilde, L; Barrio, M; Becerra, J J; Quintela, L A; Herradón, P G

2003-02-01

The addition of 0.5% (v/v) of Equex STM Paste (Nova Chemical Sales, Scituate Inc., MA, USA), whose active ingredient is sodium dodecyl sulphate (SDS), to a Tris-egg yolk extender was demonstrated to improve the longevity of frozen-thawed dog spermatozoa during in vitro incubation at 38 degrees C. The aim of the first experiment was to compare the effects of two SDS-containing compounds, Equex STM Paste and Equex Pasta (Minitüb, Tiefenbach, Germany), when added to a Tris-egg yolk based extender, on the post-thaw longevity of dog spermatozoa, as well as on the intracellular Ca2+ concentration of spermatozoa, during post-thaw incubation at 38 degrees C. The post-thaw sperm survival and longevity, as well as the quality of the sperm movement, were significantly better when using Equex STM Paste. Such prolonged sperm longevity, however, was associated to a higher intracellular Ca2+ concentration in a large subpopulation of the live spermatozoa. A second experiment was aimed to evaluate the effects of sperm dilution immediately post-thaw with a Tris buffer containing glucose or fructose. The two Tris buffers were no different for any of the sperm parameters studied. The aim of a third experiment was to evaluate the sperm longevity, motility patterns and intracellular Ca2+ concentration of cryopreserved dog spermatozoa during post-thaw incubation in capacitating conditions [canine capacitating medium (CCM) with or without 5 microg/ml of heparin]. Heparin had no significant effects on any of the sperm parameters evaluated. During the first 8 h of incubation, the majority of the live spermatozoa had a high intracellular Ca2+ content. However, after 8-10 h of incubation, it had significantly declined. The highest proportion of fast motile sperm, and the highest curvilinear velocity, average path velocity and amplitude of lateral head displacement for the total motile sperm were observed during the 2-4-h incubation period. It was concluded that: (a) the addition of 0.5% (v/v) of Equex STM Paste to a Tris-egg yolk based extender significantly improved the post-thaw longevity of dog spermatozoa, but the same concentration of Equex Pasta had no significant beneficial effects; (b) sperm dilution after thawing with a Tris buffer containing glucose or fructose made no difference in post-thaw sperm longevity; (c) the addition of 5 microg/ml of heparin to CCM had no significant capacitating effects on frozen-thawed dog spermatozoa.

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

PubMed

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 < 0.0001) and extender (P = 0.10) and time (P < 0.0001) for VIA. Experiment 2 evaluated the effect of temperature and time of thawing on in vitro fertility at intervals after thawing. Straws (0.5 mL) from different 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 < 0.0001) and an effect of time of evaluation (P < 0.0001) on MOT and ACR. Thawing at 70 °C for 20 seconds reduced (P < 0.05) MOT, VIA, and ACR compared to other treatments. Experiment 3 tested the effects of storage temperature and time after thawing using 20 ejaculates. Samples were thawed, diluted, and allotted to storage at 17 °C, 26 °C, or 37 °C with evaluation at 2, 6, 12, and 24 hours. There was a storage temperature and time effect and an interaction for MOT and VIA (P < 0.0001). Storage at 17 °C and 26 °C increased (P < 0.05) MOT over all times (38.5%) compared to 37 °C (26%), whereas MOT was reduced at intervals. Viability was also greatest with 17 °C and 26 °C compared to 37 °C and was also affected by time and decreased with time. These results indicate that FTS can be held at 17 °C or 26 °C for up to 2 hours before use and would allow for preparation of multiple doses. These data suggest in vitro fertility of FTS is affected by extenders, thawing, and storage. Copyright © 2015 Elsevier Inc. All rights reserved.

Geophysical imaging and thermal modeling of subsurface morphology and thaw evolution of discontinuous permafrost

NASA Astrophysics Data System (ADS)

McClymont, Alastair F.; Hayashi, Masaki; Bentley, Laurence R.; Christensen, Brendan S.

2013-09-01

our current understanding of permafrost thaw in subarctic regions in response to rising air temperatures, little is known about the subsurface geometry and distribution of discontinuous permafrost bodies in peat-covered, wetland-dominated terrains and their responses to rising temperature. Using electrical resistivity tomography, ground-penetrating radar profiling, and thermal-conduction modeling, we show how the land cover distributions influence thawing of discontinuous permafrost at a study site in the Northwest Territories, Canada. Permafrost bodies in this region occur under forested peat plateaus and have thicknesses of 5-13 m. Our geophysical data reveal different stages of thaw resulting from disturbances within the active layer: from widening and deepening of differential thaw features under small frost-table depressions to complete thaw of permafrost under an isolated bog. By using two-dimensional geometric constraints derived from our geophysics profiles and meteorological data, we model seasonal and interannual changes to permafrost distribution in response to contemporary climatic conditions and changes in land cover. Modeling results show that in this environment (1) differences in land cover have a strong influence on subsurface thermal gradients such that lateral thaw dominates over vertical thaw and (2) in accordance with field observations, thaw-induced subsidence and flooding at the lateral margins of peat plateaus represents a positive feedback that leads to enhanced warming along the margins of peat plateaus and subsequent lateral heat conduction. Based on our analysis, we suggest that subsurface energy transfer processes (and feedbacks) at scales of 1-100 m have a strong influence on overall permafrost degradation rates at much larger scales.

Metagenomic analysis of a permafrost microbial community reveals a rapid response to thaw

USGS Publications Warehouse

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

2011-01-01

Permafrost contains an estimated 1672????????Pg carbon (C), an amount roughly equivalent to the total currently contained within land plants and the atmosphere. This reservoir of C is vulnerable to decomposition as rising global temperatures cause the permafrost to thaw. During thaw, trapped organic matter may become more accessible for microbial degradation and result in greenhouse gas emissions. Despite recent advances in the use of molecular tools to study permafrost microbial communities, their response to thaw remains unclear. Here we use deep metagenomic sequencing to determine the impact of thaw on microbial phylogenetic and functional genes, and relate these data to measurements of methane emissions. Metagenomics, the direct sequencing of DNA from the environment, allows the examination of whole biochemical pathways and associated processes, as opposed to individual pieces of the metabolic puzzle. Our metagenome analyses reveal that during transition from a frozen to a thawed state there are rapid shifts in many microbial, phylogenetic and functional gene abundances and pathways. After one week of incubation at 5 ??C, permafrost metagenomes converge to be more similar to each other than while they are frozen. We find that multiple genes involved in cycling of C and nitrogen shift rapidly during thaw. We also construct the first draft genome from a complex soil metagenome, which corresponds to a novel methanogen. Methane previously accumulated in permafrost is 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. ?? 2011 Macmillan Publishers Limited. All rights reserved.

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

PubMed

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.

Embolism Formation during Freezing in the Wood of Picea abies1

PubMed Central

Mayr, Stefan; Cochard, Hervé; Améglio, Thierry; Kikuta, Silvia B.

2007-01-01

Freeze-thaw events can cause embolism in plant xylem. According to classical theory, gas bubbles are formed during freezing and expand during thawing. Conifers have proved to be very resistant to freeze-thaw induced embolism, because bubbles in tracheids are small and redissolve during thawing. In contrast, increasing embolism rates upon consecutive freeze-thaw events were observed that cannot be explained by the classical mechanism. In this study, embolism formation during freeze-thaw events was analyzed via ultrasonic and Cryo-scanning electron microscope techniques. Twigs of Picea abies L. Karst. were subjected to up to 120 freeze-thaw cycles during which ultrasonic acoustic emissions, xylem temperature, and diameter variations were registered. In addition, the extent and cross-sectional pattern of embolism were analyzed with staining experiments and Cryo-scanning electron microscope observations. Embolism increased with the number of freeze-thaw events in twigs previously dehydrated to a water potential of −2.8 MPa. In these twigs, acoustic emissions were registered, while saturated twigs showed low, and totally dehydrated twigs showed no, acoustic activity. Acoustic emissions were detected only during the freezing process. This means that embolism was formed during freezing, which is in contradiction to the classical theory of freeze-thaw induced embolism. The clustered pattern of embolized tracheids in cross sections indicates that air spread from a dysfunctional tracheid to adjacent functional ones. We hypothesize that the low water potential of the growing ice front led to a decrease of the potential in nearby tracheids. This may result in freezing-induced air seeding. PMID:17041033

Transformation of Upland Water and Carbon Dynamics by Thawing Permafrost in the Alaskan Interior

NASA Astrophysics Data System (ADS)

Ewing, S. A.; Paces, J. B.; O'Donnell, J. A.; Kanevskiy, M. Z.; Shur, Y.; Jorgenson, M. T.; Harden, J.; Aiken, G. R.; Striegl, R.

2009-05-01

Large arctic rivers can provide an integrated signal of regional permafrost thaw and associated carbon dynamics. A long-term (30-y) decrease in dissolved organic carbon (DOC) and increase in dissolved inorganic carbon in the Yukon River Basin (YRB) suggest increased flow through mineral soils as a result of permafrost thaw. We used U series isotopes to test for the influence of thaw on soil and surface waters in small upland catchments at two sites within the YRB. In natural waters, 234U/238U activity ratios exceed 1.00 (secular equilibrium) as a function of water-rock contact time. Previous work has shown that in major YRB rivers, seasonally and spatially variable 234U/238U ratios could indicate both groundwater inputs and permafrost thaw, with ratios ranging from 1.1 to 2.6. We show that 234U/238U ratios in soil and surface water from these small catchments span the range of values observed in the major rivers, and indicate greater influence of older water where the mineral soil and underlying sediment facilitate drainage and permafrost degradation. Analysis of deep, ice-rich loess permafrost cores (2-10 m) reveals that thaw of Pleistocene ice can release high concentrations of DOC (>1000 ppm) and ammonium in thaw waters. The age and chemical composition of these waters allows for improved prediction of downstream carbon dynamics upon thaw. Field observation of hillslope soil sequences indicates that both topography and mineral substrate influence the effects of thaw on water and carbon dynamics in small catchments.

The transcriptional response of microbial communities in thawing Alaskan permafrost soils

PubMed Central

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

The effect of cryopreservation on goat semen characteristics related to sperm freezability.

PubMed

Dorado, J; Muñoz-Serrano, A; Hidalgo, M

2010-08-01

Seminal quality parameters were used to evaluate the effect of freeze-thawing procedure on goat sperm characteristics, and to relate possible changes in sperm parameters to cryopreservation success. Semen samples (n=110) were frozen with TRIS and milk-based extenders and thawed. Sperm quality parameters (motility, morphology and acrosome) were compared between fresh and frozen-thawed samples. Sperm freezability was judged by classifying the semen samples as "suitable" or "not suitable" according to the sperm quality parameters assessed before and after thawing. Fertility data was obtained after cervical insemination with frozen semen doses. The ejaculates were grouped into two categories according to their fertility results. In experiment 1, significant differences were found between semen extenders (P<0.001), bucks (P<0.05) and ejaculates within the same male (P<0.05) in terms of sperm quality. There was no seasonal effect (P>0.05) on the majority of the sperm parameters assessed after thawing. Moreover, significant differences (P<0.001) in semen parameters assessed in fresh semen and frozen-thawed samples were found between groups. The effect of the freeze-thawing procedure on sperm quality parameters was also different (P<0.05) between extenders within the same group. The number of sperm quality parameters that had changed after cryopreservation was lower in "suitable" semen samples before and after thawing. In experiment 2, no differences (P>0.05) in semen parameters assessed in fresh semen and frozen-thawed samples were found between groups. The effect of freezing and thawing on sperm quality parameters were different (P<0.05) between extenders within the same group. Only mean beat cross frequency (BCF) values were significantly higher (P<0.05) in TRIS diluted samples that led to successful pregnancies after artificial insemination. In conclusion, CASA-derived motility parameters, together with traditional semen assessment methods, give valuable information on sperm quality before and after freezing. Therefore, the identification of ejaculates as "good" or "bad" based on fresh and post-thaw semen parameters studied in the present experiment were good indicators of goat semen freezability, although the fertilizing capacity of frozen-thawed goat spermatozoa are not revealed by this quality study. Copyright (c) 2010 Elsevier B.V. All rights reserved.

The Canadian Experiment for Freeze/Thaw in 2012 or 2013 CanEx-FT12 or FT13

NASA Technical Reports Server (NTRS)

Belair, Stephane; Bernier, Monique; Colliander, Andreas; Jackson, Thomas; McDonald, Kyle; Walker, Anne

2011-01-01

General objectives of the experiment are: Pre-launch Calibration/Validation of SMAP Freeze/Thaw products and retrieval algorithms and rehearsal for Soil Moisture Active-Passive (SMAP) post launch validation. The basis of the radar freeze-thaw measurement is the large shift in dielectric constant and backscatter (dB) between predominantly frozen & thawed conditions. The Dielectric constant of liquid water varies with frequency, whereas that of pure ice is constant

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

PubMed

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 < 0.05) when the thawing extender was supplemented with 12.5, 25 and 50 mg CLC/500 × 10(6) sperm. Nevertheless, motility decreased (p < 0.05) when the concentration of CLC exceeded 12.5 mg CLC/500 × 10(6) sperm. In conclusion, our results suggest that the supplementation of thawing extenders with CLC improves sperm viability and reduces acrosome damage after freezing/thawing. © 2014 Blackwell Verlag GmbH.

Climate Change and Thawing Permafrost in Two Iñupiaq Communities of Alaska's Arctic: Observations, Implications, and Resilience

NASA Astrophysics Data System (ADS)

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.

Freezing/Thawing without Cryoprotectant Damages Native but not Decellularized Porcine Renal Tissue

PubMed Central

Poornejad, Nafiseh; Frost, Timothy S; Scott, Daniel R; Elton, Brinden B; Reynolds, Paul R; Roeder, Beverly L; Cook, Alonzo D

2015-01-01

abstract Whole organ decellularization of porcine renal tissue and recellularization with a patient's own cells would potentially overcome immunorejection, which is one of the most significant problems with allogeneic kidney transplantation. However, there are obstacles to achieving this goal, including preservation of the decellularized extracellular matrix (ECM), identifying the proper cell types, and repopulating the ECM before transplantation. Freezing biological tissue is the best option to avoid spoilage; however, it may damage the structure of the tissue or disrupt cellular membranes through ice crystal formation. Cryoprotectants have been used to repress ice formation during freezing, although cell toxicity can still occur. The effect of freezing/thawing on native (n = 10) and decellularized (n = 10) whole porcine kidneys was studied without using cryoprotectants. Results showed that the elastic modulus of native kidneys was reduced by a factor of 22 (P < 0.0001) by freezing/thawing or decellularization, while the elastic modulus for decellularized ECM was essentially unchanged by the freezing/thawing process (p = 0.0636). Arterial pressure, representative of structural integrity, was also reduced by a factor of 52 (P < 0.0001) after freezing/thawing for native kidneys, compared to a factor of 43 (P < 0.0001) for decellularization and a factor of 4 (P < 0.0001) for freezing/thawing decellularized structures. Both freezing/thawing and decellularization reduced stiffness, but the reductions were not additive. Investigation of the microstructure of frozen/thawed native and decellularized renal tissues showed increased porosity due to cell removal and ice crystal formation. Orcein and Sirius staining showed partial damage to elastic and collagen fibers after freezing/thawing. It was concluded that cellular damage and removal was more responsible for reducing stiffness than fibril destruction. Cell viability and growth were demonstrated on decellularized frozen/thawed and non-frozen samples using human renal cortical tubular epithelial (RCTE) cells over 12 d. No adverse effect on the ability to recellularize after freezing/thawing was observed. It is recommended that porcine kidneys be frozen prior to decellularization to prevent contamination, and after decellularization to prevent protein denaturation. Cryoprotectants may still be necessary, however, during storage and transportation after recellularization. PMID:25730294

Isolation of a Soluble Component of Plasmodium berghei Which Induces Immunity in Rats

PubMed Central

Grothaus, G. D.; Kreier, J. P.

1980-01-01

Soluble material was obtained from sonically freed plasmodiae by three procedures. Two procedures, cryo-impacting and freeze-thawing, were evaluated for their ability to disrupt the parasites and release soluble material. The soluble materials obtained by these procedures were compared to materials washed from the surfaces of sonically freed parasites. Between 35 and 40% of the total parasite protein was solubilized by freeze-thawing or cryo-impacting. One cycle of freeze-thawing released nearly as much protein as could be released by this method, and additional cycles of freeze-thawing had little additional effect. Cryo-impacting solubilized only a small amount of protein in addition to that which was released by the cycle of freeze-thawing inherent in the procedure. Reductions in the packed cell volume of the material remaining after freeze-thawing or cryo-impacting indicate that the insoluble fragments are broken into smaller pieces as treatment is extended. Electron microscopy of 30-s cryo-impacted and three-times freeze-thawed parasites revealed membrane fragments similar in appearance. Patterns obtained by polyacrylamide gel electrophoresis of the soluble material from freeze-thawed and cryo-impacted parasites were also similar, and approximately 13 protein bands were demonstrated. The material washed from the surfaces of the free parasites, on the other hand, resolved into only two to four major bands on the gel columns. In immunization studies, the soluble and insoluble fractions obtained by freeze-thawing or cryo-impacting and the material washed from the surfaces of the parasites all stimulated a protective immune response. On the basis of the amount of protein required to stimulate roughly comparable immunity, the soluble fraction obtained by freeze-thawing or cryo-impacting free parasites was about twice as potent an immunogen as was the insoluble fraction. The material obtained by gentle washing of the freed parasites was approximately 20 times as potent an immunogen as were the freed parasites and about 7 times as potent as the soluble material obtained by freeze-thawing or cryo-impacting. Images Fig. 1 Fig. 2 PMID:6991439

Influence of freeze-thaw events on carbon dioxide emission from soils at different moisture and land use.

PubMed

Kurganova, Irina; Teepe, Robert; Loftfield, Norman

2007-02-19

The repeated freeze-thaw events during cold season, freezing of soils in autumn and thawing in spring are typical for the tundra, boreal, and temperate soils. The thawing of soils during winter-summer transitions induces the release of decomposable organic carbon and acceleration of soil respiration. The winter-spring fluxes of CO2 from permanently and seasonally frozen soils are essential part of annual carbon budget varying from 5 to 50%. The mechanisms of the freeze-thaw activation are not absolutely clear and need clarifying. We investigated the effect of repeated freezing-thawing events on CO2 emission from intact arable and forest soils (Luvisols, loamy silt; Central Germany) at different moisture (65% and 100% of WHC). Due to the measurement of the CO2 flux in two hours intervals, the dynamics of CO2 emission during freezing-thawing events was described in a detailed way. At +10 degrees C (initial level) in soils investigated, carbon dioxide emission varied between 7.4 to 43.8 mg C m-2h-1 depending on land use and moisture. CO2 flux from the totally frozen soil never reached zero and amounted to 5 to 20% of the initial level, indicating that microbial community was still active at -5 degrees C. Significant burst of CO2 emission (1.2-1.7-fold increase depending on moisture and land use) was observed during thawing. There was close linear correlation between CO2 emission and soil temperature (R2 = 0.86-0.97, P < 0.001). Our investigations showed that soil moisture and land use governed the initial rate of soil respiration, duration of freezing and thawing of soil, pattern of CO2 dynamics and extra CO2 fluxes. As a rule, the emissions of CO2 induced by freezing-thawing were more significant in dry soils and during the first freezing-thawing cycle (FTC). The acceleration of CO2 emission was caused by different processes: the liberation of nutrients upon the soil freezing, biological activity occurring in unfrozen water films, and respiration of cold-adapted microflora.

Freeze-thaw performance testing of whole concrete railroad ties.

DOT National Transportation Integrated Search

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...

Influence of Thawing Methods and Storage Temperatures on Bacterial Diversity, Growth Kinetics, and Biogenic Amine Development in Atlantic Mackerel.

PubMed

Onyango, S; Palmadottir, H; Tómason, T; Marteinsson, V T; Njage, P M K; Reynisson, E

2016-11-01

Limited knowledge is currently available on the influence of fish thawing and subsequent storage conditions on bacterial growth kinetics, succession, and diversity alongside the production of biogenic amines. This study aimed to address these factors during the thawing and subsequent storage of mackerel. Thawing was either done fast in 18°C water for 2 h or slowly at 30°C overnight. Subsequent storage was at 30°C (ambient) for 36 h and 2 to 5°C (refrigerated) for 12 days. The cultivation methods used were total viable counts, hydrogen sulfide-producing bacteria, and Pseudomonas . Maximum growth rate, population density, and lag time were fitted on the counts using the Baranyi model. The bacterial diversity and succession were based on sequencing of 16S rRNA amplicons, and biogenic amines were quantified on high-pressure liquid chromatography-UV. The results show that lag time of hydrogen sulfide-producing bacteria was significantly affected by both thawing methods, and further, the interaction between thawing and storage significantly affected the maximum growth rate of these bacteria. However, the maximum growth rate of Pseudomonas was higher during refrigerated storage compared with storage at ambient temperature. Total viable counts showed longer lag time and reduced growth rate under refrigerated storage. Higher bacterial diversity was correlated to slow thawing and storage at ambient temperature compared with slow thawing and refrigerated storage. Overall, Acinetobacter and Psychrobacter genera were the dominant bacterial populations. The amine levels were low and could not be differentiated along the thawing and storage approaches, despite a clear increase in bacterial load, succession, and diversity. This corresponded well with the low abundance of biogenic amine-producing bacteria, with the exception of the genus Proteus , which was 8.6% in fast-thawed mackerel during storage at ambient temperature. This suggests that the decarboxylation potential is dependent on both microbial load and microbial community structure.

Temperature sensitivity of methanogenesis in a thermokarst lake sediment core

NASA Astrophysics Data System (ADS)

Heslop, J. K.; Walter Anthony, K. M.; Grosse, G.; Anthony, P.; Bondurant, A.

2016-12-01

Little is known about temperature sensitivity of permafrost organic carbon (OC) mineralization over time scales of years to centuries following thaw. Due to their formation and thaw histories, taliks (thaw bulbs) beneath thermokarst lakes provide a unique natural laboratory from which to examine how permafrost thawed in saturated anaerobic conditions responds to changes in temperature following long periods of time since thaw. We anaerobically incubated samples from a 590 cm thermokarst lake sediment core near Fairbanks, Alaska at four temperatures (0, 3, 10, and 25 ºC) bracketing observed talik temperatures. We show that since initial thaw 400 yr BP CH4 production shifts from being most sensitive to at lower (0-3 ºC; Q10-EC=1.15E7) temperatures to being most sensitive at higher (10-25 ºC; Q10-EC=67) temperatures. Frozen sediments collected from beneath the talik, thawed at the commencement of the incubation, had significant (p ≤ 0.05) increases in CH4 production rates at lower temperatures but did not show significant CH4 production rate increases at higher temperatures (10-25 ºC). We hypothesize the thawing of sediments removed a major barrier to C mineralization, leading to rapid initial permafrost C mineralization and preferential mineralization of the most biolabile OC compounds. In contrast, sediments which had been thawed beneath the lake for longer periods of time did not experience statistically significant increases in CH4 production at lower temperatures (0-10 ºC), but had high temperature sensitivities at higher temperatures (10-25 ºC). We believe these rate increases are due to warmer temperatures in the experimental incubations crossing activation energy thresholds, allowing previously recalcitrant fractions of OC to be utilized, and/or the presence of different microbial communities adapted to thawed sediments. Recently-deposited sediments at shallow depths in the lake core experienced increases in CH4 production across all incubation temperatures (Q10-ST=4.4).

Influence of freeze-thawing on hyaluronic acid binding of human spermatozoa.

PubMed

Nijs, Martine; Creemers, Eva; Cox, Annemie; Janssen, Mia; Vanheusden, Elke; Castro-Sanchez, Yovanna; Thijs, Herbert; Ombelet, Willem

2009-08-01

Mature human spermatozoa have at least three specific hyaluronic acid (HA) binding proteins present on their sperm membrane. These receptors play a role in the acrosome reaction, hyaluronidase activity, hyaluronan-mediated motility and sperm-zona and sperm-oolemmal binding. Cryopreservation of spermatozoa can cause ultrastructural and even molecular damage. The aim of this study was to investigate if HA binding receptors of human spermatozoa remain functional after freeze-thawing. Forty patients were enrolled in the study. Semen samples were analysed before and after cryopreservation. Parameters analysed included concentration, motility, morphology and hyaluronan binding. Samples were frozen in CBS straws using a glycerol-glucose-based cryoprotectant. HA binding was studied using the sperm-hyaluronan binding assay. Freeze-thawing resulted in a significant decline in motility: the percentage of motile spermatozoa reduced from 50.6 to 30.3% (P < 0.001). HA binding properties of frozen-thawed spermatozoa remained unchanged after the freeze-thawing process: 68.5 +/- 17.1% spermatozoa of the neat sample were bound to HA, as were 71.3 +/- 20.4 of the frozen-thawed sample. This study indicates that freeze-thawing did not alter the functional hyaluronan binding sites of mature motile spermatozoa, and therefore will not alter their fertilizing potential.

Effect of wildfire and fireline construction on the annual depth of thaw in a black spruce permafrost forest in interior Alaska: a 36-year record of recovery

Treesearch

Leslie A. Viereck; Nancy R. Werdin-Pfisterer; Phyllis C. Adams; Kenji Yoshikawa

2008-01-01

Maximum thaw depths were measured annually in an unburned stand, a heavily burned stand, and a fireline in and adjacent to the 1971 Wickersham fire. Maximum thaw in the unburned black spruce stand ranged from 36 to 52 cm. In the burned stand, thaw increased each year to a maximum depth of 302 cm in 1995. In 1996, the entire layer of seasonal frost remained, creating a...

Dog Sperm Cryopreservation in Glucose-Fructose or Sucrose Supplemented Glycerol-Free Tris: Effect of Post-Thaw Incubation Time on Gene Expression Related to Apoptosis and Motility.

PubMed

Rahman, M A; Park, S H; Yu, I J

　　OBJECTIVE: The study evaluated the effects of glucose-fructose or sucrose supplementation in glycerol-free Tris (GFT) solution on motility, viability, the level of reactive oxygen species (ROS), as well as the level of apoptosis (BAX and BCL2) and motility (SMCP)-related gene expression of dog spermatozoa. Spermatozoa (5×10 7 sperm/ml) were cryopreserved in GFT containing 86 mM glucose and 86 mM fructose (GF-GFT) or 100 mM sucrose (S-GFT). Progressive motility, viability, ROS (H 2 O 2 ) level and mRNA gene expression of spermatozoa were evaluated 0 h, 3 h or 6 h post-thaw at 24°C. The motility of spermatozoa cryopreserved in GF-GFT was increased throughout the post-thaw incubation time. The motility of spermatozoa cryopreserved in S-GFT was increased at 3 h of post-thaw incubation. The sperm ROS level in the GF-GFT group was inconsistent during the post-thaw incubation time; however, the ROS level in the S-GFT group was gradually increased with progression of the post-thaw incubation period. The post-thaw incubation had no substantial effect on the mRNA expression of the BAX, BCL2, and SMCP genes of spermatozoa in both the GF-GFT and S-GFT groups. The supplementation of glucose and fructose improves progressive sperm motility during 6 h of post-thaw incubation while maintaining similar sperm viability. The addition of GF to GFT for cryopreservation and post-thaw incubation would yield more functional spermatozoa for future assisted reproduction practices.

Improvement of post-thaw sperm survivals using liquid nitrogen vapor in a spermcasting oyster Ostrea angasi.

PubMed

Hassan, Md Mahbubul; Li, Xiaoxu; Qin, Jian G

2017-10-01

Low survival of cryopreserved sperm impedes the application of cryopreservation technique in spermcasting oyster species. This study developed a simple method of liquid nitrogen vapor freezing to improve post-thaw sperm survival in the spermcasting oyster Ostrea angasi. The results indicate that the permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG) were non-toxic to sperm up to 20% concentration and 90 min exposure whereas methanol at 10% or higher was toxic to sperm for any exposure over 30 min. Among the treatments with permeable cryoprotectants, 15% EG produced the highest post-thaw sperm motility. Sperm motility was further improved by the addition of non-permeable cryoprotectants (trehalose and glucose), with 15% EG + 0.2 M trehalose resulting in the highest post-thaw sperm motility among all the combinations evaluated. The durations of 20, 30 and 60 min equilibrations produced a higher post-thaw sperm motility and plasma membrane integrity (PMI) than 10 min. Higher post-thaw motility and PMI were achieved by freezing sperm at the 8 cm height from the liquid nitrogen surface than at the 2, 4, 6, 10 or 12 cm height. Holding sperm for 10 min in liquid nitrogen vapor produced higher post-thaw motility and PMI than for 2, 5 or 20 min. The cryopreservation protocol developed in this study improved both post-thaw motility and PMI of O. angasi sperm at least 15% higher than those cryopreserved using programmable freezing method. Liquid nitrogen vapor freezing might have greater applicability in improving post-thaw sperm quality of spermcasting oyster species. Copyright © 2017 Elsevier Inc. All rights reserved.

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

PubMed

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.

Colloid-facilitated mobilization of metals by freeze-thaw cycles.

PubMed

Mohanty, Sanjay K; Saiers, James E; Ryan, Joseph N

2014-01-21

The potential of freeze-thaw cycles to release colloids and colloid-associated contaminants into water is unknown. We examined the effect of freeze-thaw cycles on the mobilization of cesium and strontium in association with colloids in intact cores of a fractured soil, where preferential flow paths are prevalent. Two intact cores were contaminated with cesium and strontium. To mobilize colloids and metal cations sequestered in the soil cores, each core was subjected to 10 intermittent wetting events separated by 66 h pauses. During the first five pauses, the cores were dried at room temperature, and during last five pauses, the cores were subjected to 42 h of freezing followed by 24 h of thawing. In comparison to drying, freeze-thaw cycles created additional preferential flow paths through which colloids, cesium, and strontium were mobilized. The wetting events following freeze-thaw intervals mobilized about twice as many colloids as wetting events following drying at room temperature. Successive wetting events following 66 h of drying mobilized similar amounts of colloids; in contrast, successive wetting events after 66 h of freeze-thaw intervals mobilized greater amounts of colloids than the previous one. Drying and freeze-thaw treatments, respectively, increased and decreased the dissolved cesium and strontium, but both treatments increased the colloidal cesium and strontium. Overall, the freeze-thaw cycles increased the mobilization of metal contaminants primarily in association with colloids through preferential flow paths. These findings suggest that the mobilization of colloid and colloid-associated contaminants could increase when temperature variations occur around the freezing point of water. Thus, climate extremes have the potential to mobilize contaminants that have been sequestered in the vadose zone for decades.

Pentoxifylline added to freezing or post-thaw extenders does not improve the survival or in vitro fertilising capacity of boar spermatozoa.

PubMed

Gil, María A; Hernandez, Marta; Roca, Jordi; Almiñana, Carmen; Lucas, Xiomara; Cuello, Cristina; Vazquez, Juan M; Martínez, Emilio A

2010-03-01

This study evaluated whether pentoxifylline added to freezing and thawing extenders influenced the function of boar spermatozoa. In Experiment 1, pooled ejaculated sperm-rich fractions were frozen in 0.5 ml straws after dilution in extender supplemented with pentoxifylline to a final concentration of 0, 2, 4, 8, 16 or 32 mM. The addition of 4, 8, 16 and 32 mM pentoxifylline to the freezing extender significantly decreased the progressive and total motility of spermatozoa. The percentage of viable spermatozoa with intact acrosomes as well as the penetration rate and the efficiency of fertilisation were significantly lower in pentoxifylline-treated groups compared with the untreated control. In Experiment 2, a pool of three straws with 'good' post-thaw sperm quality parameters and another three straws with 'poor' sperm quality were diluted in extender with 0, 1, 2, 4, 8, 16 or 32 mM pentoxifylline. Post-thaw samples with both 'good' and 'poor' sperm quality with 0, 2, 4, 8 and 16 mM were used to assess IVF parameters. The addition of pentoxifylline to post-thaw extender did not improve the post-thaw motility or viability of spermatozoa compared with the control. The in vitro penetration was higher (P<0.05) than the control for oocytes fertilised with spermatozoa that were thawed and incubated in extender with 4, 8 and 16 mM pentoxifylline. However, no differences were observed in the efficiency of fertilisation. We conclude that pentoxifylline, as a supplement added to the freezing extender, has a deleterious effect and that it does not improve the survival or in vitro fertilising efficiency of frozen-thawed boar spermatozoa when added after thawing.

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.

PubMed

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.

ROCK inhibitor Y-27632 increases thaw-survival rates and preserves stemness and differentiation potential of human Wharton's jelly stem cells after cryopreservation.

PubMed

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

2010-12-01

The ROCK inhibitor Y-27632 inhibits apoptosis and increases proliferation of frozen-thawed cells. We examined the role of Y-27632 on human umbilical cord Wharton's jelly stem cells (hWJSCs) for (1) thaw-survival (2) proliferation and (3) preservation of stemness and differentiation potential after cryopreservation. hWJSCs were allotted to 4 groups [Gp I: Untreated hWJSC controls; Gp II: Pretreatment with Y-27632 (10 μM) for 24 h before freezing; Gp III: Y-27632 (10 μM) in freezing medium and Gp IV: Pretreatment with Y-27632 (10 μM) for 24 h and inclusion in freezing medium]. All groups were frozen using a rapid freezing method and stored at -196°C in liquid nitrogen for 90 days before evaluation for apoptosis, cell proliferation, stemness and differentiation. After thawing, Groups II, III and IV showed improved cell attachment, increased thaw-survival (live/dead cell counts) and increased cell proliferation (Trypan blue and MTT assay) compared to controls. CD marker stemness profiles, morphology and normal karyotypes were maintained in the treatment groups after thawing and there was no obvious evidence of apoptosis (Annexin V-FITC and TUNEL assays). After thawing, qRT-PCR demonstrated up-regulation of the anti-apoptotic BCL2 gene and down-regulation of the pro-apoptotic BAX gene and cell cycle regulators (P53 and P21) in the treatment groups. Treated frozen-thawed hWJSCs from all groups differentiated into a neuronal phenotype (neuronal morphology and expression of GFAP, β-3 tubulin and SOX2). Increased thaw-survival and retention of stemness and differentiation potential in hWJSCs following cryopreservation is useful for their storage in cord blood banks for future regenerative medicine purposes.

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

PubMed

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.

Actin Cytoskeletal Disruption following Cryopreservation Alters the Biodistribution of Human Mesenchymal Stromal Cells In Vivo

PubMed Central

Chinnadurai, Raghavan; Garcia, Marco A.; Sakurai, Yumiko; Lam, Wilbur A.; Kirk, Allan D.; Galipeau, Jacques; Copland, Ian B.

2014-01-01

Summary Mesenchymal stromal cells have shown clinical promise; however, variations in treatment responses are an ongoing concern. We previously demonstrated that MSCs are functionally stunned after thawing. Here, we investigated whether this cryopreservation/thawing defect also impacts the postinfusion biodistribution properties of MSCs. Under both static and physiologic flow, compared with live MSCs in active culture, MSCs thawed from cryopreservation bound poorly to fibronectin (40% reduction) and human endothelial cells (80% reduction), respectively. This reduction correlated with a reduced cytoskeletal F-actin content in post-thaw MSCs (60% reduction). In vivo, live human MSCs could be detected in murine lung tissues for up to 24 hr, whereas thawed MSCs were undetectable. Similarly, live MSCs whose actin cytoskeleton was chemically disrupted were undetectable at 24 hr postinfusion. Our data suggest that post-thaw cryopreserved MSCs are distinct from live MSCs. This distinction could significantly affect the utility of MSCs as a cellular therapeutic. PMID:25068122

Cultures of Holder-pasteurized donor human milk after use in a neonatal intensive care unit.

PubMed

Cohen, Ronald S; Huang, Chien-Fang Riva; Xiong, Sean C; Sakamoto, Pauline

2012-08-01

Current guidelines state that human milk, once thawed, should be kept in a refrigerator for only 24 hours. We cultured Holder-pasteurized donor human milk (DHM) after thawing and refrigeration under clinical conditions. Bottles of pasteurized DHM were thawed and used in a regional level 3 neonatal intensive care unit (NICU) in standard clinical fashion and kept refrigerated when not in use. Once no longer needed clinically, aliquots were cultured for bacteria. In total, 30 bottles were returned for culture; six were excluded from analysis because human milk fortifier had been added, and two had been left out of the refrigerator. The remaining 22 bottles were culture-negative after having been thawed for 7-122 hours. DHM without additives was culture-negative for 24 hours or longer after thawing and routine NICU handling. These data indicate that unfortified Holder-pasteurized DHM handled appropriately and refrigerated remains sterile for 24 hours after thawing and perhaps longer. Further study is needed to confirm this.

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

PubMed

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

2016-05-01

This work examines the effects of subsequent cycles of freezing-thawing on giant panda (Ailuropoda melanoleuca) sperm morphometry and function, and assesses whether density-gradient centrifugation (DGC) can increase the number of freezing-thawing cycles this sperm can withstand. A sperm sample was collected by electroejaculation from a mature giant panda and subjected to five freezing-thawing cycles. Although repeated freezing-thawing negatively affected (P < 0.05) sperm motility and membrane integrity, in both nonselected and DCG-selected sperm samples, >60% of the sperm cells in both treatments showed acrosome integrity even after the fifth freezing cycle. In fresh semen, the sperm head length was 4.7 μm, the head width 3.6 μm, area 14.3 μm(2) and perimeter length 14.1 μm. The present results suggest that giant panda sperm trends to be resistant to repeated freezing-thawing, even without DGC selection. © 2015 Blackwell Verlag GmbH.

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

PubMed Central

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

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

PubMed

Jongsutjarittam, Nisachon; Charoenrein, Sanguansri

2013-09-12

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

[Cryopreservation of mouse embryos in ethylene glycol-based solutions: a search for the optimal and simple protocols].

PubMed

Luo, Ming-Jiu; Liu, Na; Miao, De-Qiang; Lan, Guo-Cheng; Suo-Feng; Chang, Zhong-Le; Tan, Jing-He

2005-09-01

Although ethylene glycol (EG) has been widely used for embryo cryopreservation in domestic animals, few attempts were made to use this molecule to freeze mouse and human embryos. In the few studies that used EG for slow-freezing of mouse and human embryos, complicated protocols for human embryos were used, and the protocols need to be simplified. Besides, freezing mouse morula with EG as a cryoprotectant has not been reported. In this paper, we studied the effects of embryo stages, EG concentration, duration and procedure of equilibration, sucrose supplementation and EG removal after thawing on the development of thawed mouse embryos, using the simple freezing and thawing procedures for bovine embryos. The blastulation and hatching rates (81.92% +/- 2.24% and 68.56% +/- 2.43%, respectively) of the thawed late compact morulae were significantly (P < 0.05) higher than those of embryos frozen-thawed at other stages. When mouse late compact morulae were frozen with different concentrations of EG, the highest rates of blastocyst formation and hatching were obtained with 1.8mol/L EG. The blastulation rate was significantly higher when late morulae were equilibrated in 1.8 mol/L EG for 10 min prior to freezing than when they were equilibrated for 30 min, and the hatching rate of embryos exposed to EG for 10 min was significantly higher than that of embryos exposed for 20 and 30 min. Both rates of blastocyst formation and hatching obtained with two-step equilibration were higher (P < 0.05) than with one-step equilibration in 1.8 mol/L EG. Addition of sucrose to the EG-based solution had no beneficial effects. On the contrary, an increased sucrose level (0.4 mol/L) in the solution impaired the development of the frozen-thawed embryos. In contrast, addition of 0.1 mol/L sucrose to the propylene glycol (PG)-based solution significantly improved the development of the frozen-thawed embryos. Elimination of the cryoprotectant after thawing did not improve the development of the thawed embryos. The cell numbers were less (P < 0.05) in blastocysts developed from the thawed morulae than in the in vivo derived ones. In summary, embryo stage, EG concentration, duration and procedure of equilibration and sucrose supplementation had marked effects on development of the thawed mouse embryos, and a protocol for cryopreservation of mouse embryos is recommended in which the late morulae are frozen in 1.8 mol/L EG using the simple freezing and thawing procedures of bovine embryos after a two-step equilibration and the embryos can be cultured or transferred without EG removal after thawing.

Transmembrane ion distribution during recovery from freezing in the woolly bear caterpillar Pyrrharctia isabella (Lepidoptera: Arctiidae).

PubMed

Boardman, Leigh; Terblanche, John S; Sinclair, Brent J

2011-08-01

During extracellular freezing, solutes in the haemolymph are concentrated, resulting in osmotic dehydration of the cells, which must be reversed upon thawing. Here, we used freeze tolerant Pyrrharctia isabella (Lepidoptera: Arctiidae) larvae to examine the processes of ion redistribution after thawing. To investigate the effect of the intensity of cold exposure on ion redistribution after thawing, we exposed caterpillars to -14°C, -20°C or -30°C for 35h. To investigate the effect of duration of cold exposure on ion redistribution after thawing, we exposed the caterpillars to -14°C for up to 6 weeks while sampling several time points. The concentrations of Na(+), K(+), Mg(2+) and Ca(2+) were measured after thawing in the haemolymph, fat body, muscle, midgut tissue and hindgut tissue. Being frozen for long durations (>3 weeks) or at low temperatures (-30°C) both result in 100% mortality, although different ions and tissues appear to be affected by each treatment. Both water distribution and ion content changes were detected after thawing, with the largest effects seen in the fat body and midgut tissue. Magnesium homeostasis appears to be vital for post-freeze survival in these larvae. The movement of ions during thawing lagged behind the movement of water, and ion homeostasis was not restored within the same time frame as water homeostasis. Failure to regain ion homeostasis after thawing is therefore implicated in mortality of freeze tolerant insects. Copyright © 2011 Elsevier Ltd. All rights reserved.

Methane bubbling from Siberian thaw lakes as a positive feedback to climate warming.

PubMed

Walter, K M; Zimov, S A; Chanton, J P; Verbyla, D; Chapin, F S

2006-09-07

Large uncertainties in the budget of atmospheric methane, an important greenhouse gas, limit the accuracy of climate change projections. Thaw lakes in North Siberia are known to emit methane, but the magnitude of these emissions remains uncertain because most methane is released through ebullition (bubbling), which is spatially and temporally variable. Here we report a new method of measuring ebullition and use it to quantify methane emissions from two thaw lakes in North Siberia. We show that ebullition accounts for 95 per cent of methane emissions from these lakes, and that methane flux from thaw lakes in our study region may be five times higher than previously estimated. Extrapolation of these fluxes indicates that thaw lakes in North Siberia emit 3.8 teragrams of methane per year, which increases present estimates of methane emissions from northern wetlands (< 6-40 teragrams per year; refs 1, 2, 4-6) by between 10 and 63 per cent. We find that thawing permafrost along lake margins accounts for most of the methane released from the lakes, and estimate that an expansion of thaw lakes between 1974 and 2000, which was concurrent with regional warming, increased methane emissions in our study region by 58 per cent. Furthermore, the Pleistocene age (35,260-42,900 years) of methane emitted from hotspots along thawing lake margins indicates that this positive feedback to climate warming has led to the release of old carbon stocks previously stored in permafrost.

Freeze-thaw and high-voltage discharge allow macromolecule uptake into ileal brush-border vesicles

DOE Office of Scientific and Technical Information (OSTI.GOV)

Donowitz, M.; Emmer, E.; McCullen, J.

1987-06-01

High-voltage discharge or one cycle of freeze-thawing are shown to transiently permeabilize rabbit ileal brush-border membrane vesicles to macromolecules. Uptake of the radiolabeled macromolecule dextran, mol wt 70,000, used as a marker for vesicle permeability, was determined by a rapid filtration technique, with uptake defined as substrate associated with the vesicle and releasable after incubation of vesicles with 0.1% saponin. Dextran added immediately after electric shock (2000 V) or at the beginning of one cycle of freeze-thawing was taken up approximately eightfold compared with control. ATP also was taken up into freeze-thawed vesicles, whereas there was no significant uptake intomore » control vesicles. The increase in vesicle permeability was reversible, based on Na-dependent D-glucose uptake being decreased when studied 5 but not 15 min after electric shock, and was not significantly decreased after completion of one cycle of freeze-thawing. In addition, adenosine 3',5'-cyclic monophosphate and Ca/sup 2 +/-calmodulin-dependent protein kinase activity were similar in control vesicles and vesicles exposed to high-voltage discharge or freeze-thawing. Also, vesicles freeze-thawed with (/sup 32/P)ATP demonstrated increased phosphorylation compared with nonfrozen vesicles, while freeze-thawing did not alter vesicle protein as judged by Coomassie blue staining. These techniques should allow intestinal membrane vesicles to be used for studies of intracellular control of transport processes, for instance, studies of protein kinase regulation of transport.« less

Display stability of fresh and thawed lamb supplemented with vitamin E or sprayed with an antioxidant borage seed extract.

PubMed

Bellés, Marc; Alonso, Verónica; Roncalés, Pedro; Beltrán, Jose A

2018-06-01

The commercialization of thawed lamb packaged in modified atmosphere and maintained on display could serve as an alternative capable of satisfying the requirements of both customers and distributors. However, previous studies have suggested that lipid oxidation may accelerate post-thawing because peroxidation occurs during frozen storage, thereby leading to rapid and severe secondary lipid oxidation. The addition of an antioxidant compound either in the lamb diet or in the packaged meat could resolve this problem. Therefore, the present study aimed to compare the effect of dietary vitamin E (1000 mg of dl-α-tocopheryl acetate per kg of basal diet) and the spraying of borage seed aqueous extract (10% p/v) on the quality of fresh and thawed lamb leg chops. Both borage extract and vitamin E improved colour (as measured via instrumental and visual assessment of colour) and lipid stability (thiobarbituric acid reactive substances) of fresh and thawed lamb throughout display, although neither of them had any antimicrobial effect. Freezing/thawing accelerated bone marrow darkening and reduced redness but delayed microbial growth. Both of these antioxidant strategies would be very profitable for the preservation of lamb meat, allowing thawed meat packaged in a modified atmosphere to be commercialized. However, additional studies should be carried out to determine how bone darkening in thawed chops can be avoided. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

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

PubMed Central

Jang, Seok-Joon; Rokugo, Keitetsu; Park, Wan-Shin; Yun, Hyun-Do

2014-01-01

This paper provides experimental results to investigate the mechanical properties of sustainable strain-hardening cement composite (2SHCC) for infrastructures after freeze-thaw actions. To improve the sustainability of SHCC materials in this study, high energy-consumptive components—silica sand, cement, and polyvinyl alcohol (PVA) fibers—in the conventional SHCC materials are partially replaced with recycled materials such as recycled sand, fly ash, and polyethylene terephthalate (PET) fibers, respectively. To investigate the mechanical properties of green SHCC that contains recycled materials, the cement, PVA fiber and silica sand were replaced with 10% fly ash, 25% PET fiber, and 10% recycled aggregate based on preliminary experimental results for the development of 2SHCC material, respectively. The dynamic modulus of elasticity and weight for 2SHCC material were measured at every 30 cycles of freeze-thaw. The effects of freeze-thaw cycles on the mechanical properties of sustainable SHCC are evaluated by conducting compressive tests, four-point flexural tests, direct tensile tests and prism splitting tests after 90, 180, and 300 cycles of rapid freeze-thaw. Freeze-thaw testing was conducted according to ASTM C 666 Procedure A. Test results show that after 300 cycles of freezing and thawing actions, the dynamic modulus of elasticity and mass loss of damaged 2SHCC were similar to those of virgin 2SHCC, while the freeze-thaw cycles influence mechanical properties of the 2SHCC material except for compressive behavior. PMID:28788522

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

PubMed

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

2018-07-01

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

Geotextile reinforcement of low-bearing capacity soils: comparison of two design methods applicable to thawing soils

DOT National Transportation Integrated Search

1999-06-01

Thawing fine-grained soils are often saturated and have extremely low bearing capacity. Geosynthetics are used to reinforce unsurfaced roads on weak, saturated soils and therefore are good candidates for use in stabilization of thawing soils. To stab...

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

DOT National Transportation Integrated Search

2015-05-01

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

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

DOT National Transportation Integrated Search

2015-05-01

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

Assessment of the sensitivity of radar backscatter to seasonal snow and vegetation thaw dynamics in a boreal ecosystem

NASA Technical Reports Server (NTRS)

McDonald, K. C.; Qualls, B.; Hardy, J.

2002-01-01

We examine the sensitivity of ERS-1 C-band synthetic aperture radar (SAR) backscatter to springtime snow and vegetation thaw dynamics for boreal forest stands within the BOREAS Southern Study Area (SSA) in Canada during the 1994 winter-spring thaw transition.

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

USDA-ARS?s Scientific Manuscript database

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...

Measurement of the pore size distribution of limestone aggregates in concrete pavement cores : phase I.

DOT National Transportation Integrated Search

2012-04-01

Freeze-thaw damage is one of the common forms of distress for concrete pavements in Kansas. D-Cracking is a form of : freeze-thaw damage caused by aggregates with poor freeze-thaw durability. It is believed that pores in the aggregates below : 10 m...

9 CFR 381.125 - Special handling label requirements.

Code of Federal Regulations, 2011 CFR

2011-01-01

... immediate containers for products that are frozen during distribution and intended to be thawed prior to or... frozen. Thaw in refrigerator or microwave. (Any portion of this statement that is in conflict with the product's specific handling instructions may be omitted, e.g., instructions to cook without thawing.) (A...

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

DOT National Transportation Integrated Search

2016-11-22

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

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

NASA Astrophysics Data System (ADS)

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

2018-03-01

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

Rapid Permafrost Carbon Degradation at the Land-Ocean Interface

NASA Astrophysics Data System (ADS)

Tanski, G.

2015-12-01

Climate change has a strong impact on permafrost coasts in the Arctic. With increasing air and water temperatures, the ice-rich unlithified permafrost coasts will thaw and erode at a greater pace. Organic carbon that has been stored for thousands of years is mobilized and degrades on its way to the ocean. The objective of this study is to investigate to what extent permafrost carbon degrades after thawing before it enters the ocean in a retrogressive thaw slump. A slump located on Herschel Island (Yukon Territory, Canada) was sampled systematically along transects from the permafrost headwall to the coastline. Concentrations of particulate and dissolved organic carbon (POC and DOC) as well as its stable carbon isotopes (δ13C-POC and δ13C-DOC) were measured and compared in frozen deposits and in thawed sediments. Ammonium, nitrite and nitrate were also analyzed in order to identify and understand the carbon metabolization mechanisms taking place during slump activity. Our results show that major portions of permafrost carbon are metabolized right after thawing. Ammonium concentrations are highest in areas where thawed permafrost material directly accumulates. We suggest that before entering the nearshore zone permafrost organic carbon and nitrogen is subject to major degradation and metabolization. This makes permafrost coasts and retrogressive thaw slumps degradation hotspots at the land-ocean-interface.

Development of the Soil Moisture Active Passive (SMAP) radiometer derived landscape freeze/thaw product

NASA Astrophysics Data System (ADS)

Colliander, A.; Xu, X.; Dunbar, R. S.; Derksen, C.; Kim, Y.; Kimball, J. S.

2016-12-01

A baseline SMAP mission objective was to determine the land surface binary freeze/thaw (FT) state for northern (>45°N) regions with 80% spatial classification accuracy at 3 km resolution and 2-day average intervals. These requirements were initially achieved from the SMAP radar until the sensor failed in July 2015. The FT algorithm is now transitioning to using SMAP radiometer inputs. The main compromises of this change are a coarse (36 km) radiometer footprint, enhanced noise and potential FT signal degradation from seasonal vegetation biomass, soil moisture and surface inundation changes. The new daily passive FT product (L3_FT_P) is based on the same seasonal threshold algorithm as the radar derived product (L3_FT_A): instantaneous SMAP measurements are compared to reference signatures acquired during seasonal frozen and thawed states. Instead of radar inputs, the normalized polarization ratio (NPR) is calculated from SMAP radiometer measurements. The L3_FT_P algorithm is applied using NPR inputs, whereby NPR decreases and increases are associated with respective landscape freezing and thawing. A lower NPR under frozen conditions is due to smaller V-pol brightness temperature increases and larger H-pol increases. Using in situ measurements from core validation sites, the temporal behavior of backscatter and NPR measurements were evaluated during the spring 2015 radar and radiometer overlap period. The transition from frozen to thawed states produced a NPR response similar in timing and magnitude to the radar response, resulting in similar freeze to thaw seasonal transition dates. While the post-thaw radar backscatter consistently remained at elevated values relative to the frozen state, the NPR drifted downwards following the main thaw transition (due to de-polarization of the scene), which may introduce false freeze classification errors. Both radar and radiometer results tended to lead observed soil thawing due to strong sensitivity of the microwave retrievals to wet snow. Continued analysis of SMAP radiometer measurements will help to identify different landscape components of the SMAP freeze-thaw temporal signal.

Comparative evaluation of Nabi and Beltsville extenders for cryopreservation of rooster semen.

PubMed

Nabi, Mohammad Mahdi; Kohram, Hamid; Zhandi, Mahdi; Mehrabani-Yeganeh, Hassan; Sharideh, Hossein; Zare-Shahaneh, Ahmad; Esmaili, Vahid

2016-02-01

Two experiments were conducted to evaluate the new rooster semen freezing extender which is containing a low level of glycerol and soybean lecithin as an alternative protective agent in the extender. The aim of the first experiment was to evaluate a new extender for freeze-thawing rooster semen known as "Nabi" extender compared to Beltsville. Second experiment was also performed to determine whether the Nabi extender has negative reactions on fertilization after artificial insemination (AI) or no. In the first experiment, post-thaw motion parameters, mitochondrial function and sperm apoptosis were analyzed using Sperm Class Analyzer (SCA), rhodamine-123 and Annexin-V, respectively for frozen-thawed semen in Nabi and Beltsville extender. Results showed that total motility, progressive motility, velocity parameters (VCL, VSL, VAP, LIN and STR) and live spermatozoa with active mitochondria were significantly higher in Nabi compare to Beltsville extender (P < 0.01). Also, the percentages of post-thawed live and early apoptotic spermatozoa were significantly higher in Nabi compared to Beltsville extender (14.46 ± 0.95 vs. 19.27 ± 0.95 and 14.83 ± 4.51 vs. 39.27 ± 4.51, respectively). For apoptotic spermatozoa, the percentages of post-thawed late apoptotic spermatozoa were significantly lower in Nabi (29.66 ± 3.11) compared to Beltsville extender (69.07 ± 3.11), but the type of extender had no effect on the percentages of post-thawed necrotic spermatozoa. In the second experiment, 20 broiler breeder hens (Ross 308) were inseminated with thawed semen using the new freezing diluents or fresh semen for determination of fertility rate. Fertility rate with thawed semen (with Nabi extender) was lower compared to fresh semen (by approximately 8% points). It can be concluded that Nabi extender would improve post-thawed rooster sperm in vitro quality compared to Beltsville extender. The fertility rates of insemination in hens with freeze-thaw sperm were comparable with fresh sperm. Copyright © 2015 Elsevier Inc. All rights reserved.

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

PubMed

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 < 0.05) field fertility for the group of straws associated with the longest interval from thawing to AI. However, the results of the laboratory experiment were unable to explain the findings of the field experiment. Sperm width:length ratio of morphometric analysis was the single sperm characteristic with a significant interaction between sire and straw group (P = 0.02). It was concluded that sequence of insemination after simultaneous thawing of 10 semen straws can differently affect 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.

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

PubMed

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.

The Adsorption and Desorption of Pb(2+) and Cd(2+) in Freeze-Thaw Treated Soils.

PubMed

Li, Linhui; Ma, Jincai; Xu, Meng; Li, Xu; Tao, Jiahui; Wang, Guanzhu; Yu, Jitong; Guo, Ping

2016-01-01

Adsorption and desorption are important processes that influence the potential toxicity and bioavailability of heavy metals in soils. However, information regarding adsorption and desorption behavior of heavy metals in soils subjected to freeze-thaw cycles is poorly understood. In the current study, the effect of freeze-thaw cycles with different freezing temperature (-15, -25, -35°C) on soil properties was investigated. Then the adsorption and desorption behavior of Pb(2+) and Cd(2+) in freeze-thaw treated soils was studied. The adsorption amounts of Pb(2+) and Cd(2+) in freeze-thaw treated soils were smaller than those in unfrozen soils (p < 0.05), due to the fact that pH, cation exchange capacity, organic matter content, free iron oxide content, and CaCO3 content in freeze-thaw treated soils were smaller than those in unfrozen soils. The adsorption amounts of Pb(2+) and Cd(2+) in soils treated with lower freezing temperatures were higher than those in soils treated with higher freezing temperatures. Desorption percentages of Pb(2+) and Cd(2+) in unfrozen soils were smaller than those in freeze-thaw treated soils (p < 0.05). The desorption percentages of Pb(2+) and Cd(2+) were smaller in soils treated with lower freezing temperatures than those in soils treated with higher freezing temperatures. The results obtained highlight the change of the adsorption and desorption behavior of typical heavy metals in freeze-thaw treated soils located in seasonal frozen soils zone in northeast China.

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

NASA Astrophysics Data System (ADS)

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

2018-01-01

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

Microbial functional diversity covaries with permafrost thaw-induced environmental heterogeneity in tundra soil.

PubMed

Yuan, Mengting M; Zhang, Jin; Xue, Kai; Wu, Liyou; Deng, Ye; Deng, Jie; Hale, Lauren; Zhou, Xishu; He, Zhili; Yang, Yunfeng; Van Nostrand, Joy D; Schuur, Edward A G; Konstantinidis, Konstantinos T; Penton, Christopher R; Cole, James R; Tiedje, James M; Luo, Yiqi; Zhou, Jizhong

2018-01-01

Permafrost soil in high latitude tundra is one of the largest terrestrial carbon (C) stocks and is highly sensitive to climate warming. Understanding microbial responses to warming-induced environmental changes is critical to evaluating their influences on soil biogeochemical cycles. In this study, a functional gene array (i.e., geochip 4.2) was used to analyze the functional capacities of soil microbial communities collected from a naturally degrading permafrost region in Central Alaska. Varied thaw history was reported to be the main driver of soil and plant differences across a gradient of minimally, moderately, and extensively thawed sites. Compared with the minimally thawed site, the number of detected functional gene probes across the 15-65 cm depth profile at the moderately and extensively thawed sites decreased by 25% and 5%, while the community functional gene β-diversity increased by 34% and 45%, respectively, revealing decreased functional gene richness but increased community heterogeneity along the thaw progression. Particularly, the moderately thawed site contained microbial communities with the highest abundances of many genes involved in prokaryotic C degradation, ammonification, and nitrification processes, but lower abundances of fungal C decomposition and anaerobic-related genes. Significant correlations were observed between functional gene abundance and vascular plant primary productivity, suggesting that plant growth and species composition could be co-evolving traits together with microbial community composition. Altogether, this study reveals the complex responses of microbial functional potentials to thaw-related soil and plant changes and provides information on potential microbially mediated biogeochemical cycles in tundra ecosystems. © 2017 John Wiley & Sons Ltd.

Effects of straw mulching on soil evaporation during the soil thawing period in a cold region in northeastern China

NASA Astrophysics Data System (ADS)

Fu, Qiang; Yan, Peiru; Li, Tianxiao; Cui, Song; Peng, Li

2018-04-01

To study the effect of straw mulching on soil water evaporation, it is necessary to measure soil water evaporation under different conditions of straw mulching during the soil thawing period. A field experiment was conducted in winter, and soil evaporation was measured using a microlysimeter on bare land (LD) and 4500 (GF4500), 9000 (GF9000) and 13500 kg/hm2 (GF13500) straw mulch. The influence of different quantities of straw mulch on soil water evaporation during the thawing period was analyzed using the Mallat algorithm, statistical analysis and information cost function. The results showed that straw mulching could delay the thawing of the surface soil by 3-6 d, decrease the speed at which the surface soil thaws by 0.40-0.80 cm/d, delay the peak soil liquid water content, increase the soil liquid water content, reduce the cumulative evaporation by 2.70-7.40 mm in the thawing period, increase the range of soil evaporation by 0.04-0.10 mm in the early stage of the thawing period, and reduce the range of soil evaporation by 0.25-0.90 mm in the late stage of the thawing period. Straw mulching could reduce the range of and variation in soil evaporation and can reduce the effect of random factors on soil evaporation. When the amount of straw mulch exceeded 9000 kg/hm2, the effect of increasing the amount of straw mulch on daily soil water evaporation was small.

Influence of vertical and lateral heat transfer on permafrost thaw, peatland landscape transition, and groundwater flow

USGS Publications Warehouse

Kurylyk, Barret L.; Masaki, Masaki; Quinton, William L.; McKenzie, Jeffrey M.; Voss, Clifford I.

2016-01-01

Recent climate change has reduced the spatial extent and thickness of permafrost in many discontinuous permafrost regions. Rapid permafrost thaw is producing distinct landscape changes in the Taiga Plains of the Northwest Territories, Canada. As permafrost bodies underlying forested peat plateaus shrink, the landscape slowly transitions into unforested wetlands. The expansion of wetlands has enhanced the hydrologic connectivity of many watersheds via new surface and near-surface flow paths, and increased streamflow has been observed. Furthermore, the decrease in forested peat plateaus results in a net loss of boreal forest and associated ecosystems. This study investigates fundamental processes that contribute to permafrost thaw by comparing observed and simulated thaw development and landscape transition of a peat plateau-wetland complex in the Northwest Territories, Canada from 1970 to 2012. Measured climate data are first used to drive surface energy balance simulations for the wetland and peat plateau. Near-surface soil temperatures simulated in the surface energy balance model are then applied as the upper boundary condition to a three-dimensional model of subsurface water flow and coupled energy transport with freeze-thaw. Simulation results demonstrate that lateral heat transfer, which is not considered in many permafrost models, can influence permafrost thaw rates. Furthermore, the simulations indicate that landscape evolution arising from permafrost thaw acts as a positive feedback mechanism that increases the energy absorbed at the land surface and produces additional permafrost thaw. The modeling results also demonstrate that flow rates in local groundwater flow systems may be enhanced by the degradation of isolated permafrost bodies.

Shifts of tundra bacterial and archaeal communities along a permafrost thaw gradient in Alaska.

PubMed

Deng, Jie; Gu, Yunfu; Zhang, Jin; Xue, Kai; Qin, Yujia; Yuan, Mengting; Yin, Huaqun; He, Zhili; Wu, Liyou; Schuur, Edward A G; Tiedje, James M; Zhou, Jizhong

2015-01-01

Understanding the response of permafrost microbial communities to climate warming is crucial for evaluating ecosystem feedbacks to global change. This study investigated soil bacterial and archaeal communities by Illumina MiSeq sequencing of 16S rRNA gene amplicons across a permafrost thaw gradient at different depths in Alaska with thaw progression for over three decades. Over 4.6 million passing 16S rRNA gene sequences were obtained from a total of 97 samples, corresponding to 61 known classes and 470 genera. Soil depth and the associated soil physical-chemical properties had predominant impacts on the diversity and composition of the microbial communities. Both richness and evenness of the microbial communities decreased with soil depth. Acidobacteria, Verrucomicrobia, Alpha- and Gamma-Proteobacteria dominated the microbial communities in the upper horizon, whereas abundances of Bacteroidetes, Delta-Proteobacteria and Firmicutes increased towards deeper soils. Effects of thaw progression were absent in microbial communities in the near-surface organic soil, probably due to greater temperature variation. Thaw progression decreased the abundances of the majority of the associated taxa in the lower organic soil, but increased the abundances of those in the mineral soil, including groups potentially involved in recalcitrant C degradation (Actinomycetales, Chitinophaga, etc.). The changes in microbial communities may be related to altered soil C sources by thaw progression. Collectively, this study revealed different impacts of thaw in the organic and mineral horizons and suggests the importance of studying both the upper and deeper soils while evaluating microbial responses to permafrost thaw. © 2014 John Wiley & Sons Ltd.

Fresh-Frozen Plasma: Ordering Patterns and Utilization in the Operating Rooms of a Tertiary Referral Hospital.

PubMed

Meyer, Matthew J; Dzik, Walter H; Levine, Wilton C

2017-02-01

Blood product transfusion is the most commonly performed hospital procedure. Intraoperative blood product utilization varies between institutions and anesthesiologists. In the United States in 2011, nearly 4 million plasma units were transfused. A retrospective analysis of intraoperative plasma ordering patterns and utilization (thawing and transfusing) was performed at a tertiary, academic hospital between January 2015 and March 2016. Over 15 months, 46,002 operative procedures were performed. In 1540 of them, plasma was thawed or transfused: 8297 plasma units were thawed and 3306 of those units were transfused. These 3306 plasma units were transfused in 749 cases with a median of 2 plasma units (interquartile range, 2-4) transfused. The percentage of average monthly procedures with plasma thawed and none transfused was 51.3% (confidence interval, 49.0%-53.6%). The cardiac surgery service requested the greatest number of plasma units to be thawed (2143) but only transfused 712 (33.2%) of them. Of all plasma units not transfused, 45% were generated by procedures with 1 to 4 units of plasma thawed; 95.7% of these units were thawed as even integers (ie, 2, 4). For operative procedures, far more plasma was thawed than was transfused and this practice occurred across surgical specialties and anesthesiologists. Considering the plasma that was not transfused, 45% occurred in procedures with 4 or fewer units of plasma requested suggesting these low-volume requests were a primary source of potential waste. Further studies are needed to examine associations between plasma utilization and clinical outcomes.

Multi-omics of permafrost, active layer and thermokarst bog soil microbiomes

NASA Astrophysics Data System (ADS)

Hultman, Jenni; Waldrop, Mark P.; Mackelprang, Rachel; David, Maude M.; McFarland, Jack; Blazewicz, Steven J.; Harden, Jennifer; Turetsky, Merritt R.; McGuire, A. David; Shah, Manesh B.; Verberkmoes, Nathan C.; Lee, Lang Ho; Mavrommatis, Kostas; Jansson, Janet K.

2015-05-01

Over 20% of Earth's terrestrial surface is underlain by permafrost with vast stores of carbon that, once thawed, may represent the largest future transfer of carbon from the biosphere to the atmosphere. This process is largely dependent on microbial responses, but we know little about microbial activity in intact, let alone in thawing, permafrost. Molecular approaches have recently revealed the identities and functional gene composition of microorganisms in some permafrost soils and a rapid shift in functional gene composition during short-term thaw experiments. However, the fate of permafrost carbon depends on climatic, hydrological and microbial responses to thaw at decadal scales. Here we use the combination of several molecular `omics' approaches to determine the phylogenetic composition of the microbial communities, including several draft genomes of novel species, their functional potential and activity in soils representing different states of thaw: intact permafrost, seasonally thawed active layer and thermokarst bog. The multi-omics strategy reveals a good correlation of process rates to omics data for dominant processes, such as methanogenesis in the bog, as well as novel survival strategies for potentially active microbes in permafrost.

Multi-omics of permafrost, active layer and thermokarst bog soil microbiomes.

PubMed

Hultman, Jenni; Waldrop, Mark P; Mackelprang, Rachel; David, Maude M; McFarland, Jack; Blazewicz, Steven J; Harden, Jennifer; Turetsky, Merritt R; McGuire, A David; Shah, Manesh B; VerBerkmoes, Nathan C; Lee, Lang Ho; Mavrommatis, Kostas; Jansson, Janet K

2015-05-14

Over 20% of Earth's terrestrial surface is underlain by permafrost with vast stores of carbon that, once thawed, may represent the largest future transfer of carbon from the biosphere to the atmosphere. This process is largely dependent on microbial responses, but we know little about microbial activity in intact, let alone in thawing, permafrost. Molecular approaches have recently revealed the identities and functional gene composition of microorganisms in some permafrost soils and a rapid shift in functional gene composition during short-term thaw experiments. However, the fate of permafrost carbon depends on climatic, hydrological and microbial responses to thaw at decadal scales. Here we use the combination of several molecular 'omics' approaches to determine the phylogenetic composition of the microbial communities, including several draft genomes of novel species, their functional potential and activity in soils representing different states of thaw: intact permafrost, seasonally thawed active layer and thermokarst bog. The multi-omics strategy reveals a good correlation of process rates to omics data for dominant processes, such as methanogenesis in the bog, as well as novel survival strategies for potentially active microbes in permafrost.

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

NASA Astrophysics Data System (ADS)

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.

Response of changes in seasonal soil freeze/thaw state to climate change from 1950 to 2010 across china

NASA Astrophysics Data System (ADS)

Peng, Xiaoqing; Frauenfeld, Oliver W.; Cao, Bin; Wang, Kang; Wang, Huijuan; Su, Hang; Huang, Zhe; Yue, Dongxia; Zhang, Tingjun

2016-11-01

Variations in seasonal soil freeze/thaw state are important indicators of climate change and influence ground temperature, hydrological processes, surface energy, and the moisture balance. Previous studies mainly focused on the active layer and permafrost, while seasonally frozen ground research in nonpermafrost regions has received less attention. In this study, we investigate the response of changes in seasonal soil freeze/thaw state to changes in air temperatures by combining observations from more than 800 stations with gridded mean monthly air temperature data across China. The results show that mean annual air temperature (MAAT) increased statistically significantly by 0.29 ± 0.03°C/decade from 1967 to 2013, with greater warming on the Qinghai-Tibetan Plateau. There is a statistically significant decrease in the freeze/thaw cycle (FTC) at 0.39 ± 0.05 cycles/decade. In addition, there are strong negative correlations between FTC and MAAT. Estimating the soil freeze/thaw state classification based on the number of days in the month, we find that changes of mean annual area extent of seasonal soil freeze/thaw state decreased significantly for completely frozen (CF) ground, while the area extent of partially frozen (PF) and unfrozen (UF) ground both increased. Changes in mean monthly area extent of seasonal soil freeze/thaw state indicate that the extent of CF and UF area was decreasing and increasing, respectively. But for the extent of PF areas, both increasing and decreasing trends were observed. Quantifying the spatial pattern of the seasonal soil freeze/thaw, we find that CF and PF areas are located in northern China and the Tibetan Plateau from December to March, and UF areas are located in southern China. The variations of mean annual area extent departure of soil freeze/thaw states are consistent with MAAT changes in different land cover types across China.

Colloid centrifugation of fresh stallion semen before cryopreservation decreased microorganism load of frozen-thawed semen without affecting seminal kinetics.

PubMed

Guimarães, T; Lopes, G; Pinto, M; Silva, E; Miranda, C; Correia, M J; Damásio, L; Thompson, G; Rocha, A

2015-01-15

Freezability of equine semen may be influenced by microorganism population of semen. The objective of this study was to verify the effect of single-layer density gradient centrifugation (SLC) of fresh semen before cryopreservation on semen's microbial load (ML) and sperm cells kinetics after freezing-thawing. For that, one ejaculate was collected from 20 healthy stallions and split into control (C) samples (cryopreserved without previous SLC) and SLC samples (subjected to SLC). Semen cryopreservation was performed according to the same protocol in both groups. Microbial load of each microorganism species and total microbial load (TML) expressed in colony-forming units (CFU/mL) as well as frozen-thawed sperm kinetics were assessed in both groups. Additional analysis of the TML was performed, subdividing the frozen-thawed samples in "suitable" (total motility ≥ 30%) and "unsuitable" (total motility < 30%) semen for freezing programs, and comparing the C and SLC groups within these subpopulations. After thawing, SLC samples had less (P < 0.05) TML (88.65 × 10(2) ± 83.8 × 10(2) CFU/mL) than C samples (155.69 × 10(2) ± 48.85 × 10(2) CFU/mL), mainly due to a reduction of Enterococcus spp. and Bacillus spp. A relationship between post-thaw motility and SLC effect on ML was noted, as only in samples with more than 30% total motility was ML reduced (P < 0.05) by SLC (from 51.33 × 10(2) ± 33.26 × 10(2) CFU/mL to 26.68 × 10(2) ± 12.39 × 10(2) CFU/mL in "suitable" frozen-thawed semen vs. 240.90 × 10(2) ± 498.20 × 10(2) to 139.30 × 10(2) ± 290.30 × 10(2) CFU/mL in "unsuitable" frozen-thawed semen). The effect of SLC on kinetics of frozen-thawed sperm cells was negligible. Copyright © 2015 Elsevier Inc. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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.

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

PubMed

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.

Using Distributed Fiber Optic Sensing to Monitor Large Scale Permafrost Transitions: Preliminary Results from a Controlled Thaw Experiment

NASA Astrophysics Data System (ADS)

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 moisture measurements, hydrologic measurements, and multi-angle photogrammetry. This unusually dense combination of measurement techniques provides an excellent opportunity to characterize the geophysical signatures of permafrost thaw in a controlled environment.

Histologic and ultrastructural evaluation of fresh and frozen-thawed human ovarian xenografts in nude mice.

PubMed

Nisolle, M; Casanas-Roux, F; Qu, J; Motta, P; Donnez, J

2000-07-01

To compare histologic and ultrastructural characteristics of fresh and frozen-thawed human ovarian cortical tissue grafted into nude mice. Experimental prospective study. An academic research environment. Ovarian biopsy specimens were obtained from 13 women undergoing laparoscopy for tubal ligation or infertility. Forty nude mice. A minilaparotomy was performed to place fresh and frozen-thawed ovarian grafts subcutaneously (sc) or intraperitoneally (ip). Removal of the ovarian grafts was performed at 24 days. [1] the follicular population, [2] fibrosis, [3] vascularization of the grafted tissue, and [4] ultrastructural evaluation. A greater fibrosis relative surface area was noted in frozen-thawed transplanted tissue than in fresh transplants. Regardless of this fibrosis, a similar follicular density was observed in fresh and frozen-thawed ovarian tissue 24 days after transplantation. Active angiogenesis was proved by both immunohistochemical study of the vascular endothelial growth factor and morphometric study of the vascular network. Normal ultrastructural characteristics were noted in frozen-thawed ovarian biopsies. Angiogenesis allows implantation of the graft even if it has been cryopreserved and thawed similarly to implantation of fresh tissue. The greater fibrosis observed in grafts after cryopreservation and implantation does not seem to affect the primordial and primary ovocyte population and their ultrastructural characteristics, but further studies must be conducted to prove that after cryopreservation and transplantation, ovocytes may achieve full maturation and fertilization.

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

PubMed Central

Vutyavanich, Teraporn; Lattiwongsakorn, Worashorn; Piromlertamorn, Waraporn; Samchimchom, Sudarat

2012-01-01

In this study, we compared the effects of repeated freezing/thawing of human sperm by our in-house method of rapid freezing with slow programmable freezing. Sperm samples from 11 normozoospermic subjects were processed through density gradients and divided into three aliquots: non-frozen, rapid freezing and slow programmable freezing. Sperm in the rapid freezing group had better motility and viability than those in the slow freezing group (P<0.01) after the first, second and third cycles of freezing/thawing, but there was no difference in morphology. In the second experiment, rapid freezing was repeated three times in 20 subjects. The samples from each thawing cycle were evaluated for DNA fragmentation using the alkaline comet assay. DNA fragmentation began to increase considerably after the second cycle of freezing/thawing, but to a level that was not clinically important. In the third experiment, rapid freezing was done repeatedly in 10 subjects, until no motile sperm were observed after thawing. The median number of repeated freezing/thawing that yielded no motile sperm was seven (range: 5–8, mean: 6.8). In conclusion, we demonstrated that repeated freezing/thawing of processed semen using our rapid freezing method gave better results than standard slow programmable freezing. This method can help maximize the usage of precious cryopreserved sperm samples in assisted reproduction technology. PMID:23064685

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

PubMed

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.

Soil CO2 production in upland tundra where permafrost is thawing

Treesearch

Hanna Lee; Edward A.G. Schuur; Jason G. Vogel

2010-01-01

Permafrost soils store nearly half of global soil carbon (C), and therefore permafrost thawing could lead to large amounts of greenhouse gas emissions via decomposition of soil organic matter. When ice-rich permafrost thaws, it creates a localized surface subsidence called thermokarst terrain, which changes the soil microenvironment. We used soil profile CO2...

Physiological changes in red spruce seedlings during a simulated winter thaw

Treesearch

P.G. Schaberg; J.B. Shane; G.J. Hawley; G.R. Strimbeck; D.H. DeHayes; P.F. Cali; J.R. Donnelly

1996-01-01

We evaluated net photosynthesis, respiration, leaf conductance, xylem pressure potential (XPP) and cold hardiness in red spruce (Picea rubens Sarg.) seedlings exposed to either a continuous thaw (CT) or a daytime thaw with freezing nights (FN) for 8 days during mid-winter. Physiological differences between CT and FN seedlings were evident for all...

Long-term CO2 production following permafrost thawing

Treesearch

B. Elberling; A. Michelsen; C. Schadel; E.A.G. Schuur; H.H. Christiansen; L. Berg; M.P. Tamstorf; C. Sigsgaard

2013-01-01

Thawing permafrost represents a poorly understood feedback mechanism of climate change in the Arctic, but with a potential impact owing to stored carbon being mobilized1–5. We have quantified the long-term loss of carbon (C) from thawing permafrost in Northeast Greenland from 1996 to 2008 by combining repeated sediment sampling to assess changes...

The changing spatio-temporal dynamics of thaw lake development, Seward Peninsula, Alaska.

NASA Astrophysics Data System (ADS)

Cooper, Michael; Rees, Gareth; Bartsch, Annett

2014-05-01

Contemporary anthropogenic climatic warming is having an accelerated, and more pronounced effect upon Arctic regions than any other environment on Earth. Increased surface temperatures have led to widespread permafrost degradation and a shift in dynamics. One landscape manifestation of localised permafrost decay, seen to be ubiquitous across low-lying tundra regions of Alaska, Canada and Siberia, is the thermokarst lake - or 'thaw' lake. These features are seen to be truly dynamic, with a relatively rapid evolution and decay. The exact impacts of climatic perturbation on thaw lake development are in contention; however, recent studies have suggested an increased vulnerability of these features, owing to the susceptibility of the fundamental processes of initiation, expansion and drainage to climatic variation. It is often hypothesised that with current trends, thaw lakes will see a net increase in expansion rate, and areal extent, with a potential for increased drainage events. Increased permafrost thaw and thermokarst activity has also led to shifts in biogeochemical cycles, leading to an amplified release from large carbon reservoirs currently sequestered within permafrost. An example of carbon release exhibited from thaw lakes is that of methane ebullition (gas bubble formation); this has been theorised to have the potential to initiate a major positive climatic feedback leading to a continued rise in global temperatures. Due to the remote nature and large area over which these landforms occur, remotely sensed data has been widely used in order to both accurately classify features and measure change over spatially large and great temporal extents. As well as studies interpreting data collected in the visible and near-infrared spectra, studies have recently made use of radar or microwave products in order to capture imagery avoiding adverse atmospheric conditions, most notably cloud cover. Data from Envisat ASAR operating in Wide Swath Mode was acquired for this study region; however, the core of this research relied upon the analysis of the changing lake morphology using visible and near-infrared spectra from MODIS and Landsat products. This research explored: (1) intra-annual variability of freeze-thaw cycles and resultant effects on thaw lake development; and (2) the spatio-temporal trends and changing dynamism of thaw lake activity. Research presented here within suggests that although climatic trends do indeed influence widespread changes within thaw lake characteristics, site-specific phenomena of sediment type and ice-content and fluvial activity also play integral roles. Understanding and observing changing spatio-temporal dynamics, particularly on an intra-annual basis, has helped to gather more information concerning complex lake processes, and increase the understanding of permafrost decay and thaw lake development.

Nitrogen availability drives priming effect by altering microbial carbon-use efficiency after permafrost thaw

NASA Astrophysics Data System (ADS)

Chen, L.; Liu, L.; Zhang, Q.; Mao, C.; Liu, F.; Yang, Y.

2017-12-01

Enhanced vegetation growth can potentially aggravate soil C loss by accelerating the decomposition of soil organic matter (SOM) ("priming effect"), thereby reinforcing the positive C-climate feedback in permafrost ecosystems. However, the degree to which priming effect alters permafrost C dynamics is expected to be modified by nitrogen (N) availability after permafrost thaw. Despite this recognition, experimental evidence for the linkage between priming effect and post-thaw N availability is still lacking. Particularly, the microbial mechanisms involved remain unknown. Here, using a thermokarst-induced natural N gradient combined with an isotope-labeled glucose and N addition experiment, we presented a strong linkage between soil N availability and priming effect in Tibetan permafrost. We observed that the magnitude of priming effect along the thaw gradient was negatively associated with soil total dissolved nitrogen (TDN) concentration. This negative effect of post-thaw N availability was further proved by a sharply reduced priming effect following mineral N supply. These two lines of evidence jointly illustrated that the priming effect along the thaw chronosequence was controlled by N availability, supporting the `N mining theory'. In contrast to the prevailing assumption, this N-regulated priming effect was independent from changes in C- or N-acquiring enzyme activities, but positively associated with the change in metabolic quotients (△SOM-qCO2), highlighting that decreased microbial metabolism efficiency rather than increased enzyme activities account for greater priming effect under reduced N availability. Taken together, these findings demonstrate that C dynamics in melting permafrost largely depends on post-thaw N availability due to its effect of retarding SOM mineralization. This C-N interaction and the relevant microbial metabolic efficiency should be considered in Earth System Models for a better understanding of soil C dynamics after permafrost thaw.

Freezing Injury in Onion Bulb Cells

PubMed Central

Palta, Jiwan P.; Levitt, Jacob; Stadelmann, Eduard J.

1977-01-01

Onion (Allium cepa L.) bulbs were subjected for 12 days to either a moderate freeze (−4 C) or a severe freeze (−11 C). They were then thawed slowly over ice. During 7 to 12 days following the thaw, the injury progressed with time in the severely frozen bulbs, but appeared completely repaired in the moderately frozen bulbs. This was shown by the following post-thawing changes. Infiltration of the intercellular spaces increased from 80 to 90% to 100% after the severe freeze, and decreased from 30 to 50% to zero after the moderate freeze. All of the cells were alive immediately after thawing whether the freeze was moderate or severe. Corresponding to the infiltration results 7 to 12 days later, many to most were dead following the severe freeze, all were alive following the moderate freeze. The conductivity of the effusate from pieces of bulb tissue increased after the severe freezing, and decreased after the moderate freezing. The concentration of K+, total solutes, and sugars in the effusate paralleled the conductivity changes. Neither the pH of the effusate nor the permeability of the cells (as long as cells were living) to water was changed following either the severe or the moderate freezes. Some treatments of the thawed tissue following the severe freeze halted the progress of injury. The above results indicate that the semipermeable properties of the cell are uninjured but that the ion and sugar transport mechanism is damaged by freezing. Most likely the primary injury is to the active transport mechanism involved in their transport. It must be concluded that the final injury following freezing and thawing cannot be evaluated from the degree of infiltration or the conductivity of the effusate immediately after thawing, since injury may progress or recede following the thawing. PMID:16660101

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

PubMed

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.

Effect of cryoprotective diluent and method of freeze-thawing on survival and acrosomal integrity of ram spermatozoa.

PubMed

Pontbriand, D; Howard, J G; Schiewe, M C; Stuart, L D; Wildt, D E

1989-08-01

A multifactorial study analyzed the effects of freezing method, cryoprotective diluent, semen to diluent ratio, and thawing velocity on post-thaw motility, progressive status, and acrosomal integrity of ram spermatozoa. Although semen to diluent ratio (1:3 vs 1:6, v/v) had no effect (P greater than 0.05), overall post-thaw spermatozoal viability was highly dependent on freezing method and cryoprotectant. Improved results were obtained by freezing semen in 0.5-ml French straws compared to dry ice pelleting. Manually freezing straws 5 cm above liquid nitrogen (LN2) was comparable to cooling straws in an automated, programmable LN2 unit. Of the two cryoprotective diluents tested, BF5F (containing the surfactant component sodium and triethanolamine lauryl sulfate) yielded approximately 50% fewer (P less than 0.05) spermatozoa with loose acrosomal caps compared to TEST. Thawing straws in a water bath at a higher velocity (60 degrees C for 8 sec) had no effect (P greater than 0.05) on spermatozoal motility, progressive status ratings, or acrosomal integrity when compared to a lower rate (37 degrees C for 20 sec). For the TEST group, thawing pellets in a dry, glass culture tube promoted (P less than 0.05) percentage sperm motility at 3 and 6 hr post-thawing, but for BF5F diluted semen this approach decreased the % of spermatozoa with normal apical ridges. The results suggest that the poor fertility rates often experienced using thawed ram semen likely result not only from reduced sperm motility, but also from compromised ultrastructural integrity. This damage is expressed by an increased loosening of the acrosomal cap, a factor which appears insensitive to freezing method but markedly influenced by the cryoprotective properties of the diluents tested.

Optimizing the Protocol for Pulmonary Cryoablation: A Comparison of a Dual- and Triple-Freeze Protocol

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hinshaw, J. Louis, E-mail: jhinshaw@uwhealth.or; Littrup, Peter J.; Durick, Nathan

2010-12-15

The purpose of this study was to compare a double freeze-thaw protocol to a triple freeze-thaw protocol for pulmonary cryoablation utilizing an in vivo porcine lung model. A total of 18 cryoablations were performed in normal porcine lung utilizing percutaneous technique with 9 each in a double- (10-5-10) and triple-freeze (3-3-7-7-5) protocol. Serial noncontrast CT images were obtained during the ablation. CT imaging findings and pathology were reviewed. No imaging changes were identified during the initial freeze cycle with either protocol. However, during the first thaw cycle, a region of ground glass opacity developed around the probe with both protocols.more » Because the initial freeze was shorter with the triple freeze-thaw protocol, the imaging findings were apparent sooner with this protocol (6 vs. 13 min). Also, despite a shorter total freeze time (15 vs. 20 min), the ablation zone identified with the triple freeze-thaw protocol was not significantly different from the double freeze-thaw protocol (mean diameter: 1.67 {+-} 0.41 cm vs. 1.66 {+-} 0.21 cm, P = 0.77; area: 2.1 {+-} 0.48 cm{sup 2} vs. 1.99 {+-} 0.62 cm{sup 2}, P = 0.7; and circularity: 0.95 {+-} 0.04 vs. 0.96 {+-} 0.03, P = 0.62, respectively). This study suggests that there may be several advantages of a triple freeze-thaw protocol for pulmonary cryoablation, including earlier identification of the imaging findings associated with the ablation, the promise of a shorter procedure time or larger zones of ablation, and theoretically, more effective cytotoxicity related to the additional freeze-thaw cycle.« less

Changes in peat chemistry associated with permafrost thaw increase greenhouse gas production

PubMed Central

Hodgkins, Suzanne B.; Tfaily, Malak M.; McCalley, Carmody K.; Logan, Tyler A.; Crill, Patrick M.; Saleska, Scott R.; Rich, Virginia I.; Chanton, Jeffrey P.

2014-01-01

Carbon release due to permafrost thaw represents a potentially major positive climate change feedback. The magnitude of carbon loss and the proportion lost as methane (CH4) vs. carbon dioxide (CO2) depend on factors including temperature, mobilization of previously frozen carbon, hydrology, and changes in organic matter chemistry associated with environmental responses to thaw. While the first three of these effects are relatively well understood, the effect of organic matter chemistry remains largely unstudied. To address this gap, we examined the biogeochemistry of peat and dissolved organic matter (DOM) along a ∼40-y permafrost thaw progression from recently- to fully thawed sites in Stordalen Mire (68.35°N, 19.05°E), a thawing peat plateau in northern Sweden. Thaw-induced subsidence and the resulting inundation along this progression led to succession in vegetation types accompanied by an evolution in organic matter chemistry. Peat C/N ratios decreased whereas humification rates increased, and DOM shifted toward lower molecular weight compounds with lower aromaticity, lower organic oxygen content, and more abundant microbially produced compounds. Corresponding changes in decomposition along this gradient included increasing CH4 and CO2 production potentials, higher relative CH4/CO2 ratios, and a shift in CH4 production pathway from CO2 reduction to acetate cleavage. These results imply that subsidence and thermokarst-associated increases in organic matter lability cause shifts in biogeochemical processes toward faster decomposition with an increasing proportion of carbon released as CH4. This impact of permafrost thaw on organic matter chemistry could intensify the predicted climate feedbacks of increasing temperatures, permafrost carbon mobilization, and hydrologic changes. PMID:24711402

Improvement of common carp (Cyprinus carpio) sperm cryopreservation using a programable freezer.

PubMed

Bernáth, Gergely; Żarski, Daniel; Kása, Eszter; Staszny, Ádám; Várkonyi, Levente; Kollár, Tímea; Hegyi, Árpád; Bokor, Zoltán; Urbányi, Béla; Horváth, Ákos

2016-10-01

The applicability of a programmable freezer for the increased-scale cryopreservation of common carp sperm was investigated. The effect of different equilibration times, cryopreservation methods, extenders, dilution ratios, activating solutions on the post-thaw motility of common carp sperm was investigated. The suitable post-thaw storage time-interval as well as fertilizing capacity of cryopreserved sperm was also examined. The motility, curvilinear velocity (VCL) and straightness (STR) values did not decrease significantly during 60min of equilibration neither in equilibrated nor thawed groups. Motility parameters of thawed sperm were similar using a conventional cryopreservation technique using a polystyrene box [motility (33%), VCL (47μm/s) and STR (88%)] and a programmable freezer: [motility (32%), VCL (54μm/s) and STR (89%)]. The highest motility and VCL was measured with a sugar based extender (grayling extender) at a ratio 1:9 (motility: 52%, VCL: 76μm/s) and 1:20 (motility: 49%, VCL: 76μm/s). The activating solution for cyprinids (ASC) could prolong sperm movement up for 2min. A storage time of six hours following thawing did not have a significant effect on the motility parameters of thawed carp sperm. Agglutination was observed during cryopreservation of an elevated volume of sperm whereas motility 47%, VCL 62μm/s and STR 91% were measured after thawing. Fertilization rate with thawed sperm (32%) was significantly lower compared to the control group (73%). According to our results, the developed method using a programmable freezer is suitable for the cryopreservation of elevated number of straws. However, carp sperm agglutination during freezing may have a negative effect on the fertilizing capacity. Copyright © 2016 Elsevier Inc. All rights reserved.

The quality of great scallop (Pecten maximus) sperm after thawing.

PubMed

Suquet, Marc; Gourtay, Clémence; Donval, Anne; Le Goïc, Nelly; Quere, Claudie; Malo, Florent; Le Grand, Jaqueline; Ratiskol, Dominique; Mingant, Christian; Fauvel, Christian

2016-04-01

Most publications devoted to the cryopreservation of mollusc sperm have focused on the definition of technical protocols, avoiding the description of sperm quality after thawing. The present study investigated the effects of cryopreservation on sperm quality in the great scallop. Wild scallop were fished during the natural spawning period and conditioned in the hatchery before use. Sperm samples were obtained after intragonadal injection of serotonin and cryopreserved using a previously published protocol. Sperm quality was assessed using a panel of four parameters: sperm motility characteristics, using a computer assisted sperm analysis plugin with Image J, intracellular ATP content using an ATP-Lite kit, sperm integrity, using flow cytometry and sperm morphology, using transmission electron microscopy. For each parameter, fresh (control) and thawed spermatozoa were compared. A significant decrease of both the percentage of motile spermatozoa (reduction: 75%) and sperm swimming speed (86%) were observed for thawed sperm compared with fresh sperm. The percentage of living spermatozoa, as assessed using flow cytometry, was significantly lower for thawed sperm (72.4±2.5%) compared with fresh sperm (86.4±1.1). However, no significant difference of intracellular sperm ATP content was observed between fresh and thawed sperm. Post thawing, while some spermatozoa showed little or no morphological differences compared with fresh sperm, others had undergone drastic changes, including swelling of the plasma membrane, structural alterations of the chromatin and damage to mitochondria. In conclusion, the descriptive parameters studied in the present work showed that the quality of thawed great scallop sperm was lower than that of fresh cells but was still sufficient for use in aquaculture programs and sperm cryobanking for this species. Copyright © 2016 Elsevier Inc. All rights reserved.

Permafrost Thaw Induces Methane Oxidation in Transitional Thaw Stages in a Subarctic Peatland

NASA Astrophysics Data System (ADS)

Perryman, C. R.; Kashi, N. N.; Malhotra, A.; McCalley, C. K.; Varner, R. K.

2015-12-01

Rising temperatures in the subarctic are accelerating permafrost thaw and increasing methane (CH4) emissions from subarctic peatlands. Methanotrophs in these peatlands can consume/oxidize CH4, potentially mitigating CH4 emissions in these peatlands. Oxidation rates can exceed 90% of CH4 production in some settings, depending on O2 and CH4 availability and environmental conditions. Malhotra and Roulet identified 10 thaw stages in Stordalen Mire near Abisko, Sweden (68°21'N,18°49'E ) with variable vegetation, environmental conditions, and associated CH4 emissions. We investigated potential methane oxidation rates across these thaw stages. Peat cores were extracted from two depths at each stage and incubated in 350ml glass jars at in situ temperatures and CH4 concentrations. Headspace samples were collected from each incubation jar over a 48-hour period and analyzed for CH4 concentration using flame ionization detection gas chromatography (GC-FID). Oxidation rates ranged from <0.1 to 17 μg of CH4 per gram of dry biomass per day. Water table depth and pore water pH were the strongest environmental correlates of oxidation (sample size = 56, p < 0.001). The highest potential oxidation rates were observed in collapsing palsa sites and recently collapsed sedge-dominated open water sites near palsa mounds. Our results suggest that permafrost thaw induces high CH4 oxidation rates by creating conditions ideal for both methanogenic and methanotrophic microbial communities. Our results also reinforce the importance of incorporating transitional thaw stages in landscape level carbon budgets of thawing peatlands emphasized by Malhotra and Roulet. Forthcoming microbial analysis and stable isotope analysis will further elucidate the factors controlling methane oxidation rates at Stordalen Mire.

Characteristics of ultrasonic acoustic emissions from walnut branches during freeze-thaw-induced embolism formation.

PubMed

Kasuga, Jun; Charrier, Guillaume; Uemura, Matsuo; Améglio, Thierry

2015-04-01

Ultrasonic acoustic emission (UAE) methods have been applied for the detection of freeze-thaw-induced embolism formation in water conduits of tree species. Until now, however, the exact source(s) of UAE has not been identified especially in angiosperm species, in which xylem tissues are composed of diverse types of cells. In this study, UAE was recorded from excised branches of walnut (Juglans regia cv. Franquette) during freeze-thaw cycles, and attempts were made to characterize UAEs generated by cavitation events leading to embolism formation according to their properties. During freeze-thaw cycles, a large number of UAEs were generated from the sample segments. However, the cumulative numbers of total UAE during freeze-thawing were not correlated with the percentage loss of hydraulic conductivity after thawing, suggesting that the sources of UAE were not only cavitation leading to embolism formation in vessels. Among the UAEs, cumulative numbers of UAEs with absolute energy >10.0 fJ strongly correlated with the increase in percentage loss of hydraulic conductivity. The high absolute energy of the UAEs might reflect the formation of large bubbles in the large lumen of vessels. Therefore, UAEs generated by cavitation events in vessels during freeze-thawing might be distinguished from other signals according to their magnitudes of absolute energy. On the other hand, the freezing of xylem parenchyma cells was followed by a certain number of UAEs. These results indicate the possibility that UAE methods can be applied to the detection of both freeze-thaw-induced embolism and supercooling breakdown in parenchyma cells in xylem. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

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

PubMed

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.

Longer thaw seasons increase nitrogen availability for leaching during fall in tundra soils

DOE PAGES

Treat, Claire C.; Wollheim, Wilfred M.; Varner, Ruth K.; ...

2016-06-15

Climate change has resulted in warmer soil temperatures, earlier spring thaw and later fall freeze-up, resulting in warmer soil temperatures and thawing of permafrost in tundra regions. While these changes in temperature metrics tend to lengthen the growing season for plants, light levels, especially in the fall, will continue to limit plant growth and nutrient uptake. We conducted a laboratory experiment using intact soil cores with and without vegetation from a tundra peatland to measure the effects of late freeze and early spring thaw on carbon dioxide (CO 2) exchange, methane (CH 4) emissions, dissolved organic carbon (DOC) and nitrogenmore » (N) leaching from soils. We compared soil C exchange and N production with a 30 day longer seasonal thaw during a simulated annual cycle from spring thaw through freeze-up and thaw. Across all cores, fall N leaching accounted for similar to 33% of total annual N loss despite significant increases in microbial biomass during this period. Nitrate(NO 3 -) leaching was highest during the fall (5.33 ± 1.45 mgNm -2 d -1) following plant senescence and lowest during the summer (0.43 ± 0.22 mg Nm -2 d -1). In the late freeze and early thaw treatment, we found 25% higher total annual ecosystem respiration but no significant change in CH 4 emissions or DOC loss due to high variability among samples. The late freeze period magnified N leaching and likely was derived from root turnover and microbial mineralization of soil organic matter coupled with little demand from plants or microbes. Furthermore, large N leaching during the fall will affect N cycling in low-lying areas and streams and may alter terrestrial and aquatic ecosystem nitrogen budgets in the arctic.« less

Monitoring Freeze-Thaw States in the Pan-Arctic: Application of Microwave Remote Sensing to Monitoring Hydrologic and Ecological Processes

NASA Astrophysics Data System (ADS)

McDonald, K. C.; Kimball, J. S.

2004-12-01

The transition of the landscape between predominantly frozen and non-frozen conditions in seasonally frozen environments impacts climate, hydrological, ecological and biogeochemical processes profoundly. Satellite microwave remote sensing is uniquely capable of detecting and monitoring a range of related biophysical processes associated with the measurement of landscape freeze/thaw status. We present the development, physical basis, current techniques and selected hydrological applications of satellite-borne microwave remote sensing of landscape freeze/thaw states for the terrestrial cryosphere. Major landscape hydrological processes embracing the remotely-sensed freeze/thaw signal include timing and spatial dynamics of seasonal snowmelt and associated soil thaw, runoff generation and flooding, ice breakup in large rivers and lakes, and timing and length of vegetation growing seasons and associated productivity and trace gas exchange. Employing both active and passive microwave sensors, we apply a selection of temporal change classification algorithms to examine a variety of hydrologic processes. We investigate contemporaneous and retrospective applications of the QuikSCAT scatterometer, and the SSM/I and SMMR radiometers to this end. Results illustrate the strong correspondence between regional thawing, seasonal ice break up for rivers, and the springtime pulse in river flow. We present the physical principles of microwave sensitivity to landscape freeze/thaw state, recent progress in applying these principles toward satellite remote sensing of freeze/thaw processes over broad regions, and potential for future global monitoring of this significant phenomenon of the global cryosphere. This work was carried out at the Jet Propulsion Laboratory, California Institute of Technology, and at the University of Montana, Missoula, under contract to the National Aeronautics and Space Administration.

Assessment of gamete quality for the eastern oyster (Crassostrea virginica) by use of fluorescent dyes

USGS Publications Warehouse

Paniagua-Chavez, C. G.; Jenkins, J.; Segovia, M.; Tiersch, T.R.

2006-01-01

Evaluation of sperm motility is the single most widely used parameter to determine semen quality in mammals and aquatic species. While a good indicator for fresh sperm viability, post-thaw motility is not always effective at predicting fertilizing ability. Techniques using fluorescent dyes can assess functionality of mammalian sperm, but have not been widely applied in aquatic organisms. The eastern oyster Crassostrea virginica is an important mollusk in the United States, and cryopreservation protocols have been developed to preserve sperm and larvae to assist research and hatchery production. In this study, protocols were developed to assess sperm cell membrane integrity and mitochondrial function by flow cytometry and to assess viability of eggs by fluorescence microscopy. The fluorescent dyes SYBR 14 and propidium iodide (PI) (to assess membrane integrity) and rhodamine 123 (R123) (to assess mitochondrial membrane potential) were used to evaluate the quality of thawed oyster sperm previously cryopreserved with different cryoprotectant and thawing treatments. Membrane integrity results were correlated with motility of thawed sperm and mitochondrial membrane potential with fertilizing ability. Fluorescein diacetate (FDA) was used to assess cytotoxicity of cryoprotectant solutions and post-thaw damage to oyster eggs. The results indicated that membrane integrity (P = 0.004) and thawing treatments (P = 0.04), and mitochondrial membrane potential (P = 0.0015) were correlated with motility. Fertilizing ability was correlated with cryoprotectant treatments (P = 0.0258) and with mitochondrial membrane potential (P = 0.001). The dye FDA was useful in indicating structural integrity of fresh and thawed eggs. Exposure of eggs, without freezing, to dimethyl sulfoxide yielded higher percentages of stained eggs and fertilization rate than did exposure to propylene glycol (P = 0.002). Thawed eggs were not stained with FDA (<1%) and larvae were not produced. ?? 2006 Elsevier Inc. All rights reserved.

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

PubMed

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; p<0.0001) or high-density lipoprotein (r=0.93; p<0.0001). The TLX 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.

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

NASA Astrophysics Data System (ADS)

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.

The effect of freeze-thawing on magnetic resonance imaging T2* of freshly harvested bovine patellar tendon

PubMed Central

Pownder, Sarah L.; Shah, Parina H.; Potter, Hollis G.

2015-01-01

Background Analysis of fresh specimens in research studies is ideal; however, it is often necessary to freeze samples for evaluation at a later time. Limited evaluation of the effect of freeze-thawing of tendon tissue samples on inherent magnetic resonance imaging (MRI) parameters, such as ultrashort echo time (UTE) T2* values, have been performed to date. Methods This study performed UTE MRI on 14 bovine patellar tendons at harvest and after four consecutive freeze-thaw cycles. Results Results demonstrated a small but significant reduction (12%) in tendon T2* values after the first freeze thaw cycle, but not after successive cycles. Tendons from juvenile animals with open physis had a significant reduction of T2* following a single freeze thaw cycle, P<0.0001. Conclusions The results of this study emphasize the importance of using uniform tendon storage protocols when using UTE MRI in preclinical models. PMID:26029639

Change in Ice Thaw Dates for Selected U.S. Lakes, 1905-2015

EPA Pesticide Factsheets

This figure shows the change in the ice-off date, or date of ice thawing and breakup, for 14 U.S. lakes during the period from 1905 to 2015. All of the lakes have red circles with negative numbers, which represent earlier thaw dates. Larger circles indicate larger changes. For more information: www.epa.gov/climatechange/science/indicators

Three-Dimensional Microstructure of Biological Tissues during Freezing and Thawing

NASA Astrophysics Data System (ADS)

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.

Review of Thawing Time Prediction Models Depending
on Process Conditions and Product Characteristics

PubMed Central

Kluza, Franciszek; Spiess, Walter E. L.; Kozłowicz, Katarzyna

2016-01-01

Summary Determining thawing times of frozen foods is a challenging problem as the thermophysical properties of the product change during thawing. A number of calculation models and solutions have been developed. The proposed solutions range from relatively simple analytical equations based on a number of assumptions to a group of empirical approaches that sometimes require complex calculations. In this paper analytical, empirical and graphical models are presented and critically reviewed. The conditions of solution, limitations and possible applications of the models are discussed. The graphical and semi--graphical models are derived from numerical methods. Using the numerical methods is not always possible as running calculations takes time, whereas the specialized software and equipment are not always cheap. For these reasons, the application of analytical-empirical models is more useful for engineering. It is demonstrated that there is no simple, accurate and feasible analytical method for thawing time prediction. Consequently, simplified methods are needed for thawing time estimation of agricultural and food products. The review reveals the need for further improvement of the existing solutions or development of new ones that will enable accurate determination of thawing time within a wide range of practical conditions of heat transfer during processing. PMID:27904387

Leachate Properties and Cadmium Migration Through Freeze-thaw Treated Soil Columns.

PubMed

Xu, Meng; Zheng, Yue; Chen, Weiwei; Mao, Na; Guo, Ping

2017-01-01

Soil column leaching experiments were conducted to study the effects of multiple freeze-thaw cycles on the vertical migration of cadmium (Cd). Three Cd-spiked leaching solutions of different properties were derived from snowmelt, sludge, and straw, designated as B, W and J, respectively. The leaching solutions varied in dissolved organic matter (DOM) concentrations in the order of J > W > B. Changes in leachate properties and Cd concentration were observed. The results showed that pH values of all the leachate solutions through freeze-thaw treated soil columns were higher than those of leachates through unfrozen soils. However, electrical conductivity (EC) values decreased compared with leachates in unfrozen treated soil columns. Although the concentrations of DOM in leachate solutions had no evident differences between the freeze-thaw and unfrozen treated soil columns, the concentrations of DOM in the leachate solutions B, W and J were different. Freeze-thaw cycles resulted in increased concentrations of Cd in the leachate solutions in the order J > W > B, and promoted a deeper migration of Cd in the soil columns. Thus, it was shown that freeze-thaw cycles may increase the risk of groundwater pollution by Cd.

Changes to Extender, Cryoprotective Medium, and In Vitro Fertilization Improve Zebrafish Sperm Cryopreservation.

PubMed

Matthews, Jennifer L; Murphy, Joy M; Carmichael, Carrie; Yang, Huiping; Tiersch, Terrence; Westerfield, Monte; Varga, Zoltan M

2018-01-25

Sperm cryopreservation is a highly efficient method for preserving genetic resources. It extends the reproductive period of males and significantly reduces costs normally associated with maintenance of live animal colonies. However, previous zebrafish (Danio rerio) cryopreservation methods have produced variable outcomes and low post-thaw fertilization rates. To improve post-thaw fertilization rates after cryopreservation, we developed a new extender and cryoprotective medium (CPM), introduced quality assessment (QA), determined the optimal cooling rate, and improved the post-thaw in vitro fertilization process. We found that the hypertonic extender E400 preserved motility of sperm held on ice for at least 6 h. We implemented QA by measuring sperm cell densities with a NanoDrop spectrophotometer and sperm motility with computer-assisted sperm analysis (CASA). We developed a CPM, RMMB, which contains raffinose, skim milk, methanol, and bicine buffer. Post-thaw motility indicated that the optimal cooling rate in two types of cryogenic vials was between 10 and 15°C/min. Test thaws from this method produced average motility of 20% ± 13% and an average post-thaw fertilization rate of 68% ± 16%.

Permafrost thaw strongly reduces allowable CO2 emissions for 1.5°C and 2°C

NASA Astrophysics Data System (ADS)

Kechiar, M.; Gasser, T.; Kleinen, T.; Ciais, P.; Huang, Y.; Burke, E.; Obersteiner, M.

2017-12-01

We quantify how the inclusion of carbon emission from permafrost thaw impacts the budgets of allowable anthropogenic CO2 emissions. We use the compact Earth system model OSCAR v2.2 which we expand with a permafrost module calibrated to emulate the behavior of the complex models JSBACH, ORCHIDEE and JULES. When using the "exceedance" method and with permafrost thaw turned off, we find budgets very close to the CMIP5 models' estimates reported by IPCC. With permafrost thaw turned on, the total budgets are reduced by 3-4%. This corresponds to a 33-45% reduction of the remaining budget for 1.5°C, and a 9-13% reduction for 2°C. When using the "avoidance" method, however, permafrost thaw reduces the total budget by 3-7%, which corresponds to reductions by 33-56% and 56-79% of the remaining budget for 1.5°C and 2°C, respectively. The avoidance method relies on many scenarios that actually peak below the target whereas the exceedance method overlooks the carbon emitted by thawed permafrost after the temperature target is reached, which explains the difference. If we use only the subset of scenarios in which there is no net negative emissions, the permafrost-induced reduction in total budgets rises to 6-15%. Permafrost thaw therefore makes the emission budgets strongly path-dependent. We also estimate budgets of needed carbon capture in scenarios overshooting the temperature targets. Permafrost thaw strongly increases these capture budgets: in the case of a 1.5°C target overshot by 0.5°C, which is in line with the Paris agreement, about 30% more carbon must be captured. Our conclusions are threefold. First, inclusion of permafrost thaw systematically reduces the emission budgets, and very strongly so if the temperature target is overshot. Second, the exceedance method, that is the only one that complex models can follow, only partially accounts for the effect of slow non-linear processes such as permafrost thaw, leading to overestimated budgets. Third, the newfound strong path-dependency of the budgets renders the concept more delicate to use. For instance, using a budget that implicitly assumes a large development of negative emission technologies may lead to missing the target if these are not as scalable as anticipated.

Partial Recovery of Mitochondrial Function of Vitrified Porcine MII Stage Oocytes During Post-Thaw Incubation.

PubMed

Dai, J J; Yang, J H; Zhang, S S; Niu, Y F; Chen, Y N; Wu, C F; Zhang, D F

　　The survival of porcine oocytes is still very low after cryopreservation. To investigate whether and when the mitochondrial function of vitrified porcine oocytes could be recovered post-thaw. Mitochondrial potential, ROS level, ATP content, apoptotic rate, caspase activity, and parthenogenetics developmental ability of thawed porcine oocytes were measured after culture in vitro for 0, 1, 2 or 4 h. Mitochondrial potential after 2 h and 4 h post-thaw culture were 1.19 and 1.26, significantly lower than that of fresh oocytes but much higher than the groups cultured for 0 h and 1 h (P<0.05). Cryopreservation increased the ROS level in oocytes considerably, which decreased only after 2 to 4 h incubation following thaw. ATP content increased gradually over time and recovered to the level comparable to that of fresh oocytes after 4 h. Pan caspase levels increased after cryopreservation and reached the highest level at 1 h incubation. Thereafter it decreased to a low value, but still higher than fresh oocytes. Oocytes showing an early apoptotic event decreased upon 2 to 4 h incubation. The parthenogenetic cleavage and blastocyst rates were the highest (19.8% and 5.6%) after 2 h incubation. The recovery of mitochondrial function could complete after 2 to 4 h post-thaw incubation. Post-thaw incubation for 2 to 4 h reduced apoptotic events and improved parthenogenetic developmental ability of vitrified porcine MII stage oocytes.

Cooled semen for fixed-time artificial insemination in beef cattle.

PubMed

Borges-Silva, Juliana C; Silva, Márcio R; Marinho, Daniel B; Nogueira, Eriklis; Sampaio, Deiler C; Oliveira, Luiz Orcírio F; Abreu, Urbano G P; Mourão, Gerson B; Sartori, Roberto

2016-06-01

This study evaluated the use of cooled semen in a fixed-time artificial insemination (FTAI) program compared with frozen-thawed semen to improve pregnancy rates in beef cattle. Ejaculates of three bulls were collected and divided into two treatments: (1) frozen-thawed semen and (2) cooled semen. Egg-yolk extender without glycerol was used for the cooled semen treatment. Straws (25×10 6 spermatozoa) were submitted to cooling for preservation at 5°C for 24h, after which FTAI was performed. Nelore cows (n=838) submitted to FTAI were randomly inseminated using frozen-thawed semen or cooled semen. There was a 20% increase in the pregnancy per AI (P AI -1 ) using cooled semen compared with frozen-thawed semen (59.9±4.7 vs 49.4±5.0%; P<0.005). There was no difference in P AI -1 among the bulls (P=0.40). The frozen-thawed semen had fewer functional spermatozoa than did the cooled semen when evaluated by sperm motility (61.7 vs 81.0%), slow thermoresistance test (41.7 vs 66.7%) and hypoosmotic swelling test (38.3 vs 53.7%; P<0.05). The percentage of sperm abnormalities did not differ between the freeze-thawing and cooling processes (18.6 vs 22.1%; P>0.05). Because there was less damage to spermatozoa and improvement in P AI -1 , the use of cooled semen instead of frozen-thawed semen is an interesting approach to increase reproductive efficiency in cattle submitted to a FTAI protocol.

Factors affecting storage of Slovak native rabbit semen in the gene bank.

PubMed

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.

Freezing and thawing or freezing, thawing, and aging effects on beef tenderness.

PubMed

Grayson, A L; King, D A; Shackelford, S D; Koohmaraie, M; Wheeler, T L

2014-06-01

The objective of this study was to determine the effect of freezing and thawing or freezing and thawing with an additional aging period after frozen storage on the tenderness of longissimus lumborum (LL) and semitendinosus (ST) steaks relative to aged, fresh steaks. Left-side LL and ST (n = 35 each) were obtained from U.S. Select carcasses classified at the grading stand by the U.S. Meat Animal Research Center visible and near-infrared spectroscopy tenderness system to have predicted slice shear force greater than 16.5 kg at 14 d postmortem. At 2 d postmortem, 2.54 cm thick steaks were cut from each muscle and assigned to 1 of the following treatments: 2 d fresh (2FRESH), 2 d freeze + thaw (2FREEZE), 2 d freeze + thaw + 12 d age (2FREEZE+12AGE), 14 d fresh (14FRESH), 14 d freeze + thaw (14FREEZE), 14 d freeze + thaw + 14 d age (14FREEZE+14AGE), and 28 d fresh (28FRESH). Steaks assigned to a freezing treatment were frozen at -26°C for 30 d before thawing/cooking or thawing with an additional aging period at 2°C. Slice shear force for LL and ST was lower (P < 0.01) for 2FREEZE (27.4 and 24.5 kg) and 14FREEZE (22.4 and 22.4 kg) compared to 2FRESH (33.0 and 29.2 kg) and 14FRESH (25.3 and 25.5 kg), respectively. Slice shear force for LL and ST was lower (P < 0.01) for 2FREEZE+12AGE (17.8 and 20.8 kg) and 14FREEZE+14AGE (14.6 and 19.0 kg) compared to 14FRESH (25.3 and 25.5 kg) and 28FRESH (18.7 and 21.7 kg), respectively. Desmin degradation for LL was not different (P > 0.05) between 2FREEZE (21.0%) and 2FRESH (14.6%) or between 14FREEZE (40.4%) and 14FRESH (38.4%); however, desmin degradation was higher (P < 0.06) in 2FREEZE+12AGE (46.7%) and 14FREEZE+14AGE (71.1%) when compared to 14FRESH (38.4%) and 28FRESH (60.5%), respectively. Cooking loss for LL was higher (P < 0.01) in 2FREEZE+12AGE (15.2%) compared to 14FRESH (14.0%) but was not different (P > 0.05) between 14FREEZE+14AGE (15.0%) and 28FRESH (14.3%). Freezing and thawing or a combination of freezing, thawing, and aging resulted in increased tenderness for LL and ST steaks when compared to fresh steaks with the same aging time. These results indicate freezing could be incorporated into normal commercial product distribution processes to improve the consistency of meat tenderness. Researchers who freeze steaks before tenderness assessment should be aware and acknowledge that freezing affects tenderness data.

[Standardization of cryopreservation process of Saccharomyces boulardii yeasts for usage in collections and banks of industrial microorganism strains].

PubMed

Gurina, T M; Vysekantsev, I P; Babinets, O M

2013-01-01

New implementation principle of freeze-thawing during cryopreservation of microorganisms, initiation of the process of crystal formation at cooling and thawing stage with the controlled rate of heating was experimentally substantiated. This allows increasing the cell viability, guaranteeing their equal number in each preparation, decreasing the contamination risk of the samples at thawing stage.

Influences of Land Use/Cover Types on Nitrous Oxide Emissions during Freeze-Thaw Periods from Waterlogged Soils in Inner Mongolia.

PubMed

Lu, Zedong; Du, Rui; Du, Pengrui; Qin, Saisai; Liang, Zongmin; Li, Ziming; Wang, Yaling; Wang, Yanfen

2015-01-01

Nitrous oxide emissions during freeze/thaw periods contribute significantly to annual soil N2O emissions budgets in middle- and high-latitude areas; however, the freeze/thaw-related N2O emissions from waterlogged soils have hardly been studied in the Hulunber Grassland, Inner Mongolia. For this study, the effects of changes in land use/cover types on N2O emissions during freeze-thaw cycles were investigated to more accurately quantify the annual N2O emissions from grasslands. Soil cores from six sites were incubated at varying temperature (ranging from -15 to 10°C) to simulate freeze-thaw cycles. N2O production rates were low in all soil cores during freezing periods, but increased markedly after soil thawed. Mean rates of N2O production differed by vegetation type, and followed the sequence: Leymus chinensis (LC) and Artemisia tanacetifolia (AT) steppes > LC steppes ≥ Stipa baicalensis (SB) steppes. Land use types (mowing and grazing) had differing effects on freeze/thaw-related N2O production. Grazing significantly reduced N2O production by 36.8%, while mowing enhanced production. The production of N2O was related to the rate at which grassland was mowed, in the order: triennially (M3) > once annually (M1) ≥ unmown (UM). Compared with the UM control plot, the M3 and M1 mowing regimes enhanced N2O production by 57.9% and 13.0% respectively. The results of in situ year-round measurements showed that large amounts of N2O were emitted during the freeze-thaw period, and that annual mean fluxes of N2O were 9.21 μg N2O-N m-2 h-1 (ungrazed steppe) and 6.54 μg N2O-N m-2 h-1 (grazed steppe). Our results further the understanding of freeze/thaw events as enhancing N2O production, and confirm that different land use/cover types should be differentiated rather than presumed to be equivalent, regarding nitrous oxide emission. Even so, further research involving multi-year and intensive measurements of N2O emission is still needed.

Enhanced bioremediation of nutrient-amended, petroleum hydrocarbon-contaminated soils over a cold-climate winter: The rate and extent of hydrocarbon biodegradation and microbial response in a pilot-scale biopile subjected to natural seasonal freeze-thaw temperatures.

PubMed

Kim, Jihun; Lee, Aslan Hwanhwi; Chang, Wonjae

2018-01-15

A pilot-scale biopile field experiment for nutrient-amended petroleum-contaminated fine-grained soils was performed over the winter at a cold-climate site. The rate and extent of hydrocarbon biodegradation and microbial responses were determined and corresponded to the on-site soil phase changes (from unfrozen to partially frozen, deeply frozen, and thawed) associated with natural seasonal freeze-thaw conditions. Treated and untreated biopiles were constructed (~3500kg each) on an open outdoor surface at a remediation facility in Saskatoon, Canada. The treated biopile received N-P-K-based nutrient and humate amendments before seasonal freezing. Real-time field monitoring indicated significant unfrozen water content in the treated and untreated biopiles throughout the freezing period, from the middle of November to early March. Unfrozen water was slightly more available in the treated biopile due to the aqueous nutrient supply. Soil CO 2 production and O 2 consumption in the treated biopile were generally greater than in the untreated biopile. Total removal percentages for F2 (>C10-C16), F3 (>C16-C34), and total petroleum hydrocarbons (TPH) in the treated biopile were 57, 58, and 58%, respectively, of which 26, 39, and 33% were removed during seasonal freezing and early thawing between November to early March. F3 degradation largely occurred during freezing while F2 hydrocarbons were primarily removed during thawing. Biomarker-based hydrocarbon analyses confirmed enhanced biodegradation in the treated biopile during freezing. The soil treatment increased the first-order rate constants for F2, F3, and TPH degradation by a factor of 2 to 7 compared to the untreated biopile. Shifts in bacterial community appeared in both biopiles as the biopile soils seasonally froze and thawed. Increased alkB1 gene copy numbers in the treated biopile, especially in the partially thawed phase during early thawing, suggest extended hydrocarbon biodegradation to the seasonal freeze-thaw season, due to the nutrients supplied prior to seasonal freezing. Copyright © 2017 Elsevier B.V. All rights reserved.

The effect of permafrost thaw on short- and long-term carbon accumulation in permafrost mires

NASA Astrophysics Data System (ADS)

Olid, Carolina; Klaminder, Jonatan; Monteux, Sylvain; Johansson, Margareta; Dorrepaal, Ellen

2017-04-01

Permafrost stores twice as much carbon (C) as is currently present in the atmosphere. During recent years, warmer temperatures in the Arctic has caused rapid thawing of permafrost, which have dramatically altered permafrost C storage by increasing both microbial decomposition and plant productivity. Although current research focuses on the effects of climate change on these two processes, there are still no scientific consensus about the magnitude or even the direction of future C feedbacks from permafrost ecosystems. Field manipulation experiments have been widely used during the last decade to improve our knowledge about the net effects of permafrost thaw in the permafrost C storage. However, due to the slow response (decades) of permafrost ecosystems to environmental changes and the short-time nature of these experiments (usually shorter than 5-9 years), there are still concerns when attempting to extrapolate the results to predict long term effects. In addition, measurements are mostly taken exclusively during the summer season, without taking into account inter-annual variability in C fluxes and underestimating microbial activity throughout the cold season. The need to develop a comprehensive understanding of C fluxes over the entire year and at long temporal scales sets the basis of this study. This study aims to quantify the effects of permafrost thawing in permafrost C fluxes using a 12 years permafrost thaw experiment in northern Sweden. Our aims were to quantify the effect of permafrost thaw in both decomposition and primary production in active layer and newly thawed permafrost, and its implications for the C balance. Based on previous observations, we hypothesized that 1) soil decomposition rates were higher in manipulated thaw plots. However, 2) the observed increase in nutrients availability and the higher presence of vascular plants after thawing stimulate primary production, which compensates to some extent the increased C losses by respiration. To validate these hypotheses, an empirically based peat development model was applied to peat cores collected from manipulated thaw (n=6) and control (n=6) plots. Short- (decades) and long-(centuries) term C accumulation rates were estimated using a combined 210Pb and 14C chronology. In contrast to previous studies, our approach is long term and allows applying an empirical mass balance to evaluate depth-explicit changes in C inputs, C losses, and net C accumulation rates in response to permafrost thaw. Comparing shallow versus deep soil C does not only reflect short versus long-term C dynamics but also shows how the responses vary depending on different soil conditions. Our goal is to provide insights to better understand permafrost C dynamics to improve theoretical and predictive climate impact models.

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

PubMed

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

2007-06-01

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

Retained functional integrity of bull spermatozoa after double freezing and thawing using PureSperm density gradient centrifugation.

PubMed

Maxwell, W M C; Parrilla, I; Caballero, I; Garcia, E; Roca, J; Martinez, E A; Vazquez, J M; Rath, D

2007-10-01

The main aim of this study was to compare the motility and functional integrity of bull spermatozoa after single and double freezing and thawing. The viability and morphological integrity of spermatozoa selected by PureSperm density gradient centrifugation after cryopreservation of bovine semen in two commercial extenders (Experiment 1) and the function of bull spermatozoa before and after a second freezing and thawing assisted by PureSperm selection (Experiment 2) were examined. On average, 35.8 +/- 12.1% of sperm loaded onto the PureSperm density gradient were recovered after centrifugation. In Experiment 1, post-thaw motility and acrosome integrity were higher for spermatozoa frozen in Tris-egg yolk extender than in AndroMed, whether the assessments were made immediately after thawing [80.4 +/- 12.7 vs 47.6 +/- 19.0% motile and 78.8 +/- 8.3 vs 50.1 +/- 19.5% normal apical ridge (NAR), p < 0.05] or after preparation on the gradient (83.3 +/- 8.6 vs 69.4 +/- 15.9% motile and 89.5 +/- 7.2 vs 69.1 +/- 11.4% NAR, p < 0.05). For semen frozen in Tris-egg yolk extender, selection on the PureSperm gradient did not influence total motility but significantly improved the proportion of acrosome-intact spermatozoa. After the gradient, both the total motility and percentage of normal acrosomes increased for spermatozoa frozen in AndroMed (Minitüb Tiefenbach, Germany). In Experiment 2, there was no difference in sperm motility after the first and second freeze-thawing (82.9 +/- 12.7 vs 68.8 +/- 18.7%). However, the proportion of acrosome-intact spermatozoa was significantly improved by selection through the PureSperm gradient, whether measured by phase contrast microscopy (78.9 +/- 9.7 vs 90.4 +/- 4.0% NAR, p < 0.05) or flow cytometry (53.4 +/- 11.7 vs 76.3 +/- 6.0% viable acrosome-intact spermatozoa, p < 0.001). The improvement in the percentage of spermatozoa with normal acrosomes was maintained after resuspension in the cooling extender and cooling to 4 degrees C (88.2 +/- 6.2) and after re-freezing and thawing (83.6 +/- 6.56% NAR). However, flow cytometric assessment of the sperm membranes revealed a decline in the percentage of viable spermatozoa with intact membranes after the second freezing and thawing compared with after gradient centrifugation (76.3 +/- 6.0% vs 46.6 +/- 6.6%, p < 0.001) to levels equivalent to those obtained after the first round of freeze-thawing (53.4 +/- 11.7% viable acrosome-intact spermatozoa). Sperm movement characteristics assessed by computer-assisted analysis were unaffected in the population selected on the PureSperm gradients but declined after cooling of the selected and extended spermatozoa to 4 degrees C. There was no further change in these kinematic measurements after the cooled spermatozoa had undergone the second round of freeze-thawing. These results demonstrate that bull semen can be frozen and thawed, followed by a second freeze-thawing cycle of a population of spermatozoa selected by PureSperm, with retained motility and functional integrity. This points to the possibility of using double frozen spermatozoa in bovine artificial insemination programmes and to the potential benefits of PureSperm density gradient centrifugation for the application of cryopreserved bull spermatozoa to other biotechnological procedures such as flow cytometric sex sorting followed by re-freezing and thawing.

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

PubMed

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

2014-01-01

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

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

PubMed

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.

Colour Changes in Meat of Foals as Affected by Slaughtering Age and Post-thawing Time

PubMed Central

De Palo, P.; Maggiolino, A.; Centoducati, P.; Tateo, A.

2012-01-01

The aim of the present work was to investigate how colour changes of foal meat can vary after thawing out in relation to the slaughtering age of the horses and to the post-thawing time. Eighteen Italian Heavy Draught Horse (IHDH) foals were used for the trial. They were subdivided in three groups according to their slaughtering age (6, 11 and 18 months). Two different surfaces were investigated for each sample: a fresh cut surface (daily renewed cutting surface: DRCS), and not-renewed cutting surface (NRCS). The redness of both investigated surfaces increased with slaughtering age (p<0.01). Moreover, this parameter decreased during post-thawing time (p<0.01) only on the NRCS, probably due to the myoglobin oxidation processes. Colour is an important visual cue denoting perceived quality by consumers. So, by a chromatic perspective the thawed meat of IHDH foals slaughtered at 6 and 11 months proved to be that which best meets the market requirements. PMID:25049544

Effect of Fast Freeze-Thaw Cycles on Mechanical Properties of Ordinary-Air-Entrained Concrete

PubMed Central

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

2014-01-01

Freezing-thawing resistance is a very significant characteristic for concrete in severe environment (such as cold region with the lowest temperature below 0°C). In this study, ordinary-air-entrained (O-A-E) concrete was produced in a laboratory environment; the compressive strength, cubic compressive strength of C50, C40, C30, C25, and C20 ordinary-air-entrained concrete, tensile strength, and cleavage strength of C30 ordinary-air-entrained concrete were measured after fast freeze-thaw cycles. The effects of fast freeze-thaw cycles on the mechanical properties (compressive strength and cleavage strength) of ordinary-air-entrained concrete materials are investigated on the basis of the experimental results. And the concise mathematical formula between mechanical behavior and number of fast freeze-thaw cycles was established. The experiment results can be used as a reference in design, maintenance, and life prediction of ordinary-air-entrained concrete structure (such as dam, offshore platform, etc.) in cold regions. PMID:24895671

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

PubMed

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.

Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples.

PubMed

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.

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

PubMed

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.

Effects of soy hull pectin and insoluble fiber on physicochemical and oxidative characteristics of fresh and frozen/thawed beef patties.

PubMed

Kim, Hyun-Wook; Miller, Danika K; Lee, Yong Jae; Kim, Yuan H Brad

2016-07-01

The objective of this study was to determine the effects of pectin and insoluble fiber isolated from soy hulls on water-holding capacity (WHC), texture, color, and lipid oxidation of fresh and frozen/thawed beef patties. Beef patties were formulated with no dietary fiber (control), 1% soy hull pectin, insoluble fiber, or their mixture (1:1), respectively. The addition of soy hull pectin significantly decreased display weight loss and increased cook yield of both fresh and frozen/thawed beef patties. In addition, no significant difference in hardness between fresh and frozen/thawed beef patties was observed for all dietary fiber treatments. However, incorporation of insoluble soy hull fiber decreased color and lipid oxidation stabilities of both fresh and frozen/thawed beef patties. Our results indicate that the incorporation of soy hull pectin could be an effective non-meat ingredient to minimize water loss and hardness defects of frozen beef patties. Copyright © 2016 Elsevier Ltd. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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.

Limited contribution of permafrost carbon to methane release from thawing peatlands

NASA Astrophysics Data System (ADS)

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 (<2 g CH4 m-2 yr-1). Instead, fluxes were driven by anaerobic decomposition of recent C inputs. We conclude that thaw-induced changes in surface wetness and wetland area, rather than the anaerobic decomposition of previously frozen C, may determine the effect of permafrost thaw on CH4 emissions from northern peatlands.

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

PubMed

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

2014-06-01

Sugar surfactants with different alkyl chain lengths and sugar head groups were compared for their protein-stabilizing effect during freeze-thawing and freeze-drying. Six enzymes, different in terms of tolerance against inactivation because of freeze-thawing and freeze-drying, were used as model proteins. The enzyme activities that remained after freeze-thawing and freeze-drying in the presence of a sugar surfactant were measured for different types and concentrations of sugar surfactants. Sugar surfactants stabilized all of the tested enzymes both during freeze-thawing and freeze-drying, and a one or two order higher amount of added sugar surfactant was required for achieving protein stabilization during freeze-drying than for the cryoprotection. The comprehensive comparison showed that the C10-C12 esters of sucrose or trehalose were the most effective through the freeze-drying process: the remaining enzyme activities after freeze-thawing and freeze-drying increased at the sugar ester concentrations of 1-10 and 10-100 μM, respectively, and increased to a greater extent than for the other surfactants at higher concentrations. Results also indicate that, when a decent amount of sugar was also added, the protein-stabilizing effect of a small amount of sugar ester through the freeze-drying process could be enhanced. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

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

PubMed

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.

Freeze concentration of proteins in Antarctic krill wash water

NASA Astrophysics Data System (ADS)

Qi, Xiangming; Xu, Jing; Zhao, Kuo; Guo, Hui; Ma, Lei

2017-12-01

Water-washing removes fluoride from Antarctic krill but produces large volumes of wash water containing water- soluble proteins and fluoride. The freeze concentration method was tested to determine if it could be used to recover water-soluble proteins while leaving the fluoride in solution. After freezing and thawing the wash water, protein and fluoride contents of the thawed fractions were determined to explore the melting regularity of components in the wash water. The highest concentration factors of protein and fluoride were obtained after 80 min of thawing, such as 1.48 ± 0.06 and 1.35 ± 0.04 times, respectively. The free amino-nitrogen (FAN) content and sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern results revealed that the highest concentrations of all ingredients were obtained after 80 min of the process. The degree of hydrolysis of all fractions from the thawing process fluctuated in a narrow range around 12% during the entire process, indicating that the thawing order did not change with various proteins or time during the entire thawing course. These results demonstrate that the freeze concentration method can be used to concentrate protein solutions, even those with fluoride. It was concluded that condensation was achieved and no ingredient could be separated, regardless of fluoride, amino acids, or different proteins in the water.

Impact of Delayed Infusion Time in Umbilical Cord Blood Transplantation.

PubMed

Mitchell, Richard; Wagner, John E; Brunstein, Claudio; Cao, Qing; McKenna, David H; Verneris, Michael R

2017-05-01

In umbilical cord blood (UCB) transplantation, UCB units are typically thawed, washed, and infused into the patient as rapidly as possible. In some instances there is a delay in the time from the unit thaw and wash procedure to infusion into the patient. Therefore, we examined the effect of thaw duration time on engraftment outcomes in 567 patients undergoing UCB transplantation. With a range of 32 to 523 minutes, a prolonged thaw duration had no obvious effect on the incidence of neutrophil engraftment or time to recovery. This was true for recipients of single UCB transplantation (incidence: 97% versus 93%, P = .13; time to neutrophil recovery: 21 days versus 21 days, P = .32; and platelet recovery: 79% versus 78%, P = .48), and similar results were observed in double UCB transplantation (time to neutrophil engraftment: 20 days versus 19 days, P = .71). However, there was a trend toward better platelet recovery in recipients of double UCB transplants with prolonged thaw duration (HR, 1.28; P = .06). In conclusion, this study demonstrates prolonged thaw duration has no detrimental effect on engraftment after single or double UCB transplantation. Copyright © 2017 The American Society for Blood and Marrow Transplantation. Published by Elsevier Inc. All rights reserved.

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

PubMed

Wang, Jing; Fu, Rongbing; Xu, Zhen

2017-08-01

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

Controls on methane released through ebullition in peatlands affected by permafrost degradation

USGS Publications Warehouse

Klapstein, Sara J.; Turetsky, Merritt R.; McGuire, A. David; Harden, Jennifer W.; Czimczik, C.I.; Xu, Xiaomei; Chanton, J.P.; Waddington, James Michael

2014-01-01

Permafrost thaw in peat plateaus leads to the flooding of surface soils and the formation of collapse scar bogs, which have the potential to be large emitters of methane (CH4) from surface peat as well as deeper, previously frozen, permafrost carbon (C). We used a network of bubble traps, permanently installed 20 cm and 60 cm beneath the moss surface, to examine controls on ebullition from three collapse bogs in interior Alaska. Overall, ebullition was dominated by episodic events that were associated with changes in atmospheric pressure, and ebullition was mainly a surface process regulated by both seasonal ice dynamics and plant phenology. The majority (>90%) of ebullition occurred in surface peat layers, with little bubble production in deeper peat. During periods of peak plant biomass, bubbles contained acetate-derived CH4 dominated (>90%) by modern C fixed from the atmosphere following permafrost thaw. Post-senescence, the contribution of CH4 derived from thawing permafrost C was more variable and accounted for up to 22% (on average 7%), in the most recently thawed site. Thus, the formation of thermokarst features resulting from permafrost thaw in peatlands stimulates ebullition and CH4 release both by creating flooded surface conditions conducive to CH4 production and bubbling as well as by exposing thawing permafrost C to mineralization.

Spaceborne Microwave Remote Sensing of Seasonal Freeze-Thaw Processes in the Terrestrial High Latitudes: Relationships with Land-Atmosphere CO2 exchange

NASA Technical Reports Server (NTRS)

McDonald, Kyle C.; Kimball, John S.; Zhao, Maosheng; Njoku, Eni; Zimmermann, Reiner; Running, Steven W.

2004-01-01

Landscape transitions between seasonally frozen and thawed conditions occur each year over roughly 50 million square kilometers of Earth's Northern Hemisphere. These relatively abrupt transitions represent the closest analog to a biospheric and hydrologic on/off switch existing in nature, affecting surface meteorological conditions, ecological trace gas dynamics, energy exchange and hydrologic activity profoundly. We utilize time series satellite-borne microwave remote sensing measurements from the Special Sensor Microwave Imager (SSM/I) to examine spatial and temporal variability in seasonal freeze/thaw cycles for the pan-Arctic basin and Alaska. Regional measurements of spring thaw timing are derived using daily brightness temperature measurements from the 19 GHz, horizontally polarized channel, separately for overpasses with 6 AM and 6 PM equatorial crossing times. Spatial and temporal patterns in regional freeze/thaw dynamics show distinct differences between North America and Eurasia, and boreal forest and Arctic tundra biomes. Annual anomalies in the timing of thawing in spring also correspond closely to seasonal atmospheric CO2 concentration anomalies derived from NOAA CMDL arctic and subarctic monitoring stations. Classification differences between AM and PM overpass data average approximately 5 days for the region, though both appear to be effective surrogates for monitoring annual growing seasons at high latitudes.

Spaceborne microwave remote sensing of seasonal freeze-thaw processes in theterrestrial high l atitudes : relationships with land-atmosphere CO2 exchange

NASA Technical Reports Server (NTRS)

McDonald, Kyle C.; Kimball, John S.; Zhao, Maosheng; Njoku, Eni; Zimmermann, Reiner; Running, Steven W.

2004-01-01

Landscape transitions between seasonally frozen and thawed conditions occur each year over roughly 50 million square kilometers of Earth's Northern Hemisphere. These relatively abrupt transitions represent the closest analog to a biospheric and hydrologic on/off switch existing in nature, affecting surface meteorological conditions, ecological trace gas dynamics, energy exchange and hydrologic activity profoundly. We utilize time series satellite-borne microwave remote sensing measurements from the Special Sensor Microwave Imager (SSM/I) to examine spatial and temporal variability in seasonal freeze/thaw cycles for the pan-Arctic basin and Alaska. Regional measurements of spring thaw timing are derived using daily brightness temperature measurements from the 19 GHz, horizontally polarized channel, separately for overpasses with 6 AM and 6 PM equatorial crossing times. Spatial and temporal patterns in regional freeze/thaw dynamics show distinct differences between North America and Eurasia, and boreal forest and Arctic tundra biomes. Annual anomalies in the timing of thawing in spring also correspond closely to seasonal atmospheric CO2 concentration anomalies derived from NOAA CMDL arctic and subarctic monitoring stations. Classification differences between AM and PM overpass data average approximately 5 days for the region, though both appear to be effective surrogates for monitoring annual growing seasons at high latitudes.

Increased nitrous oxide emissions from Arctic peatlands after permafrost thaw

PubMed Central

Marushchak, Maija E.; Lamprecht, Richard E.; Jackowicz-Korczyński, Marcin; Lindgren, Amelie; Mastepanov, Mikhail; Granlund, Lars; Christensen, Torben R.; Tahvanainen, Teemu; Martikainen, Pertti J.; Biasi, Christina

2017-01-01

Permafrost in the Arctic is thawing, exposing large carbon and nitrogen stocks for decomposition. Gaseous carbon release from Arctic soils due to permafrost thawing is known to be substantial, but growing evidence suggests that Arctic soils may also be relevant sources of nitrous oxide (N2O). Here we show that N2O emissions from subarctic peatlands increase as the permafrost thaws. In our study, the highest postthaw emissions occurred from bare peat surfaces, a typical landform in permafrost peatlands, where permafrost thaw caused a fivefold increase in emissions (0.56 ± 0.11 vs. 2.81 ± 0.6 mg N2O m−2 d−1). These emission rates match those from tropical forest soils, the world’s largest natural terrestrial N2O source. The presence of vegetation, known to limit N2O emissions in tundra, did decrease (by ∼90%) but did not prevent thaw-induced N2O release, whereas waterlogged conditions suppressed the emissions. We show that regions with high probability for N2O emissions cover one-fourth of the Arctic. Our results imply that the Arctic N2O budget will depend strongly on moisture changes, and that a gradual deepening of the active layer will create a strong noncarbon climate change feedback. PMID:28559346

Do thawing and warming affect the integrity of human milk?

PubMed

Handa, D; Ahrabi, A F; Codipilly, C N; Shah, S; Ruff, S; Potak, D; Williams, J E; McGuire, M A; Schanler, R J

2014-11-01

To evaluate the integrity of the human milk (pH, bacterial counts, host defense factors and nutrients) subjected to thawing, warming, refrigeration and maintenance at room temperature. Mothers in the neonatal intensive care unit donated freshly expressed milk. A baseline sample was stored at -80 °C and the remainder of the milk was divided and stored for 7 days at -20 °C. The milk was then subjected to two methods of thawing and warming: tepid water and waterless warmer. Thawed milk also was refrigerated for 24 h prior to warming. Lastly, warmed milk was maintained at room temperature for 4 h to simulate a feeding session. Samples were analyzed for pH, bacterial colony counts, total fat and free fatty acids, and the content of protein, secretory IgA and lactoferrin. Data were analyzed by repeated-measures analysis of variance and paired t test. There were no differences between processing methods and no changes in fat, protein, lactoferrin and secretory immunoglobulin A with processing steps. Milk pH and bacterial colony counts declined while free fatty acids rose with processing. Refrigeration of thawed milk resulted in greater declines in pH and bacteria and increases in free fatty acids. Bacterial colony counts and free fatty acids increased with maintenance at room temperature. The integrity of the milk was affected similarly by the two thawing and warming methods. Thawing and warming change the integrity of previously frozen human milk, but not adversely. Concerns about maintaining warmed milk at room temperature need to be explored.

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

PubMed

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.

Balancing risk and benefit: maintenance of a thawed Group A plasma inventory for trauma patients requiring massive transfusion.

PubMed

Mehr, Chelsea R; Gupta, Rajan; von Recklinghausen, Friedrich M; Szczepiorkowski, Zbigniew M; Dunbar, Nancy M

2013-06-01

Transfusion of plasma and red blood cell (RBC) units in a balanced ratio approximating 1:1 has been shown in retrospective studies to be associated with improved outcomes for trauma patients. Our low-volume rural trauma center uses a trauma-activated transfusion algorithm. Plasma is thawed upon activation to avoid wastage. However, the time required for plasma thawing has made achievement of a 1:1 ratio early in resuscitation challenging. In this study, the time required for plasma thawing is characterized, and a potential solution is proposed. A retrospective chart study of 38 moderately and massively transfused (≥6 U in the first 24 hours) trauma patients admitted from January 2008 to March 2012 was performed. We evaluated the time required to dispense plasma and the number of RBCs dispensed before plasma in these patients. The average time between the dispense of RBCs and plasma was 26 minutes (median, 28; range, 0-48 minutes). The average number of RBCs dispensed before plasma was 8 U (median, 7 U; range, 0-24 U). Nearly one third of massively transfused patients had 10 RBCs or greater dispensed before plasma was available. There exists the potential for delayed plasma availability owing to time required for thawing, which may compromise the ability to provide balanced plasma to RBC transfusion to trauma patients. Maintenance of a thawed Group AB plasma inventory may not be operationally feasible for rural centers with low trauma volumes. Use of a thawed Group A plasma inventory is a potential alternative to ensure rapid plasma availability. Therapeutic study, level V.

The effect of two pre-cryopreservation single layer colloidal centrifugation protocols in combination with different freezing extenders on the fragmentation dynamics of thawed equine sperm DNA

PubMed Central

2012-01-01

Background Variability among stallions in terms of semen cryopreservation quality renders it difficult to arrive at a standardized cryopreservation method. Different extenders and processing techniques (such us colloidal centrifugation) are used in order to optimize post-thaw sperm quality. Sperm chromatin integrity analysis is an effective tool for assessing such quality. The aim of the present study was to compare the effect of two single layer colloidal centrifugation protocols (prior to cryopreservation) in combination with three commercial freezing extenders on the post-thaw chromatin integrity of equine sperm samples at different post-thaw incubation (37°C) times (i.e., their DNA fragmentation dynamics). Results Post-thaw DNA fragmentation levels in semen samples subjected to either of the colloidal centrifugation protocols were significantly lower (p<0.05) immediately after thawing and after 4 h of incubation at 37°C compared to samples that underwent standard (control) centrifugation. The use of InraFreeze® extender was associated with significantly less DNA fragmentation than the use of Botu-Crio® extender at 6 h of incubation, and than the use of either Botu-Crio® or Gent® extender at 24 h of incubation (p<0.05). Conclusions These results suggest that single layer colloidal centrifugation performed with extended or raw semen prior to cryopreservation reduces DNA fragmentation during the first four hours after thawing. Further studies are needed to determine the influence of freezing extenders on equine sperm DNA fragmentation dynamics. PMID:23217215

Comparison of cook loss, shear force, and sensory descriptive profiles of boneless skinless white meat cooked from a frozen or thawed state.

PubMed

Zhuang, Hong; Savage, Elizabeth M

2013-11-01

Four replications were conducted to compare quality measurements, cook loss, shear force, and sensory quality attributes of cooked boneless skinless white meat, broiler breast fillets (pectoralis major) prepared directly from a frozen state or prepared from a thawed state. In each replication, fresh broiler fillets (removed from carcasses 6-8 h postmortem) were procured from a local commercial processing plant and stored in a -20°C freezer until use. On the sensory evaluation date, fillets were cooked to an endpoint temperature of 78°C either directly from the frozen state (thawing during cooking) or after the frozen samples were thawed in a refrigerator (2°C) overnight (thawing before cooking). Cook loss and Warner-Bratzler (WB) shear force were used as indicators for instrumental quality measurements. Sensory quality measurements were conducted by trained descriptive panelists using 0 to 15 universal intensity scales for 8 texture and 10 flavor attributes. Results show that there were no differences (P > 0.05) in measurements for sensory descriptive flavor attributes of cooked fillets between the 2 sample thawing methods, indicating that the sensory flavor profiles of both methods were similar to each other. However, WB shear force (36.98 N), cook loss (21.2%), sensory texture attributes of cohesiveness (intensity score was 5.59), hardness (5.14), rate of breakdown (5.50), and chewiness (5.21) of the breast fillets cooked directly from the frozen state were significantly higher (P < 0.05) than those of the breast meat cooked after being thawed (30.56 N, 19.0%, 5.19, 4.78, 5.29, and 5.02, respectively). These results indicate that cookery directly from frozen boneless skinless white meat can result in different measurement values of cook loss, shear force, and sensory descriptive texture attributes compared with cookery after frozen fillets are thawed.

Thermokarst lake methanogenesis along a complete talik profile

USGS Publications Warehouse

Heslop, J.K.; Walter Anthony, K.M.; Sepulveda-Jauregui, A.; Martinez-Cruz, K.; Bondurant, A.; Grosse, G.; Jones, Miriam C.

2015-01-01

Thermokarst (thaw) lakes emit methane (CH4) to the atmosphere formed from thawed permafrost organic matter (OM), but the relative magnitude of CH4 production in surface lake sediments vs. deeper thawed permafrost horizons is not well understood. We assessed anaerobic CH4 production potentials from various depths along a 590 cm long lake sediment core that captured the entire sediment package of the talik (thaw bulb) beneath the center of an interior Alaska thermokarst lake, Vault Lake, and the top 40 cm of thawing permafrost beneath the talik. We also studied the adjacent Vault Creek permafrost tunnel that extends through ice-rich yedoma permafrost soils surrounding the lake and into underlying gravel. Our results showed CH4 production potentials were highest in the organic-rich surface lake sediments, which were 151 cm thick (mean ± SD: 5.95 ± 1.67 μg C–CH4 g dw−1 d−1; 125.9 ± 36.2 μg C–CH4 g C−1org d−1). High CH4 production potentials were also observed in recently thawed permafrost (1.18 ± 0.61 μg C–CH4g dw−1 d−1; 59.60± 51.5 μg C–CH4 g C−1org d−1) at the bottom of the talik, but the narrow thicknesses (43 cm) of this horizon limited its overall contribution to total sediment column CH4 production in the core. Lower rates of CH4 production were observed in sediment horizons representing permafrost that has been thawing in the talik for a longer period of time. No CH4 production was observed in samples obtained from the permafrost tunnel, a non-lake environment. Our findings imply that CH4production is highly variable in thermokarst lake systems and that both modern OM supplied to surface sediments and ancient OM supplied to both surface and deep lake sediments by in situ thaw and shore erosion of yedoma permafrost are important to lake CH4 production.

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

NASA Astrophysics Data System (ADS)

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

2017-12-01

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

Snowmelt and Surface Freeze/Thaw Timings over Alaska derived from Passive Microwave Observations using a Wavelet Classifier

NASA Astrophysics Data System (ADS)

Steiner, N.; McDonald, K. C.; Dinardo, S. J.; Miller, C. E.

2015-12-01

Arctic permafrost soils contain a vast amount of organic carbon that will be released into the atmosphere as carbon dioxide or methane when thawed. Surface to air greenhouse gas fluxes are largely dependent on such surface controls as the frozen/thawed state of the snow and soil. Satellite remote sensing is an important means to create continuous mapping of surface properties. Advances in the ability to determine soil and snow freeze/thaw timings from microwave frequency observations improves upon our ability to predict the response of carbon gas emission to warming through synthesis with in-situ observation, such as the 2012-2015 Carbon in Arctic Reservoir Vulnerability Experiment (CARVE). Surface freeze/thaw or snowmelt timings are often derived using a constant or spatially/temporally variable threshold applied to time-series observations. Alternately, time-series singularity classifiers aim to detect discontinuous changes, or "edges", in time-series data similar to those that occur from the large contrast in dielectric constant during the freezing or thaw of soil or snow. We use multi-scale analysis of continuous wavelet transform spectral gradient brightness temperatures from various channel combinations of passive microwave radiometers, Advanced Microwave Scanning Radiometer (AMSR-E, AMSR2) and Special Sensor Microwave Imager (SSM/I F17) gridded at a 10 km posting with resolution proportional to the observational footprint. Channel combinations presented here aim to illustrate and differentiate timings of "edges" from transitions in surface water related to various landscape components (e.g. snow-melt, soil-thaw). To support an understanding of the physical basis of observed "edges" we compare satellite measurements with simple radiative transfer microwave-emission modeling of the snow, soil and vegetation using in-situ observations from the SNOw TELemetry (SNOTEL) automated weather stations. Results of freeze/thaw and snow-melt timings and trends are reported for Alaska and the North-West Canadian Arctic for the period 2002 to 2015.

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

USGS Publications Warehouse

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 (<100) yr, where soils were warmer than at more recently thawed sites (14 to 21 yr; emitting 1.37 ± 0.67 mg CH4–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 CH4–C m−2 d−1 in July). Carbon lost via CH4 efflux during the growing season at these intermediate age sites was 8% of uptake by net ecosystem exchange. Our results provide evidence that CH4 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 CH4 emission, CO2 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 CH4 emission to the atmosphere that must be considered for a complete understanding of high latitude CH4 dynamics.

Prognostic value of a pre-freeze hypo-osmotic swelling test on the post-thaw quality of dog semen.

PubMed

Karger, S; Geiser, B; Grau, M; Burfeind, O; Heuwieser, W; Arlt, S P

2016-03-01

Throughout cryopreservation, sperm are exposed to major osmotic challenges. Only intact membranes of sperm cells are able to regulate these volumetric changes, which can be determined by the hypo-osmotic swelling test (HOS test). Correlations between the HOS test and conventional semen variables are inconsistent. Therefore, the objectives of this study were (1) to examine relationships between HOS test results and standard semen variables before freezing and after thawing and (2) to evaluate the prognostic value of the HOS assessments on post-thaw quality of dog semen. Semen of 35 dogs was collected and analyzed before freezing and after thawing following a 7-day freeze-thaw interval. Conventional semen variables such as sperm cell motility, membrane integrity morphology were evaluated and the HOS test was conducted with results from this test being recorded. In fresh semen the HOS test was positively correlated with progressive motility of sperm cells: r=0.52, sperm cell membrane integrity: r=0.50 and normal sperm cell morphology: r=0.46 (P<0.05). In frozen-thawed semen, the data obtained with the HOS test were positively correlated with progressive sperm cell motility: r=0.67 and membrane integrity: r=0.86 (P<0.05). The data obtained with the HOS test in fresh semen were positively correlated with sperm cell membrane integrity: r=0.50 normal sperm cell morphology: r=0.55 and data from the HOS test (r=0.43; P<0.05) with frozen-thawed semen. For the prediction of individual cryopreservation capacity, results from assessment of the fresh semen variables of good and poor semen quality were statistically compared. Based on these results, it is not possible to predict the quality of frozen-thawed dog semen using the HOS test. Copyright © 2016 Elsevier B.V. All rights reserved.

The Impact of Freeze-Thaw Cycles on Epinephrine.

PubMed

Beasley, Heather; Ng, Pearlly; Wheeler, Albert; Smith, William R; McIntosh, Scott E

2015-12-01

Epinephrine is the first-line medical treatment for anaphylaxis, a life-threatening allergic syndrome. To treat anaphylaxis, backcountry recreationalists and guides commonly carry epinephrine autoinjectors. Epinephrine may be exposed to cold temperatures and freezing during expeditions. An epinephrine solution must contain 90% to 115% of the labeled epinephrine amount to meet United States Pharmacopeia standards. The purpose of this study was to determine whether freeze-thaw cycles alter epinephrine concentrations in autoinjectors labeled to contain 1.0 mg/mL epinephrine. A further objective was to determine whether samples continued to meet United States Pharmacopeia concentration standards after freeze-thaw cycles. Epinephrine from 6 autoinjectors was extracted and divided into experimental and control samples. The experimental samples underwent 7 consecutive 12-hour freeze cycles followed by 7 12-hour thaw cycles. The control samples remained at an average temperature of 23.1°C for the duration of the study. After the seventh thaw cycle, epinephrine concentrations were measured using a high-performance liquid chromatography assay with mass spectrometry detection. The mean epinephrine concentration of the freeze-thaw samples demonstrated a statistically significant increase compared with the control samples: 1.07 mg/mL (SD ± 8.78; 95% CI, 1.04 to 1.11) versus 0.96 mg/mL (SD ± 6.81; 95% CI, 0.94 to 0.99), respectively. The maximal mean epinephrine concentration in the experimental freeze-thaw group was 1.12 mg/mL, which still fell within the range of United States Pharmacopeia standards for injectables (0.90 to 1.15 mg/mL). Although every attempt should be made to prevent freezing of autoinjectors, this preliminary study demonstrates that epinephrine concentrations remain within 90% to 115% of 1.0 mg/mL after multiple freeze-thaw cycles. Copyright © 2015 Wilderness Medical Society. Published by Elsevier Inc. All rights reserved.

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

PubMed

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 < .01); post-thawed: 51.07 ± 2.05 and 52.72 ± 1.96 (p < .01)] and progressive motility [fresh: 76.00 ± 0.58 and 78.25 ± 0.65 (p < .01); post-thawed: 40.61 ± 2.01 and 39.88 ± 2.01 (p < .01)], plasma membrane integrity [fresh: 91.60 ± 0.58 and 89.85 ± 0.59 (p < .01); post-thawed: 56.99 ± 1.86 and 54.39 ± 1.86 (p < .01)] and functionality [fresh: 75.40 ± 0.42 and 77.90 ± 0.96 (p < .01); post-thawed: 45.69 ± 1.71 and 46.35 ± 1.71 (p < .01)] was noted for both Ch-50 and Ch-75, respectively, groups compared to control group. Despite no significant change in mitochondrial activity, fertility rate of post-thawed spermatozoa was significantly improved in all Chrysin-fed groups compared to Ch-0 group. In conclusion, oral Chrysin administration to roosters could ameliorate cryopreservation-induced impairment of sperm quality and fertility rate. © 2017 Blackwell Verlag GmbH.

Effects of permafrost thaw on nitrogen availability and plant nitrogen acquisition in Interior Alaska

NASA Astrophysics Data System (ADS)

Finger, R.; Euskirchen, E. S.; Turetsky, M.

2013-12-01

The degradation of ice-rich permafrost, which covers a large portion of Interior Alaska, typically leads to thermokarst and increases in soil saturation. As a result, conifer peat plateaus degrade and are often replaced by wet collapse scar bogs. This state change results in profound changes in regional hydrology, biogeochemical cycling, and plant community composition. Preliminary data suggest that permafrost thaw can increase surface soil inorganic nitrogen (IN) concentrations but it is still unknown whether these changes in nutrient availability are short-lived (pulse releases) and whether or not they impact collapse scar vegetation composition or productivity, particularly as collapse scars undergo succession with time-after-thaw. Therefore we are currently examining changes in plant community composition, N availability and plant N acquisition along three thermokarst gradients in Interior Alaska. Each gradient is comprised of a forested permafrost peat plateau, adjacent ecotones experiencing active permafrost degradation (including a collapsing forest canopy and a saturated moat), and a collapse scar bog where permafrost has completely degraded. We predicted that IN concentrations would be highest along the active thaw margin, and lowest in the peat plateau. We also predicted that IN concentrations would be positively related to shifts in vegetation community composition, nutrient use efficiency (NUE) and tissue 15N concentrations. Preliminary results have shown that IN concentrations increase in newer collapse scar features as well as with thaw depth. Our data also show a shift from feather moss and ericaceous shrub-dominate understories in the permafrost plateau to Sphagnum and sedge dominated thaw ecotone and bog communities. Further successional development of the collapse scar bog results in the reintroduction of small evergreen and deciduous shrubs as the peat mat develops. Over time, collapse scar succession and peat accumulation appears to lead to progressive N limitations, resulting in the dominance of plants with higher NUE. This likely has implications for plant litter quality, and could inhibit decomposition processes. We are collecting additional data to compare species-level NUE and nutrient resorption efficiency. We also will measure δ15N of aboveground plant organs, roots, soil, and pore water to explore sources of plant N, which we expect will influenced rooting depth as permafrost thaws as well as differences in mycorrhizal associations along our thaw gradient. Because thawing permafrost soils are anticipated to mobilize large amounts of N from soils, our results will improve our understanding of how permafrost thaw influences vegetation and soil N pools, soil N availability, and plant nutrition.

Development of SMAP Mission Cal/Val Activities

NASA Technical Reports Server (NTRS)

Colliander, A.; Jackson, T.; Kimball, J.; Cosh, M.; Spencer, M.; Entekhabi, D.; Njoku, E.; ONeill, P.

2010-01-01

The Soil Moisture Active Passive (SMAP) mission is a NASA directed mission to map global land surface soil moisture and freeze-thaw state. Instrument and mission details are shown. The key SMAP soil moisture product is provided at 10 km resolution with 0.04cubic cm/cubic cm accuracy. The freeze/thaw product is provided at 3 km resolution and 80% frozen-thawed classification accuracy. The full list of SMAP data products is shown.

Evaluation of the effects of a freeze/thaw environment on cellular glass

NASA Technical Reports Server (NTRS)

Frickland, P.; Cleland, E.; Hasegawa, T.

1981-01-01

Using the evaluation criteria of water vapor permeability and conformability, a protective butylrubber/silicone conformal coating system was selected for use on Foamglas substrates in a freeze/thaw environment. The selection of a specific freeze/thaw cycle which closely models field conditions is discussed. A sampling plan is described which allows independent evaluation of the effects of conformal coatings, cycle number and location within the environmental chamber. The results of visual examination, measurement of density, modulus of rupture and Young's modulus are reported. Based upon statistical evaluation of the experimental results, it is concluded that no degradation in mechanical properties of either coated or uncoated Foamglas occurred within the duration of the test (53 freeze/thaw cycles).

Mapping freeze/thaw boundaries with SMMR data

NASA Technical Reports Server (NTRS)

Zuerndorfer, B. W.; England, A. W.; Dobson, M. C.; Ulaby, F. T.

1989-01-01

Nimbus 7 Scanning Multichannel Microwave Radiometer (SMMR) data are used to map daily freeze/thaw patterns in the upper Midwest for the Fall of 1984. The combination of a low 37 GHz radiobrightness and a negative 10.7, 18, and 37 GHz spectral gradient, Partial Derivative of Tb with Respect to f, appears to be an effective discriminant for classifying soil as frozen or thawed. The 37 GHz emissivity is less sensitive to soil moisture than are the lower frequency emissivities so that the 37 GHz radiobrightness appears to track soil surface temperature relatively well. The negative gradient for frozen ground is a consequence of volume scatter darkening at shorter microwave wavelengths. This shorter wavelength darkening is not seen in thawed moist soils.

Sorted clastic stripes, lobes and associated gullies in high-latitude craters on Mars: Landforms indicative of very recent, polycyclic ground-ice thaw and liquid flows

NASA Astrophysics Data System (ADS)

Gallagher, C.; Balme, M. R.; Conway, S. J.; Grindrod, P. M.

2011-01-01

Self-organised patterns of stone stripes, polygons, circles and clastic solifluction lobes form by the sorting of clasts from fine-grained sediments in freeze-thaw cycles. We present new High Resolution Imaging Science Experiment (HiRISE) images of Mars which demonstrate that the slopes of high-latitude craters, including Heimdal crater - just 25 km east of the Phoenix Landing Site - are patterned by all of these landforms. The order of magnitude improvement in imaging data resolution afforded by HiRISE over previous datasets allows not only the reliable identification of these periglacial landforms but also shows that high-latitude fluviatile gullies both pre- and post-date periglacial patterned ground in several high-latitude settings on Mars. Because thaw is inherent to the sorting processes that create these periglacial landforms, and from the association of this landform assemblage with fluviatile gullies, we infer the action of liquid water in a fluvio-periglacial context. We conclude that these observations are evidence of the protracted, widespread action of thaw liquids on and within the martian regolith. Moreover, the size frequency statistics of superposed impact craters demonstrate that this freeze-thaw environment is, at least in Heimdal crater, less than a few million years old. Although the current martian climate does not favour prolonged thaw of water ice, observations of possible liquid droplets on the strut of the Phoenix Lander may imply significant freezing point depression of liquids sourced in the regolith, probably driven by the presence of perchlorates in the soil. Because perchlorates have eutectic temperatures below 240 K and can remain liquid at temperatures far below the freezing point of water we speculate that freeze-thaw involving perchlorate brines provides an alternative "low-temperature" hypothesis to the freeze-thaw of more pure water ice and might drive significant geomorphological work in some areas of Mars. Considering the proximity of Heimdal crater to the Phoenix Landing Site, the presence of such hydrated minerals might therefore explain the landforms described here. If this is the case then the geographical distribution of martian freeze-thaw landforms might reflect relatively high temperatures (but still below 273 K) and the locally elevated concentration of salts in the regolith.

Numerical study on the thawing process of biological tissue induced by laser irradiation.

PubMed

Zhou, Jianhua; Liu, Jing; Yu, Aibing

2005-06-01

Most of the laser applications in medicine and biology involve thermal effects. The laser-tissue thermal interaction has therefore received more and more attentions in recent years. However, previous works were mainly focused on the case of laser heating on normal tissues (37 degrees C or above). To date, little is known on the mechanisms of laser heating on the frozen biological tissues. Several latest experimental investigations have demonstrated that lasers have great potentials in tissue cryopreservation. But the lack of theoretical interpretation limits its further application in this area. The present paper proposes a numerical model for the thawing of biological tissues caused by laser irradiation. The Monte Carlo approach and the effective heat capacity method are, respectively, employed to simulate the light propagation and solid-liquid phase change heat transfer. The proposed model has four important features: (1) the tissue is considered as a nonideal material, in which phase transition occurs over a wide temperature range; (2) the solid phase, transition phase, and the liquid phase have different thermophysical properties; (3) the variations in optical properties due to phase-change are also taken into consideration; and (4) the light distribution is changing continually with the advancement of the thawing fronts. To this end, 15 thawing-front geometric configurations are presented for the Monte Carlo simulation. The least-squares parabola fitting technique is applied to approximate the shape of the thawing front. And then, a detailed algorithm of calculating the photon reflection/refraction behaviors at the thawing front is described. Finally, we develop a coupled light/heat transport solution procedure for the laser-induced thawing of frozen tissues. The proposed model is compared with three test problems and good agreement is obtained. The calculated results show that the light reflectance/transmittance at the tissue surface are continually changing with the progression of the thawing fronts and that lasers provide a new heating method superior to conventional heating through surface conduction because it can achieve a uniform volumetric heating. Parametric studies are performed to test the influences of the optical properties of tissue on the thawing process. The proposed model is rather general in nature and therefore can be applied to other nonbiological problems as long as the materials are absorbing and scattering media.

Use of a probabilistic neural network to reduce costs of selecting construction rock

USGS Publications Warehouse

Singer, Donald A.; Bliss, James D.

2003-01-01

Rocks used as construction aggregate in temperate climates deteriorate to differing degrees because of repeated freezing and thawing. The magnitude of the deterioration depends on the rock's properties. Aggregate, including crushed carbonate rock, is required to have minimum geotechnical qualities before it can be used in asphalt and concrete. In order to reduce chances of premature and expensive repairs, extensive freeze-thaw tests are conducted on potential construction rocks. These tests typically involve 300 freeze-thaw cycles and can take four to five months to complete. Less time consuming tests that (1) predict durability as well as the extended freeze-thaw test or that (2) reduce the number of rocks subject to the extended test, could save considerable amounts of money. Here we use a probabilistic neural network to try and predict durability as determined by the freeze-thaw test using four rock properties measured on 843 limestone samples from the Kansas Department of Transportation. Modified freeze-thaw tests and less time consuming specific gravity (dry), specific gravity (saturated), and modified absorption tests were conducted on each sample. Durability factors of 95 or more as determined from the extensive freeze-thaw tests are viewed as acceptable—rocks with values below 95 are rejected. If only the modified freeze-thaw test is used to predict which rocks are acceptable, about 45% are misclassified. When 421 randomly selected samples and all four standardized and scaled variables were used to train aprobabilistic neural network, the rate of misclassification of 422 independent validation samples dropped to 28%. The network was trained so that each class (group) and each variable had its own coefficient (sigma). In an attempt to reduce errors further, an additional class was added to the training data to predict durability values greater than 84 and less than 98, resulting in only 11% of the samples misclassified. About 43% of the test data was classed by the neural net into the middle group—these rocks should be subject to full freeze-thaw tests. Thus, use of the probabilistic neural network would meanthat the extended test would only need be applied to 43% of the samples, and 11% of the rocks classed as acceptable would fail early.

Canonical correlation analysis to identify the semen characteristics used to forecast the freeze survival of Curimba (Prochilodus lineatus) spermatozoa.

PubMed

Carvalho, A F S; Murgas, L D S; Ferreira-Machad, M R; Andrade, E S; Felizardo, V O; Allaman, I B; de Paula, F G

　　OBJECTIVE: To identify which sperm characteristics were able to predict more accurately the quality of curimba (Prochilodus lineatus) semen upon freezing using canonical correlation analysis. Eleven fish breeders with initial mean weight of 705.21 ± 111 g were used. For cryopreservation, 200 µL of semen were taken from each animal and diluted in the cryoprotectant solution (10% dimethyl sulfoxide and 5% Beltsville Thawing Solution Minitub) in a 1:4 ratio and placed into 0.5-mL straws. Sperm characteristics (motility, sperm abnormalities, total antioxidant activity and lipid peroxidation) were evaluated. A randomized block design with duplicate samples per treatment (fresh and frozen semen) was used. The block factor was the animals, and the experimental unit the ejaculates. Canonical correlation was used to evaluate the association between sperm characteristics of fresh semen and thawed semen. There was a significant association (P = 0.10) among the variables measured in fresh semen with the variables measured in thawed semen, and 78.6% of the difference observed in the thawed semen can be attributed to variation of variables measured in fresh semen. Sperm motility, motility duration and antioxidant activity of the thawed semen showed an inverse relationship with those of the fresh semen; whereas the minor sperm abnormalities, major sperm abnormalities and lipid peroxidation showed a direct relationship with those of the fresh semen. Only the rate and motility duration of the thawed semen presented high correlation (-0.63 and -0.73, respectively) with the canonical variable represented by the sperm characteristics of fresh semen. The rate and motility duration of fresh semen may be used to predict the quality of the thawed sperm in Prochilodus lineatus.

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

PubMed

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.

Comparison of fertility results after vaginal insemination using different thawing procedures and packages for frozen ram semen

PubMed Central

Paulenz, Heiko; Ådnøy, Tormod; Söderquist, Lennart

2007-01-01

Background The effect of different thawing procedures for ram semen frozen in minitubes and mini straws on the fertility of sheep was tested in a field trial. Methods Altogether, 719 Norwegian Crossbred ewes, aged between six months and six-and-a-half years from 8 farms, were inseminated vaginally in natural oestrus with frozen-thawed semen. Minitubes were thawed at 70°C for 8 sec (T70) and mini straws either at 50°C for 9 sec (S50) or at 35°C for 12 sec (S35). Results Vaginal insemination with 200 × 106 spermatozoa resulted in 25-days non-return rates of 63.2, 59.6, and 62.5% (overall 61.8%), respectively, and lambing rates of 56.8, 55.0, and 59.2% (overall 57.0%), respectively. No significant effect on fertility (as 25-days non-return- or lambing rate) was seen for straw type/thawing temperature (P = 0.5/0.5), but semen filled in mini straws and thawed at 35°C resulted numerically in the highest lambing rate (59.2%). A significant effect was, however, seen for farmer (P = >0.0001/>0.0001) and ram (P = 0.009/0.002). Moreover, age of the ewes had a significant effect on the NR rate (0.007), but not on lambing rate (P = 0.2). Conclusion A vaginal deposition of frozen ram semen containing approximately 200 × 106 spermatozoa, filled in mini straws and thawed at 35°C is a simplified technique that under field conditions and used on a do-it-yourself regime gives acceptable lambing rates in Norway. PMID:17903246

Effect of dilution in sperm maturation media and time of storage on sperm motility and fertilizing capacity of cryopreserved semen of sex-reversed female rainbow trout.

PubMed

Judycka, Sylwia; Ciereszko, Andrzej; Dobosz, Stefan; Zalewski, Tomasz; Dietrich, Grzegorz J

2017-05-01

Masculinized females, also called neomales or sex-reversed females have a male phenotype but retain the female genotype (XX). Therefore, all spermatozoa produced in their functional testes carry an X chromosome, which is desired for the production of all-female rainbow trout populations. Semen of sex-reversed female rainbow trout is of low quality and in vitro maturation is required, which includes dilution of sperm suspensions with specially formulated maturation solutions. The aim of this study was to determine the effect of dilution in different maturation media on sperm quality (sperm motility characteristics and fertilizing capacity) of frozen/thawed sperm of sex-reversed female rainbow trout. The effect of time of post-thaw storage (0, 15, 60 and 120min) on semen quality was also tested. Sperm motility parameters and fertilization rate at the eyed and hatching stages were assessed for post-thaw semen diluted in different media. The cryopreservation procedure resulted in high post-thaw sperm motility of about 57% and did not differ from fresh semen. Unexpectedly, maturation media decreased sperm activation capacity immediately after dilution; however, sperm motility increased over time. Fertilization rates of frozen/thawed semen were high (71-87%) and did not differ significantly between experimental variants at any of tested periods of storage. Our results demonstrated that the effect of the maturation media on frozen/thawed sperm is different from that of fresh sperm. The progressive increase in post-thaw sperm motility in maturation media can potentially be applied to routine hatchery practice. Copyright © 2016 Elsevier Inc. All rights reserved.

Paleoecological inferences of recent alluvial damming of a lake basin due to retrogressive permafrost thaw slumping

NASA Astrophysics Data System (ADS)

Quinlan, R.; Delaney, S.; Lamoureux, S. F.; Kokelj, S. V.; Pisaric, M. F.

2014-12-01

Expected climate impacts of future warming in the Arctic include thawing of permafrost landscapes in northern latitudes. Thawing permafrost is expected to have major consequences on hydrological dynamics, which will affect the limnological conditions of Arctic lakes and ponds. In this study we obtained a sediment core from a small lake (informally named "FM1") near Fort McPherson, Northwest Territories, Canada, with a large retrogressive thaw slump (nearly 1 kilometre in diameter) within its catchment. A radiocarbon date from the base of the FM1 sediment core suggests the lake formed between 990-1160 Cal AD. The analysis of aerial photographs indicate the thaw slump initiated between 1970-1990, and sediment geochemistry analysis indicated major changes in sediment content at 54-cm sediment core depth. Analyses of subfossil midge (Chironomidae) fossils inferred that, pre-slump, lake FM1 was shallow with a large bog or wetland environment, with midge assemblages dominated by taxa such as Limnophyes and Parametriocnemus. Post-thaw midge assemblages were dominated by subfamily Chironominae (Tribe Tanytarsini and Tribe Chironomini) taxa, and the appearance of deepwater-associated taxa such as Sergentia suggests that lake FM1 deepened, possibly as a result of alluvial damming from slump materials washing into the lake near its outlet. Most recent stratigraphic intervals infer a reversion back to shallower conditions, with a slight recovery of bog or wetland-associated midge taxa, possibly due to rapid basin infilling from increased deposition rates of catchment-derived materials. Results emphasize that there may be a variety of different outcomes to Arctic lake and pond ecosystems as a result of permafrost thawing, contingent on system-specific characteristics such as slump location relative to the lake basin, and relative inflow and outflow locations within the lake basin.

Evaluation of ebselen supplementation on cryopreservation medium in human semen

PubMed Central

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

2014-01-01

Background: 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. Objective: 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. Materials and Methods: 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. Results: 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. Conclusion: These results suggest that the addition of ebselen to cryopreservation medium doesnot improve post-thaw parameters and DNA fragmentation of sperm. PMID:24976819

Evaluation of ebselen supplementation on cryopreservation medium in human semen.

PubMed

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.

Long-term comparison of Kuparuk Watershed active layer maps, northern Alaska, USA

NASA Astrophysics Data System (ADS)

Nyland, K. E.; Queen, C.; Nelson, F. E.; Shiklomanov, N. I.; Streletskiy, D. A.; Klene, A. E.

2017-12-01

The active layer, or the uppermost soil horizon that thaws seasonally, is among the most dynamic components of the permafrost system. Evaluation of the thickness and spatial variation of the active layer is critical to many components of Arctic research, including climatology, ecology, environmental monitoring, and engineering. In this study we mapped active-layer thickness (ALT) across the 22,278 sq. km Kuparuk River basin on Alaska's North Slope throughout the summer of 2016. The Kuparuk River extends from the Brooks Range through the Arctic Foothills and across the Arctic Coastal Plain physiographic provinces, and drains into the Beaufort Sea. Methodology followed procedures used to produce an ALT map of the basin in 1995 accounting for the effects of topography, vegetation, topoclimate, and soils, using the same spatial sampling scheme for direct ALT and temperature measurement at representative locations and relating these parameters to vegetation-soil associations. A simple semi-empirical engineering solution was used to estimate thaw rates for the different associations. An improved lapse-rate formulation and a higher-resolution DEM were used to relate temperature to elevation. Three ALT maps were generated for the 2016 summer, combining measured thaw depth, temperature records, the 25 m ArcticDEM, high resolution remote sensed data, empirical laps rates, and a topoclimatic index through the thaw solution. These maps were used to track the spatial progression of thaw through the 2016 summer season and estimate a total volume of thawed soil. Maps produced in this study were compared to the 1995 map to track areas of significant geographic changes in patterns of ALT and total volume of thawed soil.

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

NASA Technical Reports Server (NTRS)

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.

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

PubMed

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

2017-09-01

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

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

PubMed

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 < 0.05). Fructose was inferior for protecting sperm during cryopreservation when compared to sorbitol and glucose (P < 0.05). Although the viability, motility and acrosome integrity of sperm cryopreserved with a glucose-containing extender did not significantly differ from sperm frozen in the 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.

Cost-effectiveness of single versus double embryo transfer in IVF in relation to female age.

PubMed

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.

Resilience of norovirus GII.4 to freezing and thawing: implications for virus infectivity.

PubMed

Richards, Gary P; Watson, Michael A; Meade, Gloria K; Hovan, Gregory L; Kingsley, David H

2012-12-01

Genogroup II.4 norovirus (NoV) remains the predominant NoV strain in food- and water-borne outbreaks. Capsid integrity as well as viral RNA persistence were determined for GII.4 NoV by real-time RT-PCR after 1-14 freeze/thaw (F/T) cycles (-80 °C/+22 °C) or after -80 °C storage for up to 120 days. In both cases, capsid integrity and viral RNA titers remained stable. RNase was exogenously added after 1-14 F/T cycles, but did not alter the amount of genomic NoV RNA detected, indicating that capsids remained intact. Presumptive NoV infectivity was evaluated in functional studies by a porcine gastric mucin binding assay. Viruses frozen and thawed up to 14× bound similarly to porcine mucin, suggesting no reduction in virus infectivity. Overall, this study shows that a) NoV particles retain their integrity for at least 14 F/T cycles, b) long-term (120 day) frozen storage does not decrease NoV RNA titers, and c) capsid binding to receptor-like glycoprotein moieties remains unaltered after 14 F/T cycles. This work indicates that freezing and thawing of foods or beverages would not be a practical processing intervention to reduce NoV contamination. Likewise, repeated freezing and thawing, as might be encountered during winter months, is not expected to inactivate NoV in the environment. Results do show that laboratory samples destined for molecular biological analyses or for use as positive controls may be repeatedly frozen and thawed without any anticipated reduction in NoV RNA titers. This study documents the cryostability of NoV capsids and RNA to freezing and thawing and to the possible retention of virus infectivity.

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.

PubMed

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).

Extending the Season for Concrete Construction and Repair. Phase II - Defining Engineering Parameters

DTIC Science & Technology

2006-04-01

dosages, seemed to improve the freeze –thaw durability of concrete. Phase II found what appears to be a maximum dosage after which freeze –thaw...durability becomes a concern. That is because cement hydration can only create a finite amount of space to absorb these chemicals. Thus, for freeze ...protection, admixture dosages should be designed according to water content as specified in Phase I, while, for freeze –thaw durability, admixture dosages

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

PubMed

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

2015-03-03

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

Clinically applied procedures for human ovarian tissue cryopreservation result in different levels of efficacy and efficiency.

PubMed

Bastings, Lobke; Westphal, Johan R; Beerendonk, Catharina C M; Bekkers, Ruud L M; Zusterzeel, Petra L M; Hendriks, Jan C M; Braat, Didi D M; Peek, Ronald

2016-12-01

Different protocols are being used worldwide for the cryopreservation of human ovarian tissue for fertility preservation purposes. The efficiency and efficacy of the majority of these protocols has not been extensively evaluated, possibly resulting in sub-optimally cryopreserved ovarian tissue. To address the impact of this issue, we assessed the effects of two clinically successful human ovarian tissue slow-freezing cryopreservation procedures on the quality of the cryopreserved tissue. To differentiate between cryopreservation ( C ) versus thawing ( T ) related effects, four combinations of these two (A and B) very different cryopreservation/thawing protocols (A C A T , A C B T , B C A T , B C B T ) were studied. Before and after cryopreservation and thawing, the percentage of living and morphologically normal follicles, as well as the overall tissue viability, was assessed. Our experiments revealed that the choice of the cryopreservation protocol noticeably affected the overall tissue viability and percentage of living follicles, with a higher viability after protocol B C when compared to A C . No statistically significant differences in tissue viability were observed between the two thawing protocols, but thawing protocol B T required considerably more human effort and materials than thawing protocol A T . Tissue morphology was best retained using the B C A T combination. Our results indicate that extensive and systematical evaluation of clinically used protocols is warranted.

Effects of cryopreservation on microbial-contaminated cord blood.

PubMed

Clark, Pamela; Trickett, Annette; Saffo, Sandra; Stark, Damien

2014-03-01

Cord blood units (CBUs) are associated with significant risk of exposure to microbial contamination during collection and processing; however, the survival of bacteria within a CBU is poorly understood. This study aimed to determine whether contaminating organisms in CBU survive the cryopreservation, frozen storage, and subsequent thawing conditions before infusion. A total of 134 CBUs rejected from banking due to known contamination were thawed and rescreened using blood culture bottles (BacT/ALERT, bioMérieux). An additional 61 fresh CBUs were deliberately spiked with a range of microbial organisms and evaluated both before freeze and after thaw. Microbial contaminants were detected after thaw in 63% of stored contaminated CBUs and 85% of spiked CBUs. Postthaw organism detection in spiked cord blood (CB) was higher in adult culture bottles (80%) than pediatric culture bottles (61%). Twenty percent of spiked organisms, particularly Bacillus subtilis, Escherichia coli, Clostridium sporogenes, and Propionibacterium acnes, were not detected in prefreeze samples but were detectable after thaw. This study demonstrates that the majority of contaminating organisms isolated in a prefreeze sample of CB have the ability to survive cryopreservation, frozen storage, and thawing. Further, CBUs reported as microbial free may contain microbial contamination, which could result in transplantation of contaminated CB and be potentially deleterious to a patient. © 2013 The Sydney Children's Hospital Network. Transfusion © 2013 American Association of Blood Banks.

Association of Vitamin E with Rapid Thawing on Goat Semen

PubMed Central

Penitente-Filho, Jurandy Mauro; Oliveira, Fabrício Albani; Jimenez, Carolina Rodriguez; Dias, Júlio César Oliveira; Oliveira, Gisele Dias; Silveira, Renata Gomes; Silveira, Camila Oliveira; Torres, Ciro Alexandre Alves

2014-01-01

The aim of this study was to evaluate the effects of vitamin E associated with rapid thawing on cryopreserved goat semen. Two bucks were used and eight ejaculates per animal were collected using artificial vagina. Semen was diluted with the following treatments: BIOXCELL (control), BIOXCELL + Equex (sodium lauryl sulphate) and BIOXCELL + vitamin E 100 μM. Semen was packaged into 0.25 mL straws and cooled at 5°C for 1 hour. Freezing was performed in liquid nitrogen vapor (−155°C) during 15 minutes. Then, the straws were immersed in liquid nitrogen (−196°C). Straws were thawed at 38°C/60 seconds or at 60°C/7 seconds with immediate sperm analysis. Hypoosmotic swelling test was performed adding a 20 μL aliquot of thawed semen to 1 mL of hypoosmotic solution (100 mOsm·Kg−1) followed by incubation during 60 minutes in water bath (38°C). Vitamin E did not affect any studied parameters (P > 0.05). Nevertheless, defrosting rate of 60°C/7 seconds improved sperm membrane functional integrity (P < 0.05). Current knowledge about goat semen cryopreservation is not sufficient to ensure high post-thawing recovery rates; thus, this study brings important data about using antioxidants and different thawing rates on cryopreservation process. PMID:24955428

Genes Upregulated in Winter Wheat (Triticum aestivum L.) during Mild Freezing and Subsequent Thawing Suggest Sequential Activation of Multiple Response Mechanisms.

PubMed

Skinner, Daniel Z

2015-01-01

Exposing fully cold-acclimated wheat plants to a mild freeze-thaw cycle of -3 °C for 24h followed by +3 °C for 24 or 48 h results in dramatically improved tolerance of subsequent exposure to sub-freezing temperatures. Gene enrichment analysis of crown tissue from plants collected before or after the -3 °C freeze or after thawing at +3 °C for 24 or 48 h revealed that many biological processes and molecular functions were activated during the freeze-thaw cycle in an increasing cascade of responses such that over 150 processes or functions were significantly enhanced by the end of the 48 h, post-freeze thaw. Nearly 2,000 individual genes were upregulated more than 2-fold over the 72 h course of freezing and thawing, but more than 70% of these genes were upregulated during only one of the time periods examined, suggesting a series of genes and gene functions were involved in activation of the processes that led to enhanced freezing tolerance. This series of functions appeared to include extensive cell signaling, activation of stress response mechanisms and the phenylpropanoid biosynthetic pathway, extensive modification of secondary metabolites, and physical restructuring of cell membranes. By identifying plant lines that are especially able to activate these multiple mechanisms it may be possible to develop lines with enhanced winterhardiness.

Delayed soil thawing affects root and shoot functioning and growth in Scots pine.

PubMed

Repo, Tapani; Lehto, Tarja; Finér, Leena

2008-10-01

In boreal regions, soil can remain frozen after the start of the growing season. We compared relationships between root characteristics and water relations in Scots pine (Pinus sylvestris L.) saplings subjected to soil frost treatments before and during the first week of the growing period in a controlled environment experiment. Delayed soil thawing delayed the onset of sap flow or totally blocked it if soil thawing lagged the start of the growing period by 7 days. This effect was reflected in the electrical impedance of needles and trunks and in the relative electrolyte leakage of needles. Prolonged soil frost reduced or completely inhibited root growth. In unfrozen soil, limited trunk sap flow was observed despite unfavorable aboveground growing conditions (low temperature, low irradiance, short photoperiod). Following the earliest soil thaw, sap flow varied during the growing season, depending on light and temperature conditions, phenological stage of the plant and the amount of live needles in the canopy. The results suggest that delayed soil thawing can reduce tree growth, and if prolonged, it can be lethal.

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

PubMed

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.

Shrinkage void formation and its effect on freeze and thaw processes of lithium and lithium-fluoride for space applications

NASA Astrophysics Data System (ADS)

Yang, Jae-Young; El-Genk, M. S.

1991-07-01

The effects of shrinkage void forming during freezing of lithium and lithium-fluoride on subsequent thaw processes are investigated using a numerical scheme that is based on a single (solid/liquid) cell approach. Results show that a void forming at the wall appreciably reduces the solid-liquid interface velocity, during both freeze and thaw, and causes a substantial rise in the wall temperature during thaw. However, in the case of Li, the maximum wall temperature was much lower than the melting temperature of PWC-11, which is used as the structure material in the SP-100 system. Hence, it is concluded that a formation of hot spots is unlikely during the startup or restart of the SP-100 system.

Shrinkage void formation and its effect on freeze and thaw processes of lithium and lithium-fluoride for space applications

NASA Technical Reports Server (NTRS)

Yang, Jae Y.; El-Genk, Mohamed S.

1991-01-01

The effects of shrinkage void forming during freezing of lithium and lithium fluoride on subsequent thaw processes are investigated using a numerical scheme that is based on a single (solid/liquid) cell approach. Results show that a void forming at the wall appreciably reduces the solid-liquid interface velocity, during both freeze and thaw, and causes a substantial rise in the wall temperature during thaw. However, in the case of Li, the maximum wall temperature was much lower than the melting temperature of PWC-11, which is used as the structure material in the SP-100 system. Hence, it is concluded that a formation of hot spots is unlikely during the startup or restart of the SP-100 system.

Thermokarst and thaw-related landscape dynamics -- an annotated bibliography with an emphasis on potential effects on habitat and wildlife

USGS Publications Warehouse

Jones, Benjamin M.; Amundson, Courtney L.; Koch, Joshua C.; Grosse, Guido

2013-01-01

Permafrost has warmed throughout much of the Northern Hemisphere since the 1980s, with colder permafrost sites warming more rapidly (Romanovsky and others, 2010; Smith and others, 2010). Warming of the near-surface permafrost may lead to widespread terrain instability in ice-rich permafrost in the Arctic and the Subarctic, and may result in thermokarst development and other thaw-related landscape features (Jorgenson and others, 2006; Gooseff and others, 2009). Thermokarst and other thaw-related landscape features result from varying modes and scales of permafrost thaw, subsidence, and removal of material. An increase in active-layer depth, water accumulation on the soil surface, permafrost degradation and associated retreat of the permafrost table, and changes to lake shores and coastal bluffs act and interact to create thermokarst and other thaw-related landscape features (Shur and Osterkamp, 2007). There is increasing interest in the spatial and temporal dynamics of thermokarst and other thaw-related features from diverse disciplines including landscape ecology, hydrology, engineering, and biogeochemistry. Therefore, there is a need to synthesize and disseminate knowledge on the current state of near-surface permafrost terrain. The term "thermokarst" originated in the Russian literature, and its scientific use has varied substantially over time (Shur and Osterkamp, 2007). The modern definition of thermokarst refers to the process by which characteristic landforms result from the thawing of ice-rich permafrost or the melting of massive ice (van Everdingen, 1998), or, more specifically, the thawing of ice-rich permafrost and (or) melting of massive ice that result in consolidation and deformation of the soil surface and formation of specific forms of relief (Shur, 1988). Jorgenson (2013) identifies 23 distinct thermokarst and other thaw-related features in the Arctic, Subarctic, and Antarctic based primarily on differences in terrain condition, ground-ice volume, 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.

Observation-based modelling of permafrost carbon fluxes with accounting for deep carbon deposits and thermokarst activity

NASA Astrophysics Data System (ADS)

Schneider von Deimling, T.; Grosse, G.; Strauss, J.; Schirrmeister, L.; Morgenstern, A.; Schaphoff, S.; Meinshausen, M.; Boike, J.

2015-06-01

High-latitude soils store vast amounts of perennially frozen and therefore inert organic matter. With rising global temperatures and consequent permafrost degradation, a part of this carbon stock will become available for microbial decay and eventual release to the atmosphere. We have developed a simplified, two-dimensional multi-pool model to estimate the strength and timing of future carbon dioxide (CO2) and methane (CH4) fluxes from newly thawed permafrost carbon (i.e. carbon thawed when temperatures rise above pre-industrial levels). We have especially simulated carbon release from deep deposits in Yedoma regions by describing abrupt thaw under newly formed thermokarst lakes. The computational efficiency of our model allowed us to run large, multi-centennial ensembles under various scenarios of future warming to express uncertainty inherent to simulations of the permafrost carbon feedback. Under moderate warming of the representative concentration pathway (RCP) 2.6 scenario, cumulated CO2 fluxes from newly thawed permafrost carbon amount to 20 to 58 petagrams of carbon (Pg-C) (68% range) by the year 2100 and reach 40 to 98 Pg-C in 2300. The much larger permafrost degradation under strong warming (RCP8.5) results in cumulated CO2 release of 42 to 141 Pg-C and 157 to 313 Pg-C (68% ranges) in the years 2100 and 2300, respectively. Our estimates only consider fluxes from newly thawed permafrost, not from soils already part of the seasonally thawed active layer under pre-industrial climate. Our simulated CH4 fluxes contribute a few percent to total permafrost carbon release yet they can cause up to 40% of total permafrost-affected radiative forcing in the 21st century (upper 68% range). We infer largest CH4 emission rates of about 50 Tg-CH4 per year around the middle of the 21st century when simulated thermokarst lake extent is at its maximum and when abrupt thaw under thermokarst lakes is taken into account. CH4 release from newly thawed carbon in wetland-affected deposits is only discernible in the 22nd and 23rd century because of the absence of abrupt thaw processes. We further show that release from organic matter stored in deep deposits of Yedoma regions crucially affects our simulated circumpolar CH4 fluxes. The additional warming through the release from newly thawed permafrost carbon proved only slightly dependent on the pathway of anthropogenic emission and amounts to about 0.03-0.14 °C (68% ranges) by end of the century. The warming increased further in the 22nd and 23rd century and was most pronounced under the RCP6.0 scenario, adding 0.16 to 0.39 °C (68% range) to simulated global mean surface air temperatures in the year 2300.

Observation-based modelling of permafrost carbon fluxes with accounting for deep carbon deposits and thermokarst activity

NASA Astrophysics Data System (ADS)

Schneider von Deimling, T.; Grosse, G.; Strauss, J.; Schirrmeister, L.; Morgenstern, A.; Schaphoff, S.; Meinshausen, M.; Boike, J.

2014-12-01

High-latitude soils store vast amounts of perennially frozen and therefore inert organic matter. With rising global temperatures and consequent permafrost degradation, a part of this carbon store will become available for microbial decay and eventual release to the atmosphere. We have developed a simplified, two-dimensional multi-pool model to estimate the strength and timing of future carbon dioxide (CO2) and methane (CH4) fluxes from newly thawed permafrost carbon (i.e. carbon thawed when temperatures rise above pre-industrial levels). We have especially simulated carbon release from deep deposits in Yedoma regions by describing abrupt thaw under thermokarst lakes. The computational efficiency of our model allowed us to run large, multi-centennial ensembles under various scenarios of future warming to express uncertainty inherent to simulations of the permafrost-carbon feedback. Under moderate warming of the representative concentration pathway (RCP) 2.6 scenario, cumulated CO2 fluxes from newly thawed permafrost carbon amount to 20 to 58 petagrammes of carbon (Pg-C) (68% range) by the year 2100 and reach 40 to 98 Pg-C in 2300. The much larger permafrost degradation under strong warming (RCP8.5) results in cumulated CO2 release of 42-141 and 157-313 Pg-C (68% ranges) in the years 2100 and 2300, respectively. Our estimates do only consider fluxes from newly thawed permafrost but not from soils already part of the seasonally thawed active layer under preindustrial climate. Our simulated methane fluxes contribute a few percent to total permafrost carbon release yet they can cause up to 40% of total permafrost-affected radiative forcing in the 21st century (upper 68% range). We infer largest methane emission rates of about 50 Tg-CH4 year-1 around the mid of the 21st century when simulated thermokarst lake extent is at its maximum and when abrupt thaw under thermokarst lakes is accounted for. CH4 release from newly thawed carbon in wetland-affected deposits is only discernible in the 22nd and 23rd century because of the absence of abrupt thaw processes. We further show that release from organic matter stored in deep deposits of Yedoma regions does crucially affect our simulated circumpolar methane fluxes. The additional warming through the release from newly thawed permafrost carbon proved only slightly dependent on the pathway of anthropogenic emission and amounts about 0.03-0.14 °C (68% ranges) by end of the century. The warming increased further in the 22nd and 23rd century and was most pronounced under the RCP6.0 scenario with adding 0.16-0.39 °C (68% range) to simulated global mean surface air temperatures in the year 2300.

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

NASA Technical Reports Server (NTRS)

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

1994-01-01

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 borel and arctic regions thus affecting the carbon budget and the interannual variability fo regional carbon fluxes.

Spring photosynthetic onset and net CO2 uptake in Alaska triggered by landscape thawing.

PubMed

Parazoo, Nicholas C; Arneth, Almut; Pugh, Thomas A M; Smith, Ben; Steiner, Nicholas; Luus, Kristina; Commane, Roisin; Benmergui, Josh; Stofferahn, Eric; Liu, Junjie; Rödenbeck, Christian; Kawa, Randy; Euskirchen, Eugenie; Zona, Donatella; Arndt, Kyle; Oechel, Walt; Miller, Charles

2018-04-24

The springtime transition to regional-scale onset of photosynthesis and net ecosystem carbon uptake in boreal and tundra ecosystems are linked to the soil freeze-thaw state. We present evidence from diagnostic and inversion models constrained by satellite fluorescence and airborne CO 2 from 2012 to 2014 indicating the timing and magnitude of spring carbon uptake in Alaska correlates with landscape thaw and ecoregion. Landscape thaw in boreal forests typically occurs in late April (DOY 111 ± 7) with a 29 ± 6 day lag until photosynthetic onset. North Slope tundra thaws 3 weeks later (DOY 133 ± 5) but experiences only a 20 ± 5 day lag until photosynthetic onset. These time lag differences reflect efficient cold season adaptation in tundra shrub and the longer dehardening period for boreal evergreens. Despite the short transition from thaw to photosynthetic onset in tundra, synchrony of tundra respiration with snow melt and landscape thaw delays the transition from net carbon loss (at photosynthetic onset) to net uptake by 13 ± 7 days, thus reducing the tundra net carbon uptake period. Two global CO 2 inversions using a CASA-GFED model prior estimate earlier northern high latitude net carbon uptake compared to our regional inversion, which we attribute to (i) early photosynthetic-onset model prior bias, (ii) inverse method (scaling factor + optimization window), and (iii) sparsity of available Alaskan CO 2 observations. Another global inversion with zero prior estimates the same timing for net carbon uptake as the regional model but smaller seasonal amplitude. The analysis of Alaskan eddy covariance observations confirms regional scale findings for tundra, but indicates that photosynthesis and net carbon uptake occur up to 1 month earlier in evergreens than captured by models or CO 2 inversions, with better correlation to above-freezing air temperature than date of primary thaw. Further collection and analysis of boreal evergreen species over multiple years and at additional subarctic flux towers are critically needed. © 2018 John Wiley & Sons Ltd.

Changes to the Carbon and Energy fluxes in a Northern Peatland with Thawing Permafrost

NASA Astrophysics Data System (ADS)

Harder, S. R.; Roulet, N. T.; Crill, P. M.; Strachan, I. B.

2017-12-01

The maintenance of thaw of high carbon density landscapes in the permafrost region ultimately depends of how the energy balance is partitioned as temperatures and precipitation change, yet there are comparatively few energy balance studies, especially in peatlands that contain permafrost. While permafrost peatlands are currently net sinks of carbon, as Arctic temperatures rise and permafrost thaws, the future of these ecosystems and their capacity for carbon uptake is in question. Since 2012 we have been measuring the spatially integrated CO2, energy and water vapour fluxes from the Stordalen peatland (68°22'N, 19°03'E) using eddy covariance (EC). The Stordalen peatland is a heterogeneous peatland in the discontinuous permafrost zone where permafrost thaw is actively occurring, resulting in large changes to the landscape from year to year. Areas where permafrost is present are elevated by up to 1.5 m compared to the areas where permafrost has thawed causing differences in water table depth, peat temperatures, snow distribution, vegetation community and therefore in the carbon and energy fluxes. Our EC tower is located on the edge of a permafrost peat plateau (or palsa) where one fetch measures fluxes from an area underlain by permafrost and the other fetch sees the portion of the peatland where the permafrost has thawed. Within each sector, we have an array of soil temperature and water content sensors to determine the physical characteristics of each fetch. Extensive vegetation surveys (based on plant functional types or PFTs) have also been conducted to run a footprint analysis on the flux data to complete a comparative analysis of the magnitude and variability of the carbon and energy exchanges from PFT. The footprint analysis allows us to explain the difference in energy and carbon fluxes by examining the ecological, biogeochemical and physical characteristics within each footprint. We see distinctly different energy partitioning between the fetches containing intact permafrost and those where the permafrost has thawed: the evaporative efficiency is higher and the Bowen ration lower for the thawed fetches. Our results also show differences in the carbon fluxes depending on the tower footprint.

Thermokarst lake methanogenesis along a complete talik profile

DOE PAGES

Heslop, J. K.; Walter Anthony, K. M.; Sepulveda-Jauregui, A.; ...

2015-07-24

Here, thermokarst (thaw) lakes emit methane (CH 4) to the atmosphere formed from thawed permafrost organic matter (OM), but the relative magnitude of CH 4 production in surface lake sediments vs. deeper thawed permafrost horizons is not well understood. We assessed anaerobic CH 4 production potentials from various depths along a 590 cm long lake sediment core that captured the entire sediment package of the talik (thaw bulb) beneath the center of an interior Alaska thermokarst lake, Vault Lake, and the top 40 cm of thawing permafrost beneath the talik. We also studied the adjacent Vault Creek permafrost tunnel thatmore » extends through ice-rich yedoma permafrost soils surrounding the lake and into underlying gravel. Our results showed CH 4 production potentials were highest in the organic-rich surface lake sediments, which were 151 cm thick (mean ± SD: 5.95 ± 1.67 μg C–CH 4 g dw –1 d –1; 125.9 ± 36.2 μg C–CH 4 g C −1 org d –1). High CH 4 production potentials were also observed in recently thawed permafrost (1.18 ± 0.61 μg C–CH 4g dw –1 d –1; 59.60± 51.5 μg C–CH 4 g C −1 org d –1) at the bottom of the talik, but the narrow thicknesses (43 cm) of this horizon limited its overall contribution to total sediment column CH 4 production in the core. Lower rates of CH 4 production were observed in sediment horizons representing permafrost that has been thawing in the talik for a longer period of time. No CH 4 production was observed in samples obtained from the permafrost tunnel, a non-lake environment. Our findings imply that CH 4 production is highly variable in thermokarst lake systems and that both modern OM supplied to surface sediments and ancient OM supplied to both surface and deep lake sediments by in situ thaw and shore erosion of yedoma permafrost are important to lake CH 4 production.« less

Thermokarst lake methanogenesis along a complete talik profile

DOE Office of Scientific and Technical Information (OSTI.GOV)

Heslop, J. K.; Walter Anthony, K. M.; Sepulveda-Jauregui, A.

Here, thermokarst (thaw) lakes emit methane (CH 4) to the atmosphere formed from thawed permafrost organic matter (OM), but the relative magnitude of CH 4 production in surface lake sediments vs. deeper thawed permafrost horizons is not well understood. We assessed anaerobic CH 4 production potentials from various depths along a 590 cm long lake sediment core that captured the entire sediment package of the talik (thaw bulb) beneath the center of an interior Alaska thermokarst lake, Vault Lake, and the top 40 cm of thawing permafrost beneath the talik. We also studied the adjacent Vault Creek permafrost tunnel thatmore » extends through ice-rich yedoma permafrost soils surrounding the lake and into underlying gravel. Our results showed CH 4 production potentials were highest in the organic-rich surface lake sediments, which were 151 cm thick (mean ± SD: 5.95 ± 1.67 μg C–CH 4 g dw –1 d –1; 125.9 ± 36.2 μg C–CH 4 g C −1 org d –1). High CH 4 production potentials were also observed in recently thawed permafrost (1.18 ± 0.61 μg C–CH 4g dw –1 d –1; 59.60± 51.5 μg C–CH 4 g C −1 org d –1) at the bottom of the talik, but the narrow thicknesses (43 cm) of this horizon limited its overall contribution to total sediment column CH 4 production in the core. Lower rates of CH 4 production were observed in sediment horizons representing permafrost that has been thawing in the talik for a longer period of time. No CH 4 production was observed in samples obtained from the permafrost tunnel, a non-lake environment. Our findings imply that CH 4 production is highly variable in thermokarst lake systems and that both modern OM supplied to surface sediments and ancient OM supplied to both surface and deep lake sediments by in situ thaw and shore erosion of yedoma permafrost are important to lake CH 4 production.« less

Effect of sex-sorting and cryopreservation on the post-thaw sperm quality of Iberian red deer spermatozoa.

PubMed

Anel-López, L; García-Álvarez, O; Parrilla, I; Del Olmo, D; Maroto-Morales, A; Fernandez-Santos, M R; Ortiz, J A; Soler, A J; Martínez, E M; Vazquez, J M; Garde, J J

2017-02-01

This study investigated the effect of sex-sorting and cryopreservation on post-thaw characteristics and fertility of red deer (Cervus elaphus) sperm for the first time. Semen was collected by electroejaculation from 10 mature stags during the breeding season, and each ejaculate split into four experimental groups: Bulk sorted spermatozoa, sorted but not sexed (BSS); sorted high purity X-spermatozoa (XSS); sorted high purity Y-spermatozoa (YSS); and, control non-sorted spermatozoa (NS). Following, all samples were frozen over liquid nitrogen. Two straws per stag and sample type were analyzed immediately post-thaw and following a 2-h incubation period at 37 °C. Post-thaw total motility (TM) as assessed by CASA was not different (P < 0.05) among NS, BSS and YSS sperm. For XSS, post-thaw TM was lower (39%, P < 0.05) than that for NS (54%) or BSS (50%), but similar (P > 0.05) to that of YSS (47%) sperm. The percentage of apoptotic spermatozoa as assessed by PI/YO-PRO-1 and flow cytometry analysis, was higher (17%, P ≤ 0.05) for XSS sperm than NS (12%), BSS (13%) and YSS (14%) sperm. Following incubation there were no differences (P > 0.05) in TM or percent apoptosis among treatments. Post-thaw chromatin stability calculated as the DNA fragmentation index (%DFI) was similar among treatments; following incubation %DFI increased in all except YSS, which displayed the lowest value (P < 0.05). Artificial insemination of synchronized hinds yielded 44, 52 and 62% delivery rates for YSS, NS and standard frozen-thawed sperm, respectively (P < 0.05). Notably, 93 and 55% of fawns born were males for the YSS and NS spermatozoa, respectively (P < 0.05). In summary, Y-sorted sperm displayed acceptable post-thaw sperm evaluation parameters and the expected offspring sex ratio. More studies are needed to understand the source of sperm damage that may compromise the fertility of Y-sorted red deer sperm. Copyright © 2016 Elsevier Inc. All rights reserved.

Combining terrestrial, air-, and space-borne remote sensing for permafrost thaw subsidence change detection in Arctic Alaska

NASA Astrophysics Data System (ADS)

Günther, F.; Grosse, G.; Ulrich, M.; Nitze, I.; Sachs, T.; Jones, B. M.

2017-12-01

The unique feature of permafrost in the Arctic is the presence of a large amount of ice below the earth surface. Thermal degradation and subsequent permafrost destabilization causes thaw subsidence and thermokarst development. Because these processes are difficult to detect due to the lack of timely and accurate elevation datasets they have received not much attention, despite their potentially global significance through the permafrost carbon feedback. Thanks to remote sensing pioneering works in Alaska and Siberia, widespread thaw subsidence has been documented and is increasingly perceived as a potentially widespread permafrost landscape response to contemporary climate change. Clearly, however, detailed local inventories are required to calibrate regional long and short-term assessments for measuring surface deformation due to permafrost thaw. The objective of our study is to analyze time series of repeat terrestrial, air-, and space borne laser scanning (rLiDAR) for quantification of land surface lowering due to permafrost thaw, which is poorly resolved in terms of recent landscape development in the Arctic. Our work aims at finding commonalities and differences of change or no change on ground-ice-rich primary surfaces that are preserved as uplands, which cover 15 to 20% of the Teshekpuk Lake Special Area on the Arctic Coastal Plain of northern Alaska. Our approach focuses on quantifying modern thaw subsidence and thermokarst rates with high spatial resolution data over several decades as well as high temporal resolution data of inter-annual intervals. Multi-annual measurements of rLiDAR over Arctic Alaska have been made by aircraft in 2016 and in 2015+2017 through on-site surveys during field expeditions. These in situ data serve as a basis for large scale surface change assessments using time series of photogrammetrically derived elevation data from very high resolution historical aerial photographs and modern satellite imagery. The synergistic data fusion approach enhances permafrost degradation monitoring and better resolves surface deformation associated with thaw subsidence. The novel datasets also provide insights into previously unrecognized patterns of rapid permafrost thaw and related interconnections.

In situ permafrost thaw due to climate change drives holistic microbial community shifts with implications for methane cycling

NASA Astrophysics Data System (ADS)

Mondav, Rhiannon; McCalley, Carmody; Hodgkins, Suzanne; Rich, Virginia; Frolking, Steve; Saleska, Scott; Barnes, Andrew; Chanton, Jeff; Crill, Patrick

2014-05-01

Thawing permafrost is a potentially significant source of radiative forcing feedback due to increased emissions of methane, a biogenic greenhouse gas (GHG). This study investigated changes in the microbial community along a permafrost thaw gradient at Stordalen Mire, Sweden using 16S rRNA gene amplicon and metagenomic methods. In situ measurements of geochemical parameters, including CH4 and C isotopes, enabled linkage of community dynamics to significant shifts in C balance. The thaw gradient ranged from intact at a palsa (low productivity and GHG emissions), through partially thawed in a bog (high productivity, low GHG emissions) to a completely thawed fen (high productivity and GHG emissions). Microbial assemblages in both the palsa and fen were highly diverse (in both richness and evenness), consistent with climax communities. The microbial community in the bog had distinctly lower diversity, characteristic of ecosystem disturbance. The palsa community was dominated by Acidobacteria and Proteobacteria, as is typical of a range of soils including permafrost. Methanogens dominated both the bog and fen and were most abundant within the zone of water table fluctuation. Inferring methanogens' production pathway from phylogeny showed a shift from mostly hydrogenotrophic methanogens in the bog towards acetotrophic methanogens in the fen. This corroborated porewater and flux emitted CH4 and CO2 carbon isotopic 13C signatures of CH4 and CO2. The fen, where the highest CH4 flux was recorded, was significantly richer in methanogenic archaea. A novel archaea, Candidatus Methanoflorens stordalenmirensis, was present at up to 70% relative abundance in the bog, enabling recovery of a population genome. The genome (and associated metaproteome) of 'M. stordalenmirensis' indicates that hydrogenotrophic methane production is its main energy conservation pathway. 'Methanoflorens' may be an indicator species of permafrost thaw, it is globally ubiquitous, and appears a major contributor to global methane production. Our results revealed a distinct difference in the microbial community structure and membership at each site, which can be directly associated with increasing methane emission and thaw state.

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

PubMed

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 < 0.0001). There were significant differences in sperm GSH content between different 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.

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

PubMed

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 when planning experimental protocols. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Illuminating Geochemical Controls of Methane Oxidation Along a Gradient of Permafrost Thaw

NASA Astrophysics Data System (ADS)

Perryman, C. R.; Kashi, N.; McCalley, C. K.; Malhotra, A.; Giesler, R.; Varner, R.

2017-12-01

Increases in annual mean temperature in the subarctic have accelerated the thaw of organic-rich permafrost peatlands, exacerbating methane (CH4) production from microbial decomposition of peat deposits and subsequent CH4 emissions. Methanotrophic bacteria may oxidize/consume upwards of 90% of produced CH4 in some settings, pending substrate availability and environmental conditions. Redox chemistry may also control the rate of CH4 oxidation in thawing permafrost areas, particularly redox potential (Eh) and the availability of oxygen (O2) and other terminal electron receptors. We investigated potential CH4 oxidation rates across a permafrost thaw gradient in Stordalen Mire (68°21'N,18°49'E) near Abisko, Sweden. Methane oxidation rates for sites from thawing and collapsed palsa, semi-wet Sphagnum, and open-water sedge sites were determined through laboratory incubations. Peat cores were extracted from two depths at each site and incubated at in situ temperatures and CH4 concentrations. Headspace samples were collected over a 48-hour period and analyzed for CH4 concentration using flame ionization detection gas chromatography (GC-FID). Dissolved O2, Eh, and dissolved CH4 were measured in sites with porewater. Oxidation rates ranged from <0.1 to 19 μg of CH4 per gram of dry biomass per day. Eh remained positive (41.6 to 316.8 mV) with available dissolved O2 (0.3 - 5.2 mg/L) in all measurement locations down to 20cm, indicating in situ aerobic CH4 oxidation is viable across these environments. Potential CH4 oxidation rates increased with increasing dissolved CH4 concentration. Highest potential CH4 oxidation rates were found in open-water sedge sites. Eh and dissolved O2 were lowest at these sites, suggesting that methanotrophs with low-O2 demand may populate sedge areas. Furthermore, potential CH4 oxidation rates were higher at depth than at the surface in thawing palsa, suggesting CH4 oxidation may mitigate CH4 production triggered by warming in these actively thawing environments. Forthcoming elemental analyses of peat and pore water will further elucidate trends and geochemical controls of CH4 oxidation rates in thawing permafrost areas.

Post-thaw amendment of cryopreserved sperm for use in artificial insemination of a viviparous fish, the green swordtail Xiphophorus helleri

PubMed Central

Dong, Qiaoxiang; Huang, Changjiang; Tiersch, Terrence R.

2017-01-01

Sperm cryopreservation protocols have been developed for live-bearers such as the green swordtail Xiphophorus helleri and the platyfish Xiphophorus couchianus. Despite the high post-thaw motility (~75%) obtained in both species, the requirements of sperm storage within the female reproductive tract coupled with the process of internal fertilization place functional demands upon cryopreserved sperm samples far beyond those of oviparous species. The purpose of this study was to facilitate the artificial insemination process with cryopreserved sperm of X. helleri through evaluation of parameters related to sperm quality after thawing. Specifically, this study evaluated the effects on motility for fresh and thawed sperm samples of centrifugation (for concentration of sperm and washing for removal of cryoprotectant), ionic composition, and additions of glucose and fetal bovine serum (FBS) in extender solutions. Centrifugation at 1000 ×g for 10 min at 4 °C was found to have no adverse effects on sperm motility of fresh samples, and for cryopreserved samples, the removal of glycerol by washing yielded higher and longer post-thaw motility (e.g., 168 h vs. 48 h for the controls). Suspension of fresh sperm samples in magnesium-free Hanks’ balanced salt solution (HBSS) did not affect motility; however, HBSS prepared with the absence of potassium or calcium, and the use of unsupplemented saline (NaCl alone) as extenders significantly reduced sperm motility. The presence of glucose in HBSS yielded higher and longer motility for fresh and thawed samples, but addition of glucose at greater than 2 g/L were unnecessary. Addition of 20% FBS prior to freezing was found to increase the post-thaw motility significantly compared to control treatment with 14% glycerol alone. Also addition of 20% FBS after thawing and centrifugation was found to induce the formation of sperm bundles, which may be beneficial for internal fertilization success. In conclusion, concentration of sperm and the removal of cryoprotectant (through centrifugation), and the addition of 20% FBS in the extender is recommended for future insemination trials with cryopreserved samples. PMID:29269962

Potential weathering by freeze-thaw action in alpine rocks in the European Alps during a nine year monitoring period

NASA Astrophysics Data System (ADS)

Kellerer-Pirklbauer, Andreas

2017-11-01

A quantification of rock weathering by freeze-thaw processes in alpine rocks requires at least rock temperature data in high temporal resolution, in high quality, and over a sufficient period of time. In this study up to nine years of rock temperature data (2006-2015) from eleven rock monitoring sites in two of the highest mountain ranges of Austria were analyzed. Data were recorded at a half-hourly or hourly logging interval and at rock depths of 3, 10, and 30-40 cm. These data have been used to quantify mean conditions, ranges, and relationships of the potential near-surface weathering by freeze-thaw action considering volumetric-expansion of ice and ice segregation. For the former, freeze-thaw cycles and effective freeze-thaw cycles for frost shattering have been considered. For the latter, the intensity and duration of freezing events as well as time within the 'frost cracking window' have been analyzed. Results show that the eleven sites are in rather extreme topoclimatic positions and hence represent some of the highest and coolest parts of Austria and therefore the Eastern Alps. Only four sites are presumably affected by permafrost. Most sites are influenced by a long-lasting seasonal snow cover. Freeze-thaw cycles and effective freeze-thaw cycles for frost shattering are mainly affecting the near-surface and are unimportant at few tens of centimeters below the rock surface. The lowest temperatures during freezing events and the shortest freezing events have been quantified at all eleven monitoring sites very close to the surface. The time within the frost cracking window decreases in most cases from the rock surface inwards apart from very cold years/sites with very low temperatures close to the surface. As shown by this study and predicted climate change scenarios, assumed warmer rock temperature conditions in the future at alpine rock walls in Austria will lead to less severe freezing events and to shorter time periods within the frost-cracking window. Statistical correlation analyses showed furthermore that the longer the duration of the seasonal snow cover, the fewer are freeze-thaw cycles, the fewer are effective freeze-thaw cycles, the longer is the mean and the maximum duration of freezing events, and the lower is the mean annual ground temperature. The interaction of the winter snow cover history and the winter thermal regime has a complex effect on the duration of the frost cracking window but also on the number of freeze-thaw cycles as shown by a conceptual model. Predicted future warmer and snow-depleted winters in the European Alps will therefore have a complex impact on the potential weathering of alpine rocks by frost action which makes potential weathering predictions difficult. Neglecting rock moisture and rock properties in determining rock weathering limits the usefulness of solely rock temperature data. However, rock temperature data allow getting an estimate about potential weathering by freeze-thaw action which is often substantially more than previously known.

Greenhouse gas release from arctic permafrost: positive feedback to climate warming (Invited)

NASA Astrophysics Data System (ADS)

Walter Anthony, K. M.; Zimov, S. A.

2009-12-01

The release of carbon (C) in the form of greenhouse gases from thawing permafrost is one of the most likely and important positive feedbacks from the land to the atmosphere in a warmer world. Perennially frozen ground, known as permafrost, covers 20 percent of the Earth’s land surface. Recent accounting for C stored as far as 80m beneath the surface in permafrost (950 billion tons) more than doubles previous inventory estimates and is comparable to the current atmospheric CO2 burden of 750 billion tons. Permafrost organic C accumulated over tens of thousands of years. In its frozen state this C is sequestered from the atmosphere, mitigating climate warming. Long term borehole from Siberia and North America attest that permafrost is thawing. A third to half of permafrost is now within a degree to a degree and a half of thawing. In places where permafrost temperature crosses the critical 0°C threshold, ice melts causing thermokarst (ground surface collapse). Thermokarst features such as sink holes, pits, slope failure, mud flows, and the formation, expansion, and drainage of thaw lakes are widespread, up to 90% of the land area in some areas of the Arctic. Dating of features revealed that this process has been going on for the past 10,000 years, since the Earth entered the most recent interglacial warm period. However, satellite records during the past 55 years suggest that permafrost thaw in some regions is accelerating. What will happen to the climate as the rest of the permafrost thaws? When permafrost thaws, organic C is made available to microbes, which rapidly degrade it, producing greenhouse gases such as CO2 and methane (CH4, 25 times the global warming potential of CO2 over 100 years). A particularly important region for greenhouse gas emissions is the Siberian Yedoma Ice Complex (10^6 km2), a Pliestocene-aged permafrost type that contains roughly half of the Arctic’s permafrost C stock. Based on patterns of yedoma degradation during the present interglacial period, estimates of the amount of C remaining in permafrost today, long term field measurements, laboratory incubation experiments, and mass balance calculations of the efficiency of CH4 production from thawed permafrost, we predict that at least 50 billion tons of CH4 (equivalent to 10 times the current atmospheric methane burden) will escape from thermokarst lakes in Siberia as yedoma thaws. More CH4 will be released from the remainder of arctic lakes. Under current projections of warming and thaw in the Arctic (7-8 deg C by 2100), thermokarst will release 0.1-0.2 billion tons CH4 yr-1 by 2100, an order of magnitude more than its current source strength, adding another 20-40% of all human and natural sources of CH4 to the atmosphere. Frozen soils which thaw under aerobic conditions will produce CO2 with projected emissions of ~0.5-1.0 billion tons C yr-1, constituting approximately 10% of modern anthropogenic emission.

Computed parameters : freeze/thaw monograph for LTPP

DOT National Transportation Integrated Search

1998-01-01

To study the effect of freezing/thawing conditions on pavement performance, data from three electrical resistivity (ER) measurements (resistivity, resistance, and voltage) are collected approximately every month, every other year at selected Seasonal...

Thrombin generation potential and clot-forming capacity of thawed fresh-frozen plasma, plasma frozen within 24 h and solvent/detergent-treated plasma (octaplasLG® ), during 5-day storage at 1-6°C.

PubMed

Heger, A; Neisser-Svae, A; Trawnicek, L; Triulzi, D

2018-04-23

To enable rapid availability of plasma in emergency situations, the shelf-life of thawed fresh-frozen plasma (FFP) has been extended from 24 h to 5 days. The aim of this study was to evaluate the thrombin generation (TG) potential and clot-forming ability during 5 days of refrigerated storage of thawed FFP, plasma frozen within 24 h and solvent/detergent-treated plasma octaplasLG ® . During storage for 5 days, TG capacity decreased significantly over time, and rotational thromboelastometry showed significantly prolonged clotting times. However, the stability studies confirmed comparable in vitro haemostatic potentials of all three thawed plasma products at day 5. © 2018 International Society of Blood Transfusion.

Membrane damage during dilution, cooling and freezing-thawing of boar spermatozoa packaged in plastic bags.

PubMed

Ortman, K; Rodriguez-Martinez, H

1994-02-01

The objective of the present investigation was to determine the degree of membrane damage in boar spermatozoa during the course of a cryopreservation procedure, including thawing. Ejaculates from four fertile Swedish Yorkshire boars were frozen under controlled conditions in Teflon-plastic bags (2.5 ml) using 3% glycerol as cryoprotectant. Membrane integrity was monitored using supravital fluorescent dyes and confirmed by scanning electron microscopy. The results show that cooling to +5 degrees C significantly affected the permeability of the plasmalemma. Supercooling (-6 degrees C) during the freezing program did not further deteriorate the integrity of the spermatozoal membrane. The thawing procedure however, dramatically increased the frequency of spermatozoa with damaged plasma membranes. Thus particular attention must be paid on designing a better thawing procedure when dealing with boar semen frozen in plastic bags.

Cryopreservation of boar semen by egg yolk-based extenders containing lactose or fructose is better than sorbitol.

PubMed

Chanapiwat, Panida; Kaeoket, Kampon; Tummaruk, Padet

2012-03-01

The present study determined the effect of different types of sugars (lactose, fructose, glucose and sorbitol) used in egg yolk-based extender on the post-thawed boar semen quality. Twenty-two ejaculates from 6 fertility-proven Yorkshire boars were cryopreserved by liquid nitrogen vapor method. Sperm motility, viability, acrosome integrity and intact functional plasma membrane were determined at 0, 2 and 4 hr after thawing. It was found that the lactose-based extender resulted in a higher percentage of post-thawed sperm motility, viability, intact acrosome and functional plasma membrane than sorbitol-based extender (P<0.05) and fructose-based extender yielded a higher post-thawed sperm motility and viability than sorbitol-based extender (P<0.05). It could be concluded that sorbitol was not an effective sugar for the cryopreservation in boar semen.

Impact of freezing and thawing on the quality of meat: review.

PubMed

Leygonie, Coleen; Britz, Trevor J; Hoffman, Louwrens C

2012-06-01

This comprehensive review describes the effects of freezing and thawing on the physical quality parameters of meat. The formation of ice crystals during freezing damages the ultrastructure and concentrates the solutes in the meat which, in turn, leads to alterations in the biochemical reactions that occur at the cellular level and influence the physical quality parameters of the meat. The quality parameters that were evaluated are moisture loss, protein denaturation, lipid and protein oxidation, colour, pH, shear force and microbial spoilage. Additionally mechanisms employed to mitigate the effects of freezing and thawing were also reviewed. These include the use of novel methods of freezing and thawing, ante and post mortem antifreeze protein inclusion and vitamin E supplementation, brine injection and modified atmospheric packaging. Copyright © 2012 Elsevier Ltd. All rights reserved.

Application of Satellite SAR Imagery in Mapping the Active Layer of Arctic Permafrost

NASA Technical Reports Server (NTRS)

Li, Shu-Sun; Romanovsky, V.; Lovick, Joe; Wang, Z.; Peterson, Rorik

2003-01-01

A method of mapping the active layer of Arctic permafrost using a combination of conventional synthetic aperture radar (SAR) backscatter and more sophisticated interferometric SAR (INSAR) techniques is proposed. The proposed research is based on the sensitivity of radar backscatter to the freeze and thaw status of the surface soil, and the sensitivity of INSAR techniques to centimeter- to sub-centimeter-level surface differential deformation. The former capability of SAR is investigated for deriving the timing and duration of the thaw period for surface soil of the active layer over permafrost. The latter is investigated for the feasibility of quantitative measurement of frost heaving and thaw settlement of the active layer during the freezing and thawing processes. The resulting knowledge contributes to remote sensing mapping of the active layer dynamics and Arctic land surface hydrology.

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

PubMed Central

Arora, Rajeev; Palta, Jiwan P.

1991-01-01

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

Effects of Freeze-Thawing and Intravenous Infusion on Mesenchymal Stromal Cell Gene Expression.

PubMed

Hoogduijn, Martin J; de Witte, Samantha F H; Luk, Franka; van den Hout-van Vroonhoven, Mirjam C G N; Ignatowicz, Lech; Catar, Rusan; Strini, Tanja; Korevaar, Sander S; van IJcken, Wilfred F J; Betjes, Michiel G H; Franquesa, Marcella; Moll, Guido; Baan, Carla C

2016-04-15

Mesenchymal stromal cells (MSC) are increasingly used as an investigative therapeutic product for immune disorders and degenerative disease. Typically, MSC are isolated from human tissue, expanded in culture, and cryopreserved until usage. The safety and efficacy of MSC therapy will depend on the phenotypical and functional characteristics of MSC. The freeze-thawing procedure may change these characteristics. Furthermore, the cells encounter a microenvironment after administration that may impact their properties. It has been demonstrated that the majority of MSC localize to the lungs after intravenous infusion, making this the site to study the effects of the in vivo milieu on administered MSC. In this study, we investigated the effect of freeze-thawing and the mouse lung microenvironment on human adipose tissue-derived MSC. There were effects of freeze-thawing on the whole genome expression profile of MSC, although the effects did not exceed interdonor differences. There were no major changes in the expression of hemostatic regulators on transcriptional level, but significantly increased expression of procoagulant tissue factor on the surface of thawed adipose MSC, correlating with increased procoagulant activity of thawed cells. Exposure for 2 h to the lung microenvironment had a major effect on MSC gene expression and affected several immunological pathways. This indicates that MSC undergo functional changes shortly after infusion and this may influence the efficacy of MSC to modulate inflammatory responses. The results of this study demonstrate that MSC rapidly alter in response to the local milieu and disease-specific conditions may shape MSC after administration.

[Effect of different cryopreservation time on quality of umbilical cord blood cells].

PubMed

Huang, Lu; Song, Gui-Qi; Wu, Yun; Wang, Jian

2013-02-01

This study was aimed to explore the effect of different cryopreservation time on recovery rate of cord blood stem cells, and analyze the influence of cord blood cells after thawing on the engraftment speed of cord blood cells in patients. 20 cord blood units were stored at -196°C for 1 - 10 years. The cell viability, content of total nucleated cell (TNC), CD34(+) cells and the colony forming units of granulocyte/macrophage (CFU-GM) were assessed after thawing, the impact of cell recovery on engraftment speed in patients was analyzed. The results showed that as compared with data provided by Umbilical Cord Blood Bark, the different cryopreservation time had no effect on yield of cord blood stem cells after thawing. The cell viability was (92.75 ± 2.55)% after thawing, the yields of TNC, CD34(+) cells and CFU-GM were 89.9%, 84.8% and 84.3%, compared with that of pre-freezing, their differences were statistically significant (P = 0.000), however, loss of cells had no effect on the time of neutrophils and platelets engraftment. The TNC and CD34(+)cell count after thawing correlated closely with that of pre-freezing (r = 0.954 and r = 0.931, P = 0.000), but CFU-GM content poorly correlated with that (r = 0.285, P = 0.223). It is concluded that cryopreservation and thawing process can damage the cord blood stem cells, leading to cell loss, but not affect transplant results.

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

PubMed

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

2015-12-01

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

Freezing-thawing and sub-sampling influence the marination performance of chicken breast meat.

PubMed

Bowker, B; Zhuang, H

2017-09-01

Vacuum-tumbling marination is often used to improve the yield and quality of whole or portioned broiler breast fillets. The relationship between the marination performance of whole Pectoralis major muscles and breast fillet sub-samples is not well understood. The objective was to determine the effects of sub-sampling and freezing-thawing on the marination performance and cook loss of broiler breast meat. Paired right and left breast fillets were marinated as whole fillets or sub-samples (cranial and mid-caudal portions). Samples were marinated at 48 h postmortem (fresh) or stored at -20°C and then thawed prior to marination (frozen-thawed). Samples were vacuum-tumbled in 20% wt/wt brine (5% NaCl, 3% STP) and weighed pre-marination, during marination (15, 30, and 45 min), and 24 h post-marination. Samples were then cooked to 75°C for determination of cook loss. Marinade uptake was greater in caudal sub-samples than intact fillets and cranial sub-samples after 15 min of marination (P < 0.0001). After 30 min, marinade uptake was greater in caudal sub-samples and intact fillets than cranial sub-samples (P < 0.05). After 45 min, marinade uptake for fresh samples was greatest in intact fillets and lowest in cranial sub-samples. For frozen-thawed samples, marinade uptake at 45 min was greater in caudal sub-samples and intact fillets than cranial sub-samples (P < 0.0001). Marinade uptake in sub-samples at 30 min was greater in frozen-thawed versus fresh fillets (P < 0.05). Differences in marinade retention were not observed. Cook loss was similar between fresh and frozen-thawed samples but was greater in sub-samples compared to intact fillets (P < 0.0001). Correlations between marinade uptake in intact fillets and cranial sub-samples were greater in fresh (r = 0.64 to 0.78) than frozen-thawed samples (r = 0.39 to 0.59). Correlations between marinade uptake in intact fillets and caudal sub-samples were greater in frozen-thawed (r = 0.79 to 0.82) than fresh samples (r = 0.46 to 0.63). Data suggest that the relationships between marination performance of whole breast fillets and fillet sub-samples are dependent upon prior sample handling and intra-fillet sampling location. Published by Oxford University Press on behalf of Poultry Science Association 2017.

Effect of alpha-lipoic acid on boar spermatozoa quality during freezing-thawing.

PubMed

Shen, Tao; Jiang, Zhong-Liang; Li, Cong-Jun; Hu, Xiao-Chen; Li, Qing-Wang

2016-04-01

Alpha-lipoic acid (ALA) is known to be a natural antioxidant. The aim of the present study was to evaluate the cryoprotective effect of ALA on the motility of boar spermatozoa and its antioxidant effect on boar spermatozoa during freezing-thawing. Different concentrations (2.0, 4.0, 6.0, 8.0 or 10.0 mg/ml) of ALA were added to the extender used to freeze boar semen, and the effects on the quality and endogenous antioxidant enzyme activities of frozen-thawed spermatozoa were assessed. The results indicated that the addition of ALA to the extender resulted in a higher percentage of motile spermatozoa post-thaw (P < 0.05). The activities of superoxide dismutase, lactate dehydrogenase, glutamic-oxaloacetic transaminase and catalase improved after adding ALA to the extender (P < 0.05). Artificial insemination results showed that pregnancy rate and litter size were significantly higher at 6.0 mg/ml in the ALA group than in the control group (P < 0.05). In conclusion, ALA conferred a cryoprotective capacity to the extender used for boar semen during the process of freezing-thawing, and the optimal concentration of ALA for the frozen extender was 6.0 mg/ml.

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

PubMed

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 (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. 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.

Identifying Hydrologic Flowpaths on Arctic Hillslopes Using Electrical Resistivity and Self Potential

NASA Astrophysics Data System (ADS)

Voytek, E.; Rushlow, C. R.; Godsey, S.; Singha, K.

2015-12-01

Shallow subsurface flow is a dominant process controlling hillslope runoff generation, soil development, and solute reaction and transport. Despite their importance, the location and geometry of flowpaths are difficult to determine. In arctic environments, shallow subsurface flowpaths are limited to a thin zone of seasonal thaw above continuous permafrost, which is traditionally assumed to mimic to surface topography. Here we use a combined approach of electrical resistivity imaging (ERI) and self-potential measurements (SP) to map shallow subsurface flowpaths in and around water tracks, drainage features common to arctic hillslopes. ERI measurements delineate thawed zones in the subsurface that control flowpaths, while SP is sensitive to groundwater flow. We find that areas of low electrical resistivity in the water tracks are deeper than manual thaw depth estimates and variations from surface topography. This finding suggests that traditional techniques significantly underestimate active layer thaw and the extent of the flowpath network on arctic hillslopes. SP measurements identify complex 3-D flowpaths in the thawed zone. Our results lay the groundwork for investigations into the seasonal dynamics, hydrologic connectivity, and climate sensitivity of spatially distributed flowpath networks on arctic hillslopes.

Impact of Buffer, Protein Concentration and Sucrose Addition on the Aggregation and Particle Formation during Freezing and Thawing.

PubMed

Hauptmann, Astrid; Podgoršek, Katja; Kuzman, Drago; Srčič, Stanko; Hoelzl, Georg; Loerting, Thomas

2018-03-19

This study addresses the effect of freezing and thawing on a therapeutic monoclonal antibody (mAb) solution and the corresponding buffer formulation. Particle formation, crystallization behaviour, morphology changes and cryo-concentration effects were studied after varying the freezing and thawing rates, buffer formulation and protein concentration. The impact of undergoing multiple freeze/thaw (FT)-cycles at controlled and uncontrolled temperature rates on mAb solutions was investigated in terms of particle formation. Physicochemical characteristics were analysed by Differential Scanning Calorimetry whereas morphology changes are visualized by cryomicroscopy measurements. Micro Flow Imaging, Archimedes and Dynamic Light Scattering were used to investigate particle formation. Data retrieved in the present study emphasizes the damage caused by multiple FT-cyles and the need for sucrose as a cryoprotectant preventing cold-crystallization specifically at high protein concentrations. Low protein concentrations cause an increase of micron particle formation. Low freezing rates lead to a decreased particle number with increased particle diameter. The overall goal of this research is to gain a better understanding of the freezing and thawing behaviour of mAb solutions with the ultimate aim to optimize this process step by reducing the unwanted particle formation, which also includes protein aggregates.

Role of ice structuring proteins on freezing-thawing cycles of pasta sauces.

PubMed

Calderara, Marianna; Deorsola, Fabio A; Bensaid, Samir; Fino, Debora; Russo, Nunzio; Geobaldo, Francesco

2016-12-01

The freezing of the food is one of the most important technological developments for the storage of food in terms of quality and safety. The aim of this work was to study the role of an ice structuring protein (ISP) on freezing-thawing cycles of different solutions and commercial Italian pasta sauces. Ice structuring proteins were related to the modification of the structure of ice. The results showed that the freezing time of an aqueous solution containing the protein was reduced to about 20% with respect to a pure water solution. The same effect was demonstrated in sugar-containing solutions and in lipid-containing sauces. The study proved a specific role of ISP during thawing, inducing a time decrease similar to that of freezing and even more important in the case of tomato-based sauces. This work demonstrated the role of ISP in the freezing-thawing process, showing a significant reduction of processing in the freezing and thawing phase by adding the protein to pure water and different sugar-, salt- and lipid-containing solutions and commercial sauces, with considerable benefits for the food industry in terms of costs and food quality.

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

NASA Astrophysics Data System (ADS)

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.

Seminal plasma applied post-thawing affects boar sperm physiology: a flow cytometry study.

PubMed

Fernández-Gago, Rocío; Domínguez, Juan Carlos; Martínez-Pastor, Felipe

2013-09-01

Cryopreservation induces extensive biophysical and biochemical changes in the sperm. In the present study, we used flow cytometry to assess the capacitation-like status of frozen-thawed boar spermatozoa and its relationship with intracellular calcium, assessment of membrane fluidity, modification of thiol groups in plasma membrane proteins, reactive oxygen species (ROS) levels, viability, acrosomal status, and mitochondrial activity. This experiment was performed to verify the effect of adding seminal plasma on post-thaw sperm functions. To determine these effects after cryopreservation, frozen-thawed semen from seven boars was examined after supplementation with different concentrations of pooled seminal plasma (0%, 10%, and 50%) at various times of incubation from 0 to 4 hours. Incubation caused a decrease in membrane integrity and an increase in acrosomal damage, with small changes in other parameters (P > 0.05). Although 10% seminal plasma showed few differences with 0% (ROS increase at 4 hours, P < 0.05), 50% seminal plasma caused important changes. Membrane fluidity increased considerably from the beginning of the experiment, and ROS and free thiols in the cell surface increased by 2 hours of incubation. By the end of the experiment, viability decreased and acrosomal damage increased in the 50% seminal plasma samples. The addition of 50% of seminal plasma seems to modify the physiology of thawed boar spermatozoa, possibly through membrane changes and ROS increase. Although some effects were detrimental, the stimulatory effect of 50% seminal plasma could favor the performance of post-thawed boar semen, as showed in the field (García JC, Domínguez JC, Peña FJ, Alegre B, Gonzalez R, Castro MJ, Habing GG, Kirkwood RN. Thawing boar semen in the presence of seminal plasma: effects on sperm quality and fertility. Anim Reprod Sci 2010;119:160-5). Copyright © 2013 Elsevier Inc. All rights reserved.

Effect of controlled and uncontrolled rate of cooling, prior to controlled rate of freezing, on motion characteristics and acrosomal integrity of cryopreserved ram spermatozoa.

PubMed

Joshi, Anil; Kumar, Davendra; Naqvi, S M K; Maurya, V P

2008-12-01

A programmable cell freezer provides ideal cryobiological conditions for controlled-rate cooling and freezing of ram spermatozoa. The purpose of this study was to investigate the effects of controlled (Group 1) and uncontrolled (Group 2) cooling conditions prior to programmable freezing of ram semen on post-thaw sperm motion characteristics and acrosomal integrity of ram spermatozoa. Semen samples of good initial motility obtained from adult Malpura rams were pooled, diluted to 1 × 10(9) spermatozoa per milliliter with Egg yolk-TEST-glycerol extender, and packaged in 0.25 mL straws. Straws representing Group 1 were cooled in a programmable cell freezer from 25°C to 5°C at the rate of -0.15°C per minute followed by a holding time of 2 h for equilibration, while straws of Group 2 were allowed to cool slowly up to 5°C and equilibrate for 2 h in the cold cabinet. After equilibration, straws of Group 2 were also loaded in the cell freezer for freezing straws of both the treatment groups simultaneously from 5°C to -125°C at the rate of -25°C per minute. Thawing of straws was done at 50°C for 10 s and the quality of frozen-thawed spermatozoa was objectively assessed by using sperm motility analyzer. Thawed samples were also evaluated for acrosomal integrity after staining the dried semen smears with Giemsa stain. The average post-thaw motility of straws was significantly higher (P < 0.05) in samples frozen after controlled cooling, compared with samples frozen after uncontrolled rate of cooling. The percent of spermatozoa with normal acrosome was also significantly (P < 0.05) higher in Group 1, compared to Group 2. The results indicate that controlled-rate cooling has a significant effect on post-thaw motility and acrosomal integrity of frozen-thawed ram spermatozoa, compared to uncontrolled-rate cooling prior to programmable freezing.

Potential Arctic tundra vegetation shifts in response to changing temperature, precipitation and permafrost thaw

NASA Astrophysics Data System (ADS)

van der Kolk, Henk-Jan; Heijmans, Monique M. P. D.; van Huissteden, Jacobus; Pullens, Jeroen W. M.; Berendse, Frank

2016-11-01

Over the past decades, vegetation and climate have changed significantly in the Arctic. Deciduous shrub cover is often assumed to expand in tundra landscapes, but more frequent abrupt permafrost thaw resulting in formation of thaw ponds could lead to vegetation shifts towards graminoid-dominated wetland. Which factors drive vegetation changes in the tundra ecosystem are still not sufficiently clear. In this study, the dynamic tundra vegetation model, NUCOM-tundra (NUtrient and COMpetition), was used to evaluate the consequences of climate change scenarios of warming and increasing precipitation for future tundra vegetation change. The model includes three plant functional types (moss, graminoids and shrubs), carbon and nitrogen cycling, water and permafrost dynamics and a simple thaw pond module. Climate scenario simulations were performed for 16 combinations of temperature and precipitation increases in five vegetation types representing a gradient from dry shrub-dominated to moist mixed and wet graminoid-dominated sites. Vegetation composition dynamics in currently mixed vegetation sites were dependent on both temperature and precipitation changes, with warming favouring shrub dominance and increased precipitation favouring graminoid abundance. Climate change simulations based on greenhouse gas emission scenarios in which temperature and precipitation increases were combined showed increases in biomass of both graminoids and shrubs, with graminoids increasing in abundance. The simulations suggest that shrub growth can be limited by very wet soil conditions and low nutrient supply, whereas graminoids have the advantage of being able to grow in a wide range of soil moisture conditions and have access to nutrients in deeper soil layers. Abrupt permafrost thaw initiating thaw pond formation led to complete domination of graminoids. However, due to increased drainage, shrubs could profit from such changes in adjacent areas. Both climate and thaw pond formation simulations suggest that a wetter tundra can be responsible for local shrub decline instead of shrub expansion.

Comparative cryopreservation of avian spermatozoa: benefits of non-permeating osmoprotectants and ATP on turkey and crane sperm cryosurvival.

PubMed

Blanco, Juan M; Long, Julie A; Gee, George; Wildt, David E; Donoghue, Ann M

2011-02-01

A comparative approach was used to evaluate the cryosurvival of turkey and crane sperm frozen in a dimethylacetamide (DMA) cryodiluent supplemented with osmoprotectants and ATP. A range (6-26%) of DMA concentrations was used alone or in combination with ATP (30, 60 or 118mM) or one of the following osmoprotectants: (1) sucrose (turkey, 8.0%; crane, 5.0%); (2) 5.0% sucrose and 5.0% trehalose; or (3) betaine hydrochloride (0.1, 0.2 or 0.4mM). The viability of thawed sperm was assessed using the nigrosin-eosin stain and sperm motility was determined using the hanging-drop technique. For semen frozen only with DMA, post-thaw sperm motility was greatest (P<0.05) for the 6.0%, 10.0% and 18% concentrations, regardless of species. Turkey sperm frozen with the sucrose/trehalose combination had greater (P<0.05) post-thaw motility for all DMA treatments compared to DMA alone. The lowest concentration of the osmoprotectant betaine hydrochloride substantially improved turkey sperm viability post-thaw in all treatments compared to DMA alone (P<0.05). The post-thaw motility of crane sperm was improved (P<0.05) with a combination of 18.0%, 24.0% or 26.0% DMA and 30mM ATP. Moreover, in the presence of osmoprotectants, crane sperm motility decreased as the osmoprotectant concentration increased. The lowest concentration of ATP also improved crane sperm viability post-thaw, especially for DMA concentrations 18% or greater. The combination of sucrose and trehalose improved (P<0.05) crane sperm viability only with 6% and 10% DMA. These data affirm that there are avian-specific differences in sperm survival after cryopreservation and suggest that post-thaw survival can be enhanced by including species-based osmoprotectant/ATP combinations in a cryodiluent where DMA is the cryoprotectant. Published by Elsevier B.V.

Rhesus monkey sperm cryopreservation with TEST-yolk extender in the absence of permeable cryoprotectant.

PubMed

Dong, Qiaoxiang; Correa, Liane M; VandeVoort, Catherine A

2009-02-01

Recently, there has been increased interest in ultra-rapid freezing with mammalian spermatozoa, especially for vitrification in the absence of cryoprotectants. Sperm cryopreservation in non-human primates has been successful, but the use of frozen-thawed sperm in standard artificial insemination (AI) remains difficult, and removal of permeable cryoprotectant may offer opportunities for increased AI success. The present study intended to explore the possibility of freezing rhesus monkey sperm in the absence of permeable cryoprotectants. Specifically, we evaluated various factors such as presence or absence of egg yolk, the percentage of egg yolk in the extenders, and the effect of cooling and thawing rate on the success of freezing without permeable cryoprotectants. Findings revealed that freezing with TEST in the absence of egg yolk offers little protection (<15% post-thaw motility). Egg yolk of 40% or more in TEST resulted in decreased motility, while egg yolk in the range of 20-30% yielded the most motile sperm. Cooling at a slow rate (29 degrees C/min) reduced post-thaw motility significantly for samples frozen with TEST-yolk alone, but had no effect for controls in the presence of glycerol. Similarly, slow thawing in room temperature air is detrimental for freezing without permeable cryoprotectant (<2% motility). In addition to motility, the ability of sperm to capacitate based on an increase in intracellular calcium levels upon activation with cAMP and caffeine suggested no difference between fresh and frozen-thawed motile sperm, regardless of treatment. In summary, the present study demonstrates that ejaculated and epididymal sperm from rhesus monkeys can be cryopreserved with TEST-yolk (20%) in the absence of permeable cryoprotectant when samples were loaded in a standard 0.25-mL straw, cooled rapidly in liquid nitrogen vapor at 220 degrees C/min, and thawed rapidly in a 37 degrees C water bath. This study also represents the first success of freezing without permeable cryoprotectant in non-human primates.

Soil thaw effects on river discharge recessions of a subarctic catchment

NASA Astrophysics Data System (ADS)

Ploum, Stefan; Lyon, Steve; Teuling, Ryan; van der Velde, Ype

2017-04-01

Thawing permafrost in circumpolar regions is likely to change subsurface hydrology. In high latitude areas continuous permafrost is expected to partially thaw leading to sporadic permafrost with deeper groundwater flow paths. Moreover, freeze-thaw cycles of the shallow subsurface are likely to increase. River discharge recession analysis can be particularly useful to understand the hydrological effects of a thawing Arctic. Here we examine river discharge recessions of the Abiskojokka, a 560 km2 watershed with sporadic permafrost, using a river discharge record of 30 years (1985 - 2015). Snow observation records were used to separate river recessions in snowmelt and snowfree periods. We found significant differences between recessions during the snowmelt and snowfree seasons. During the snowmelt, recessions were close to linear (b=1.11), while during the snowfree period, recessions were more non-linear (b=1.54). Typically, non-linearity has been found to increase with discharge magnitude, while we observed the opposite (snowfree periods tend to have lower discharges than the snowmelt periods). We explain these contrasting results by hypothesizing that increased connectivity (increasing magnitude and number of water flow paths) between groundwater and stream leads to higher non-linearity. In temperate catchments without frozen soils, connectivity tends to increase with increasing discharge. In contrast, in Arctic systems, where soils are frozen, connectivity between groundwater and stream is limited. Therefore, thawing of frozen soils is expected to increase connectivity and thus non-linearity of river discharges. We tested this hypothesis with a detailed analysis of all spring flood recessions. Years with cold soil temperatures (b=1.08) and years with a below median snowpack depth were found to have progressively linear slopes (b=1.08 and 1.01 respectively). On the other hand, years with warm soil conditions show increasingly non-linear recessions (b=1.67). Although limited in spatial extent, these results further support our connectivity hypothesis, which predicts increasing non-linearity of river discharges (higher discharge peaks and lower low flows under the same precipitation regime) as permafrost thaws.

Feasibility of Steel Fiber-Reinforced Rubberized Concrete in Cold Regions for High Volume Intersections

NASA Astrophysics Data System (ADS)

Abou Eid, Mahear A.

There are many challenges faced with the use of Portland Cement Concrete (PCC) in cold regions, but with the inclusion of new technologies such as steel fibers and recycled tire crumb rubber efficient construction may be possible. Research was conducted on a modified concrete material that included both steel fibers and crumb rubber. The composite material was called Steel Fiber-Reinforced Rubberized Concrete (SFRRC). The objective of this investigation was to provide evidence showing that SFRRC can reduce tire rutting compared to asphaltic pavement. In addition, the research showed that the SFRRC could withstand freeze-thaw cycles and increase service life of roadways. Several tests were performed to determine the characteristics of the material. Freeze-thaw testing was performed to determine compressive strength loss and visual deterioration of the material. Wheel tracker rut testing was performed both with the standard steel wheel and with a modified studded rubber tire to determine plastic deformation and rut resistance. An experimental test slab was cast in place on a public approach to observe the construction procedures, the effects of studded tire wear and the frost actions in cold region conditions. Based on freeze-thaw and wheel tracker test results and observations of the experimental test slab, the SFRRC material shows viability in cold regions for resisting freeze-thaw actions. The freeze-thaw testing resulted in increased compressive strength after 300 freeze-thaw cycles and very low deterioration of material compared to standard PCC. The wheel tracker testing resulted in very low plastic deformation and minor material rutting with use of the studded rubber tire. The test slab showed very minor surface wear, no freeze-thaw cracking and no rutting after one winter of use. It is recommended that further testing of the material be conducted by means of a large-scale trial section. This would provide information with respect to cost analysis and integration of the material into the Department of Transportation construction program.

Ecosystem recovery: a neglected factor in greenhouse gas emission from permafrost degradation.

NASA Astrophysics Data System (ADS)

van Huissteden, J.; Mi, Y.; Gallagher, A.; Budishchev, A.

2012-04-01

It is estimated that northern soils hold nearly twice as much carbon as the atmosphere. Permafrost degradation caused by a warming climate will destabilize this carbon store. Part of this carbon will enter the atmosphere as CO2 or CH4, contributing to a positive feedback on climate warming. However, a neglected factor is the recovery of ecosystems after permafrost thaw. Modeling of thaw lake expansion and drainage has shown that thaw lake expansion by climatic warming is strongly limited by lake drainage. Thaw lakes are drained or filled in with sediment, followed by recolonization by generally productive wetland ecosystems. Decomposition of soil carbon also releases nutrients, enhancing vegetation recolonization in types of permafrost degradation features. Examples from the Kytalyk/Chokurdagh research site in the Indigirka lowlands of northeastern Siberia illustrate that ecosystem recovery after localized permafrost degradation may effectively counteract carbon loss. The research site is located in a drained Early Holocene thaw lake basin, and is presently a greenhouse gas sink during the growing season. Formation of thaw ponds has increased strongly recently. Although fresh ponds may be emitting CO2 and CH4, they are rapidly invaded by vegetation which decreases net greenhouse gas emission, although the ponds continue to be a source of CH4. Areas of intense mass wasting by permafrost slides are colonized by a productive pioneer vegetation, contributing to stabilization of the soil and enhancing CO2 uptake. It is therefore essential that not only the greenhouse gas emission related to permafrost degradation is quantified, but also the carbon sinks and recovery rates. Paleo-environmental and geomorphological studies may help to quantify recovery processes, in particular those processes that leave their trace in the sedimentary record. For instance Pleistocene and younger thaw lake deposits in Europe and Siberia may provide information on carbon loss and carbon storage in successive stages of development.

Flow cytometric sorting of fresh and frozen-thawed spermatozoa in the western lowland gorilla (Gorilla gorilla gorilla).

PubMed

O'Brien, J K; Stojanov, T; Crichton, E G; Evans, K M; Leigh, D; Maxwell, W M C; Evans, G; Loskutoff, N M

2005-08-01

We adapted flow cytometry technology for high-purity sorting of X chromosome-bearing spermatozoa in the western lowland gorilla (Gorilla gorilla gorilla). Our objectives were to develop methodologies for liquid storage of semen prior to sorting, sorting of liquid-stored and frozen-thawed spermatozoa, and assessment of sorting accuracy. In study 1, the in vitro sperm characteristics of gorilla ejaculates from one male were unchanged (P > 0.05) after 8 hr of liquid storage at 15 degrees C in a non-egg yolk diluent (HEPES-buffered modified Tyrode's medium). In study 2, we examined the efficacy of sorting fresh and frozen-thawed spermatozoa using human spermatozoa as a model for gorilla spermatozoa. Ejaculates from one male were split into fresh and frozen aliquots. X-enriched samples derived from both fresh and frozen-thawed human semen were of high purity, as determined by fluorescence in situ hybridization (FISH; 90.7%+/-2.3%, overall), and contained a high proportion of morphologically normal spermatozoa (86.0%+/-1.0%, overall). In study 3, we processed liquid-stored semen from two gorillas for sorting using a modification of methods for human spermatozoa. The sort rate for enrichment of X-bearing spermatozoa was 7.3+/-2.5 spermatozoa per second. The X-enriched samples were of high purity (single-sperm PCR: 83.7%) and normal morphology (79.0%+/-3.9%). In study 4 we examined frozen-thawed gorilla semen, and the sort rate (8.3+/-2.9 X-bearing sperm/sec), purity (89.7%), and normal morphology (81.4%+/-3.4%) were comparable to those of liquid-stored semen. Depending on the male and the type of sample used (fresh or frozen-thawed), 0.8-2.2% of gorilla spermatozoa in the processed ejaculate were present in the X-enriched sample. These results demonstrate that fresh or frozen-thawed gorilla spermatozoa can be flow cytometrically sorted into samples enriched for X-bearing spermatozoa. Copyright 2005 Wiley-Liss, Inc.

Improved spring load restriction guidelines using mechanistic analysis

DOT National Transportation Integrated Search

2000-07-01

This project used research to develop more effective criteria for placement and removal of spring load restrictions (SLR). Researchers investigated a method that uses a thawing index equation based on air temperatures to predict thawing events. Resul...

Sensitivity of boreal forest carbon balance to soil thaw

USGS Publications Warehouse

Goulden, M.L.; Wofsy, S.C.; Harden, J.W.; Trumbore, S.E.; Crill, P.M.; Gower, S.T.; Fries, T.; Daube, B.C.; Fan, S.-M.; Sutton, D.J.; Bazzaz, A.; Munger, J.W.

1998-01-01

We used eddy covariance; gas-exchange chambers; radiocarbon analysis; wood, moss, and soil inventories; and laboratory incubations to measure the carbon balance of a 120-year-old black spruce forest in Manitoba, Canada. The site lost 0.3 ?? 0.5 metric ton of carbon per hectare per year (ton C ha-1 year-1) from 1994 to 1997, with a gain of 0.6 ?? 0.2 ton C ha-1 year-1 in moss and wood offset by a loss of 0.8 ?? 0.5 ton C ha-1 year-1 from the soil. The soil remained frozen most of the year, and the decomposition of organic matter in the soil increased 10-fold upon thawing. The stability of the soil carbon pool (~150 tons C ha-1) appears sensitive to the depth and duration of thaw, and climatic changes that promote thaw are likely to cause a net efflux of carbon dioxide from the site.

Comparison of Mitochondrial Function in Boar and Bull Spermatozoa Throughout Cryopreservation Based on JC-1 Staining.

PubMed

Hu, C H; Zhuang, X J; Wei, Y M; Zhang, M; Lu, S S; Lu, Y Q; Yang, X G; Lu, K H

Poor reproductivity hampers the commercialization of cryopreserved boar semen. This study was to determine the differences in the sperm mitochondrial function between boar and bull semen at different cryopreservation stages. Boar and bull fresh, equilibrated, and frozen-thawed spermatozoa were evaluated for mitochondrial function using JC-1 under a fluorescent microscope. Bull and boar percentage of spermatozoa staining green (PSSG) showed no difference between fresh and equilibrated semen (P> 0.05). However, frozen-thawed bull and boar semen demonstrated significantly higher PSSG (P < 0.01) than fresh and equilibrated semen. Frozen-thawed boar semen represented a significantly higher PSSG (P < 0.01) than bull semen. Negative cryopreservation influence on boar and bull spermatozoa was not significantly produced by pre-freezing procedures, but rather by freezing and thawing. Cryopreservation has more pronounced negative effects on boar than on bull spermatozoa, which partly explains lagged commercialization of frozen boar semen.

Method for Differentiation between Fresh and Frozen-thawed Fish

NASA Astrophysics Data System (ADS)

Kitamikado, Manabu; Yoshioka, Keiko

In Japan fresh fish has a much higher market price than that for frozen-thawed fish. However, a large number of frozen-thawed fish are sold without being differentiated from fresh fish. We discuss here the differentiation methods described in literatures and our works in the search for such a method. We used the opacity of crystalline lens and the destruction of red blood cells as the index for the differentiation, in addition to the activity of neutral β-N-acetylglucosaminidase in blood. Thus, a fluorometric method and a rapid paper test method were developed based on measurement of the activity of this enzyme. This enzyme, found in fish red blood cells, was inactive in intact cells but was activated when cells were disrupted by freezing, and thawing. Both methods were applicable for testing most commom edible fish prior to filleting and required about 20 min using a UV-lamp.

Antioxidant Effect of Xanthan Gum on Ram Sperm after Freezing and Thawing.

PubMed

Gastal, G DA; Silva, E F; Mion, B; Varela Junior, A S; Rosa, C E; Corcini, C D; Mondadori, R G; Vieira, A D; Bianchi, I; Lucia, T

Xanthan gum is used as thickener in media to preserve food products, having cryoprotectant and antioxidant properties that may be relevant for sperm cryopreservation. To evaluate the effects of adding xanthan gum to freezing extenders on post-thawing quality and oxidant activity of ram sperm. Ejaculates from seven rams extended TRIS-egg yolk-glycerol were split in three treatments including xanthan gum (0.15%; 0.20%; and 0.25%) and a control with no xanthan gum. After thawing, motility and production of reactive oxygen species (ROS) with 0.20% and 0.25% xanthan gum were lower than for the control (P < 0.05), but mitochondrial functionality and integrity of membrane, acrosome and DNA did not differ (P > 0.05). Xanthan gum at 0.20% and 0.25% may be an efficient antioxidant for frozen-thawed ram sperm, due to the reduction in ROS production.

Freeze-thaw durability of concrete: Ice formation process in pores

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cai, H.; Liu, X.

1998-09-01

Freeze-thaw durability of concrete is of great importance to hydraulic structures in cold areas. Study of ice formation process in concrete pores is necessary to evaluate the damages in concrete caused by freezing. In this paper, freezing of pore solution in concrete exposed to a freeze-thaw cycle is studied by following the change of concrete electrical conductivity with freezing temperatures. Concretes were subjected to freeze-thaw cycles with temperature varying between {minus}0 C and {minus}20 C. In the freezing process, the changing rate of concrete electrical conductivity obviously decreases at about {minus}10 C, indicating that more pore solution in concrete freezesmore » above {minus}10 C than below {minus}10C. According to Powers` static hydraulic pressure hypothesis, it is thought that frost damage mainly occurs between 0 C and {minus}100 C. To ordinary concrete, frost damages below {minus}10 C are negligible.« less

Freeze-thaw durability of microwave cured air-entrained concrete

DOE Office of Scientific and Technical Information (OSTI.GOV)

Pheeraphan, T.; Leung, C.K.Y.

1997-03-01

The strength development of concrete can be greatly accelerated by curing with microwave energy. Microwave curing can therefore be beneficial to construction operations such as concrete precasting and repair. To provide freeze-thaw durability for infrastructure applications, air entrainment has to be introduced. In this investigation, the freeze-thaw resistance of microwave cured air-entrained concrete is measured, and compared to that of air-entrained concrete under normal curing. Their compressive strength at 14 days and air-void characteristics are also measured and compared. The test results indicate that microwave curing can impair the freeze-thaw durability of high w/c concrete but not for low w/cmore » concrete. Also, under microwave curing, the decrease in strength due to air entrainment becomes more significant. Based on these observations, it is recommended that for microwave cured air-entrained concrete, a low w/c ratio should be employed.« less

Enhancing the Properties of Conductive Polymer Hydrogels by Freeze-Thaw Cycles for High-Performance Flexible Supercapacitors.

PubMed

Li, Wanwan; Lu, Han; Zhang, Ning; Ma, Mingming

2017-06-14

We report that a postsynthesis physical process (freeze-thaw cycles) can reform the microstructure of conductive polymer hydrogels from clustered nanoparticles to interconnected nanosheets, leading to enhanced mechanical and electrochemical properties. The polyaniline-poly(vinyl alcohol) hydrogel after five freeze-thaw cycles (PPH-5) showed remarkable tensile strength (16.3 MPa), large elongation at break (407%), and high electrochemical capacitance (1053 F·g -1 ). The flexible supercapacitor based on PPH-5 provided a large capacitance (420 mF·cm -2 and 210 F·g -1 ) and high energy density (18.7 W·h·kg -1 ), whose robustness was demonstrated by its 100% capacitance retention after 1000 galvanostatic charge-discharge cycles or after 1000 mechanical folding cycles. The outstanding performance enables PPH-5 based supercapacitor as a promising power device for flexible electronics, which also demonstrates the merit of freeze-thaw cycles for enhancing the performance of functional hydrogels.

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

PubMed

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

2000-01-01

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

A requirement for sucrose in xylem sap flow from dormant maple trees.

PubMed

Johnson, R W; Tyree, M T; Dixon, M A

1987-06-01

The response of excised stem segments of several tree species to freezing and thawing cycles was studied. All species studied (Thuja occidentalis, Fagus grandifolia, and Betula papyrifera) except maple (Acer spp.) exuded sap while freezing and absorbed on thawing. Maple stems absorbed sap while freezing and exuded sap during the thaw only when sucrose was present in the vessel solution. Increased concentration of sucrose in the vessel sap led to increased exudation. In the absence of sucrose, maple stems absorbed sap on thawing. The presence of sucrose enhanced sap absorption during freezing cycles in maples. In general, large sugars, disaccharides and larger, could substitute for sucrose in the maple exudation response while sugar hexoses could not. The results are discussed in relation to the O'Malley-Milburn model (1983 Can J Bot 61: 3100-3106) of sap flow in maples.

A Requirement for Sucrose in Xylem Sap Flow from Dormant Maple Trees 1

PubMed Central

Johnson, Robert W.; Tyree, Melvin T.; Dixon, Michael A.

1987-01-01

The response of excised stem segments of several tree species to freezing and thawing cycles was studied. All species studied (Thuja occidentalis, Fagus grandifolia, and Betula papyrifera) except maple (Acer spp.) exuded sap while freezing and absorbed on thawing. Maple stems absorbed sap while freezing and exuded sap during the thaw only when sucrose was present in the vessel solution. Increased concentration of sucrose in the vessel sap led to increased exudation. In the absence of sucrose, maple stems absorbed sap on thawing. The presence of sucrose enhanced sap absorption during freezing cycles in maples. In general, large sugars, disaccharides and larger, could substitute for sucrose in the maple exudation response while sugar hexoses could not. The results are discussed in relation to the O'Malley-Milburn model (1983 Can J Bot 61: 3100-3106) of sap flow in maples. Images Fig. 2 PMID:16665468

High preservation of DNA standards diluted in 50% glycerol.

PubMed

Schaudien, Dirk; Baumgärtner, Wolfgang; Herden, Christiane

2007-09-01

Standard curves are important tools in real-time quantitative polymerase chain reaction (PCR) to precisely analyze gene expression patterns under physiologic and pathologic conditions. Handling of DNA standards often implies multiple cycles of freezing and thawing that might affect DNA stability and integrity. This in turn might influence the reliability and reproducibility of quantitative measurements in real-time PCR assays. In this study, 3 DNA standards such as murine tumor necrosis factor (TNF) alpha, interferon (IFN) gamma, and kainat-1 receptor were diluted in 50% glycerol or water after 1, 4, and 16 cycles of freezing and thawing and amplified copy numbers after real-time PCR were compared. The standards diluted in water showed a reduction to 83%, 55%, and 50% after 4 cycles, to 24%, 5%, and 4% after 16 cycles for kainat-1 receptor, TNFalpha, and IFNgamma standards, respectively, when compared with a single cycle of freezing and thawing. Interestingly, all cDNA samples diluted in 50% glycerol were amplified in comparable copy numbers even after 16 cycles of freezing and thawing. The effect of the standards undergoing different cycles of freezing and thawing on sample values was demonstrated by amplifying cDNA obtained from Borna disease virus infected and noninfected TNF-transgenic mice brain. This revealed significant differences of measured cDNA copy numbers using water-diluted DNA standards. In contrast, sample values did not vary using glycerol-diluted standards that were frozen and thawed for 16 times. In conclusion, glycerol storage of DNA standards represents a suitable tool for the accurate and reproducible quantification of cDNA samples in real-time PCR analysis.

The effects of different levels of catalase and superoxide dismutase in modified Beltsville extender on rooster post-thawed sperm quality.

PubMed

Amini, Mahmood Reza; Kohram, Hamid; Zare-Shahaneh, Ahmad; Zhandi, Mahdi; Sharideh, Hossein; Nabi, Mohammad Mehdi

2015-06-01

Oxidative damage of sperm by means of reactive oxygen species generated by the cellular components of semen is one of the main reasons for decreased sperm motility and fertility during the freeze-thawing process. This study was conducted to determine the influence of catalase (CAT) and superoxide dismutase (SOD) on rooster sperm motility, viability and MDA level after freezing and thawing. Semen samples from 10 sexually-mature Ross 308 breeder roosters were collected and pooled, divided into nine equal parts and diluted with modified Beltsville extender containing no antioxidants (control), or supplemented with 50, 100, 200 and 300 μg/mL CAT, or 50, 100, 200 and 300 U/mL SOD. After thawing, sperm motility and motion parameters were assessed using a CASA system. Sperm viability and MDA level were assessed by eosin-nigrosin and MDA test, respectively. The results of this experiment showed that the extender supplemented with 100 and 200 μg CAT, and 50 U SOD had the highest sperm motility (P<0.05) in sperm motility. Also, addition 100, 200 and 300 μg CAT, and 50 U SOD can improve significantly viability after freeze-thaw. Extender supplemented with 100 μg CAT had significantly lower MDA level compared to control and 300 μg CAT. In conclusion, the results of the present study demonstrate that addition of CAT (100 μg/mL) and SOD (50 U/mL) independently have beneficial effect on quality of post-thawed rooster semen. Copyright © 2015 Elsevier Inc. All rights reserved.

High severity experimental burns in Siberian larch forests increase permafrost thaw and larch tree regeneration

NASA Astrophysics Data System (ADS)

Alexander, H. D.; Davydov, S.; Zimov, N.; Mack, M. C.

2013-12-01

Global change models predict increased fire activity in boreal forests as climate warms and dries. We hypothesized that fire-driven decreases in soil organic layer (SOL) depth will (1) increase permafrost thaw by reducing the insulating capacity of the SOL and (2) improve seedbed conditions for tree regeneration. Over time, these changes will lead to altered patterns of above- and belowground carbon (C) accumulation. To test these hypotheses, we conducted plot-level experimental burns in July 2012 in a low-density, mature larch stand near the Northeast Science Station in Cherskii, Siberia. Dried fuels of naturally occurring vegetation were added to plots to achieve four burn severity treatments based on residual SOL depths: control, low (> 8 cm), moderate (5-8 cm), and high severity (2-5 cm). Pre-fire and during two growing seasons post-fire, we measured thaw depth, soil moisture, and soil temperature to determine severity effects on permafrost thaw. We also sowed larch seeds in fall 2012 and quantified germination rates the following growing season. By 1 wk post-fire, thaw depth was 15-25 cm deeper in plots burned at high severity (55 cm) compared to other treatments (30-40 cm). These differences in thaw depth with burn severity were maintained during the subsequent growing season and were associated with increased soil temperature and moisture. Larch regeneration was 10x higher on severely burned plots than those unburned. Our findings highlight the potential for increased fire severity to degrade permafrost and alter successional dynamics and patterns of C accumulation.

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

NASA Astrophysics Data System (ADS)

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.

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

PubMed

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

2016-10-01

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

Modification of membrane cholesterol and its impact on frozen-thawed chicken sperm characteristics.

PubMed

Partyka, Agnieszka; Bonarska-Kujawa, Dorota; Sporniak, Marta; Strojecki, Maciej; Niżański, Wojciech

2016-10-01

This study was conducted to determine the changes in chicken sperm plasma membranes fluidity and polarity as lipid packing arrangement induced by cholesterol-loaded cyclodextrin (CLC) and 2-hydroxypropyl-β-cyclodextrin (HBCD) and how sperm cryopreservation outcomes are improved by these changes. Treatment with 2 mg HBCD supported the highest (P < 0.01) percentage of viable spermatozoa compared with the control and CLCs groups after cryopreservation. The percentage of post-thaw progressive and rapid sperm motility was highest in 2 mg HBCD (P < 0.01). After thawing, sperm treated with 1 or 2 mg CLC showed the highest anisotropy at 5, 21, 25 and 40°C (P < 0.01). At 25°C, the lowest anisotropy was observed in the thawed semen from the control group. The highest value (P < 0.01) of generalized polarization (GP) (0.5) at 5°C was observed in the 1 mg CLC treated sample. After 2 h of incubation, the highest percentage of viable spermatozoa was observed in the HBCD group in relation to the other treatments (P < 0.01). Exposure to 1 mg or 2 mg of CLC significantly decreased the percentage of live spermatozoa after thawing (P < 0.01). In conclusion, HBCD appears to play a role in the modification of sperm membranes, increasing their fluidity and preventing them against membrane phase transition to gel, thus minimizing freezing-thaw sperm damage. HBCD treatment enhances chicken sperm viability and motility after cryopreservation and subsequent storage. This novel procedure may be useful for improving the technology for cryopreservation of fowl spermatozoa.

Freeze/thaw conditions at periglacial landforms in Kapp Linné, Svalbard, investigated using field observations, in situ, and radar satellite monitoring

NASA Astrophysics Data System (ADS)

Eckerstorfer, M.; Malnes, E.; Christiansen, H. H.

2017-09-01

In periglacial landscapes, snow dynamics and microtopography have profound implications of freeze-thaw conditions and thermal regime of the ground. We mapped periglacial landforms at Kapp Linné, central Svalbard, where we chose six widespread landforms (solifluction sheet, nivation hollow, palsa and peat in beach ridge depressions, raised marine beach ridge, and exposed bedrock ridge) as study sites. At these six landforms, we studied ground thermal conditions, freeze-thaw cycles, and snow dynamics using a combination of in situ monitoring and C-band radar satellite data in the period 2005-2012. Based on these physical parameters, the six studied landforms can be classified into raised, dry landforms with minor ground ice content and a thin, discontinuous snow cover and into wet landforms with high ice content located in the topographical depressions in-between with medium to thick snow cover. This results in a differential snow-melting period inferred from the C-band radar satellite data, causing the interseasonal and interlandform variability in the onset of ground surface thawing once the ground becomes snow free. Therefore, variability also exists in the period of thawed ground surface conditions. However, the length of the season with thawed ground surface conditions does not determine the mean annual ground surface temperature, it only correlates well with the active layer depths. From the C-band radar satellite data series, measured relative backscatter trends hint toward a decrease in snow cover through time and a more frequent presence of ice layers from mid-winter rain on snow events at Kapp Linné, Svalbard.

Successful vitrification of human amnion-derived mesenchymal stem cells.

PubMed

Moon, Jeong Hee; Lee, Jung Ryeol; Jee, Byung Chul; Suh, Chang Suk; Kim, Seok Hyun; Lim, Hyun Jung; Kim, Hae Kwon

2008-08-01

A cryopreservation protocol for human amnion-derived mesenchymal stem cells (HAMs) is required because these cells cannot survive for long periods in culture. The aim of this study was to determine whether vitrification is a useful freezing method for storage of HAMs. HAMs were cryopreserved using vitrification method. The morphology and viability of thawed HAMs was evaluated by Trypan Blue staining. The expression of several embryonic stem cell (ESC) markers was evaluated using flow cytometry, RT-PCR and immunocytochemistry. Von Kossa, Oil Red O and Alcian Blue staining were used to asses the differentiation potential of thawed HAMs. The post-thawing viability of HAMs was 84.3 +/- 3.2% (Mean +/- SD, n = 10). The thawed HAMs showed morphological characteristics indistinguishable from the non-vitrified fresh HAMs. The expression of surface antigens (strong positive for CD44, CD49d, CD59, CD90, CD105 and HLA-ABC; weak positive for HLA-G; negative for CD31, CD34, CD45, CD106, CD117 and HLA-DR) and the expression of ESC markers [CK18, fibroblast growth factor-5, GATA-4, neural cell adhesion molecule, Nestin, Oct-4, stem cell factor, HLA-ABC, Vimentin, bone morphogenetic protein (BMP) 4, hepatocyte nuclear factor 4 alpha (HNF-4 alpha), Pax-6, alpha-fetoprotein, Brachyury, BMP-2, TRA-1-60, stage-specific embryonic antigen (SSEA-3, SSEA-4)] were maintained in the vitrified-thawed HAMs. The thawed HAMs retained ability to differentiate into osteoblasts, adipocytes and chondrocytes under appropriate culture conditions. Our results suggest that vitrification is a reliable and effective method for cryopreservation of HAMs.

Comparison of the clinical outcomes between fresh blastocyst and vitrified-thawed blastocyst transfer.

PubMed

Ku, Pei-Yun; Lee, Robert Kuo-Kuang; Lin, Shyr-Yeu; Lin, Ming-Huei; Hwu, Yuh-Ming

2012-12-01

To compare the clinical outcomes between fresh and vitrified-thawed day 5 blastocyst transfers. Retrospective case control study. Tertiary referral center. Patients 38 years of age or less who underwent IVF/ICSI cycles with fresh or frozen-thawed blastocysts transferred from June 1, 2009 to November 30, 2011 Vitrification and thawing of day 5 blastocysts using the Cryotop method. (Kitazato BioPharma Co., Ltd., Fuji city, Shizuoka, Japan) Clinical pregnancy rate, implantation rate, ongoing pregnancy rate, and multiple pregnancy rates. Of the 118 cycles in the fresh transfer group, 234 blastocysts were transferred. The clinical pregnancy rate was 66.1 % and implantation rate was 50.9 %. The ongoing pregnancy rate was 56.8 % and the rates for singleton and twin pregnancies were 53.7 % and 44.8 %. Of the 59 cycles in the vitrified-thawed group, 111 blastocysts were transferred. The clinical pregnancy rate was 59.3 % and implantation rate was 43.2 %. The ongoing pregnancy rate was 47.5 % and the rates for singleton and twin pregnancies were 60.7 % and 39.3 %. The clinical pregnancy rate, implantation rate and ongoing pregnancy rate did not differ significantly between the two groups. The implantation rates were not significantly different between the fresh and the vitrified-thawed groups. Thus, single embryo transfer may be considered in fresh cycles to decrease multiple pregnancy rates. The surplus embryos should be vitrified for the frozen embryo transfer to improve the cumulative pregnancy rate.

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

PubMed

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.

Stabilisation and dewatering of primary sludge using ferrate(VI) pre-treatment followed by freeze-thaw in simulated drainage beds.

PubMed

Diak, James; Örmeci, Banu

2018-06-15

This study evaluated the ability of potassium ferrate(VI) and freeze-thaw to stabilise and dewater primary sludge. Potassium ferrate(VI) additions of 0.5 and 5.0 g/L were used as a pre-treatment prior to freeze-thaw. Samples were frozen at -10, -20 and -30 °C, and were kept frozen for 1, 8 and 15 days. The samples were subsequently thawed at room temperature in a setup which allowed meltwater to be separated from the sludge cake via gravity drainage. The meltwater was characterised in terms of fecal coliform, soluble chemical oxygen demand (COD), soluble proteins, soluble carbohydrates, pH and turbidity. The sludge cake was characterised in terms of fecal coliform, total solids (TS) and volatile solids (VS). Freeze-thaw with gravity meltwater drainage reduced the sludge volume by up to 79%. After being frozen for only 1 day, the concentrations of fecal coliform in many of the primary sludge samples were reduced to <1000 MPN/g dry solids (DS), representing >3-log inactivation in some cases. However, pre-treatment of the primary sludge with ≤5.0 g/L potassium ferrate(VI) resulted in significant increases in soluble proteins, soluble carbohydrates, and sCOD, and reduced the effectiveness of stand-alone freeze-thaw. Follow-up experiments using higher doses ranging from 5.1 to 24.9 g/L of potassium ferrate(VI) demonstrated that >5-log inactivation of fecal coliform in raw primary sludge can be achieved within 15 min using 15 g/L of potassium ferrate(VI), and the resulting concentration of fecal coliform in the sludge was 1023 MPN/g DS. Pre-treatment with 22.0 g/L of potassium ferrate(VI), followed by freeze-thaw, with only 3 days frozen, reduced the concentration of fecal coliform to below the detection limit in the meltwater and the sludge cake. This demonstrates that potassium ferrate(VI) and freeze-thaw offers the flexibility to adjust the ferrate(VI) dose to meet treatment requirements for land application, and can be used as a stand-alone sludge treatment technology for primary sludge that achieves both treatment and dewatering. Copyright © 2017 Elsevier Ltd. All rights reserved.

Laboratory performance of pervious concrete subjected to deicing salts and freeze-thaw.

DOT National Transportation Integrated Search

2015-06-01

Significant research and development have occurred for pervious concrete, but its acceptance in : cold climates is still limited. Vulnerability to freeze-thaw and salt exposure has led to uncertainty : about its long-term performance. Additionally, t...

Thaw weakening and load restriction practices on low volume roads

DOT National Transportation Integrated Search

2000-06-01

Low volume roads subjected to seasonal freezing are highly susceptible to damage from traffic during midwinter and spring thaws. Such traffic-induced damage can be minimized by a variety of design methods; however, most are not economically feasible....

Comparison of two freeze-thaw apparatus.

DOT National Transportation Integrated Search

1982-01-01

The purpose of this study was to compare the results of rapid freezing and thawing tests conducted on machine A with results from machine B, which is intended to replace the aging machine A. Concrete samples were prepared to attain levels of resistan...

Assessing cryogenic testing of aggregates for concrete pavements

DOT National Transportation Integrated Search

1995-02-01

Damage to concrete pavements caused by freeze-thaw deterioration of concrete aggregate remains a serious problem. Current tests for determining an aggregate's freeze-thaw durability can take up to 70 days to perform and results from these tests don't...

Aggregate Freezing-Thawing Performance Using the Iowa Pore Index : final report.

DOT National Transportation Integrated Search

2016-10-01

In cold climates, the use of non-durable aggregate leads to premature pavement deterioration due to damage caused by freezing-thawing cycles. Differentiating durable and non-durable aggregates is a crucial yet challenging task. The frost durability o...

Rapid stabilization of thawing soils For enhanced vehicle mobility: a field demonstration project

DOT National Transportation Integrated Search

1999-02-01

Thawing soil presents a formidable challenge for vehicle operations cross-country and on unsurfaced roads. To mitigate the problem, a variety of stabilization techniques were evaluated for their suitability for rapid employment to enhance military ve...

Resilient modulus characterization of Alaskan granular base materials.

DOT National Transportation Integrated Search

2010-08-01

When spring comes to cold regions, the active layer (the top few feet of soil that freezes and thaws seasonally) thaws quickly, while : deeper soil remains frozen. The active layer becomes saturated with water from snowmelt that collects atop the fro...

Startup thaw concept for the SP-100 space reactor power system

NASA Technical Reports Server (NTRS)

Kirpich, A.; Das, A.; Choe, H.; Mcnamara, E.; Switick, D.; Bhandari, P.

1990-01-01

A thaw concept for a space reactor power system which employs lithium as a circulant for both the heat-transport and the heat-rejection fluid loops is presented. An exemplary thermal analysis for a 100-kWe (i.e., SP-100) system is performed. It is shown that the design of the thaw system requires a thorough knowledge of the various physical states of the circulant throughout the system, both spatially and temporally, and that the design has to provide adequate margins for the system to avoid a structural or thermally induced damage.

Multi-Scale Process of Soil Freezing, Thawing, and Thaw-Settlement

DTIC Science & Technology

2013-02-18

constitutive model was implemented into the finite element system ABAQUS using subroutine UMAT and UEXPAN to solve boundary value problems. The model...soils, 16 Int. Conf. Soil Mech. Geotech. Eng. (12 2012) 02/15/2013 13.00 Yao Zhang, Radoslaw L. Michalowski. Thermal -mechanical constitutive modeling...freezing and thawing soils…… 19 5.1 The model …………………………………………………….. 19 5.2 Thermal -mechanical load process ……………………………. 23 5.3 Implementation

Factors in Design and Construction of a Device for Heating and Dispensing Food Components

DTIC Science & Technology

1974-09-01

thawing meth ods as ambient t emperature thawing 0 thawing at refrigerated temperatures 0 thawi ng i n running tap wa ter 0 a nd d i rect cooki ng...Nutri t iona l quali ty i s als o affected by long standing period s due t o ox i datio n and l eaching o f nutri ents in watero Cooki ng is a maj or...indicating that nutri ent ret e n t i on was approximately the same for electroni c cooki ng and good conventi onal cookingo Boiling,brai singo and

A Peltier-based freeze-thaw device for meteorite disaggregation

NASA Astrophysics Data System (ADS)

Ogliore, R. C.

2018-02-01

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

Liposome encapsulated soy lecithin and cholesterol can efficiently replace chicken egg yolk in human semen cryopreservation medium.

PubMed

Mutalik, Srinivas; Salian, Sujith Raj; Avadhani, Kiran; Menon, Jyothsna; Joshi, Haritima; Hegde, Aswathi Raju; Kumar, Pratap; Kalthur, Guruprasad; Adiga, Satish Kumar

2014-06-01

Cryopreservation of spermatozoa plays a significant role in reproductive medicine and fertility preservation. Chicken egg yolk is used as an extender in cryopreservation of human spermatozoa using glycerol egg yolk citrate (GEYC) buffered medium. Even though 50% survival of spermatozoa is generally achieved with this method, the risk of high levels of endotoxins and transmission pathogens from chicken egg yolk is a matter of concern. In the present study we attempted to establish a chemically defined cryopreservation medium which can replace the chicken egg yolk without affecting sperm survival. Ejaculates from 28 men were cryopreserved with GEYC based freezing medium or liposome encapsulated soy lecithin-cholesterol based freezing medium (LFM). The semen samples were subjected to rapid thawing after 14 days of storage in liquid nitrogen. Post-thaw analysis indicated significantly higher post-thaw motility and sperm survival in spermatozoa cryopreserved with LFM compared to conventional GEYC freezing medium. The soy lecithin and cholesterol at the ratio of 80:20 with sucrose showed the highest percentage of post-thaw motility and survival compared to the other compositions. In conclusion, chemically defined cryopreservation medium with liposome encapsulated soy lecithin and cholesterol can effectively replace the chicken egg yolk from human semen cryopreservation medium without compromising post-thaw outcome.

Effects of permafrost thaw on carbon emissions under aerobic and anaerobic environments in the Great Hing'an Mountains, China.

PubMed

Song, Changchun; Wang, Xianwei; Miao, Yuqing; Wang, Jiaoyue; Mao, Rong; Song, Yanyu

2014-07-15

The carbon (C) pool of permafrost peatland is very important for the global C cycle. Little is known about how permafrost thaw could influence C emissions in the Great Hing'an Mountains of China. Through aerobic and anaerobic incubation experiments, we studied the effects of permafrost thaw on CH4 and CO2 emissions. The rates of CH4 and CO2 emissions were measured at -10, 0 and 10°C. Although there were still C emissions below 0°C, rates of CH4 and CO2 emissions significantly increased with permafrost thaw under aerobic and anaerobic conditions. The C release under aerobic conditions was greater than under anaerobic conditions, suggesting that permafrost thaw and resulting soil environment change should be important influences on C emissions. However, CH4 stored in permafrost soils could affect accurate estimation of CH4 emissions from microbial degradation. Calculated Q10 values in the permafrost soils were significantly higher than values in active-layer soils under aerobic conditions. Our results highlight that permafrost soils have greater potential decomposability than soils of the active layer, and such carbon decomposition would be more responsive to the aerobic environment. © 2013 Elsevier B.V. All rights reserved.

Vitrification of mouse embryos using the thin plastic strip method

PubMed Central

Hur, Yong Soo; Ann, Ji Young; Maeng, Ja Young; Park, Miji; Park, Jeong Hyun; Yoon, Jung; Yoon, San Hyun; Hur, Chang Young; Lee, Won Don; Lim, Jin Ho

2012-01-01

Objective The aim of this study was to compare vitrification optimization of mouse embryos using electron microscopy (EM) grid, cryotop, and thin plastic strip (TPS) containers by evaluating developmental competence and apoptosis rates. Methods Mouse embryos were obtained from superovulated mice. Mouse cleavage-stage, expanded, hatching-stage, and hatched-stage embryos were cryopreserved in EM grid, cryotop, and TPS containers by vitrification in 15% ethylene glycol, 15% dimethylsulfoxide, 10 µg/mL Ficoll, and 0.65 M sucrose, and 20% serum substitute supplement (SSS) with basal medium, respectively. For the three groups in which the embryos were thawed in the EM grid, cryotop, and TPS containers, the thawing solution consisted of 0.25 M sucrose, 0.125 M sucrose, and 20% SSS with basal medium, respectively. Rates of survival, re-expansion, reaching the hatched stage, and apoptosis after thawing were compared among the three groups. Results Developmental competence after thawing of vitrified expanded and hatching-stage blastocysts using cryotop and TPS methods were significantly higher than survival using the EM grid (p<0.05). Also, apoptosis positive nuclei rates after thawing of vitrified expanded blastocysts using cryotop and TPS were significantly lower than when using the EM grid (p<0.05). Conclusion The TPS vitrification method has the advantages of achieving a high developmental ability and effective preservation. PMID:23346525

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

NASA Astrophysics Data System (ADS)

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.

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

PubMed

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.

Effect of frozen storage, different thawing methods and cooking processes on the survival of Salmonella spp. and Escherichia coli O157:H7 in commercially shaped beef patties.

PubMed

Manios, Stavros G; Skandamis, Panagiotis N

2015-03-01

The effect of common handling practices (i.e., freezing, thawing and cooking) of beef patties on the survival of Salmonella spp. and Escherichia coli O157:H7, was evaluated. Inoculated ground beef was stored at -22 °C for 5 and 75 days. After thawing at 4 °C/16 h, 20 °C/12 h, in microwave/22-24 min, or without prior thawing, beef patties (90 g) were shaped and cooked in oven-broiler or in pan-grill to internal temperatures of 60 °C or 71 °C. Cooking in oven-broiler was more effective compared to pan-grill, especially when cooked to 71 °C. Defrosting methods did not affect significantly (P ≥ 0.05) the survival of the pathogens during subsequent cooking. Frozen storage for 75 days enhanced the survival of E. coli O157:H7, as the pathogen survived 3.1 logCFU/g when cooked in oven-broiler at 71 °C. Results may supplement the existing guidelines for the appropriate practices, associated with freezing, thawing and cooking of patties in households or catering services. Copyright © 2014 Elsevier Ltd. All rights reserved.

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

PubMed

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.

Growth of high-elevation Cryptococcus sp. during extreme freeze-thaw cycles.

PubMed

Vimercati, L; Hamsher, S; Schubert, Z; Schmidt, S K

2016-09-01

Soils above 6000 m.a.s.l. are among the most extreme environments on Earth, especially on high, dry volcanoes where soil temperatures cycle between -10 and 30 °C on a typical summer day. Previous studies have shown that such sites are dominated by yeast in the cryophilic Cryptococcus group, but it is unclear if they can actually grow (or are just surviving) under extreme freeze-thaw conditions. We carried out a series of experiments to determine if Cryptococcus could grow during freeze-thaw cycles similar to those measured under field conditions. We found that Cryptococcus phylotypes increased in relative abundance in soils subjected to 48 days of freeze-thaw cycles, becoming the dominant organisms in the soil. In addition, pure cultures of Cryptococcus isolated from these same soils were able to grow in liquid cultures subjected to daily freeze-thaw cycles, despite the fact that the culture medium froze solid every night. Furthermore, we showed that this organism is metabolically versatile and phylogenetically almost identical to strains from Antarctic Dry Valley soils. Taken together these results indicate that this organism has unique metabolic and temperature adaptations that make it able to thrive in one of the harshest and climatically volatile places on Earth.

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

NASA Astrophysics Data System (ADS)

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

2016-06-01

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

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

PubMed Central

Pierini, Antonio; Baker, Jeanette; Armstrong, Randall; Pan, Yuqiong; Leveson-Gower, Dennis; Negrin, Robert; Meyer, Everett

2015-01-01

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. PMID:26693907

Retention of membrane charge attributes by cryopreserved-thawed sperm and zeta selection.

PubMed

Kam, Tricia L; Jacobson, John D; Patton, William C; Corselli, Johannah U; Chan, Philip J

2007-09-01

Mature sperm can be selected based on their negative zeta electrokinetic potential. The zeta selection of cryopreserved sperm is unknown. The objective was to study the effect of zeta processing on the morphology and kinematic parameters of cryopreserved-thawed sperm. Colloid-washed sperm (N = 9 cases) were cryopreserved for 24 h, thawed and diluted in serum-free medium in positive-charged tubes. After centrifugation, the tubes were decanted, serum-supplemented medium was added and the resuspended sperm were analyzed. Untreated sperm and fresh sperm served as the controls. There were improvements in strict normal morphology in fresh (11.8 +/- 0.3 versus control 8.8 +/- 0.3 %, mean +/- SEM) and thawed (8.7 +/- 0.2 versus control 5.4 +/- 0.2%) sperm after zeta processing. Percent sperm necrosis was reduced after zeta processing (66.0 +/- 0.6 versus unprocessed 74.6 +/- 0.3%). Progression decreased by 50% but not total motility after zeta processing of thawed sperm. The results suggested that the cryopreservation process did not impact the sperm membrane net zeta potential and higher percentages of sperm with normal strict morphology, acrosome integrity and reduced necrosis were recovered. The zeta method was simple and improved the selection of quality sperm after cryopreservation but more studies would be needed before routine clinical application.

Binder of Sperm Proteins protect ram spermatozoa from freeze-thaw damage.

PubMed

Pini, Taylor; Farmer, Kiri; Druart, Xavier; Teixeira-Gomes, Ana Paula; Tsikis, Guillaume; Labas, Valerie; Leahy, Tamara; de Graaf, Simon P

2018-06-01

Cryopreservation causes sub-lethal damage which limits the fertility of frozen thawed spermatozoa. Seminal plasma has been investigated as a cryoprotectant, but has yielded inconsistent results due to considerable variation in its constituents. Individual seminal plasma proteins offer an ideal alternative to whole seminal plasma, and several have been correlated with freezing success. Binder of Sperm Proteins (BSPs) are abundant ram seminal plasma proteins which have been suggested to have significant protective effects on ram spermatozoa during cold shock. This is in direct opposition to bull spermatozoa, where BSPs cause sperm deterioration during in vitro handling. We investigated the potential of BSP1 and BSP5 to prevent freezing associated damage to important functional parameters of ram spermatozoa. BSPs purified by size exclusion chromatography improved post thaw motility and penetration through artificial mucus. Highly purified BSP1 and BSP5, isolated by gelatin affinity and RP-HPLC, improved motility and membrane integrity, and reduced post thaw protein tyrosine phosphorylation. Exposure to BSP5 before freezing increased the amount of phosphatidylethanolamine on the sperm surface after thawing. Neither BSP1 nor BSP5 prevented freezing associated changes in membrane lipid disorder. These results suggest that BSPs may significantly improve freezing outcomes of ram spermatozoa. Copyright © 2018 Elsevier Inc. All rights reserved.

Freeze-thaw resistance of concrete with marginal air content

DOT National Transportation Integrated Search

2007-05-01

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

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

DOT National Transportation Integrated Search

2006-12-01

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

Concrete pavement quality control testing requirements needed for the super air meter : final report.

DOT National Transportation Integrated Search

2016-10-01

Concrete freeze-thaw durability is prominently linked to the air void system within the concrete. : Concrete pavements in Kansas undergo repetitive freeze-thaw cycles. Total air content measurements : currently used on fresh concrete do not provide a...

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

DOT National Transportation Integrated Search

1995-12-01

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

NASA Satellite Images Annual Spring Thaw, Red River, North Dakota

NASA Image and Video Library

2011-04-21

NASA Terra spacecraft shows the annual spring thaw in the upper Midwest is underway. Snow-covered ground contrasts with the dark tones of water under broken cloud cover. Along the Red River in North Dakota, floodwaters are moving northward into Canada.

Impact of freeze-thaw on liquefaction potential and dynamic properties of Mabel Creek silt.

DOT National Transportation Integrated Search

2009-02-01

"This study examines the influence of temperature rise and freeze-thaw cycles on the soil liquefaction potential. More specifically, dynamic properties and post-cyclicloading : settlement of fine-grained soils are evaluated in this study. The results...

Concrete pavement quality control testing requirements needed for the super air meter : technical summary.

DOT National Transportation Integrated Search

2016-10-01

Concrete freeze-thaw durability is prominently linked to the air void system : within the concrete. Concrete pavements in Kansas undergo repetitive : freeze-thaw cycles. Total air content measurements currently used on fresh : concrete do not provide...

Chicks produced in the Magellanic penguin (Spheniscus magellanicus) after cloacal insemination of frozen-thawed semen.

PubMed

O'Brien, Justine Kellie; Steinman, Karen J; Montano, Gisele A; Dubach, Jean M; Robeck, Todd R

2016-07-01

The in vitro and in vivo functionality of cryopreserved spermatozoa was examined over two breeding seasons in a zoological colony of Magellanic penguins (Spheniscus magellanicus). Frozen-thawed semen was inseminated into five anesthetized females, over a total of eight egg production cycles, with a different male used for each artificial insemination (AI) within each season. Females were maintained within the colony in cordoned nest sites to prevent copulation with their paired male, and were inseminated every 3-10 days until the first oviposition. Semen frozen from seven males using a straw method retained 39.8%, 25.7%, 74.0%, and 52.1% of its initial total motility, progressive motility, average path velocity, and plasma membrane integrity, respectively. Normal morphology of motile cells was reduced (P < 0.05) during freeze-thawing from 76.7% immediately prior to freezing to 65.3% post-thawing. Conceptive females received 1.6 ± 0.2 inseminations before the first oviposition, with 19.2 ± 1.6 × 10(6) motile, morphologically normal spermatozoa per insemination. Overall fertility was 53.3% (8/15 eggs), hatchability was 50.0% (4/8), and genetic analyses confirmed that all embryos and hatchlings were sired by the AI male. Fertile eggs were laid at 4.0-12.1 days after AI, indicating that frozen-thawed spermatozoa resided in the female reproductive tract for up to ∼7.2 days prior to fertilization. Results demonstrate that frozen-thawed Magellanic penguin spermatozoa are fully functional in vivo and support the use of genome banking and AI as tools for managing the sustainability of zoological penguin populations. Zoo Biol. 35:326-338, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

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

PubMed

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) (P<0.05). After recryopreservation, sorted spermatozoa frozen with the directional freezing method maintained higher (P<0.05) motility parameters over a 24-h incubation period compared to spermatozoa frozen using straws. Quality of sperm DNA of non-sorted spermatozoa, as assessed by the sperm chromatin structure assay (SCSA), was high and remained unchanged throughout freeze-thawing and incubation processes. Though a possible interaction between Hoechst 33342 and the SCSA-derived acridine orange was observed in stained and sorted samples, the proportion of sex-sorted, recryopreserved spermatozoa exhibiting denatured DNA was low (6.6±4.1%) at 6 h after the second thawing step and remained unchanged (P>0.05) at 24 h. The viability of sorted spermatozoa was higher (P<0.05) than that of non-sorted spermatozoa across all time points after recryopreservation. Collective results indicate that bottlenose dolphin spermatozoa undergoing cryopreservation, sorting and recryopreservation are of adequate quality for use in AI.

Supplementation of soybean lecithin-based semen extender by antioxidants: complementary flowcytometric study on post-thawed ram spermatozoa.

PubMed

Sharafi, Mohsen; Zhandi, Mahdi; Akbari Sharif, Abbas

2015-06-01

The purpose of the current study was to evaluate the effects of cysteine (C) and glutathione (G) on the post-thawed ram sperm quality. Collected semen samples from four mature rams were diluted with five soybean lecithin (SL)-based extenders containing: no antioxidant (SL-0), 5 mM cysteine (SL-C5), 10 mM cysteine (SL-C10), 5 mM glutathione (SL-G5) and 10 mM glutathione (SL-G10). After freeze-thawing process, motion and velocity parameters, plasma membrane integrity and functionality, morphological abnormality, lipid peroxidation, acrosomal status, mitochondria activity, and apoptosis status of post-thawed ram spermatozoa were assessed. The results showed that SL-C10 increased the total motility and plasma membrane integrity (p < 0.05) of post-thawed ram spermatozoa (55.86 ± 1.37 and 60.57 ± 1.34 %) compared to other extenders. Progressive motility was significantly higher in SL-C10 (24.71 ± 1.13 %) compared to SL-0 (20 ± 1.13 %) and SL-G10 (15 ± 1.13 %). Mitochondrial activity was significantly higher in SL-C10 (56.83 ± 2.29 %) compared to SL-G10 (38.75 ± 2.29 %). Capacitation and acrosomal status, lipid peroxidation, and the percentage of dead spermatozoa were not affected by different extenders. The percentage of live spermatozoa was higher in SL-C10 (56.33 ± 1.35 %) compared to other extenders. Also, SL-C10 resulted in a lower percentage of apoptotic spermatozoa (14.17 ± 0.53 %) compared to other extenders. The results of this study showed that supplementation of SL-based ram semen extender with 10 mM cysteine resulted in an improved quality of post-thawed ram spermatozoa.

Different patterns of metabolic cryo-damage in domestic cat (Felis catus) and cheetah (Acinonyx jubatus) spermatozoa.

PubMed

Terrell, Kimberly A; Wildt, David E; Anthony, Nicola M; Bavister, Barry D; Leibo, S P; Penfold, Linda M; Marker, Laurie L; Crosier, Adrienne E

2012-04-01

Felid spermatozoa are sensitive to cryopreservation-induced damage, but functional losses can be mitigated by post-thaw swim-up or density gradient processing methods that selectively recover motile or structurally-normal spermatozoa, respectively. Despite the importance of sperm energy production to achieving fertilization, there is little knowledge about the influence of cryopreservation or post-thaw processing on felid sperm metabolism. We conducted a comparative study of domestic cat and cheetah sperm metabolism after cryopreservation and post-thaw processing. We hypothesized that freezing/thawing impairs sperm metabolism and that swim-up, but not density gradient centrifugation, recovers metabolically-normal spermatozoa. Ejaculates were cryopreserved, thawed, and processed by swim-up, Accudenz gradient centrifugation, or conventional washing (representing the 'control'). Sperm glucose and pyruvate uptake, lactate production, motility, and acrosomal integrity were assessed. Mitochondrial membrane potential (MMP) was measured in cat spermatozoa. In both species, lactate production, motility, and acrosomal integrity were reduced in post-thaw, washed samples compared to freshly-collected ejaculates. Glucose uptake was minimal pre- and post-cryopreservation, whereas pyruvate uptake was similar between treatments due to high coefficients of variation. In the cat, swim-up, but not Accudenz processing, recovered spermatozoa with increased lactate production, pyruvate uptake, and motility compared to controls. Although confounded by differences in non-specific fluorescence among processing methods, MMP values within treatments were positively correlated to sperm motility and acrosomal integrity. Cheetah spermatozoa isolated by either selection method exhibited improved motility and/or acrosomal integrity, but remained metabolically compromised. Collectively, findings revealed a metabolically-robust subpopulation of cryopreserved cat, but not cheetah, spermatozoa, recovered by selecting for motility rather than morphology. Published by Elsevier Inc.

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

PubMed

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

2011-02-01

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shier, W.T.

Normally a freeze-thaw cycle is a very efficient method of killing mammalian cells. However, this report describes conditions that prevent killing of cultured mammalian cells by nucleated freezing at -24 degrees C. Optimal protection from cell killing at -24 degrees C was obtained in isotonic solutions containing an organic cryoprotectant such as dimethyl sulfoxide (DMSO; 10%, v/v), a saccharide such as sucrose over a broad concentration range from 50 to 150 mM, and glucose. Glycerol was also an effective cryoprotectant but other organic solvents were ineffective, although in some cases they appeared to protect cell membranes, while not protecting othermore » vital components. A wide variety of saccharide structures were effective at protecting cells from freeze-thaw killing, with trehalose being particularly effective. The degree of resistance to killing by a freeze-thaw cycle under these conditions varied widely among different cell lines. If toxicity of DMSO was responsible for this variability of cryoprotection, it must have been due to short-term, not longer term, toxicity of DMSO. Studies on the mechanism by which cells are protected from killing under these conditions indicated that neither vitrification of the medium nor the concentrating of components during freezing were involved. One model not eliminated by the mechanistic studies proposes that the organic solvent cryoprotectant component acts by fluidizing membranes under the thawing conditions, so that any holes produced by ice crystals propagating through membranes can reseal during the thawing process. In this model one of the mechanisms by which the saccharide component could act is by entering the cells and stabilizing vital intracellular components. Consistent with this, a freeze-thaw cycle promoted the uptake of labeled sucrose into cultured cells.« less

Contrasting approaches to determine the impact of permafrost thaw on C cycling in northern peatlands

NASA Astrophysics Data System (ADS)

Heffernan, L.; Estop Aragones, C.; Blodau, C.; Olefeldt, D.

2016-12-01

Northern peatlands are a globally significant pool of terrestrial carbon (C) and have acted as a major C sink since the last deglaciation. The peatland complexes found in western Canada are a mosaic of peat plateaus underlain by permafrost interspersed with permafrost free thermokarst bogs. Recent climate change has led to increased rates of thermokarst bog expansion due to permafrost thaw. In order to understand whether permafrost thaw causes net C loss or uptake, we studied both the variation in C stocks and of surface greenhouse gas fluxes along a pair of thermokarst chronosequences located in thick (>6 m) peatlands in the discontinuous permafrost zone of western Canada. Each zone of a chronosequence is characterized by permafrost conditions, including a zone where permafrost is still present (peat plateau), a zone of recent thermokarst development (dominated by Sphagnum riparium and sedges), and a mature thermokarst zone (dominated by Sphagnum fuscum and woody shrubs) where permafrost thaw occurred >50 years ago. Each zone has distinct hydrological conditions, with the recent thermokarst being much wetter. Increased wetness causes increased heat conduction and average July to November soil temperatures at 50, 100, and 200 cm were 1.6, 2.2, and 1.7 °C warmer, respectively, in the recent than in the mature thermokarst zone. This difference likely increases respiration rates at depth substantially. Our study aims to reconcile findings from other sites where investigations of C stock and greenhouse gas emissions have yielded disparate conclusions. By combining C stocks and C emissions along a thaw chronosequence, and highlighting the differences in C cycling between recently and older thawed thermokarst features, this current work aims to reconcile these contrasting views.

Evaluation of cryopreserved stallion semen from Tori and Estonian breeds using CASA and flow cytometry.

PubMed

Kavak, A; Johannisson, A; Lundeheim, N; Rodriguez-Martinez, H; Aidnik, M; Einarsson, S

2003-04-15

Methods to evaluate the quality of frozen-thawed stallion semen are still needed, particularly those considering the sperm function. The present study evaluated sperm motility, membrane and acrosome integrity and the capacitation status of frozen-thawed spermatozoa from seven Tori and six Estonian breed stallions by way of computer assisted sperm analysis (CASA), a triple fluorophore stain combination and Merocyanine 540, respectively, the latter ones using flow cytometry. Two ejaculates from each stallion were cryopreserved using the Hannover method in 0.5 ml plastic straws. Two straws per ejaculate per stallion were thawed at 37 degrees C for 30s. Motility was analysed with CASA immediately after thawing, while for flow cytometry spermatozoa were cleansed by 70:40% Percoll discontinuous density gradient separation before analysed for sperm viability, acrosome integrity (stained with SNARF, PI and FITC-PSA) and capacitation status (stained with Merocyanine 540/Yo-Pro-1). Results (as least square means) were as follows: the motility of frozen-thawed semen was 43.4% for Tori stallions and 42.3% for Estonian stallions (P>0.05). After Percoll separation 79.3% of the spermatozoa from Tori stallions had intact acrosomes and 1.7% of them showed early signs of capacitation. The same parameters for Estonian stallions were 84.5 and 2.3%, respectively. There were no statistically significant differences between breeds or ejaculates within breed for any evaluated parameter. We conclude that triple staining and flow cytometry are valuable techniques to evaluate frozen-thawed stallion spermatozoa, and that no differences in quality of frozen semen were registered between Tori and Estonian breed stallions, allowing implementation of this technology in the Estonian horse population.

Spatial considerations during cryopreservation of a large volume sample.

PubMed

Kilbride, Peter; Lamb, Stephen; Milne, Stuart; Gibbons, Stephanie; Erro, Eloy; Bundy, James; Selden, Clare; Fuller, Barry; Morris, John

2016-08-01

There have been relatively few studies on the implications of the physical conditions experienced by cells during large volume (litres) cryopreservation - most studies have focused on the problem of cryopreservation of smaller volumes, typically up to 2 ml. This study explores the effects of ice growth by progressive solidification, generally seen during larger scale cryopreservation, on encapsulated liver hepatocyte spheroids, and it develops a method to reliably sample different regions across the frozen cores of samples experiencing progressive solidification. These issues are examined in the context of a Bioartificial Liver Device which requires cryopreservation of a 2 L volume in a strict cylindrical geometry for optimal clinical delivery. Progressive solidification cannot be avoided in this arrangement. In such a system optimal cryoprotectant concentrations and cooling rates are known. However, applying these parameters to a large volume is challenging due to the thermal mass and subsequent thermal lag. The specific impact of this to the cryopreservation outcome is required. Under conditions of progressive solidification, the spatial location of Encapsulated Liver Spheroids had a strong impact on post-thaw recovery. Cells in areas first and last to solidify demonstrated significantly impaired post-thaw function, whereas areas solidifying through the majority of the process exhibited higher post-thaw outcome. It was also found that samples where the ice thawed more rapidly had greater post-thaw viability 24 h post-thaw (75.7 ± 3.9% and 62.0 ± 7.2% respectively). These findings have implications for the cryopreservation of large volumes with a rigid shape and for the cryopreservation of a Bioartificial Liver Device. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Effect of rainbow trout (Oncorhynchus mykiss) seminal plasma on the post-thaw quality of ram semen cryopreserved in a soybean lecithin-based or egg yolk-based extender.

PubMed

Ustuner, Burcu; Alcay, Selim; Toker, M Berk; Nur, Zekariya; Gokce, Elif; Sonat, Fusun Ak; Gul, Zulfiye; Duman, Muhammed; Ceniz, Cafer; Uslu, Aydın; Sagirkaya, Hakan; Soylu, M Kemal

2016-01-01

The aim of the current study was to evaluate the effects of different concentrations of rainbow trout seminal plasma (RTSP) (0.1%, 1% and 10%) in extenders containing either egg yolk or lecithin for use in Awassi ram semen cryopreservation. Pooled sperm were diluted in a two-step dilution method to a final concentration of 1/5 (semen/extender) in egg yolk or lecithin extender containing no RTSP, 0.1%, 1% or 10% RTSP (v/v). Semen samples were assessed for sperm motility, plasma membrane integrity [hypoosmotic swelling test (HOST) and Hoechst 33258] and defective acrosomes [FITC-conjugated Pisum sativum agglutinin (PSA-FITC)] at the following five time points: after dilution with extender A; after equilibration; and post-thaw at 0h, 3h and 5h. Malondialdehyde (MDA) was examined only after thawing. Freezing and thawing procedures (dilution, equilibration and post-thaw incubation at 0h, 3h and 5h) negatively affected the motility (P<0.001) and acrosome integrity (P<0.001). Additionally, freezing and thawing negatively affected the plasma membrane integrity, as determined by the HOST and Hoechst 33258 (P<0.001). The extender group affected the motility (P<0.001) and the HOST results (P<0.001). Levels of MDA in the egg yolk extender with 1% RTSP group were significantly lower than in the lecithin control group (P<0.05). In conclusion, the egg yolk extender groups that were supplemented with 10% and 1% RTSP provided greater cryoprotective effects for semen survivability during 5h incubation than the other extender groups. Copyright © 2015 Elsevier B.V. All rights reserved.

In vitro and in vivo survival of bisected sheep embryos derived from frozen-thawed unsorted, and frozen-thawed sex-sorted and refrozen-thawed ram spermatozoa.

PubMed

Morton, Katherine M; Rowe, Anthony M; Chis Maxwell, W M; Evans, Gareth

2006-04-15

Ovine IVP embryos were derived from frozen-thawed unsorted and frozen-thawed sex-sorted spermatozoa that had been refrozen and thawed. The embryos were bisected and cultured in vitro, or transferred to recipient ewes to determine their survival in vitro and in vivo. Oocyte progression to the blastocyst stage was similar for unsorted (97/232, 41.8%) and sex-sorted spermatozoa (113/286, 39.5%; P > 0.05). Embryo survival in vitro post-bisection was similar for demi-embryos derived from unsorted and sex-sorted sperm, and embryos bisected at the blastocyst and expanded blastocyst stage (P > 0.05). A higher proportion of recipient ewes were pregnant at Day 63 after transfer of two intact embryos derived from unsorted (17/21, 80.9%) than two demi-embryos derived from unsorted (5/15, 33.3%) or sex-sorted spermatozoa (7/17, 41.2%). The number of fetuses per original embryo at Day 63 did not differ among groups (unsorted intact: 23/42, 54.8%; unsorted demi: 7/15, 46.7%; sex-sorted demi: 10/17, 58.8%) and twin pregnancies were observed in all groups. Embryo survival to term was high, and was not significantly different among intact (unsorted: 22/42, 52.4%) and demi-embryos (unsorted: 4/15, 26.7%; sex-sorted spermatozoa: 7/17, 41.2%; P > 0.05). Dizygotic twins (n = 6 sets) were born after the transfer of two intact embryos derived from unsorted spermatozoa, but only singleton lambs resulted from the transfer of demi-embryos. In conclusion, bisected IVP embryos successfully developed into morphologically normal lambs. However, embryo survival to term was neither increased nor decreased by embryo bisection.

Calcium interacts with antifreeze proteins and chitinase from cold-acclimated winter rye.

PubMed

Stressmann, Maja; Kitao, Satoshi; Griffith, Marilyn; Moresoli, Christine; Bravo, León A; Marangoni, Alejandro G

2004-05-01

During cold acclimation, winter rye (Secale cereale) plants accumulate pathogenesis-related proteins that are also antifreeze proteins (AFPs) because they adsorb onto ice and inhibit its growth. Although they promote winter survival in planta, these dual-function AFPs proteins lose activity when stored at subzero temperatures in vitro, so we examined their stability in solutions containing CaCl2, MgCl2, or NaCl. Antifreeze activity was unaffected by salts before freezing, but decreased after freezing and thawing in CaCl2 and was recovered by adding a chelator. Ca2+ enhanced chitinase activity 3- to 5-fold in unfrozen samples, although hydrolytic activity also decreased after freezing and thawing in CaCl2. Native PAGE, circular dichroism, and Trp fluorescence experiments showed that the AFPs partially unfold after freezing and thawing, but they fold more compactly or aggregate in CaCl2. Ruthenium red, which binds to Ca(2+)-binding sites, readily stained AFPs in the absence of Ca2+, but less stain was visible after freezing and thawing AFPs in CaCl2. We conclude that the structure of AFPs changes during freezing and thawing, creating new Ca(2+)-binding sites. Once Ca2+ binds to those sites, antifreeze activity, chitinase activity and ruthenium red binding are all inhibited. Because free Ca2+ concentrations are typically low in the apoplast, antifreeze activity is probably stable to freezing and thawing in planta. Ca2+ may regulate chitinase activity if concentrations are increased locally by release from pectin or interaction with Ca(2+)-binding proteins. Furthermore, antifreeze activity can be easily maintained in vitro by including a chelator during frozen storage.

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

NASA Astrophysics Data System (ADS)

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.

Semen molecular and cellular features: these parameters can reliably predict subsequent ART outcome in a goat model

PubMed Central

Berlinguer, Fiammetta; Madeddu, Manuela; Pasciu, Valeria; Succu, Sara; Spezzigu, Antonio; Satta, Valentina; Mereu, Paolo; Leoni, Giovanni G; Naitana, Salvatore

2009-01-01

Currently, the assessment of sperm function in a raw or processed semen sample is not able to reliably predict sperm ability to withstand freezing and thawing procedures and in vivo fertility and/or assisted reproductive biotechnologies (ART) outcome. The aim of the present study was to investigate which parameters among a battery of analyses could predict subsequent spermatozoa in vitro fertilization ability and hence blastocyst output in a goat model. Ejaculates were obtained by artificial vagina from 3 adult goats (Capra hircus) aged 2 years (A, B and C). In order to assess the predictive value of viability, computer assisted sperm analyzer (CASA) motility parameters and ATP intracellular concentration before and after thawing and of DNA integrity after thawing on subsequent embryo output after an in vitro fertility test, a logistic regression analysis was used. Individual differences in semen parameters were evident for semen viability after thawing and DNA integrity. Results of IVF test showed that spermatozoa collected from A and B lead to higher cleavage rates (0 < 0.01) and blastocysts output (p < 0.05) compared with C. Logistic regression analysis model explained a deviance of 72% (p < 0.0001), directly related with the mean percentage of rapid spermatozoa in fresh semen (p < 0.01), semen viability after thawing (p < 0.01), and with two of the three comet parameters considered, i.e tail DNA percentage and comet length (p < 0.0001). DNA integrity alone had a high predictive value on IVF outcome with frozen/thawed semen (deviance explained: 57%). The model proposed here represents one of the many possible ways to explain differences found in embryo output following IVF with different semen donors and may represent a useful tool to select the most suitable donors for semen cryopreservation. PMID:19900288

Effect of post-thaw incubation on sperm kinematics and acrosomal integrity of ram spermatozoa cryopreserved in medium-sized French straws.

PubMed

Bag, Sadhan; Joshi, Anil; Naqvi, S M K; Mittal, J P

2004-08-01

The objectives were to assess the effect of post-thaw in vitro incubation on motion characteristics and acrosomal integrity of ram spermatozoa of native Malpura and Bharat Merino breeds maintained under a semi-arid tropical environment. Good quality semen samples of both breeds were diluted, packaged in medium-sized straws, and frozen under controlled conditions. Straws were thawed at 60 degrees C for 10s and thawed samples were incubated at 37 degrees C for 4h. Post-thaw motion characteristics and acrosomal integrity of incubated spermatozoa were assessed (by computer-aided semen analysis and Giemsa staining, respectively) just prior to incubation and at hourly intervals thereafter. There was a significant effect of incubation time on motility characteristics and the proportion of spermatozoa with normal acrosomes; 81.4% (arcsin transformed value, 65.2) of spermatozoa were motile at the start of incubation, with 47.9% (arcsin transformed value, 44.4) motile after 4h. At the corresponding times, there were normal acrosomes in 65.8 (arcsin transformed value, 54.8) and 55.7% (arcsin transformed value, 48.9) of spermatozoa, respectively. The percentage straightness of spermatozoa varied during incubation (P < 0.01). However, there was no significant change in percentage linearity, curvilinear velocity, average path velocity, straight line velocity, lateral head displacement, and beat cross frequency of spermatozoa during incubation. There were no breed variations in any motility parameters during incubation, except percentage straightness (P < 0.05), lateral head displacement (P < 0.05) and beat cross frequency (P < 0.01). That sperm motility and acrosomal morphology were very acceptable immediately post-thaw and after 4h of incubation indicated the efficacy of cryopreserving ram spermatozoa under controlled conditions in medium-sized straws.

The effects of different levels of vitamin E and vitamin C in modified Beltsville extender on rooster post-thawed sperm quality.

PubMed

Amini, Mahmood Reza; Kohram, Hamid; Zare Shahaneh, Ahmad; Zhandi, Mahdi; Sharideh, Hossein; Nabi, Mohammad Mehdi

2015-12-01

Oxidative damage of sperm by means of reactive oxygen species generated by the cellular components of semen is one of the main reason of declined motility and fertility of sperm during the freeze-thawing process. This study was conducted to determine the influence of vitamin C and vitamin E on rooster post-thawed sperm motility, viability and malondialdehyde (MDA) level. Semen samples from 10 sexually-mature Ross 308 breeder roosters were collected and pooled, divided into nine equal parts and diluted with modified Beltsville extender containing with no antioxidants (control), or containing 100 (C100), 200 (C200), 400 (C400), 800 (C800) µg/mL vitamin C, and 2 (E2), 5 (E5), 10 (E10) and 15 (E15) µg/mL vitamin E. After thawing, total and progressive sperm motility, sperm viability and semen MDA level were assessed. The results shown that C200 and E5 extenders resulted in higher total motility (p < 0.05) compared to other extenders, with exception of E10 extender. Progressive motility was higher in E5 extender (p < 0.05) compared to other extenders, with exception of C200 and E10 extenders. Also, C200 and E5 extenders resulted in higher viability of post-thawed spermatozoa (p < 0.05) compared to other extenders. Finally, the results showed that MDA level was lower in C100 and C200 extenders compared to other extenders (p < 0.05), with exception of E5 extender. In conclusion, the results of the present study demonstrate that C200 and E5 can improve the function of post-thawed rooster spermatozoa.

Evaluation of NGAL TestTM on Cobas 6000.

PubMed

Hansen, Young B L; Damgaard, Anette; Poulsen, Jørgen H

2014-01-01

Neutrophil Gelatinase-Associated Lipocalin (NGAL) is a promising biomarker for acute kidney injury (AKI). Our objectives were to evaluate the NGAL Test(TM) from Bioporto for both urine NGAL and plasma NGAL on the Cobas 6000 c501 (Roche Diagnostics, Rotkreuz, Switzerland) with matched measurements run on Hitachi 917, the method's linearity on the Cobas 6000 in urine, EDTA and Lithium-Heparin (Li-Hep), the influence of using EDTA or Li-Hep tubes and, finally, the impact of freezing and thawing on the sample. Forty matched samples of Li-Hep and EDTA plasma and 40 urine samples were analyzed for method, anticoagulant, and freeze-thaw comparisons. Linearity was assessed using high NGAL samples diluted in urine, EDTA, and Li-Hep plasma. Commercial internal controls were used for the imprecision study. The Cobas 6000 measured identically with the Hitachi 917, however, not in EDTA plasma (Median Difference = 17.50 μg/L, p < 0.0001). Freeze-thaw process reduced NGAL ((EDTA: Mean Difference = = 15.13 μg/L, p = 0.0014)(Li-Hep: Median Difference = = 6.5 μg/L, p = 0.0129)). NGAL results were higher in Li-Hep plasma than in EDTA plasma ((Non-thawed: Median Difference = = 14.5 μg/L, p < 0.0001), (Thawed: Median Difference = = 21.5 μg/L, p = 0.0003)). Linearity agreements were observed in all three specimens. Imprecision (CV%) was below 3%. The NGAL Test(TM) can be applied on the Cobas 6000 with acceptable performance, although the Cobas 6000 measured higher than the Hitachi 917 in EDTA plasma. Though clinically insignificant, we found that the freeze-thaw process had a reduced effect. NGAL results were higher in Li-Hep tubes than in EDTA tubes. Thus, for blood samples we recommend use of EDTA tubes for NGAL measurements.

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

PubMed

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.

Methane dynamics regulated by microbial community response to permafrost thaw.

PubMed

McCalley, Carmody K; Woodcroft, Ben J; Hodgkins, Suzanne B; Wehr, Richard A; Kim, Eun-Hae; Mondav, Rhiannon; Crill, Patrick M; Chanton, Jeffrey P; Rich, Virginia I; Tyson, Gene W; Saleska, Scott R

2014-10-23

Permafrost contains about 50% of the global soil carbon. It is thought that the thawing of permafrost can lead to a loss of soil carbon in the form of methane and carbon dioxide emissions. The magnitude of the resulting positive climate feedback of such greenhouse gas emissions is still unknown and may to a large extent depend on the poorly understood role of microbial community composition in regulating the metabolic processes that drive such ecosystem-scale greenhouse gas fluxes. Here we show that changes in vegetation and increasing methane emissions with permafrost thaw are associated with a switch from hydrogenotrophic to partly acetoclastic methanogenesis, resulting in a large shift in the δ(13)C signature (10-15‰) of emitted methane. We used a natural landscape gradient of permafrost thaw in northern Sweden as a model to investigate the role of microbial communities in regulating methane cycling, and to test whether a knowledge of community dynamics could improve predictions of carbon emissions under loss of permafrost. Abundance of the methanogen Candidatus 'Methanoflorens stordalenmirensis' is a key predictor of the shifts in methane isotopes, which in turn predicts the proportions of carbon emitted as methane and as carbon dioxide, an important factor for simulating the climate feedback associated with permafrost thaw in global models. By showing that the abundance of key microbial lineages can be used to predict atmospherically relevant patterns in methane isotopes and the proportion of carbon metabolized to methane during permafrost thaw, we establish a basis for scaling changing microbial communities to ecosystem isotope dynamics. Our findings indicate that microbial ecology may be important in ecosystem-scale responses to global change.

Shallow soil moisture - ground thaw interactions and controls - Part 2: Influences of water and energy fluxes

NASA Astrophysics Data System (ADS)

Guan, X. J.; Spence, C.; Westbrook, C. J.

2010-01-01

The companion paper (Guan et al., 2010) demonstrated variable interactions and correlations between shallow soil moisture and ground thaw in soil filled areas along a wetness spectrum in a subarctic Canadian Precambrian Shield landscape. From wetter to drier, these included a wetland, peatland and soil filled valley. Herein, water and energy fluxes were examined for these same subarctic study sites to discern the key controlling processes on the found patterns. Results showed the key control in variable soil moisture and frost table interactions among the sites was the presence of surface water. At the peatland and wetland sites, accumulated water in depressions and flow paths maintained soil moisture for a longer duration than at the hummock tops. These wet areas were often locations of deepest thaw depth due to the transfer of latent heat accompanying lateral surface runoff. Although the peatland and wetland sites had large inundation extent, modified Péclet numbers indicated the relative influence of external and internal hydrological processes at each site were different. Continuous inflow from an upstream lake into the wetland site caused advective and conductive thermal energies to be of equal importance to conductive ground thaw. The absence of continuous surface flow at the peatland and valley sites led to dominance of conductive thermal energy over advective energy for ground thaw. The results suggest that the modified Péclet number could be a very useful parameter to differentiate landscape components in modeling frost table heterogeneity. The calculated water and energy fluxes, and the modified Péclet number provide quantitative explanations for the shallow soil moisture-ground thaw patterns by linking them with hydrological processes and hillslope storage capacity.

Shallow soil moisture - ground thaw interactions and controls - Part 2: Influences of water and energy fluxes

NASA Astrophysics Data System (ADS)

Guan, X. J.; Spence, C.; Westbrook, C. J.

2010-07-01

The companion paper (Guan et al., 2010) demonstrated variable interactions and correlations between shallow soil moisture and ground thaw in soil filled areas along a wetness spectrum in a subarctic Canadian Precambrian Shield landscape. From wetter to drier, these included a wetland, peatland and soil filled valley. Herein, water and energy fluxes were examined for these same subarctic study sites to discern the key controlling processes on the found patterns. Results showed the presence of surface water was the key control in variable soil moisture and frost table interactions among sites. At the peatland and wetland sites, accumulated water in depressions and flow paths maintained soil moisture for a longer duration than at the hummock tops. These wet areas were often locations of deepest thaw depth due to the transfer of latent heat accompanying lateral surface runoff. Although the peatland and wetland sites had large inundation extent, modified Péclet numbers indicated the relative influence of external and internal hydrological and energy processes at each site were different. Continuous inflow from an upstream lake into the wetland site caused advective and conductive thermal energies to be of equal importance to ground thaw. The absence of continuous surface flow at the peatland and valley sites led to dominance of conductive thermal energy over advective energy for ground thaw. The results suggest that the modified Péclet number could be a very useful parameter to differentiate landscape components in modeling frost table heterogeneity. The calculated water and energy fluxes, and the modified Péclet number provide quantitative explanations for the shallow soil moisture-ground thaw patterns by linking them with hydrological processes and hillslope storage capacity.

Seismic Monitoring of Permafrost During Controlled Thaw: An Active-Source Experiment Using a Surface Orbital Vibrator and Fiber-Optic DAS Arrays

NASA Astrophysics Data System (ADS)

Dou, S.; Wood, T.; Lindsey, N.; Ajo Franklin, J. B.; Freifeld, B. M.; Gelvin, A.; Morales, A.; Saari, S.; Ekblaw, I.; Wagner, A. M.; Daley, T. M.; Robertson, M.; Martin, E. R.; Ulrich, C.; Bjella, K.

2016-12-01

Thawing of permafrost can cause ground deformations that threaten the integrity of civil infrastructure. It is essential to develop early warning systems that can identify critically warmed permafrost and issue warnings for hazard prevention and control. Seismic methods can play a pivotal role in such systems for at least two reasons: First, seismic velocities are indicative of mechanical strength of the subsurface and thus are directly relevant to engineering properties; Second, seismic velocities in permafrost systems are sensitive to pre-thaw warming, which makes it possible to issue early warnings before the occurrence of hazardous subsidence events. However, several questions remain: What are the seismic signatures that can be effectively used for early warning of permafrost thaw? Can seismic methods provide enough warning times for hazard prevention and control? In this study, we investigate the feasibility of using permanently installed seismic networks for early warnings of permafrost thaw. We conducted continuous active-source seismic monitoring of permafrost that was under controlled heating at CRREL's Fairbanks permafrost experiment station. We used a permanently installed surface orbital vibrator (SOV) as source and surface-trenched DAS arrays as receivers. The SOV is characterized by its excellent repeatability, automated operation, high energy level, and the rich frequency content (10-100 Hz) of the generated wavefields. The fiber-optic DAS arrays allow continuous recording of seismic data with dense spatial sampling (1-meter channel spacing), low cost, and low maintenance. This combination of SOV-DAS provides unique seismic datasets for observing time-lapse changes of warming permafrost at the field scale, hence providing an observational basis for design and development of early warning systems for permafrost thaw.

Winter Thaws Can Raise Ground Water Levels in Driftless Area

Treesearch

Richard S. Sartz

1967-01-01

Springflow and ground water levels both rose with winter thaws, even when the ground was frozen. A high soil water content suggests that water moved to the water table through a continuous column of soil water rather than as a wetting front

Spring thaw predictor & development of real time spring load restrictions.

DOT National Transportation Integrated Search

2011-02-01

This report summarizes the results of a study to develop a correlation between weather forecasts and the : spring thaw in order to reduce the duration of load limits on New Hampshire roadways. The study used a falling : weight deflectometer at 10 sit...

Evaluation of Canadian unconfined aggregate freeze-thaw tests for identifying nondurable aggregates : technical summary.

DOT National Transportation Integrated Search

2012-06-01

The Kansas Department of Transportation (KDOT) wants to construct durable concrete pavements with minimal maintenance needs. This goal can only be achieved by using durable aggregates that are resistant to freezing and thawing damage when used in con...

Field information links permafrost carbon to physical vulnerabilities of thawing

NASA Astrophysics Data System (ADS)

Harden, Jennifer W.; Koven, Charles D.; Ping, Chien-Lu; Hugelius, Gustaf; David McGuire, A.; Camill, Phillip; Jorgenson, Torre; Kuhry, Peter; Michaelson, Gary J.; O'Donnell, Jonathan A.; Schuur, Edward A. G.; Tarnocai, Charles; Johnson, Kristopher; Grosse, Guido

2012-08-01

Deep soil profiles containing permafrost (Gelisols) were characterized for organic carbon (C) and total nitrogen (N) stocks to 3 m depths. Using the Community Climate System Model (CCSM4) we calculate cumulative distributions of active layer thickness (ALT) under current and future climates. The difference in cumulative ALT distributions over time was multiplied by C and N contents of soil horizons in Gelisol suborders to calculate newly thawed C and N. Thawing ranged from 147 PgC with 10 PgN by 2050 (representative concentration pathway RCP scenario 4.5) to 436 PgC with 29 PgN by 2100 (RCP 8.5). Organic horizons that thaw are vulnerable to combustion, and all horizon types are vulnerable to shifts in hydrology and decomposition. The rates and extent of such losses are unknown and can be further constrained by linking field and modelling approaches. These changes have the potential for strong additional loading to our atmosphere, water resources, and ecosystems.

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

PubMed

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.

Field information links permafrost carbon to physical vulnerabilities of thawing

USGS Publications Warehouse

Harden, Jennifer W.; Koven, Charles; Ping, Chien-Lu; Hugelius, Gustaf; McGuire, A. David; Camill, P.; Jorgenson, Torre; Kuhry, Peter; Michaelson, Gary; O'Donnell, Jonathan A.; Schuur, Edward A.G.; Tamocai, Charles; Johnson, Kevin; Grosse, G.

2012-01-01

Deep soil profiles containing permafrost (Gelisols) were characterized for organic carbon (C) and total nitrogen (N) stocks to 3m depths. Using the Community Climate System Model (CCSM4) we calculate cumulative probability functions (PDFs) for active layer depths under current and future climates. The difference in PDFs over time was multiplied by C and N contents of soil horizons in Gelisol suborders to calculate newly thawed C and N, Thawing ranged from 147 PgC with 10 PgN by 2050 (representative concentration pathway RCP scenario 4.5) to 436 PgC with 29 PgN by 2100 (RCP 8.5). Organic horizons that thaw are vulnerable to combustion, and all horizon types are vulnerable to shifts in hydrology and decomposition. The rates and extent of such losses are unknown and can be further constrained by linking field and modelling approaches. These changes have the potential for strong additional loading to our atmosphere, water resources, and ecosystems.

Analytical solutions for benchmarking cold regions subsurface water flow and energy transport models: one-dimensional soil thaw with conduction and advection

USGS Publications Warehouse

Kurylyk, Barret L.; McKenzie, Jeffrey M; MacQuarrie, Kerry T. B.; Voss, Clifford I.

2014-01-01

Numerous cold regions water flow and energy transport models have emerged in recent years. Dissimilarities often exist in their mathematical formulations and/or numerical solution techniques, but few analytical solutions exist for benchmarking flow and energy transport models that include pore water phase change. This paper presents a detailed derivation of the Lunardini solution, an approximate analytical solution for predicting soil thawing subject to conduction, advection, and phase change. Fifteen thawing scenarios are examined by considering differences in porosity, surface temperature, Darcy velocity, and initial temperature. The accuracy of the Lunardini solution is shown to be proportional to the Stefan number. The analytical solution results obtained for soil thawing scenarios with water flow and advection are compared to those obtained from the finite element model SUTRA. Three problems, two involving the Lunardini solution and one involving the classic Neumann solution, are recommended as standard benchmarks for future model development and testing.

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

NASA Astrophysics Data System (ADS)

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

2018-06-01

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

The Soil Moisture Active and Passive Mission (SMAP): Science and Applications

NASA Technical Reports Server (NTRS)

Entekhabi, Dara; O'Neill, Peggy; Njoku, Eni

2009-01-01

The Soil Moisture Active and Passive mission (SMAP) will provide global maps of soil moisture content and surface freeze/thaw state. Global measurements of these variables are critical for terrestrial water and carbon cycle applications. The SMAP observatory consists of two multipolarization L-band sensors, a radar and radiometer, that share a deployable-mesh reflector antenna. The combined observations from the two sensors will allow accurate estimation of soil moisture at hydrometeorological (10 km) and hydroclimatological (40 km) spatial scales. The rotating antenna configuration provides conical scans of the Earth surface at a constant look angle. The wide-swath (1000 km) measurements will allow global mapping of soil moisture and its freeze/thaw state with 2-3 days revisit. Freeze/thaw in boreal latitudes will be mapped using the radar at 3 km resolution with 1-2 days revisit. The synergy of active and passive observations enables measurements of soil moisture and freeze/thaw state with unprecedented resolution, sensitivity, area coverage and revisit.

Disaggregating meteorites by automated freeze thaw

NASA Astrophysics Data System (ADS)

Charles, Christopher R. J.

2011-06-01

An automated freeze-thaw (AFT) instrument for disaggregating meteorites is described. Meteorite samples are immersed in 18.2 MΩ water and hermetically sealed in a clean 30 ml Teflon vial. This vial and its contents are dipped between baths of liquid nitrogen and hot water over a number of cycles by a dual-stepper motor system controlled by LabView. Uniform and periodic intervals of freezing and thawing induce multiple expansions and contractions, such that cracks propagate along natural flaws in the meteorite for a sufficient number of AFT cycles. For the CR2 chondrite NWA801, the boundaries between different phases (i.e., silicates, metal, matrix) became progressively weaker and allowed for an efficient recovery of 500 individual chondrules and chondrule fragments spanning 0.2-4.7 mm diameters after 243 AFT cycles over 103.3 h. Further FT experiments on a basalt analog showed that the time required for freezing and thawing the same number of cycles can be reduced by a factor of ˜4.

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

PubMed Central

Wei, Ran; Wang, Peng; Han, Minyi; Chen, Tianhao; Xu, Xinglian; Zhou, Guanghong

2017-01-01

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. PMID:27554358

Freeze/thaw stress induces organelle remodeling and membrane recycling in cryopreserved human mature oocytes.

PubMed

Nottola, Stefania Annarita; Albani, Elena; Coticchio, Giovanni; Palmerini, Maria Grazia; Lorenzo, Caterina; Scaravelli, Giulia; Borini, Andrea; Levi-Setti, Paolo Emanuele; Macchiarelli, Guido

2016-12-01

Our aim was to evaluate the ultrastructure of human metaphase II oocytes subjected to slow freezing and fixed after thawing at different intervals during post-thaw rehydration. Samples were studied by light and transmission electron microscopy. We found that vacuolization was present in all cryopreserved oocytes, reaching a maximum in the intermediate stage of rehydration. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates decreased following thawing, particularly in the first and intermediate stages of rehydration, whereas mitochondria-vesicle (MV) complexes augmented in the same stages. At the end of rehydration, vacuoles and MV complexes both diminished and M-SER aggregates increased again. Cortical granules (CGs) were scarce in all cryopreserved oocytes, gradually diminishing as rehydration progressed. This study also shows that such a membrane remodeling is mainly represented by a dynamic process of transition between M-SER aggregates and MV complexes, both able of transforming into each other. Vacuoles and CG membranes may take part in the membrane recycling mechanism.

Long-term anoxia and release of ancient, labile carbon upon thaw of Pleistocene permafrost

USGS Publications Warehouse

Ewing, Stephanie A.; O'Donnell, Jonathan A.; Aiken, George R.; Butler, Kenna D.; Butman, David; Windham-Myers, Lisamarie; Kanevskiy, Mikhail

2015-01-01

The fate of permafrost carbon upon thaw will drive feedbacks to climate warming. Here we consider the character and context of dissolved organic carbon (DOC) in yedoma permafrost cores from up to 20 m depth in central Alaska. We observed high DOC concentrations (4 to 129 mM) and consistent low molecular weight organic acid concentrations in three cores. We estimate a DOC production rate of 12 µmol DOC m−2 yr−1 based on model ages of up to ~200 kyr derived from uranium isotopes. Acetate C accounted for 24 ± 1% of DOC in all samples. This proportion suggests long-term anaerobiosis and is likely to influence thaw outcomes due to biolability of acetate upon release in many environments. The combination of uranium isotopes, ammonium concentrations, and calcium concentrations explained 86% of the variation in thaw water DOC concentrations, suggesting that DOC production may be related to both reducing conditions and mineral dissolution over time.

Methane emission from high-latitude (>50N) lakes: Annual cycle of climatological emissions using satellite-derived lake-ice phenology and freeze-thaw dynamics

NASA Astrophysics Data System (ADS)

Matthews, E.; Romanski, J.; Du, J.; Watts, J. D.

2017-12-01

Lakes are increasingly recognized as potentially important contributors to global methane emissions despite occupying only a few percent of Earth's ice-free land surface. More than 40% of the global lake area lies in regions of amplified warming north of 50˚N. As with wetlands, lake emissions are sensitive to interannual fluctuations in, e.g., temperature and duration of thaw season. Several estimates of CH4emission from high-latitude lakes have been published but none relies on geospatial lake distributions and satellite-based duration and timing of thaw seasons. We report on a climatology of weekly, spatially-explicit methane emissions from high-latitude lakes. Lake break-up and freeze-up dates for lakes >50km^2 were determined from a lake-ice phenology data set derived from brightness temperature (Tb) observations of space-borne Advanced Microwave Scanning Radiometer (AMSR-E/2) sensors. The lake-ice conditions for smaller lakes were estimated using an Earth System Data Record for Land Surface Freeze-Thaw State derived from Tb observations of Scanning Multichannel Microwave Radiometer (SMMR), Special Sensor Microwave Imager (SSM/I), and SSM/I Sounder (SSMIS). Climatologies encompass 2002-2015 for lake ice phenology and 1979 to 2010 for the land surface freeze-thaw state. Climatologies encompass 2003-2014 for ice phenology and 1979 to 2010 for freeze-thaw dynamics. Length and timing of typical methane-emission periods, derived from the satellite data, were integrated with daily diffusive and ebulliative methane fluxes for lake types following the work of Wik et al. (Nature, 2016) to estimate a full annual cycle of emissions from lakes >50˚N. We explored several approaches to estimate the large bursts of emissions observed over short periods during lake-ice breakup immediately prior to full lake thaw since several studies suggest that a substantial fraction of total annual emissions may occur at this time. While highly uncertain, we plan to investigate whether the modest, short-lived but annual uptick in atmospheric methane concentrations in late winter/early spring may be associated with these bursts of methane from lakes.

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

PubMed

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 sperm.

Nutrient Controls on Methane Emissions in a Permafrost Thaw Subarctic Peatland

NASA Astrophysics Data System (ADS)

Kashi, N. N.; Perryman, C. R.; Malhotra, A.; Marek, E. A.; Giesler, R.; Varner, R. K.

2015-12-01

Permafrost peatlands in northern latitudes are large reservoirs of sequestered carbon that are vulnerable to climate change. While peatlands account for a small fraction of total global land surfaces, their potential to release sequestered carbon in response to higher temperatures is of concern. Of particular relevance is the conversion of these carbon stores into methane (CH4), a strong greenhouse gas with a global warming potential 20 times greater than that of CO2 over a 100-year time frame. Here, we explore how key nutrients impact the consumption of CH4 at the Stordalen Mire in Abisko, Sweden, a discontinuous permafrost peatland with expanding thaw over the last century. Peatland CH4 emissions are highly spatially variable due to multiple emission pathways and strong dependence on several environmental factors. Among controls on CH4 emissions, such as temperature and water table depth, primary production of wetland vegetation is also a strong factor in the variability of CH4 emissions. Plant community shifts among permafrost thaw stages subsequently change nutrient cycling and availability, which in turn impacts primary production. Early stages of permafrost thaw are mosaicked with a variety of vascular plants and mosses. We analyzed potential enzymatic activities of chitinase, glucosidase, and phosphatase as proxies for organic nitrogen, carbon, and phosphorus cycling, respectively, in tandem with potential CH4 oxidation rates. In addition, stoichiometric ratios of carbon, nitrogen, and phosphorus concentrations are used to illustrate nutrient limitation controls on CH4 oxidation rates. While CH4 emissions are low throughout initial thaw stages, < 7 CH4 mg m-2 day-1, we found they had the highest rates of potential CH4 oxidation. These permafrost thaw-induced CH4 oxidation rates are 5 and 11 times higher, in the surface and depth of the peat profile respectively, than subsequent aerobic permafrost thaw stages. As CH4 emissions are low in intact permafrost peatlands, these high rates of potential CH4 oxidation indicate the importance of plant communities and the methanotrophic microbes they harbor.

The development of the cell cryopreservation protocol with controlled rate thawing.

PubMed

Gurina, Tatyana M; Pakhomov, Alexandr V; Polyakova, Anna L; Legach, Evgeniy I; Bozhok, Galyna A

2016-06-01

Thawing in the water bath is often considered as a standard procedure. The thermal history of samples thawed in this way is poorly controlled, but cryopreservation and banking of cell-based products require standardization, automation and safety of all the technological stages including thawing. The programmable freezers allow implementation of the controlled cooling as well as the controlled thawing. As the cell damage occurs during the phase transformation that takes place in the cryoprotectant medium in the process of freezing-thawing, the choice of warming rates within the temperature intervals of transformations is very important. The goal of the study was to investigate the influence of warming rates within the intervals of the phase transformations in the DMSO-based cryoprotectant medium on the cell recovery and to develop a cryopreservation protocol with controlled cooling and warming rates. The temperature intervals of phase transformations such as melting of the eutectic mixture of the cryoprotectant solution (MEMCS), melting of the eutectic salt solution (MESS), melting of the main ice mass (MMIM), recrystallization before MEMCS, recrystallization before MESS and recrystallization before MMIM were determined by thermo-mechanical analysis. The biological experiments were performed on the rat testicular interstitial cells (TIC). The highest levels of the cell recovery and metabolic activity after cryopreservation were obtained using the protocol with the high (20 °C/min) warming rate in the temperature intervals of crystallization of the eutectics as well as recrystallizations and the low (1 °C/min) warming rate in the temperature intervals of melting of the eutectics as well as MMIM. The total cell recovery was 65.3 ± 2.1 %, the recovery of the 3-beta-HSD-positive (Leydig) cells was 82.9 ± 1.8 %, the MTT staining was 32.5 ± 0.9 % versus 42.1 ± 1.7 %; 57.4 ± 2.1 % and 24.0 ± 1.1 % respectively, when compared to the thawing in the water bath.

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

PubMed

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 spermatozoa from deleterious effects of cryopreservation. The findings of the study indicated that GSH at 0.5mM can be effectively used as an additive in bull semen extender for freezing and thawing. Copyright © 2017 Elsevier B.V. All rights reserved.

Elemental composition and optical properties reveal changes in dissolved organic matter along a permafrost thaw chronosequence in a subarctic peatland

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hodgkins, Suzanne B.; Tfaily, Malak M.; Podgorski, David C.

2016-08-01

The fate of carbon stored in permafrost-zone peatlands represents a significant uncertainty in global climate modeling. Given that the breakdown of dissolved organic matter (DOM) is often a major pathway for decomposition in peatlands, knowledge of DOM reactivity under different permafrost regimes is critical for determining future climate feedbacks. To explore the effects of permafrost thaw and resultant plant succession on DOM reactivity, we used a combination of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), UV/Vis absorbance, and excitation-emission matrix spectroscopy (EEMS) to examine the DOM elemental composition and optical properties of 27 pore water samples gathered frommore » various sites along a permafrost thaw sequence in Stordalen Mire, a thawing subarctic peatland in northern Sweden. The presence of dense Sphagnum moss, a feature that is dominant in the intermediate thaw stages, appeared to be the main driver of variation in DOM elemental composition and optical properties at Stordalen. Specifically, DOM from sites with Sphagnum had greater aromaticity, higher average molecular weights, and greater O/C, consistent with a higher abundance of phenolic compounds that likely inhibit decomposition. These compounds are released by Sphagnum and may accumulate due to inhibition of phenol oxidase activity by the acidic pH at these sites. In contrast, sites without Sphagnum, specifically fully-thawed rich fens, had more saturated, more reduced compounds, which were high in N and S. Optical properties at rich fens were indicated the presence of microbially-derived DOM, consistent with the higher decomposition rates previously measured at these sites. These results indicate that Sphagnum acts as an inhibitor of rapid decomposition and CH4 release in thawing subarctic peatlands, consistent with lower rates of CO2 and CH4 production previously observed at these sites. However, this inhibitory effect may disappear if Sphagnumdominated bogs transition to more waterlogged rich fens that contain very little to no living Sphagnum. Release of this inhibition allows for higher levels of microbial activity and potentially greater CH4 release, as has been observed in these fen sites.« less

The benefits of cooling boar semen in long-term extenders prior to cryopreservation on sperm quality characteristics.

PubMed

Wasilewska, K; Zasiadczyk, Ł; Fraser, L; Mogielnicka-Brzozowska, M; Kordan, W

2016-10-01

This study investigated the effects of long-term extenders on post-thaw sperm quality characteristics following different holding times (HT) of boar semen at 17 and 10°C. Sperm-rich fractions, collected from five boars, were diluted in Androhep(®) Plus (AHP), Androstar(®) Plus (ASP), Safecell(®) Plus and TRIXcell(®) Plus (TCP) extenders. The extended semen samples were held for 2 hr at 17°C (HT 1) and additionally for 24 hr at 10°C (HT 2), after they were evaluated and frozen. CASA sperm motility and motion patterns, mitochondrial membrane potential (MMP), plasma membrane integrity (PMI) and normal apical ridge (NAR) acrosome integrity were assessed in the pre-freeze and frozen-thawed semen. The Vybrant Apoptosis Assay Kit was used to analyse the proportions of viable and plasma membrane apoptotic-like changes in spermatozoa. Results indicated that boar variability, extender and HT significantly affected the sperm quality characteristics, particularly after freezing-thawing. Differences in the pre-freeze semen were more marked in the sperm motion patterns between the HTs. Pre-freeze semen in HT 2 showed significantly higher VCL and VAP, whereas no marked effects were observed in the sperm membrane integrity and viability (YO-PRO-1(-) /PI(-) ) among the extenders. Post-thaw sperm TMOT and PMOT were significantly higher in the AHP and ASP extenders of HT 2 group, whereas VSL, VCL and VAP were markedly lower in the TCP extender. Furthermore, spermatozoa from the AHP- and ASP-extended semen of HT 2 group were characterized by higher MMP, PMI and NAR acrosome integrity following freezing-thawing. In most of the extenders, the incidence of frozen-thawed spermatozoa with apoptotic-like changes was greater in HT 1. The findings of this study indicate that holding of boar semen at 10°C for 24 hr in long-term preservation extenders modulates post-thaw sperm quality characteristics in an extender-dependent manner. These results will further contribute to the improvement in the cryopreservation technology of boar semen. © 2016 Blackwell Verlag GmbH.

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

PubMed

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 (P<0.001) in plasma membrane integrity and in mitochondrial activity in the spermatozoa frozen with Orvus ES Paste in both freezing extenders. Furthermore, spermatozoa frozen with Orvus ES Paste in both freezing extenders exhibited lower (P<0.05) motility and kinematic parameters than those frozen in the absence of Orvus ES Paste in the first freezing extender. 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 (P<0.05) over time. The deterioration in post-thaw sperm motility and kinematic parameters were concurrent with reduced sperm characteristics. It can be suggested that in the Iberian pig, the beneficial effects of Orvus ES Paste during the freezing process of spermatozoa is time dependent. The analysis of different sperm characteristics such as motility, plasma membrane integrity and mitochondrial function, determined that the extenders studied in the present experiment affected the quality of frozen-thawed semen in Iberian boar.

Early thawing after snow removal and no straw mulching accelerates organic carbon cycling in a paddy soil in Northeast China.

PubMed

Zhang, Hao; Tang, Jie; Liang, Shuang; Li, Zhaoyang; Wang, Jingjing; Wang, Sining

2018-03-01

Variations in soil organic carbon (SOC) have implications for atmospheric CO 2 concentrations and the greenhouse effect. However, the effects of snow cover and straw mulching on the variations in SOC fractions across winter remain largely unknown. In this study, soil samples were collected during different stages of winter from an in situ experiment comprising three treatments: 1) snow removal with no straw mulching (Sn-SM-); 2) snow cover with no straw mulching (SC), and; 3) snow cover with straw mulching (SC + SM+). Results showed that labile organic carbon, semi-labile organic carbon, recalcitrant organic carbon (ROC), the light fraction of organic carbon (LFOC), and easily oxidized organic carbon (EOC) contents did not vary significantly (P > .05) during the unfrozen to hard frost stages. Compared to the unfrozen stage, microbial biomass carbon (MBC) contents decreased by 519.03 mg kg -1 , 325.21 mg kg -1 , and 244.09 mg kg -1 and dissolved organic carbon (DOC) contents increased by 473.36 mg kg -1 , 348.10 mg kg -1 , and 258.89 mg kg -1  at the hard frost stage in Sn-SM-, SC, and SC + SM + treatments, respectively. Throughout all thawing stages, > 61% and 59% of SOC and ROC accumulation, respectively in the three treatments were observed in thawing stage II, indicating that higher temperatures and microbial activities in thawing stage II accelerated the inputs of SOC and ROC. ROC accumulation accounted for >65% of the SOC accumulation and the proportions of ROC in SOC increased in the three treatments during the thawing stages. SC + SM + treatment maintained lower EOC contents during thawing stages than other treatments. The observation of lowest SOC and LFOC accumulation and contents in the SC + SM + treatment during thawing stages showed that SC + SM + experienced the least inputs of SOC in the soil. Copyright © 2018 Elsevier Ltd. All rights reserved.

Elemental composition and optical properties reveal changes in dissolved organic matter along a permafrost thaw chronosequence in a subarctic peatland

NASA Astrophysics Data System (ADS)

Hodgkins, Suzanne B.; Tfaily, Malak M.; Podgorski, David C.; McCalley, Carmody K.; Saleska, Scott R.; Crill, Patrick M.; Rich, Virginia I.; Chanton, Jeffrey P.; Cooper, William T.

2016-08-01

The fate of carbon stored in permafrost-zone peatlands represents a significant uncertainty in global climate modeling. Given that the breakdown of dissolved organic matter (DOM) is often a major pathway for decomposition in peatlands, knowledge of DOM reactivity under different permafrost regimes is critical for determining future climate feedbacks. To explore the effects of permafrost thaw and resultant plant succession on DOM reactivity, we used a combination of Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), UV/Vis absorbance, and excitation-emission matrix spectroscopy (EEMS) to examine the DOM elemental composition and optical properties of 27 pore water samples gathered from various sites along a permafrost thaw sequence in Stordalen Mire, a thawing subarctic peatland in northern Sweden. The presence of dense Sphagnum moss, a feature that is dominant in the intermediate thaw stages, appeared to be the main driver of variation in DOM elemental composition and optical properties at Stordalen. Specifically, DOM from sites with Sphagnum had greater aromaticity, higher average molecular weights, and greater O/C, consistent with a higher abundance of phenolic compounds that likely inhibit decomposition. These compounds are released by Sphagnum and may accumulate due to inhibition of phenol oxidase activity by the acidic pH at these sites. In contrast, sites without Sphagnum, specifically fully-thawed rich fens, had more saturated, more reduced compounds, which were high in N and S. Optical properties at rich fens indicated the presence of microbially-derived DOM, consistent with the higher decomposition rates previously measured at these sites. These results indicate that Sphagnum acts as an inhibitor of rapid decomposition and CH4 release in thawing subarctic peatlands, consistent with lower rates of CO2 and CH4 production previously observed at these sites. However, this inhibitory effect may disappear if Sphagnum-dominated bogs transition to more waterlogged rich fens that contain very little to no living Sphagnum. Release of this inhibition allows for higher levels of microbial activity and potentially greater CH4 release, as has been observed in these fen sites.

Direct N2O Fluxes from Agroecosystems in Cold Climates: Importance of Soil Freeze-Thaw

NASA Astrophysics Data System (ADS)

Congreves, K. A.; Wagner-Riddle, C.; Abalos, D.; Ambadan, J. T.; Brown, S. E.; Tenuta, M.; Gao, X.; Amiro, B. D.; Berg, A. A.

2016-12-01

To develop effective climate change mitigation strategies and reduce N2O emissions, the global contribution of freeze-thaw cycles from croplands must be characterized; we present the first study to do so. Long-term N2O flux datasets from micrometeorological approaches were compiled from two Canadian sites (Elora ON & Glenlea MB). Measurements encompassed a total of 21-yr on 16-ha of land producing annual field crops, yielding an unprecedented record of N2O fluxes (42118 half-hourly values) at sites subjected to freeze-thaw cycles from Nov-Apr. At Elora (the warmer site) N2O flux events were associated with thaw cycles throughout Nov-Apr and the main thaw event took place between mid/end of April when air temperatures rose above 0°C and snow melted. The continental site (Glenlea) did not have significant N2O flux events during the prolonged freeze period, but had considerably higher emissions over the thaw period when soil temperature and liquid water content increased more slowly than Elora. Based on cumulative N2O emissions from both sites (Nov-Apr), emissions were closely related to freezing degree days (FDD). An exponential-to-plateau model significantly fit (p<0.0001, r= 0.72) the relationship between N2O emissions and FDD, characterizing larger N2O emissions as FDD increased (y=1.95 (1-exp-0.00852x), y=cumulative N2O-N kg ha-1 and x=FDD). To estimate the global contribution of N2O emissions from seasonally frozen croplands in the northern hemisphere, we applied the algorithm to a global map of FDD derived from three reanalysis products (ERA-Interim, MERRA-Land, GLDAS-NOAH) combined with MODIS land fraction data for croplands. Average global freeze-thaw induced N2O emissions for croplands was estimated at 1.07 Tg N, though it may range from 0.79 - 1.35 Tg N due to model error and variation. This global contribution of N2O from seasonally frozen cropland soils may be responsible for previously observed discrepancies between top-down and bottom-up approaches.

Long Term Thawing Experiments on Intact Cores of Arctic Mineral Cryosol: Implications for Greenhouse Gas Feedbacks from Global Warming

NASA Astrophysics Data System (ADS)

Onstott, T. C.; Stackhouse, B. T.; Lau, C. Y. M.; Whyte, L. G.; Pfiffner, S. M.; Vishnivetskaya, T. A.

2015-12-01

Mineral cryosols comprise >87% of Arctic tundra. Much attention has focused on high-organic carbon cryosols and how they will respond to global warming. The biogeochemical processes related to the greenhouse gas release from mineral cryosols, however, have not been fully explored. To this end, seventeen intact cores of active layer and underlying permafrost of mineral cryosol from Axel Heiberg Island, Nunavut, Canada, were subjected to 85 weeks of thawing at 4.5°C under various treatment regimes. The fluxes of CO2 and CH4 across the atmosphere-soil boundary and vertical profiles of the gas and water chemistry and the metagenomes were determined. The flux measurements were compared to those of microcosms and field measurements. The main conclusions were as follows: 1) CO2 emission rates from the intact cores do not behave in the typical fast to slow carbon pool fashion that typify microcosm experiments. The CO2 emission rates from the intact cores were much slower than those from the microcosm initially, but steadily increased with time, overtaking and then exceeding microcosm release rates after one year. 2) The increased CO2 flux from thawing permafrost could not be distinguished from that of control cores until after a full year of thawing. 3) Atmospheric CH4 oxidation was present in all intact cores regardless of whether they are water saturated or not, but after one year it had diminished to the point of being negligible. Over that same time the period the metagenomic data recorded a significant decline in the proportion of high-affinity methanotrophs. 4) Thaw slumps in the cores temporarily increased the CH4 oxidation and the CO2 emission rates. 5) The microbial community structures varied significantly by depth with methanotrophs being more abundant in above 35 cm depth than below 35 cm depth. 6) Other than the diminishment of Type II methanotrophs, the microbial community structure varied little after one week of thawing, nor even after 18 months of thaw.

Effect of Thawing Time, Cooling Rate and Boron Nutrition on Freezing Point of the Primordial Shoot in Norway Spruce Buds

PubMed Central

RÄISÄNEN, MIKKO; REPO, TAPANI; LEHTO, TARJA

2006-01-01

• Background Effects of cooling rates on bud frost hardiness have been studied but there is little information on bud responses to thawing. Since the cell wall pore size has been found to increase with boron (B) deficiency, B deficiency may affect the supercooling ability of buds in winter. • Methods The effects of duration of thawing time and rate of cooling on bud frost hardiness of Norway spruce (Picea abies) were studied in a B fertilization trial in February 2003 and March 2005. Frost hardiness of apical buds was determined by differential thermal analysis (DTA) and visual scoring of damage. • Key Results In 2003, the freezing point of primordial shoots of buds (Tf), i.e. the low-temperature exotherm (LTE), was, on average, −39 °C when buds were thawed for less than 3 h and the Tf increased to −21 °C after 18 h of thawing. During the first 4 h of thawing, the rate of dehardening was 6 °C h−1. In 2005, buds dehardened linearly from −39 °C to −35 °C at a rate of 0·7 °C h−1. In 2003, different cooling rates of 1–5 °C h−1 had a minor effect on Tf but in 2005 with slow cooling rates Tf decreased. In both samplings, at cooling rates of 2 and 1 °C h−1, Tf was slightly higher in B-fertilized than in non-fertilized trees. By contrast, at very short thawing times in 2003, Tf was somewhat lower in B-fertilized trees. • Conclusions There was little evidence of reduced frost hardiness in trees with low B status. This study showed that buds deharden rapidly when exposed to above-freezing temperatures in winter, but if cooled again they reharden more slowly. According to this study, rapid dehardening of buds has to be taken into account in assessments of frost hardiness. PMID:16464880

Genes, isotopes, and ecosystem biogeochemistry. Dissecting methane flux at the leading edge of global change

DOE Office of Scientific and Technical Information (OSTI.GOV)

Saleska, Scott; Rich, Virginia; Tyson, Gene

This project integrates across three fields (microbiology, biogeochemistry, and modeling) to understand the mechanisms of methane cycling in thawing permafrost. We have made substantial progress in each area, and in cross-cutting interdisciplinary synthesis. Large releases of CH 4 from thawing permafrost to the atmosphere, a strong positive feedback to global warming, are plausible but little is known about the controls on such release. Our project (“IsoGenie”) addresses the key question: What is the interplay of microbial communities and soil organic matter composition in the decomposition of organic C to CH 4 across a permafrost thaw gradient?

Application of Satellite SAR Imagery in Mapping the Active Layer of Arctic Permafrost

NASA Technical Reports Server (NTRS)

Zhang, Ting-Jun; Li, Shu-Sun

2003-01-01

The objective of this project is to map the spatial variation of the active layer over the arctic permafrost in terms of two parameters: (i) timing and duration of thaw period and (ii) differential frost heave and thaw settlement of the active layer. To achieve this goal, remote sensing, numerical modeling, and related field measurements are required. Tasks for the University of Colorado team are to: (i) determine the timing of snow disappearance in spring through changes in surface albedo (ii) simulate the freezing and thawing processes of the active layer and (iii) simulate the impact of snow cover on permafrost presence.

Novel and traditional traits of frozen-thawed porcine sperm related to in vitro fertilization success

USDA-ARS?s Scientific Manuscript database

Cryopreserved semen allows the use of single ejaculates for repeated analyses, potentially improving in vitro fertilization (IVF) consistency by eliminating inter-ejaculate variability observed with fresh semen. However, the freezing and thawing processes result in compromised sperm function and IVF...

Comparison of spacing factors as measured by the air-void analyzer and ASTM C457.

DOT National Transportation Integrated Search

2015-12-01

Freezing and thawing cycles will result in damage to concrete that is saturated : unless the concrete is properly entrained with small and well-dispersed air : voids. Durable concrete subject to cycles of freezing and thawing must : have an air-void ...

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

DOT National Transportation Integrated Search

1978-01-01

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

Correlation of air void parameters obtained by linear traverse with freeze-thaw durability.

DOT National Transportation Integrated Search

1983-01-01

The correlations obtainable from comparisons of the various air void parameters with the freeze-thaw durability of concretes are listed. It is shown that correlations are no better when only small voids are used than when the total void content is us...

The circuitry of ecosystem metabolism: CO2 and CH4 flux from permafrost soils

USDA-ARS?s Scientific Manuscript database

Microbial decomposition of thawed permafrost organic matter could release greenhouse gases (GHG) to the atmosphere and accelerate the carbon (C)-climate feedback. Greenhouse gas emissions from thawed permafrost are difficult to predict because they result from complex interactions between abiotic dr...

Long-Time Cooling before Cryopreservation Decreased Translocation of Phosphatidylserine (Ptd-L-Ser) in Human Ovarian Tissue

PubMed Central

2015-01-01

Objectives To translocation (externalization) of phosphatidylserine lead at least the five negative effects observed during cells cryopreservation: hypoxia, increasing of intracellular Ca2+, osmotic disruption of cellular membranes, generation of reactive oxygen species (ROS) and lipid peroxidation. The aim of this study was to test the intensiveness of the phosphatidylserine translocation immediately after thawing and after 45 d xenografting of human ovarian tissue, which was either frozen just after operative removal from patient or cooled before cryopreservation to 5°C for 24 h and then frozen. Materials and Methods Ovarian fragments from twelve patients were divided into small pieces in form of cortex with medulla, and randomly divided into the following four groups. Pieces of Group 1 (n=30) were frozen immediately after operation, thawed and just after thawing their quality was analyzed. Group 2 pieces (n=30) after operation were cooled to 5°C for 24 h, then frozen after 24 h pre-cooling to 5°C, thawed and just after thawing their quality was analyzed. Group 3 pieces (n=30) were frozen immediately after operation without pre-cooling, thawed, transplanted to SCID mice and then, after 45 d of culture their quality was analyzed. Group 4 pieces (n=30) were frozen after 24 h pre-cooling to 5°C, thawed, transplanted to SCID mice and then, after 45 d their quality was analyzed. The effectiveness of the pre-freezing cooling of tissuewas evaluated by the development of follicles (histology) and by intensiveness of translocation of phosphatidylserine (FACS with FITC-Annexin V and Propidium Iodide). Results For groups 1, 2, 3 and 4 the mean densities of follicles per 1 mm3 was 19.0, 20.2, 12.9, and 12.2, respectively (P1-2, 3-4 >0.1). For these groups, 99%, 98%, 88% and 90% preantral follicles, respectively were morphologically normal (P1-2, 3-4 >0.1). The FACS analysis showed significantly decreased intensiveness of translocation of phosphatidylserine after pre-cooling of frozen tissue (46.3% and 33.6% in Groups 2 and 4, respectively), in contrast with tissue frozen without pre-cooling (77.1% and 60.2 % in Groups 1 and 3, respectively, P1, 3-2, 4 <0.05). Conclusions Long time (24 h) cooling of ovarian tissue to 5°C before cryopreservation decreased translocation of phosphatidylserine that evidences about increases the viability of the cells in the tissue after thawing. PMID:26083026

Long-Time Cooling before Cryopreservation Decreased Translocation of Phosphatidylserine (Ptd-L-Ser) in Human Ovarian Tissue.

PubMed

Isachenko, Vladimir; Todorov, Plamen; Isachenko, Evgenia; Rahimi, Gohar; Tchorbanov, Andrey; Mihaylova, Nikolina; Manoylov, Iliyan; Mallmann, Peter; Merzenich, Markus

2015-01-01

To translocation (externalization) of phosphatidylserine lead at least the five negative effects observed during cells cryopreservation: hypoxia, increasing of intracellular Ca2+, osmotic disruption of cellular membranes, generation of reactive oxygen species (ROS) and lipid peroxidation. The aim of this study was to test the intensiveness of the phosphatidylserine translocation immediately after thawing and after 45 d xenografting of human ovarian tissue, which was either frozen just after operative removal from patient or cooled before cryopreservation to 5°C for 24 h and then frozen. Ovarian fragments from twelve patients were divided into small pieces in form of cortex with medulla, and randomly divided into the following four groups. Pieces of Group 1 (n=30) were frozen immediately after operation, thawed and just after thawing their quality was analyzed. Group 2 pieces (n=30) after operation were cooled to 5°C for 24 h, then frozen after 24 h pre-cooling to 5°C, thawed and just after thawing their quality was analyzed. Group 3 pieces (n=30) were frozen immediately after operation without pre-cooling, thawed, transplanted to SCID mice and then, after 45 d of culture their quality was analyzed. Group 4 pieces (n=30) were frozen after 24 h pre-cooling to 5°C, thawed, transplanted to SCID mice and then, after 45 d their quality was analyzed. The effectiveness of the pre-freezing cooling of tissuewas evaluated by the development of follicles (histology) and by intensiveness of translocation of phosphatidylserine (FACS with FITC-Annexin V and Propidium Iodide). For groups 1, 2, 3 and 4 the mean densities of follicles per 1 mm3 was 19.0, 20.2, 12.9, and 12.2, respectively (P1-2, 3-4 >0.1). For these groups, 99%, 98%, 88% and 90% preantral follicles, respectively were morphologically normal (P1-2, 3-4 >0.1). The FACS analysis showed significantly decreased intensiveness of translocation of phosphatidylserine after pre-cooling of frozen tissue (46.3% and 33.6% in Groups 2 and 4, respectively), in contrast with tissue frozen without pre-cooling (77.1% and 60.2 % in Groups 1 and 3, respectively, P1, 3-2, 4 <0.05). Long time (24 h) cooling of ovarian tissue to 5°C before cryopreservation decreased translocation of phosphatidylserine that evidences about increases the viability of the cells in the tissue after thawing.

Tundra ecosystem respiration is dominated by recent C inputs, masking contributions from old and more decomposed substrates

NASA Astrophysics Data System (ADS)

Mauritz, M.; Celis, G.; Ebert, C.; Hutchings, J. A.; Ledman, J.; Pegoraro, E.; Salmon, V.; Schaedel, C.; Taylor, M.; Schuur, E.

2017-12-01

Rising global temperatures and increasing soil respiration are of great concern in high latitude permafrost ecosystems where substantial amounts of carbon (C) are stabilized by cold temperatures. The isotopic δ13C and Δ14C signature of respiration can be used to determine contributions of decomposition from above- and belowground plant respiration, and different parts of the soil column because δ13C and Δ14C change with depth, reflecting new plant inputs at the surface and organic matter in later stages of decomposition at depth. We measured ecosystem respiration (Reco) δ13C from early summer thaw until the end of summer transition from net ecosystem C uptake to net C release in a warming experiment with accelerated permafrost thaw and a vegetation removal treatment and determined the effect of thaw, water table, and plant productivity on seasonal Reco δ13C. When the system was a net CO2 sink in early August and after the system switched to a source we measured Δ14C to further resolve Reco sources. Reco δ13C was most enriched in spring (-23.02 ‰) suggesting that spring thaw released winter-trapped CO2 from soil decomposition in deeper soil layers. In areas with shallow thaw depletion of Reco δ13C from spring (-22.54 ‰) to autumn (-24.54 ‰) indicates a seasonally increasing contribution from plant root respiration and surface soil decomposition, even after surface soils cooled and aboveground leaves senesced. In deeply thawed, dry areas Reco δ13C (-23.33 ‰) was significantly enriched and showed no seasonal pattern while Reco δ13C from deeply thawed, wet areas (-24.19 ‰) was significantly depleted and similarly lacked seasonal change. Reco δ 13C from vegetation free areas was depleted and remained similar all season (-25.28 ‰). Decline of Reco Δ14C between early August (Δ14C: 30 ‰) and September (Δ14C: 6.85 ‰), indicates an underlying contribution to Reco from old, deep soil sources that was masked by high plant activity and surface soil decomposition, even when vegetation was removed (Δ14C removal, Aug: 53.55 ‰; Sep: -86.9 ‰). Our findings demonstrate that the strength of seasonal patterns in Reco δ13C depend on thaw and water table heterogeneity of the landscape. We conclude that the timing of Reco δ 13C and Δ 14C measurements will strongly influence any conclusions about deep soil contributions.

Effects of arginine and trehalose on post-thawed bovine sperm quality.

PubMed

Öztürk, Caner; Güngör, Şükrü; Ataman, Mehmet Bozkurt; Bucak, Mustafa Numan; Başpinar, Nuri; Ili, Pınar; Inanç, Muhammed Enes

2017-09-01

The present study was conducted to examine the protective role of arginine and trehalose on post-thaw bull sperm and oxidative stress parameters. Five ejaculates for each bull were used in the study. Each ejaculate, split into three equal aliquots and diluted at 37 °C with base extenders containing 2 mM arginine, 25 mM trehalose and no antioxidant (control) was cooled to 5 °C and then frozen. Frozen straws were thawed in a water bath for evaluation. Supplementation of the semen extender with arginine decreased the percentages of post-thawed subjective motility (29 ± 8.21%), CASA motility (12.2 ± 5.69%) and progressive motility (3.52 ± 2.13%), compared with the controls (43 ± 2.73%, 55.4 ± 6.78% and 33.48 ± 4.14%, respectively, P < 0.05). Supplementation of the semen extender with trehalose produced a higher mitochondrial activity and sperm viability (36.3 ± 3.99% and 44.1 ± 2.18%) compared with the control (13 ± 8.15 and 31.7 ± 3.94%, respectively, P < 0.05). It was established that trehalose (95.1%) and arginine (92.8%) protect DNA integrity compared to the control (90.4%) (P < 0.05). Trehalose supplementation in semen extenders provided great benefit in terms of viability, mitochondrial activity, and intact sperm DNA on frozen-thawed bull sperm.

Effectiveness of glucose-methanol extender for cryopreservation of Huso huso spermatozoa.

PubMed

Aramli, Mohammad Sadegh; Golshahi, Karim; Nazari, Rajab Mohammad; Aramli, Salim; Banan, Ashkan

2015-11-01

The present approach was designed to evaluate the methanol-glucose extender effects on sperm cryopreservation in beluga sturgeon, Huso huso. Sperm quality was examined by measuring post-thaw sperm motility and fertilizing rate at hatching stage. We first tested the effect of glucose concentration (0, 0.10, 0.15, 0.20 and 0.30M) in a methanol extender on post-thaw sperm motility. The optimal cryopreservation conditions were found to be 0.2M glucose in the extender. Then, motility and fertilization rates of sperm cryopreserved with 0.2M glucose and 10% methanol (GM) were compared to Tris-sucrose-KCl in 10% methanol extender (TSKM). Additionally, sperm motility and fertilizing ability in relation to 15 and 30min equilibration in GM extender before and after cryopreservation were measured. Higher post-thaw sperm motility duration and percentage as well as fertilization rate were obtained with the GM extender when compared to TSKM extender. Equilibration of sperm in extender did not affect the motility quality of either fresh-diluted or frozen/thawed sperm, while fertilization rate showed a significant decline alone after 30min of post-thaw storage. Our results indicated that the use of a simple extender consisting of 0.2M glucose in 10% methanol can be an alternative cryopreservation method to those previously described for sturgeons. Copyright © 2015 Elsevier B.V. All rights reserved.

Effect of prostatic fluid on the quality of fresh and frozen-thawed canine epididymal spermatozoa.

PubMed

Korochkina, E; Johannisson, A; Goodla, Lavanya; Morrell, J M; Axner, E

2014-12-01

Canine epididymal spermatozoa have a low freeze-tolerance ability compared with ejaculated spermatozoa, which could arise from the absence of prostatic fluid (PF). Therefore, the purpose of this work was to elucidate the influence of PF on the quality of canine epididymal sperm before and after freezing. Caudae epididymides were retrieved from eight dogs after routine castration. Spermatozoa were released by slicing the tissue and were extended in either Tris solution or PF before freezing. Frozen sperm samples were thawed at 70 °C for 8 seconds in a waterbath. Sperm concentration, motility using computer-assisted sperm analysis, morphology, plasma membrane, acrosome and chromatin integrity were assessed in the fresh sperm samples (after 20 minutes incubation) and at 0 and 4 hours after thawing. Progressive motility, distance straight line, distance average path, average path velocity, curvilinear velocity, straight line velocity, straightness, linearity, wobble, and beat cross frequency were significantly increased after extraction into PF. There was a higher proportion of spermatozoa with DNA damage in the PF treatment group at 4 hours after thawing than in the Tris treatment group (15.8% vs. 6.7%, P < 0.05). These results suggest that the addition of PF to canine spermatozoa activates sperm motility in fresh spermatozoa but has a negative effect on chromatin integrity after freezing-thawing. Copyright © 2014 Elsevier Inc. All rights reserved.

New strategies of boar sperm cryopreservation: development of novel freezing and thawing methods with a focus on the roles of seminal plasma.

PubMed

Okazaki, Tetsuji; Shimada, Masayuki

2012-09-01

Cryopreservation of boar spermatozoa offers an effective means of long-term storage of important genetic material. Many researchers have investigated how to improve reproductive performance by artificial insemination (AI) using cryopreserved boar spermatozoa. Recently, we and other groups reported that high conception rates (70-80%) can be achieved by AI with frozen-thawed boar spermatozoa using a modified temperature program during freezing, or a novel cryopreservation extender to improve sperm quality (including sperm survivability, motility, membrane status and fertilization ability) after thawing, or a novel sperm infusion method, deep intra uterine insemination. However, these techniques have not yet been used for commercial pig production. The variation in sperm freezability among boars or among ejaculations in an identical boar is one of the main reasons for this problem. In our previous study, it was revealed that some components of seminal plasma have a negative effect on the freezability of boar sperm. One of these factors is bacteria-released endotoxin (lipopolysaccharide: LPS). LPS binds to Toll-like receptor-4 (TLR-4) expressed on the sperm surface, resulting in induction of apoptosis. On the other hand, seminal plasma suppresses cryo-capacitation induced by thawing stress. On the basis of these findings, we designed a novel protocol of AI using frozen-thawed boar sperm. © 2012 Japanese Society of Animal Science.

Induction of instability in water-in-oil-in-water double emulsions by freeze-thaw cycling.

PubMed

Rojas, Edith C; Papadopoulos, Kyriakos D

2007-06-19

Individual water-in-oil-in-water (W1/O/W2) double-emulsion globules loaded with fluorescently labeled bovine serum albumin (FITC-BSA) were optically monitored within cylindrical capillaries during freeze-thaw cycling. Coalescence of internal aqueous droplets (W1) and external aqueous phase (W2), termed external coalescence, was not observed before or during freezing of the oil phase (O). On the other hand, this instability mechanism was readily promoted during thawing. This realization confirms the previously suggested potential of W1/O/W2 double emulsions to trigger release upon oil thawing and demonstrates that it is a direct result of globule breakage through external coalescence. The presented results also identified a threshold in the relative W1 droplet size above which instability occurred, while smaller droplets remained unperturbed and therefore indicate that optimization of the delivery can be achieved by tuning the size of W1 droplets. In addition, we propose a possible explanation for the occurrence of instability during oil thawing and its dependence on the size of W1 droplets. Because this alternative globule-breakage mechanism simply uses temperature increase (solid-to-liquid-phase transition) as external stimulus, W1/O/W2 double-emulsion delivery systems can be easily tailored by choosing an oil phase with the appropriate phase-transition temperature.

Prospective randomized comparison of human oocyte cryopreservation with slow-rate freezing or vitrification.

PubMed

Smith, Gary D; Serafini, Paulo C; Fioravanti, Joyce; Yadid, Isaac; Coslovsky, Marcio; Hassun, Pericles; Alegretti, José Roberto; Motta, Eduardo L

2010-11-01

To compare cryopreservation of mature human oocytes with slow-rate freezing and vitrification and determine which is most efficient at establishing a pregnancy. Prospective randomized. Academically affiliated, private fertility center. Consenting patients with concerns about embryo cryopreservation and more than nine mature oocytes at retrieval were randomized to slow-rate freezing or vitrification of supernumerary (more than nine) oocytes. Oocytes were frozen or vitrified, and upon request oocytes were thawed or warmed, respectively. Oocyte survival, fertilization, embryo development, and clinical pregnancy. Patient use has resulted in 30 thaws and 48 warmings. Women's age at time of cryopreservation was similar. Oocyte survival was significantly higher following vitrification/warming (81%) compared with freezing/thawing (67%). Fertilization was more successful in oocytes vitrified/warmed compared with frozen/thawed. Fertilized oocytes from vitrification/warming had significantly better cleavage rates (84%) compared with freezing/thawing (71%) and resulted in embryos with significantly better morphology. Although similar numbers of embryos were transferred, embryos resulting from vitrified oocytes had significantly enhanced clinical (38%) pregnancy rates compared with embryos resulting from frozen oocyte (13%). Miscarriage and/or spontaneous abortion rates were similar. Our results suggest that vitrification/warming is currently the most efficient means of oocyte cryopreservation in relation to subsequent success in establishing pregnancy. Copyright © 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Influence of time, storage temperature and freeze/thaw cycles on the activity of digestive enzymes from gilthead sea bream (Sparus aurata).

PubMed

Solovyev, Mikhail; Gisbert, Enric

2016-10-01

In this study, we tested the effects of long-term storage (2 years) at -20 °C and short-term storage (several hours) in ice and freeze/thaw cycles on the activities of pancreatic, gastric and intestinal (brush border and cytosolic) digestive enzymes in a teleost fish species. The results revealed a significant lose in activity of pancreatic (trypsin, chymotrypsin, total alkaline proteases and α-amylase) and intestinal cytosolic (leucine-alanine peptidase) enzymes between 140 and 270 days of storage at -20 °C, whereas in contrast, the activity of all the assayed brush border enzymes remained constant during the first 2 years of storage at -20 °C. During short-term storage conditions, the most stable enzymes assayed were those of the enterocytes of the brush border, which did not show any change in activity after being held for 5 h in ice. Five freezing and thawing cycles did not affect the activity of the intestinal brush border enzymes and the cytosolic ones, whereas the activity of trypsin, α-amylase and bile-salt-activated lipase was significantly affected by the number of freezing and thawing cycles. No changes in pepsin activity were found in samples exposed to 1 and 2 freezing and thawing cycles.

Dehydrin gene expression provides an indicator of low temperature and drought stress: transcriptome-based analysis of barley (Hordeum vulgare L.).

PubMed

Tommasini, Livia; Svensson, Jan T; Rodriguez, Edmundo M; Wahid, Abdul; Malatrasi, Marina; Kato, Kenji; Wanamaker, Steve; Resnik, Josh; Close, Timothy J

2008-11-01

Low temperature and drought have major influences on plant growth and productivity. To identify barley genes involved in responses to these stresses and to specifically test the hypothesis that the dehydrin (Dhn) multigene family can serve as an indicator of the entire transcriptome response, we investigated the response of barley cv. Morex to: (1) gradual drought over 21 days and (2) low temperature including chilling, freeze-thaw cycles, and deacclimation over 33 days. We found 4,153 genes that responded to at least one component of these two stress regimes, about one fourth of all genes called "present" under any condition. About 44% (1,822 of 4,153) responded specifically to drought, whereas only 3.8% (158 of 4,153) were chilling specific and 2.8% (119 of 4,153) freeze-thaw specific, with 34.1% responsive to freeze-thaw and drought. The intersection between chilling and drought (31.9%) was somewhat smaller than the intersection between freeze-thaw and drought, implying an element of osmotic stress response to freeze-thaw. About 82.4% of the responsive genes were similar to Arabidopsis genes. The expression of 13 barley Dhn genes mirrored the global clustering of all transcripts, with specific combinations of Dhn genes providing an excellent indicator of each stress response. Data from these studies provide a robust reference data set for abiotic stress.

The Different Calcium+2 Intensity Profile and Quality of Oocyte and Goat Sperms after Cryopreservation

NASA Astrophysics Data System (ADS)

Ciptadi, G.; Rahayu, S.; Fatchiyah; Wahyuningsih, S.; Budiarto, A.; Nasich, M.; Putri, A. R. I.; Mudawamah, M.; Ihsan, M. N.

2018-02-01

This research aims were to study the effect of the oocyte and sperms cryopreservation of Indonesian local goat on the post-thawing quality and profile or characters of Calcium+2 intensity in relating with their fertility capacity. A study was conducted to test the freezing method and post-thawing viability both stock cells stored in the deep freezer and liquid nitrogen (-80°C of vs -196°C). A fertility test of sperms has been conducted through in vitro of sperm quality, while the oocytes cryopreserved test was done by in vitro maturation (IVM) rate (%). The profile of Calcium 2+ was performed and analysis by Confocal Laser Scanned Microscope (CLSM). The result showed that IVM rate of goat oocyte is considered lower when cryopreserved in -80°C than in -196°C. Meanwhile, sperm is considered having a good quality in 2 methods of cryopreservation with post-thawing motility > 40 % (SNI 2014). There is an important difference between Calcium intensity of fresh and post-thawing both for oocyte and spermatozoa. Calcium +2 profiles is varied individually on the peak of intensity, but it considered expressed the same profile of each fresh and post-thawing cell. In vitro fertilization test need to be performed to complete the viability and fertility competence of these sperm and oocyte freezing stocks.

Effect of Repeated Freezing and Thawing on 18 Clinical Chemistry Analytes in Rat Serum

PubMed Central

Kale, Vijay P; Patel, Sweta G; Gunjal, Prashant S; Wakchaure, Santosh U; Sundar, Rajesh S; Ranvir, Ramchandra K; Jain, Mukul R

2012-01-01

In a preclinical research laboratory, using serum samples that have been frozen and thawed repeatedly is sometimes unavoidable when needing to confirm previous results or perform additional analysis. Here we determined the effects of multiple cycles of refrigeration or freezing and thawing of rat serum at 3 temperature conditions for different storage times on clinical chemistry analytes. Serum samples obtained from adult Wistar rats were stored at 2 to 8 °C and −10 to −20 °C for as long as 72 h and at −70 °C for as long as 30 d. At different time points (24, 48, and 72 h for samples stored at 2 to 8 °C or −10 to −20 °C and 1, 7, and 30 d for samples stored at −70 °C), the samples were brought to room temperature, analyzed, and then stored again at the designated temperature. The results obtained after each storage cycle were compared with those obtained from the initial analysis of fresh samples. Of the 18 serum analytes evaluated, 14 were stable without significant changes, even after 3 freeze–thaw cycles at the tested temperature ranges. Results from this study will help researchers working with rat serum to interpret the biochemical data obtained from serum samples that have been frozen and thawed repeatedly. PMID:23043814

Motility and fertilizing ability of cryopreserved Caspian brown trout (Salmo trutta caspius) sperm: Effect of post-thaw storage time and different sperm-to-egg ratios.

PubMed

Golshahi, Karim; Shabani, Nariman; Aramli, Mohammad Sadegh; Noori, Elnaz

2015-10-01

This study was designed to test the effect of post-thaw storage time on sperm motility parameters of Caspian brown trout (n=7). Furthermore, we investigated the effect of sperm-to-egg ratios of 100,000:1, 300,000:1 and 600,000:1 on fertility of cryopreserved Caspian brown semen. Quality was assessed by measuring sperm motility parameters and fertilization rates at the eyed and hatching stages. The percentage of post-thawed sperm motility, curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH) were not affected by 60 min of storage, whereas a decrease in straight line velocity (VSL), average path velocity (VAP) and linearity (LIN) were found in cryopreserved semen. Thus, the cryopreserved sperm of Caspian brown trout could be stored up to 60 min without loss of the percentage of sperm motility. The fertilization rate was not affected by 60 min of post-thaw storage and was over 70% for sperm-to-egg ratios of both 300,000 and 600,000:1. To our knowledge, this study is the first to report the high post-thaw fertilization ability of Caspian brown trout semen at a sperm-to-egg ratio as low as 300,000:1. This procedure after scaling up can be recommended for routine Caspian brown trout sperm cryopreservation. Copyright © 2015 Elsevier Inc. All rights reserved.

Cross-tolerance between osmotic and freeze-thaw stress in microbial assemblages from temperate lakes.

PubMed

Wilson, Sandra L; Frazer, Corey; Cumming, Brian F; Nuin, Paulo A S; Walker, Virginia K

2012-11-01

Osmotic stress can accompany increases in solute concentrations because of freezing or high-salt environments. Consequently, microorganisms from environments with a high-osmotic potential may exhibit cross-tolerance to freeze stress. To test this hypothesis, enrichments derived from the sediment and water of temperate lakes with a range of salt concentrations were subjected to multiple freeze-thaw cycles. Surviving isolates were identified and metagenomes were sampled prior to and following selection. Enrichments from alkali lakes were typically the most freeze-thaw resistant with only 100-fold losses in cell viability, and those from freshwater lakes were most susceptible, with cell numbers reduced at least 100,000-fold. Metagenomic analysis suggested that selection reduced assemblage diversity more in freshwater samples than in those from saline lakes. Survivors included known psychro-, halo- and alkali-tolerant bacteria. Characterization of freeze-thaw-resistant isolates from brine and alkali lakes showed that few isolates had ice-associating activities such as antifreeze or ice nucleation properties. However, all brine- and alkali-derived isolates had high intracellular levels of osmolytes and/or appeared more likely to form biofilms. Conversely, these phenotypes were infrequent amongst the freshwater-derived isolates. These observations are consistent with microbial cross-tolerance between osmotic and freeze-thaw stresses. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Cryopreservation of bull spermatozoa alters the perinuclear theca.

PubMed

Martínez, Carmen Omega; Juárez-Mosqueda, María de Lourdes; Hernández, Jorge; Valencia, Javier

2006-11-01

The perinuclear theca (PT) is involved in several important sperm functions leading to fertilization. The objective of this study was to investigate the effect of cryopreservation of bull spermatozoa on the integrity of the PT and the relationship between PT integrity and semen characteristics. Semen from seven bulls was evaluated before and after cryopreservation, comparing the integrity of the plasma membrane (hypo-osmotic test), percentage of live and dead spermatozoa (triple stain), acrosome integrity (triple stain) and the integrity of the PT (negative stain by electron microscopy). Cryopreservation of bull semen caused substantial damage to the PT; the proportion of spermatozoa with a damaged PT was 15.2% versus 52.5% (P<0.05) in fresh versus frozen-thawed spermatozoa, respectively. Furthermore, on average, 67.4% (range, 64-72%) of fresh spermatozoa were live, compared to 53.1% (range, 49-58%) for frozen-thawed spermatozoa; there was an inverse correlation between the percentage of live spermatozoa and the percentage with damage to the PT. Although 59.1% of frozen-thawed spermatozoa had an intact acrosome, only 43.7% of them still remained alive. In frozen-thawed semen, there was a high correlation (r=0.69) between live spermatozoa with an intact acrosome and spermatozoa that maintained an intact PT. In conclusion, freezing/thawing of bull spermatozoa altered the PT and maintaining PT integrity may be necessary to maintain acrosome integrity.

The origin of shallow landslides in Moravia (Czech Republic) in the spring of 2006

NASA Astrophysics Data System (ADS)

Bíl, Michal; Müller, Ivo

2008-07-01

At the end of March 2006, the Czech Republic (CZ) witnessed a fast thawing of an unusually thick snow cover in conjunction with massive rainfall. Most watercourses suffered floods, and more than 90 shallow landslides occurred in the Moravian region of Eastern CZ, primarily in non-forested areas. This region, geologically part of the Outer Western Carpathians, is prone to landslides because the bedrock is highly erodible Mesozoic and Tertiary flysch. The available meteorological data (depth of snow, water equivalent of the snow, cumulative rainfall, air and soil temperatures) from five local weather stations were used to construct indices quantitatively describing the snow thaw. Among these, the Total Cumulative Precipitation ( TCP) combines the amount of water from both thawing snow and rainfall. This concurrence of rain and runoff from snow melt was the decisive factor in triggering the landslides in the spring. The TCP index was applied to data of snow thaw periods for the last 20 years, when no landslides were recorded. This was to establish the safe threshold of TCP without landslides. The calculated safe threshold value for the region is ca. 100 mm of water delivered to the soil during the spring thaw (corresponding to ca. 11 mm day - 1 ). In 2006, 10% of the landslides occurred under or at 100 mm of TCP. The upper value of 155 mm covered all of the landslides.

Dynamics and characteristics of soil temperature and moisture of active layer in central Tibetan Plateau

NASA Astrophysics Data System (ADS)

Zhao, L.; Hu, G.; Wu, X.; Tian, L.

2017-12-01

Research on the hydrothermal properties of active layer during the thawing and freezing processes was considered as a key question to revealing the heat and moisture exchanges between permafrost and atmosphere. The characteristics of freezing and thawing processes at Tanggula (TGL) site in permafrost regions on the Tibetan Plateau, the results revealed that the depth of daily soil temperature transmission was about 40 cm shallower during thawing period than that during the freezing period. Soil warming process at the depth above 140 cm was slower than the cooling process, whereas they were close below 140 cm depth. Moreover, the hydro-thermal properties differed significantly among different stages. Precipitation caused an obviously increase in soil moisture at 0-20 cm depth. The vertical distribution of soil moisture could be divided into two main zones: less than 12% in the freeze state and greater than 12% in the thaw state. In addition, coupling of moisture and heat during the freezing and thawing processes also showed that soil temperature decreased faster than soil moisture during the freezing process. At the freezing stage, soil moisture exhibited an exponential relationship with the absolute soil temperature. Energy consumed for water-ice conversion during the freezing process was 149.83 MJ/m2 and 141.22 MJ/m2 in 2011 and 2012, respectively, which was estimated by the soil moisture variation.

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

DOT National Transportation Integrated Search

2013-12-01

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

Vulnerability and feedbacks of permafrost to climate change

Treesearch

Guido Grosse; Vladimir Romanovsky; Torre Jorgenson; Katey Walter Anthony; Jerry Brown; Pier Paul Overduin; Alfred Wegener

2011-01-01

The effects of permafrost degradation on terrestrial and offshore environments in polar regions and on the Earth's atmosphere are significant. Field-based observations, remote sensing, and modeling document regional warming and thawing of permafrost. However, major research questions regarding vulnerability of permafrost to thawing, the projected decline in...

Three-dimensional reconstruction of frozen and thawed plant tissues from microscopic images

USDA-ARS?s Scientific Manuscript database

Histological analysis of frozen and thawed plants has been conducted for many years but the observation of individual sections only provides a 2 dimensional representation of a 3 dimensional phenomenon. Most techniques for viewing internal plant structure in 3 dimensions is either low in resolution...

Physiological responses of planting frozen and thawed Douglas-fir seedlings

Treesearch

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...

Post-thaw motility of frozen boar sperm does not predict success with in vitro fertilization

USDA-ARS?s Scientific Manuscript database

Using cryopreserved boar sperm rather than liquid semen for in vitro fertilization (IVF) allows improved IVF consistency. However, cryopreservation of boar sperm results in reduced post-thaw motility, fertilization and embryo development. Boars are often screened on an individual basis prior to use ...

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

DOT National Transportation Integrated Search

2013-12-01

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

Damage assessment, characterization, and modeling for enhanced design of concrete bridge decks in cold regions : [project brief].

DOT National Transportation Integrated Search

2015-07-01

Freeze-thaw and fatigue-type loading processes degrade concrete materials and reduce the load carrying capacity of concrete decks. Damage to concrete decks is caused by the formation of cracks and micro-cracks during fatigue and freeze-thaw cycles. T...

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

DOT National Transportation Integrated Search

2016-11-22

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

Highway concrete pavement technology development and testing. Volume 2, field evaluation of strategic highway research program (SHRP) C-203 test sites (freeze-thaw resistance)

DOT National Transportation Integrated Search

2006-08-01

Field test sections were constructed during 1992 as part of the Strategic Highway Research Program (SHRP) investigation of the frost resistance of concrete. The first freeze-thaw-related deterioration expected for pavement concrete exposed to de-icin...

21 CFR 864.9145 - Processing system for frozen blood.

Code of Federal Regulations, 2012 CFR

2012-04-01

... system for frozen blood is a device used to glycerolize red blood cells prior to freezing to minimize... thawing of red blood cells and to deglycerolize and wash thawed cells for subsequent reinfusion. (b... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Processing system for frozen blood. 864.9145...

21 CFR 864.9145 - Processing system for frozen blood.

Code of Federal Regulations, 2014 CFR

2014-04-01

... system for frozen blood is a device used to glycerolize red blood cells prior to freezing to minimize... thawing of red blood cells and to deglycerolize and wash thawed cells for subsequent reinfusion. (b... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Processing system for frozen blood. 864.9145...

21 CFR 864.9145 - Processing system for frozen blood.

Code of Federal Regulations, 2010 CFR

2010-04-01

... system for frozen blood is a device used to glycerolize red blood cells prior to freezing to minimize... thawing of red blood cells and to deglycerolize and wash thawed cells for subsequent reinfusion. (b... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Processing system for frozen blood. 864.9145...

21 CFR 864.9145 - Processing system for frozen blood.

Code of Federal Regulations, 2013 CFR

2013-04-01

... system for frozen blood is a device used to glycerolize red blood cells prior to freezing to minimize... thawing of red blood cells and to deglycerolize and wash thawed cells for subsequent reinfusion. (b... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Processing system for frozen blood. 864.9145...

21 CFR 864.9145 - Processing system for frozen blood.

Code of Federal Regulations, 2011 CFR

2011-04-01

... system for frozen blood is a device used to glycerolize red blood cells prior to freezing to minimize... thawing of red blood cells and to deglycerolize and wash thawed cells for subsequent reinfusion. (b... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Processing system for frozen blood. 864.9145...

Wooden breast condition results in reduced thaw loss in frozen-thawed broiler breast fillets

USDA-ARS?s Scientific Manuscript database

Wooden breast condition (WBC) is an emerging quality issue with broiler breast meat that significantly affects both raw and cooked meat properties. The objective of this study was to investigate the effects of WBC on meat water-holding capacity (WHC) estimated with different methods. Broiler breast ...

High risk of permafrost thaw

Treesearch

E.A.G. Schuur; B.W. Abbott; W.B. Bowden; V. Brovkin; P. Camill; J.P. Canadell; F.S. Chapin; T.R. Christensen; J.P. Chanton; P. Ciais; P.M. Crill; B.T. Crosby; C.I. Czimczik; G. Grosse; D.J. Hayes; G. Hugelius; J.D. Jastrow; T. Kleinen; C.D. Koven; G. Krinner; P. Kuhry; D.M. Lawrence; S.M. Natali; C.L. Ping; A. Rinke; W.J. Riley; V.E. Romanovsky; A.B.K. Sannel; C. Schadel; K. Schaefer; Z.M. Subin; C. Tarnocai; M. Turetsky; K. M. Walter-Anthony; C.J. Wilson; S.A. Zimov

2011-01-01

Arctic temperatures are rising fast, and permafrost is thawing. Carbon released into the atmosphere from permafrost soils will accelerate climate change, but the magnitude of this effect remains highly uncertain. Our collective estimate is that carbon will be released more quickly than models suggest, and at levels that are cause for serious concern. We calculate that...

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

PubMed Central

Braslavsky, Ido

2018-01-01

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

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

PubMed

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.

Human oocyte cryopreservation and the fate of cortical granules.

PubMed

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.

The cryoprotective effect of trehalose supplementation on boar spermatozoa quality.

PubMed

Hu, J-H; Li, Q-W; Jiang, Z-L; Yang, H; Zhang, S-S; Zhao, H-W

2009-08-01

In order to improve boar sperm quality during frozen-thawed process, the influence of the presence of trehalose on success of cryopreservation of boar sperm were investigated. We evaluated freeze-thawing tolerance of boar spermatozoa in a base cooling extender with the addition of different trehalose concentrations (0, 25, 50, 100 and 200 mm), and try to determine the optimum concentration of trehalose. We chose sperm motility, mitochondrial activity, acrosome integrity and membrane integrity as parameters to evaluate cryopreservation capacity of boar spermatozoa. We obtained the best results for 100 mm trehalose-supplemented extenders, with values of 49.89% for motility, 44.69% for mitochondrial activity, 66.52% for acrosome integrity and 44.61% for membrane integrity, while freeze-thawing tolerance diminished significantly for 200 . The synergic effect of trehalose and glycerol resulted in better cryosurvival of boar spermatozoa than that of a single cryoprotectant. In conclusion, when trehalose-supplementation was added up to 100 mm, trehalose confers a greater cryoprotective capacity to the extender, and the sperm motility, mitochondrial activity, membrane integrity and acrosome integrity parameters were significantly improved during frozen-thawed process.

Using carbon isotopes of methane from porewater to understand methane emissions across a permafrost thaw gradient

NASA Astrophysics Data System (ADS)

Varner, R. K.; McCalley, C. K.; Clarizia, P. E.; Verbeke, B. A.; Werner, S. L.; Burke, S. A.; Malhotra, A.; Rocci, K.

2016-12-01

Methane (CH4) emissions from high latitude ecosystems are controlled in part by the presence/absence of permafrost and concomitant modifications in vegetation composition. Rapid transitions in habitat impact CH4 emissions both by changing the moisture regime as well as the production and emission pathways. Measurement of the isotopic composition of CH4 in porewater in these thawed ecosystems can indicate shifts in production pathways of CH4. We measured CH4 and carbon dioxide (CO2) emission, belowground CH4 concentration and 13CH4 of porewater, vegetative type, and vascular greenness area (VGA) along a thaw gradient during summers 2012-2016 in Stordalen Mire, Sweden. Concentrations of CH4 belowground showed positive correlation with aboveground emissions. Carbon isotopic signatures of CH4 varied varied between sites with more hydrogenotrophic signatures in sites dominated by Sphagnum spp. and acetate fermentation signatures in sedge dominated sites (Carex and Eriophorum spp.). These data indicate that these ecosystems transition from thaw, their 13CH4 emissions will change and therefore need to be accounted for in global atmospheric budgets and models.

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

PubMed

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.

Observations using Phosphorus-31 nuclear magnetic resonance (31P-NMR) of structural changes in freeze-thawed hen egg yolk.

PubMed

Wakamatsu, Hiroki; Handa, Akihiro; Chiba, Kazuhiro

2018-04-01

Hen egg yolk (EY) has a complicated structure consisting of lipids and proteins, and its structure is deeply related with its functional properties. 31 P-NMR is an efficient technique to non-destructively detect the dynamic behaviour of phospholipids, the main component of bio-membranes. We determined conditions for measuring the 31 P NMR spectra of EY and identified the components. 31 P-NMR was used to detect phosvitin, inorganic phosphate, and lipoprotein as well as structural changes such as granule collapse and freeze-thaw denaturation as signal changes. Freeze-thaw denaturation generated a new denaturation peak. We separated aggregates of LDL from freeze-thawed plasma using centrifugation. TEM and 31 P-NMR observations revealed that the denaturation peak corresponded to LDL aggregates. The 31 P-NMR spectra suggested the formation of multiple forms of LDL aggregates in which the head groups of phospholipid molecules adopt a face-to-face orientation, similar to that observed following the flocculation of lipoproteins or in the lamellar-like structures of phospholipids. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

PubMed

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

2015-10-01

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

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

PubMed

Bahari, Liat; Bein, Amir; Yashunsky, Victor; Braslavsky, Ido

2018-01-01

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

Potency and stability of frozen urokinase solutions in syringes.

PubMed

Dedrick, Stephen C; Ramirez-Rico, José

2004-08-01

The stability and potency of frozen urokinase solutions in syringes were studied. To determine the stability and potency of compounded urokinase dilutions after multiple freeze-thaw cycles, a total of 160 syringes containing five urokinase concentrations (2,500, 5,000, 7,500, 12,500, and 25,000 IU/mL) were prepared. For each of the five concentrations tested, two syringes per concentration were reserved for baseline testing. The remaining 150 syringes were frozen at -30 degrees C. After 7 days, half of the syringes (group 1) were thawed at room temperature, tested, and left at room temperature for 12 hours before refreezing. The other half of the syringes (group 2) were kept frozen for 30 days. Thirty days after initial compounding, all syringes were thawed, and the samples' urokinase potency, pH, and physical appearance were evaluated. Syringes were visually inspected for color, clarity, and precipitation. Descriptive statistics were computed for each concentration group and testing day. The compounded dilutions were stable under each experimental condition, with no physical deterioration or loss of in vitro potency after two freeze-thaw cycles. The reduced waste associated with the ability to refreeze unused urokinase could substantially lower the cost of procedures such as thrombolysis after intraventricular hemorrhage and catheter clearance by as much as 95%. Dilutions of urokinase 2,500-25,000 IU/mL were stable in single-use syringes after being frozen for 7 days, thawed, and refrozen for another 23 days.

Hydrologic impacts of thawing permafrost—A review

USGS Publications Warehouse

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.

l-Cysteine improves antioxidant enzyme activity, post-thaw quality and fertility of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa.

PubMed

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 < 0.05) at pre-freezing and post-thawing in extender containing 2.0 mm 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 < 0.05). The fertility rates (59 versus 43%) were higher (P < 0.05) in buffaloes inseminated with doses containing 2.0 mm of 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.

Leveraging Subsidence in Permafrost with Remotely Sensed Active Layer Thickness (ReSALT) Products

NASA Astrophysics Data System (ADS)

Schaefer, K. M.; Chen, A.; Chen, J.; Chen, R. H.; Liu, L.; Michaelides, R. J.; Moghaddam, M.; Parsekian, A.; Tabatabaeenejad, A.; Thompson, J. A.; Zebker, H. A.; Meyer, F. J.

2017-12-01

The Remotely Sensed Active Layer Thickness (ReSALT) product uses the Interferometric Synthetic Aperture Radar (InSAR) technique to measure ground subsidence in permafrost regions. Seasonal subsidence results from the expansion of soil water into ice as the surface soil or active layer freezes and thaws each year. Subsidence trends result from large-scale thaw of permafrost and from the melting and subsequent drainage of excess ground ice in permafrost-affected soils. The attached figure shows the 2006-2010 average seasonal subsidence from ReSALT around Barrow, Alaska. The average active layer thickness (the maximum surface thaw depth during summer) is 30-40 cm, resulting in an average seasonal subsidence of 1-3 cm. Analysis of the seasonal subsidence and subsidence trends provides valuable insights into important permafrost processes, such as the freeze/thaw of the active layer, large-scale thawing due to climate change, the impact of fire, and infrastructure vulnerability. ReSALT supports the Arctic-Boreal Vulnerability Experiment (ABoVE) field campaign in Alaska and northwest Canada and is a precursor for a potential NASA-ISRO Synthetic Aperture Radar (NISAR) product. ReSALT includes uncertainties for all parameters and is validated against in situ measurements from the Circumpolar Active Layer Monitoring (CALM) network, Ground Penetrating Radar and mechanical probe measurements. Here we present examples of ReSALT products in Alaska to highlight the untapped potential of the InSAR technique to understand permafrost dynamics, with a strong emphasis on the underlying processes that drive the subsidence.

Implications of the pH and temperature of diluted, cooled boar semen on fresh and frozen-thawed sperm motility characteristics.

PubMed

Purdy, P H; Tharp, N; Stewart, T; Spiller, S F; Blackburn, H D

2010-10-15

Boar semen is typically collected, diluted and cooled for AI use over numerous days, or frozen immediately after shipping to capable laboratories. The storage temperature and pH of the diluted, cooled boar semen could influence the fertility of boar sperm. Therefore, the purpose of this study was to determine the effects of pH and storage temperature on fresh and frozen-thawed boar sperm motility end points. Semen samples (n = 199) were collected, diluted, cooled and shipped overnight to the National Animal Germplasm Program laboratory for freezing and analysis from four boar stud facilities. The temperature, pH and motility characteristics, determined using computer automated semen analysis, were measured at arrival. Samples were then cryopreserved and post-thaw motility determined. The commercial stud was a significant source of variation for mean semen temperature and pH, as well as total and progressive motility, and numerous other sperm motility characteristics. Based on multiple regression analysis, pH was not a significant source of variation for fresh or frozen-thawed boar sperm motility end points. However, significant models were derived which demonstrated that storage temperature, boar, and the commercial stud influenced sperm motility end points and the potential success for surviving cryopreservation. We inferred that maintaining cooled boar semen at approximately 16 °C during storage will result in higher fresh and frozen-thawed boar sperm quality, which should result in greater fertility. Copyright © 2010 Elsevier Inc. All rights reserved.

Reduced glutathione enhances fertility of frozen/thawed C57BL/6 mouse sperm after exposure to methyl-beta-cyclodextrin.

PubMed

Takeo, Toru; Nakagata, Naomi

2011-11-01

Sperm cryopreservation is useful for the effective storage of genomic resources derived from genetically engineered mice. However, freezing the sperm of C57BL/6 mice, the most commonly used genetic background for genetically engineered mice, considerably reduces its fertility. We previously reported that methyl-beta-cyclodextrin dramatically improved the fertility of frozen/thawed C57BL/6 mouse sperm. Recently, it was reported that exposing sperm to reduced glutathione may alleviate oxidative stress in frozen/thawed mouse sperm, thereby enhancing in vitro fertilization (IVF); however, the mechanism underlying this effect is poorly understood. In the present study, we examined the combined effects of methyl-beta-cyclodextrin and reduced glutathione on the fertilization rate of IVF with frozen/thawed C57BL/6 mouse sperm and the characteristic changes in the zona pellucida induced by reduced glutathione. Adding reduced glutathione to the fertilization medium increased the fertilization rate. Methyl-beta-cyclodextrin and reduced glutathione independently increased fertilization rates, and their combination produced the strongest effect. We found that reduced glutathione increased the amount of free thiols in the zona pellucida and promoted zona pellucida enlargement. Finally, 2-cell embryos produced by IVF with the addition of reduced glutathione developed normally and produced live offspring. In summary, we have established a novel IVF method using methyl-beta-cyclodextrin during sperm preincubation and reduced glutathione during the IVF procedure to enhance fertility of frozen/thawed C57BL/6 mouse sperm.

Assessing the toxic effects of DMSO on cord blood to determine exposure time limits and the optimum concentration for cryopreservation.

PubMed

Fry, L J; Querol, S; Gomez, S G; McArdle, S; Rees, R; Madrigal, J A

2015-08-01

Advantages of using cord blood (CB) over other sources of haematopoietic progenitor cells, such as bone marrow, include the ability to cryopreserve and bank the samples until requested for a transplant. Cryopreservation requires the addition of a cryoprotectant to prevent the formation of intracellular ice during freezing. Dimethyl sulphoxide (DMSO) is commonly used at a concentration of 10% (v/v); however, there is evidence to suggest this chemical is toxic to cells as well as to patients after infusion. The toxic effects of DMSO were assessed through cell viability and in vitro functional assays in fresh and post-thaw CB samples before determining the maximum exposure time and optimal concentration for cryopreservation. A dose-dependent toxicity of DMSO was observed in fresh samples with 40% removing all viable and functional haematopoietic progenitor cells (HPC). In fresh and post-thaw analysis, minimal toxic effect was observed when cryopreservation was delayed for up to 1 h after 10% DMSO addition. After thawing, DMSO washout was superior to dilution or unmanipulated when maintained for long periods (advantage observed 1 h after thawing). Finally, the optimum concentration for cryopreserving CB was found to be 7.5 to 10% with detrimental effects observed outside of this range. These results support the use of 7.5-10% as the optimal DMSO concentration and the maximum exposure time should be limited to <1 h prior to freezing and 30 min post-thaw. © 2015 International Society of Blood Transfusion.

Encapsulation of Nucleic Acids into Giant Unilamellar Vesicles by Freeze-Thaw: a Way Protocells May Form

NASA Astrophysics Data System (ADS)

Qiao, Hai; Hu, Na; Bai, Jin; Ren, Lili; Liu, Qing; Fang, Liaoqiong; Wang, Zhibiao

2017-12-01

Protocells are believed to consist of a lipid membrane and encapsulated nucleic acid. As the lipid membrane is impermeable to macromolecules like nucleic acids, the processes by which nucleic acids become encapsulated inside lipid membrane compartments are still unknown. In this paper, a freeze-thaw method was modified and applied to giant unilamellar vesicles (GUVs) and deoxyribonucleic acid (DNA) in mixed solution resulting in the efficient encapsulation of 6.4 kb plasmid DNA and similar length linear DNA into GUVs. The mechanism of encapsulation was followed by observing the effect of freeze-thaw temperatures on GUV morphological change, DNA encapsulation and ice crystal formation, and analyzing their correlation. Following ice crystal formation, the shape of spherical GUVs was altered and membrane integrity was damaged and this was found to be a necessary condition for encapsulation. Heating alone had no effects on DNA encapsulation, but was helpful for restoring the spherical shape and membrane integrity of GUVs damaged during freezing. These results suggested that freeze-thaw could promote the encapsulation of DNA into GUVs by a mechanism: the vesicle membrane was breached by ice crystal formation during freezing, DNA entered into damaged GUVs through these membrane gaps and was encapsulated after the membrane was resealed during the thawing process. The process described herein therefore describes a simple way for the encapsulation of nucleic acids and potentially other macromolecules into lipid vesicles, a process by which early protocells might have formed.

In vitro survival of fresh and frozen/thawed bovine demi-embryos.

PubMed

Lucas-Hahn, A; Niemann, H

1991-10-01

Three experiments were conducted to investigate the effects of type of culture medium in freshly bisected bovine embryos and the effects of agar embedding and of 1.2 propanediol (PROH) as the cryoprotectant in frozen/thawed bisected bovine embryos on development in vitro. A total of 265 bovine embryos were used as controls or were microsurgically bisected and were cultured in vitro for 48 hours and development was determined 24 and 48 hours after the onset of culture. Whitten's medium supported more (P<0.05) intact and demi-embryos to grow to expanded blastocysts (92.9 and 73.1%, respectively) compared with Ham's F10 (43.8 and 26.3%, respectively) and PBS (53.8 and 12.5%, respectively). Embedding in agar and culture in Whitten's medium resulted in a higher (P<0.05) percentage of in vitro development of frozen/thawed demi-embryos after 24 hours than the freezing of nonembedded demi-embryos (44.1 versus 19.6%, respectively). This difference disappeared, however, after a 48 hours culture period (17.6 versus 11.8%, respectively). Following freezing in PROH, survival rates of 40 and 28%, respectively after 24 hours of culture were obtained for intact and demi-embryos. The respective percentages after 48 hours were 8.6 and 16%. Since neither embedding in agar nor the use of PROH as the cryoprotectant resulted in high survival rates of frozen/thawed demi-embryos in vitro, new freezing procedures are needed to overcome the sensitivity of demi-embryos to freezing and thawing.

Development of frozen-thawed demi-embryos and production of identical twin calves of different ages.

PubMed

Seike, N; Sakai, M; Kanagawa, H

1991-02-01

The percentages of morphologically transferable embryos obtained from frozen-thawed demi-embryos which were embedded with or without agar, and from those with or without zonae pellucidae were 26.3% (5/19), 36.4% (8/22), 39.5% (15/38) and 40.0% (22/55), respectively. No significant differences were observed between these groups. Development to calves of frozen-thawed demi-embryos with or without zonae pellucidae was 25.0% (3/12) and 26.7% (4/15), respectively. There was also no significant difference between them. On the trial for production of identical twin calves of different ages, the pregnancy rates of fresh and frozen demi-embryos after transfer were 69.2% (9/13) and 11.1% (1/9), respectively. Out of 13 fresh demi-embryos and 9 frozen demi-embryos transferred, only one pair of identical twin male calves of different ages were produced. This frozen-thawed demi-embryo was stored for 43 days in liquid nitrogen before thawing and transfer. These twin calves were confirmed to be identical by blood typing. Although these calves had different birth dates, their growth rates indicated similar developmental patterns. We suggest that it is possible to produce identical twin calves of different ages. This possibility would be useful for predicting the sex, milk producing ability and progeny test of a pair of demi-embryos before a decision to transfer the other half of a pair is made.

Incipient balancing selection through adaptive loss of aquaporins in natural Saccharomyces cerevisiae populations.

PubMed

Will, Jessica L; Kim, Hyun Seok; Clarke, Jessica; Painter, John C; Fay, Justin C; Gasch, Audrey P

2010-04-01

A major goal in evolutionary biology is to understand how adaptive evolution has influenced natural variation, but identifying loci subject to positive selection has been a challenge. Here we present the adaptive loss of a pair of paralogous genes in specific Saccharomyces cerevisiae subpopulations. We mapped natural variation in freeze-thaw tolerance to two water transporters, AQY1 and AQY2, previously implicated in freeze-thaw survival. However, whereas freeze-thaw-tolerant strains harbor functional aquaporin genes, the set of sensitive strains lost aquaporin function at least 6 independent times. Several genomic signatures at AQY1 and/or AQY2 reveal low variation surrounding these loci within strains of the same haplotype, but high variation between strain groups. This is consistent with recent adaptive loss of aquaporins in subgroups of strains, leading to incipient balancing selection. We show that, although aquaporins are critical for surviving freeze-thaw stress, loss of both genes provides a major fitness advantage on high-sugar substrates common to many strains' natural niche. Strikingly, strains with non-functional alleles have also lost the ancestral requirement for aquaporins during spore formation. Thus, the antagonistic effect of aquaporin function-providing an advantage in freeze-thaw tolerance but a fitness defect for growth in high-sugar environments-contributes to the maintenance of both functional and nonfunctional alleles in S. cerevisiae. This work also shows that gene loss through multiple missense and nonsense mutations, hallmarks of pseudogenization presumed to emerge after loss of constraint, can arise through positive selection.

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

PubMed

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.

Comparison of CH4 Emission and CO2 Exchange Between 2013 and 2014 in a Subarctic Peatland

NASA Astrophysics Data System (ADS)

Clarizia, P. E.; Verbeke, B. A.; McCalley, C. K.; Werner, S. L.; Malhotra, A.; Burke, S. A.; Crill, P. M.; Varner, R. K.

2014-12-01

One of the major concerns with climate change is the potential feedback from the emission of greenhouse gases, carbon dioxide (CO2) and methane (CH4), from high latitude thawing organic soils. With increasing temperatures in Arctic regions, thawing permafrost palsas transition to wetter sedge-dominated wetlands, which account for 20-39% of global atmospheric CH4 burden. This rapid change in habitat raises the following question: how do CO2 exchange rates and CH4 emissions change along a gradient of permafrost thaw and what is the interannual variability in these fluxes? To address this question, we measured CO2 exchange, CH4 flux, vegetative type and vascular green area (VGA) along a thaw gradient during July of 2013 and 2014 in Stordalen Mire, Sweden. Environmental variables showed that 2013 and 2014 were climatically different; higher photosynthetically active radiation (PAR) and measurements of water table level and temperature showed that 2014 was warmer and drier than 2013. Warmer conditions led to higher rates of respiration and gross primary productivity (GPP), with the largest increases observed in the palsa sites, likely due to an increase in mean temperature. Methane fluxes showed a less consistent response to climate differences between years, fluxes were higher in 2014 in the mostly inundated Eriophorum angustifolium dominated site and lower in the drier Sphagnum and Eriophorum vaginatum dominated sites. Results of this study highlight the need for accounting for interannual variability when predicting greenhouse gas emissions during permafrost thaw.

Cavitation and water fluxes driven by ice water potential in Juglans regia during freeze–thaw cycles

PubMed Central

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

2016-01-01

Freeze–thaw cycles induce major hydraulic changes due to liquid-to-ice transition within tree stems. The very low water potential at the ice–liquid interface is crucial as it may cause lysis of living cells as well as water fluxes and embolism in sap conduits, which impacts whole tree–water relations. We investigated water fluxes induced by ice formation during freeze–thaw cycles in Juglans regia L. stems using four non-invasive and complementary approaches: a microdendrometer, magnetic resonance imaging, X-ray microtomography, and ultrasonic acoustic emissions analysis. When the temperature dropped, ice nucleation occurred, probably in the cambium or pith areas, inducing high water potential gradients within the stem. The water was therefore redistributed within the stem toward the ice front. We could thus observe dehydration of the bark’s living cells leading to drastic shrinkage of this tissue, as well as high tension within wood conduits reaching the cavitation threshold in sap vessels. Ultrasonic emissions, which were strictly emitted only during freezing, indicated cavitation events (i.e. bubble formation) following ice formation in the xylem sap. However, embolism formation (i.e. bubble expansion) in stems was observed only on thawing via X-ray microtomography for the first time on the same sample. Ultrasonic emissions were detected during freezing and were not directly related to embolism formation. These results provide new insights into the complex process and dynamics of water movements and ice formation during freeze–thaw cycles in tree stems. PMID:26585223

Sensitivity of airborne geophysical data to sublacustrine permafrost thaw

NASA Astrophysics Data System (ADS)

Minsley, B. J.; Wellman, T. P.; Walvoord, M. A.; Revil, A.

2014-12-01

A coupled hydrogeophysical forward and inverse modeling approach is developed to illustrate the ability of frequency-domain airborne electromagnetic (AEM) data to characterize subsurface physical properties associated with sublacustrine permafrost thaw during lake talik formation. Several scenarios are evaluated that consider the response to variable hydrologic forcing from different lake depths and hydrologic gradients. The model includes a physical property relationship that connects the dynamic distribution of subsurface electrical resistivity based on lithology as well as ice-saturation and temperature outputs from the SUTRA groundwater simulator with freeze/thaw physics. Electrical resistivity models are used to simulate AEM data in order to explore the sensitivity of geophysical observations to permafrost thaw. Simulations of sublacustrine talik formation over a 1000 year period modeled after conditions found in the Yukon Flats, Alaska, are evaluated. Synthetic geophysical data are analyzed with a Bayesian Markov chain Monte Carlo algorithm that provides a probabilistic assessment of geophysical model uncertainty and resolution. Major lithological and permafrost features are well resolved in the examples considered. The subtle geometry of partial ice-saturation beneath lakes during talik formation cannot be resolved using AEM data, but the gross characteristics of sub-lake resistivity models reflect bulk changes in ice content and can be used to determine the presence of a talik. A final example compares AEM and ground-based electromagnetic responses for their ability to resolve shallow permafrost and thaw features in the upper 1-2 m below ground.

Soil data from fire and permafrost-thaw chronosequences in upland Picea mariana stands near Hess Creek and Tok, interior Alaska

USGS Publications Warehouse

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.

Single-layer centrifugation through PureSperm® 80 selects improved quality spermatozoa from frozen-thawed dog semen.

PubMed

Dorado, J; Alcaraz, L; Gálvez, M J; Acha, D; Ortiz, I; Urbano, M; Hidalgo, M

2013-08-01

The aim of this study was to investigate whether single-layer centrifugation (SLC) with PureSperm® 80 could select good quality spermatozoa, including those with specific motility patterns, from doses of frozen dog semen. Semen from 5 dogs was collected and cryopreserved following a standard protocol. After thawing, semen samples were divided into two aliquots: one of them was used as control and the other one processed by SLC. Assessment of sperm motility (assessed by computer-assisted semen analysis), morphology (Diff-Quick staining) and viability (triple fluorescent stain of propidium iodine/isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin/Rhodamine 123), were performed on aliquots of fresh semen, frozen-thawed control and frozen-thawed SLC treated samples. A multivariate clustering procedure separated 26,051 motile spermatozoa into three subpopulations (sP): sP1 consisting of highly active but non-progressive spermatozoa (40.3%), sP2 consisting of spermatozoa with high velocity and progressive motility (30.0%), and sP3 consisting of poorly active and non-progressive spermatozoa (29.7%). SLC with PureSperm® 80 yielded sperm suspensions with improved motility, morphology, viability and acrosome integrity (P<0.001). The frozen-thawed SLC treated samples were enriched in sP2, reaching a proportion of 44.1% of the present spermatozoa. From these results, we concluded that SLC with PureSperm® 80 may be an alternative and successful method for improving the quality of frozen-thawed dog spermatozoa. Moreover, sP2 (high-speed and progressive spermatozoa) was more frequently observed after SLC. Finally, this study also demonstrated that the general motile sperm structure present in dogs remained constant despite the effect caused by either cryopreservation or separation by SLC through PureSperm® 80. Copyright © 2013 Elsevier B.V. All rights reserved.

Freeze-Thaw Cycles Effects on Soil Compaction in a Clay Loam

NASA Astrophysics Data System (ADS)

Jabro, J.; Evans, R.; Iversen, W.

2012-04-01

Inappropriate soil management practices and heavier farm machinery and equipment have led to an increase in soil compaction in the last two decades prompting increased global concern regarding the impact of soil compaction on crop production and soil quality in modern mechanized agriculture. A 3-yr comprehensive study was established to evaluate the dynamic of freeze-thaw cycles on soil compaction in a clay loam soil. Plots of frozen soils were compared with plots where soils were prevented from freezing with electrically heated blankets commonly used on concrete. Results showed that frequent freeze-thaw cycles over the winter alleviated a majority of soil compaction at the 0 - 20 cm depth. Soil penetration resistance in compacted soils was reduced by 73 and 68% over the winter at the 0 - 10 and 10 - 20 cm depths, respectively, due to dynamic effects of freeze-thaw cycles on soil structure and particles configuration. In unfrozen compacted soils, the penetration resistance was also reduced by 50 and 60% over winter at the 0 - 10 and 10 - 20 cm depths, respectively, due to the biology of soil, microbial activity, and disruptive effects of shrink-swell cycles. These results have demonstrated of how repeated freeze-thaw cycles can alleviate soil compaction, alter soil physical quality and create optimal soil conditions required for profitable growth of agricultural crops. The results from this study will save growers considerable time, money and energy currently required to alleviate soil compaction using other methods such as sub-soiling and deep tillage. We believe that Mother Nature provides ways to reverse soil compaction and improve soil structure and aggregation through the dynamic of freeze-thaw cycles that soils in Montana and other parts of the country go through each year. We concluded that the Mother Nature is the most effective and cheapest way to alleviate soil compaction.

Effect of Thaw Depth on Fluxes of CO2 and CH4 in Manipulated Arctic Coastal Tundra of Barrow, Alaska

NASA Astrophysics Data System (ADS)

Kim, Y.

2014-12-01

Changes in CO2 and CH4 emissions represent one of the most significant consequences of drastic climate change in the Arctic, by way of thawing permafrost, a deepened active layer, and decline of thermokarst lakes in the Arctic. This study conducted flux-measurements of CO2 and CH4, as well as environmental factors such as temperature, moisture, and thaw depth, as part of a water table manipulation experiment in the Arctic coastal plain tundra of Barrow, Alaska during autumn. The manipulation treatment consisted of draining, controlling, and flooding treated sections by adjusting standing water. Inundation increased CH4 emission by a factor of 4.3 compared to non-flooded sections. This may be due to the decomposition of organic matter under a limited oxygen environment by saturated standing water. On the other hand, CO2 emission in the dry section was 3.9-fold higher than in others. CH4 emission tends to increase with deeper thaw depth, which strongly depends on the water table; however, CO2 emission is not related to thaw depth. Quotients of global warming potential (GWPCO2) (dry/control) and GWPCH4 (wet/control) increased by 464 and 148 %, respectively, and GWPCH4 (dry/control) declined by 66 %. This suggests that CO2 emission in a drained section is enhanced by soil and ecosystem respiration, and CH4 emission in a flooded area is likely stimulated under an anoxic environment by inundated standing water. The findings of this manipulation experiment during the autumn period demonstrate the different production processes of CO2 and CH4, as well as different global warming potentials, coupled with change in thaw depth. Thus the outcomes imply that the expansion of tundra lakes leads the enhancement of CH4 release, and the disappearance of the lakes causes the stimulated CO2 production in response to the Arctic climate change.

Improvement of post-thawed sperm quality and fertility of Arian rooster by oral administration of d-aspartic acid.

PubMed

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.

Multiscale radar mapping of surface melt over mountain glaciers in High Mountain Asia

NASA Astrophysics Data System (ADS)

Steiner, N.; McDonald, K. C.

2017-12-01

Glacier melt dominates input for many hydrologic systems in the Himalayan Hindukush region that feed rivers that are critical for downstream ecosystems and hydropower generation in this highly populated area. Deviation in seasonal surface melt timing and duration with a changing climate has the potential to affect up to a billion people on the Indian Subcontinent. Satellite-borne microwave remote sensing has unique capabilities that allow monitoring of numerous landscape processes associated with snowmelt and freeze/thaw state, without many of the limitations in optical-infrared sensors such as solar illumination or atmospheric conditions. The onset of regional freeze/thaw and surface melting transitions determine important surface hydrologic variables like river discharge. Theses regional events are abrupt therefore difficult to observe with low-frequency observation sensors. Recently launched synthetic aperture radar (SAR) onboard the Sentinel-1 A and B satellites from the European Space Agency (ESA) provide wide-swath and high spatial resolution (50-100 m) C-Band SAR observations with observations frequencies not previously available, on the order of 8 to 16 days. The Sentinel SARs provide unique opportunity to study freeze/thaw and mountain glacier melt dynamics at process level scales, spatial and temporal. The melt process of individual glaciers, being fully resolved by imaging radar, will inform on the radiometric scattering physics associated with surface hydrology during the transition from melted to thawed state and during refreeze. Backscatter observations, along with structural information about the surface will be compared with complimentary coarse spatial resolution C-Band radar scatterometers, Advanced Scatterometer (ASCAT Met Op A+B), to understand the sub-pixel contribution of surface melting and freeze/thaw signals. This information will inform on longer-scale records of backscatter from ASCAT, 2006-2017. We present a comparison of polarimetric C-Band melt signals contained in the multi-scale backscatter and present a coincident freeze/thaw and snowmelt records from ASCAT and Sentinel-1 for the Gandaki basin, Nepal.

Durability assessments of concrete using electrical properties and acoustic emission testing

NASA Astrophysics Data System (ADS)

Todak, Heather N.

Premature damage deterioration has been observed in pavement joints throughout the Midwestern region of the United States. Over time, severe joint damage creates a transportation safety concern and the necessary repairs can be an extreme economic burden. The deterioration is due in part to freeze-thaw damage associated with fluid accumulation at the pavement joints. This very preventable problem is an indication that current specifications and construction practices for freeze-thaw durability of concrete are inadequate. This thesis serves to create a better understanding of moisture ingress, freeze-thaw damage mechanisms, and the effect of variations in mixture properties on freeze-thaw behavior of concrete. The concepts of the nick point degree of saturation, sorptivity rates, and critical degree of saturation are discussed. These factors contribute to service life, defined in this study as the duration of time a concrete element remains below levels of critical saturation which are required for damage development to initiate. A theoretical model and a simple experimental procedure are introduced which help determine the nick point for a series of 32 concrete mixtures with unique mixture proportions and air entrainment properties. This simple experimental procedure is also presented as a method to measure important electrical properties in order to establish the formation factor, a valuable measure of concrete transport properties. The results of freeze-thaw testing with acoustic emission monitoring are presented to help understand and quantify damage development in concrete specimens when conditioned to various degrees of saturation. This procedure was used to study the relationship between air entrainment properties and the critical degree of saturation. Applying the concepts of degree of saturation and sorptivity, a performance-based model is proposed as a new approach to specifications for freeze-thaw durability. Finally, a conceptual model is presented to illustrate the effect of various changes in mixture proportions and air void properties on service life.

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

USGS Publications Warehouse

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.

Spatial and temporal variations of thaw layer thickness and its controlling factors identified using time-lapse electrical resistivity tomography and hydro-thermal modeling

NASA Astrophysics Data System (ADS)

Tran, Anh Phuong; Dafflon, Baptiste; Bisht, Gautam; Hubbard, Susan S.

2018-06-01

Quantitative understanding of controls on thaw layer thickness (TLT) dynamics in the Arctic peninsula is essential for predictive understanding of permafrost degradation feedbacks to global warming and hydrobiochemical processes. This study jointly interprets electrical resistivity tomography (ERT) measurements and hydro-thermal numerical simulation results to assess spatiotemporal variations of TLT and to determine its controlling factors in Barrow, Alaska. Time-lapse ERT measurements along a 35-m transect were autonomously collected from 2013 to 2015 and inverted to obtain soil electrical resistivity. Based on several probe-based TLT measurements and co-located soil electrical resistivity, we estimated the electrical resistivity thresholds associated with the boundary between the thaw layer and permafrost using a grid search optimization algorithm. Then, we used the obtained thresholds to derive the TLT from all soil electrical resistivity images. The spatiotemporal analysis of the ERT-derived TLT shows that the TLT at high-centered polygons (HCPs) is smaller than that at low-centered polygons (LCPs), and that both thawing and freezing occur earlier at the HCPs compared to the LCPs. In order to provide a physical explanation for dynamics in the thaw layer, we performed 1-D hydro-thermal simulations using the community land model (CLM). Simulation results showed that air temperature and precipitation jointly govern the temporal variations of TLT, while the topsoil organic content (SOC) and polygon morphology are responsible for its spatial variations. When the topsoil SOC and its thickness increase, TLT decreases. Meanwhile, at LCPs, a thicker snow layer and saturated soil contribute to a thicker TLT and extend the time needed for TLT to freeze and thaw. This research highlights the importance of combination of measurements and numerical modeling to improve our understanding spatiotemporal variations and key controls of TLT in cold regions.

Development of procedures for sex-sorting frozen-thawed bovine spermatozoa.

PubMed

Underwood, S L; Bathgate, R; Maxwell, W M C; Evans, G

2009-06-01

Dairy bull sperm may be sex-sorted, frozen and used to artificially inseminate heifers with acceptable fertility if the herd is well-managed. One drawback to the technology is that donor bulls must be located within a short distance of the sorting facility in order to collect semen, which limits the number of bulls from which sorted sperm are available. A successful method used to overcome this limitation in sheep is sex-sorting from frozen-thawed semen and refreezing for artificial insemination. This technique is attractive to the dairy industry, and therefore a series of three experiments was designed to investigate the optimal methods to prepare, sex-sort and re-freeze frozen-thawed bovine sperm. Sperm were prepared for sorting by density gradient separation in either PureSperm or BoviPure, followed by staining in one of three diluents (Androhep, Bovine Sheath Fluid + 0.3% BSA or TALP buffer). Sperm were sorted and collected into Test yolk buffer, and frozen in an extender containing 0, 0.25, 0.375 or 0.5% Equex STM Paste. Frozen-thawed sperm were better orientated (p = 0.006) and had fewer damaged membranes (8.7 +/- 0.6% vs 19.5 +/- 2.4%; p = 0.003) after centrifugation in PureSperm rather than BoviPure gradients. Sperm orientation (p < 0.05) and motility (69.9 +/- 3.0 vs 55.6 +/- 4.0; p < 0.001) were highest after staining in Androhep rather than in TALP buffer. Sperm were more motile (58.2 +/- 4.7 vs 38.7 +/- 3.5; p < 0.001) and had better acrosome integrity (74.3 +/- 2.9 vs 66.8 +/- 2.0; p < 0.001) after freezing in an extender containing 0.375% Equex STM Paste than in extender without Equex. Hence, a protocol has been developed to allow frozen-thawed bull sperm to be sex-sorted with high resolution between the sexes, then re-frozen and thawed with retention of motility and acrosome integrity.

Monitoring an Induced Permafrost Warming Experiment Using ERT, Temperature, and NMR in Fairbanks, Alaska

NASA Astrophysics Data System (ADS)

Ulrich, C.; Ajo Franklin, J. B.; Ekblaw, I.; Lindsey, N.; Wagner, A. M.; Saari, S.; Daley, T. M.; Freifeld, B. M.

2016-12-01

As global temperatures continue to rise, permafrost landscapes will experience more rapid changes than other global climate zones. Permafrost thaw is a result of increased temperatures in arctic settings resulting in surface deformation and subsurface hydrology changes. From an engineering perspective, surface deformation poses a threat to the stability of existing infrastructure such as roads, utility piping, and building structures. Preemptively detecting or monitoring subsurface thaw dynamics presents a difficult challenge due to the long time scales as deformation occurs. Increased subsurface moisture content results from permafrost thaw of which electrical resistivity tomography (ERT), soil temperature, and nuclear magnetic resonance (NMR) are directly sensitive. In this experiment we evaluate spatial and temporal changes in subsurface permafrost conditions (moisture content and temperature) at a experimental heating plot in Fairbanks, AK. This study focuses on monitoring thaw signatures using multiple collocated electrical resistivity (ERT), borehole temperature, and borehole nuclear magnetic resonance (NMR) measurements. Timelapse ERT (sensitive to changes in moisture content) was inverted using collocated temperature and NMR to constrain ERT inversions. Subsurface thermal state was monitored with timelapse thermistors, sensitive to soil ice content. NMR was collected in multiple boreholes and is sensitive to changes in moisture content and pore scale distribution. As permafrost thaws more hydrogen, in the form of water, is available resulting in a changing NMR response. NMR requires the availability of liquid water in order to induce spin of the hydrogen molecule, hence, if frozen water molecules will be undetectable. In this study, the permafrost is poised close to 0oC and is mainly silt with small pore dimensions; this combination makes NMR particularly useful due to the possibility of sub-zero thaw conditions within the soil column. Overall this experiment presents a complementary suite of methods that provides feedback on subsurface permafrost state even in cases where soil texture might control unfrozen water content.

Thaw depth spatial and temporal variability at the Limnopolar Lake CALM-S site, Byers Peninsula, Livingston Island, Antarctica.

PubMed

de Pablo, M A; Ramos, M; Molina, A; Prieto, M

2018-02-15

A new Circumpolar Active Layer Monitoring (CALM) site was established in 2009 at the Limnopolar Lake watershed in Byers Peninsula, Livingston Island, Antarctica, to provide a node in the western Antarctic Peninsula, one of the regions that recorded the highest air temperature increase in the planet during the last decades. The first detailed analysis of the temporal and spatial evolution of the thaw depth at the Limnopolar Lake CALM-S site is presented here, after eight years of monitoring. The average values range between 48 and 29cm, decreasing at a ratio of 16cm/decade. The annual thaw depth observations in the 100×100 m CALM grid are variable (Variability Index of 34 to 51%), although both the Variance Coefficient and the Climate Matrix Analysis Residual point to the internal consistency of the data. Those differences could be explained then by the terrain complexity and node-specific variability due to the ground properties. The interannual variability was about 60% during 2009-2012, increasing to 124% due to the presence of snow in 2013, 2015 and 2016. The snow has been proposed here as one of the most important factors controlling the spatial variability of ground thaw depth, since its values correlate with the snow thickness but also with the ground surface temperature and unconfined compression resistance, as measured in 2010. The topography explains the thaw depth spatial distribution pattern, being related to snowmelt water and its accumulation in low-elevation areas (downslope-flow). Patterned grounds and other surface features correlate well with high thaw depth patterns as well. The edaphic factor (E=0.05842m 2 /°C·day; R 2 =0.63) is in agreement with other permafrost environments, since frozen index (F>0.67) and MAAT (<-2°C) denote a continuous permafrost existence in the area. All these characteristics provided the basis for further comparative analyses between others nearby CALM sites. Copyright © 2017 Elsevier B.V. All rights reserved.

Microbial Insights into Shifting Methane Production Potential in Thawing Permafrost

NASA Astrophysics Data System (ADS)

Crossen, K.; Wilson, R.; Raab, N.; Neumann, R.; Chanton, J.; Saleska, S. R.; Rich, V. I.

2017-12-01

Permafrost, which stores 50% of global soil carbon, is thawing rapidly due to climate change, and resident microbes are contributing to changing carbon gas emissions. Predictions of the fate of carbon in these regions is poorly constrained; however, improved, careful mapping of microbial community members influencing CO2 and CH4 emissions will help clarify the system response to continued change. In order to more fully understand connections between the microbial communities, major geochemical transformations, and CO2 and CH4 emissions, peat cores were collected from the active layers of three permafrost habitats spanning a thaw gradient (collapsed palsa, bog, and fen) at Stordalen Mire, Abisko, Sweden. Anaerobic incubations of shallow and deep subsamples from these sites were performed, with time-course characterization of the changes in microbial communities, peat geochemistry, and carbon gas production. The latter were profiled with 16S rRNA amplicon sequencing, and targeted metagenomes. The communities within each habitat and depth were statistically distinct, and changed significantly over the course of the incubations. Acidobacteria was consistently the dominant bacterial phylum in all three habitat types. With increased thaw, the relative abundance of Actinobacteria tended to decrease, while Chloroflexi and Bacteroidetes increased with thaw. The relative abundance of methanogens increased with thaw and with depth within each habitat. Over time in the incubations, the richness of the communities tended to decrease. Homoacetogenesis (CO2 + H2 -> CH3COOH) has been documented in other peatlands, and homoacetogens can influence CH4 production by interacting with methanogens, competing with hydrogenotrophs while providing substrate for acetoclasts. Modelling of microbial reaction networks suggests potential for highest homoacetogenesis rates in the collapsed palsa, which also contains the highest relative abundances of lineages taxonomically affiliated with known homoacetogens. We are working to link changes in the relative abundances of specific, differentiating lineages with observed geochemical transformations and measured carbon gas production. This work will increase our knowledge of factors influencing greenhouse gas emissions from this climatically important habitat.

Thermokarst in pingos and adjacent collapse scar bogs in interior Alaska

NASA Astrophysics Data System (ADS)

Douglas, T. A.; Turetsky, M. R.

2017-12-01

A region of discontinuous permafrost 50 kilometers southeast of Fairbanks, Alaska exhibits rapid thermokarst and landscape change. The area contains a dozen pingos (hydrolaccoliths), mounds of ice covered by earth material typically 100 meters across and 20 meters above the surrounding ground surface. The pingos have sunken craters in their centers formed through melting and collapse of an inner ice lens core. Adjacent to the pingos are collapse scar bogs in various states of formation and ice wedge terrain undergoing thaw subsidence to polygons and thermokarst mounds (baydzherakhs). With a mean annual temperature of -1 degree C the area contains warm ecosystem-protected permafrost vulnerable to thaw. We analyzed historical imagery to the 1970s to track water features in a subset of pingos. The craters have expanded over the past few decades suggesting melting and collapse of the ice cored center and potential permafrost degradation along pingo margins. Collapse scar bogs in adjacent low-elevation terrain are roughly the same size as the pingos but have little vertical elevation gradient compared to the surrounding terrain. Electrical resistivity tomography (ERT) measurements, high resolution GPS surveys, SIPRE coring, and thaw depth probing were focused along nine 400 meter transects across three of the pingos to identify relationships between geophysical properties, permafrost composition, seasonal thaw, and ecological state. A large ( 40 meters across and 20 meters thick) lens shaped region of thawed permafrost is evident in the ERT results about 10 meters below the ground surface in the center of one pingo we surveyed in detail. This is believed to be the original ice cored region of the pingo that has melted. A thin (1-5 meters thick) layer of permafrost is present above this thawed region while the rampart margins surrounding the pingo are underlain by thick (10-30 m) permafrost. The pingo and thermokarst features reside in a location where rapid permafrost thaw in response to warming or changing hydrology could provide a hot spot for landscape change, particularly given a projected climate warming of 5 degrees C over the next 80 years in the area. Their future thermal, geomorphological, and ecological states may be a harbinger for how discontinuous permafrost in the region responds to projected climate warming.

Methane production as key to the greenhouse gas budget of thawing permafrost on climate relevant time scales

NASA Astrophysics Data System (ADS)

Knoblauch, C.; Beer, C.; Liebner, S.; Schütt, A.; Grigoriev, M.; Pfeiffer, E. M.

2017-12-01

Permafrost in circum-arctic soils stores as much carbon as the global atmosphere. Permafrost thaw liberates organic matter, which is mineralized by microorganisms to carbon dioxide (CO2) and methane (CH4). The release of these greenhouse gases (GHGs) may form a positive feedback to atmospheric CO2 and CH4 concentrations and accelerate climate change. The microbial formation of CH4, which has 28 to 45 times the global warming potential (GWP) of CO2 (100 years time scale), requires anoxic conditions. Current studies indicate that permafrost thaw at the bottom of well drained (oxic) soils cause a higher formation of GHGs than in water saturated (anoxic) soils since more CO2 is formed under oxic conditions and only small amounts of CH4 are formed from permafrost organic matter under anoxic conditions. Here we show through 7-year laboratory incubations and molecular analysis of Siberian permafrost that low CH4 production from permafrost organic matter is due to the lack of active methanogens. Equal amounts of permafrost organic carbon are mineralized to CO2 and CH4 under anoxic conditions after an active methanogenic community has established. Field measurements demonstrate that recently thawed permafrost organic matter is a substantial source for CH4 if primed with surface soil. An organic carbon decomposition model, calibrated with the collected long-term incubation data, predicts a higher loss of permafrost carbon under oxic conditions but a twice as high production of CO2-C equivalents under anoxic conditions when considering a GWP of 28 for CH4. Combining these model results with observed permafrost carbon profile data, up-scaled carbon stocks and thaw depth projections suggests a global formation of 3 - 10 Pg CO2-C from thawing permafrost in oxic soils compared to 0.2 - 0.6 Pg CO2-C and 0.2- 0.8 Pg CH4-C in anoxic soils until 2100. However, based on CO2-C equivalents the GHG production in anoxic soils (2 - 9 Pg CO2-C equivalents) is similar to those in oxic soils. These findings challenge the view of a stronger permafrost carbon-climate feedback from drained soils and emphasize the importance of CH4 production in thawing permafrost.

Effects of Conversion from Boreal Forest to Arctic Steppe on Soil Communities and Ecosystem Carbon Pools

NASA Astrophysics Data System (ADS)

Han, P. D.; Natali, S.; Schade, J. D.; Zimov, N.; Zimov, S. A.

2014-12-01

The end of the Pleistocene marked the extinction of a great variety of arctic megafauna, which, in part, led to the conversion of arctic grasslands to modern Siberian larch forest. This shift may have increased the vulnerability of permafrost to thawing because of changes driven by the vegetation shift; the higher albedo of grassland and low insulation of snow trampled by animals may have decreased soil temperatures and reduced ground thaw in the grassland ecosystem, resulting in protection of organic carbon in thawed soil and permafrost. To test these hypothesized impacts of arctic megafauna, we examined an experimental reintroduction of large mammals in northeast Siberia, initiated in 1988. Pleistocene Park now contains 23 horses, three musk ox, one bison, and several moose in addition to the native fauna. The park is 16 square km with a smaller enclosure (< 1 km) where animals spend most of their time and our study was focused. We measured carbon-pools in forested sites (where scat surveys showed low animal use), and grassy sites (which showed higher use), within the park boundaries. We also measured thaw depth and documented the soil invertebrate communities in each ecosystem. There was a substantial difference in number of invertebrates per kg of organic soil between the forest (600 ± 250) and grassland (300 ± 250), though these differences were not statistically significant they suggest faster nutrient turnover in the forest or a greater proportion of decomposition by invertebrates than other decomposers. While thaw depth was deeper in the grassland (60 ± 4 cm) than in the forest (40 ± 6 cm), we did not detect differences in organic layer depth or percent organic matter between grassland and forest. However, soil in the grassland had higher bulk density, and higher carbon stocks in the organic and mineral soil layers. Although deeper thaw depth in the grassland suggests that more carbon is available to microbial decomposers, ongoing temperature monitoring will help determine whether or not differences detected in summer continue throughout the year. This study provides a preliminary understanding of the role megafauna play in carbon emissions from the arctic, and suggests that animals play a critical role in regulating permafrost thaw and the fate of carbon stored in Arctic soils.

Clinical evaluation of frozen/thawed embryo transfer following transport of oocytes and embryos

PubMed Central

2004-01-01

Background and Aims:  We evaluated the efficacy of the transport oocyte/embryo frozen/thawed embryo transfer method, in which oocytes or embryos were transported from satellite clinics to the main assisted reproductive technology (ART) center, and surplus embryos were placed in cryopreservation. Methods:  We evaluated 41 cycles in 34 patients in the transport oocyte group (TO group). In the TO group the oocytes were collected at the satellite clinics, transported to the main ART center and underwent in vitro fertilization or intracytoplasmic sperm injection. Surplus embryos were used for frozen/thawed embryo transfer. We also evaluated 17 cycles in 10 patients in the transport embryo group (TE group), where surplus embryos were transported to the main ART center and used for frozen/thawed embryo transfer; and 189 cycles in 134 patients in the center group (C group), where surplus embryos collected at the same time at the main ART center were used for frozen/thawed embryo transfer. Oocytes were transported from satellite clinics in HEPES buffered human tubal fluid (HTF) culture medium, and embryos in 30% synthetic serum substitute + HEPES buffered HTF, using a portable incubator we devised. Results:  The proportions of undamaged embryos after freeze/thawing were 47% for the C group, 46% for the TO group, and 46% for the TE group. The numbers of embryos transferred were 2.0 ± 0.7 for the C group, 2.0 ± 0.6 for the TO group, and 2.2 ± 0.4 for the TE group. The rate of embryo transfer was 63% for the C group, 68% for the TO group, and 76% for the TE group. Pregnancy rates per patient were 16% for the C group, 24% for the TO group, and 40% for the TE group. The embryo survival rates (number of embryos with ≥50% viable blastomeres/total number of embryos) were 55% for the C group, 60% for the TO group, and 54% for the TE group. No significant differences were seen between the C group and either the TO or TE groups in any of these parameters. Conclusions:  Favorable results were achieved with the transport oocyte/embryo frozen/thawed embryo transfer method, and it is suitable for widespread clinical application. (Reprod Med Biol 2004; 3: 1–8) PMID:29662379

The effect of glycerol-related osmotic changes on post-thaw motility and acrosomal integrity of ram spermatozoa.

PubMed

Fiser, P S; Fairfull, R W

1989-02-01

Ram semen, collected by artificial vagina, was diluted and processed for long-term storage as described by P. S. Fiser, L. Ainsworth, and R. W. Fairfull (Canad. J. Anim. Sci. 62, 425-428, 1982). The concentration of the cryoprotectant, glycerol, was adjusted to 4% in the diluted semen prior to freezing by a one-step addition at 30 degrees C (Method 1), by cooling the semen to 5 degrees C and addition of the glycerol gradually over 30 min (Method 2), by one-step addition of glycerol prior to equilibration for 2 hr (Method 3), or by cooling to 5 degrees C, followed by a holding period of 2 hr at 5 degrees C, and the one-step addition of glycerol just prior to freezing (Method 4). After thawing, the glycerol concentration of the semen was reduced by stepwise dilution from 4 to 0.4% over 15 or 30 min or by a one-step ten-fold dilution. The average post-thaw percentage of motile spermatozoa was significantly lower after addition of glycerol by Method 1 (39.9%) than when the glycerol was added by the other three methods (range, 44.0-46.4% averaged over the glycerol dilution). The average post-thaw percentage of intact acrosomes (61.2%), highest in semen in which the glycerol was added by Method 2, was not significantly different from those in which glycerol was added to semen by Methods 3 and 4, but it was significantly higher than that found in semen in which the glycerol was added by Method 1 (54.4%). However, when averaged over the method of glycerolation, the post-thaw percentage of motile spermatozoa (range, 43.7-44.2%) and the percentage of intact acrosomes (range, 56.8-59.5%) did not differ significantly in semen subjected to gradual decrease in glycerol concentration and diluent osmolality (over 15 and 30 min) or by a one-step, 10-fold dilution. These data indicate that post-thaw survival of spermatozoa can be influenced by the way in which glycerol is added prior to freezing. However, post-thaw spermatozoa motility and acrosomal integrity can be maintained even after a rapid decrease in glycerol concentration such as that which accompanies insemination or dilution of semen for assessment of motility.

Interaction of extender composition and freezing method for effective semen cryopreservation in the North American river otter (Lontra canadensis).

PubMed

Bateman, Helen L; Swanson, William F

2017-10-01

Semen cryopreservation and storage in genome resource banks (GRBs), in combination with artificial insemination (AI), could be invaluable for genetic management and conservation of endangered otter species. For any applied conservation benefit, effective methods for otter sperm processing and cryopreservation first must be established. In this study, our objective was to develop an effective semen cryopreservation method for the North American river otter, evaluating the effect of extender composition (i.e., glycerol concentration, Equex STM paste supplementation) and freezing protocol (timing of glycerol addition, pre-freeze cooling rate, freezing/packaging method) on post-thaw sperm motility, longevity and acrosome status. Semen was collected from 14 otters housed at 9 zoos, and following cryopreservation in an egg-yolk based extender, thawed to assess sperm motility and acrosome status immediately post-thaw and during 6 h of in vitro culture. Results indicated that extender containing 4% glycerol was preferable (p < 0.05) to 8% glycerol but the temperature/timing of extender addition containing 4% glycerol did not affect (p > 0.05) post-thaw sperm parameters. Treatments with extender containing Equex and frozen by pelleting on dry ice showed greater (p < 0.05) motility and percentage of intact acrosomes compared to treatments frozen in extender without Equex, regardless of pre-freeze cooling rate. In the absence of Equex, pelleting provided superior post-thaw sperm motility (p < 0.01) and higher (p < 0.001) percentage of sperm with intact acrosomes compared to samples frozen in straws over liquid nitrogen vapor. Results of this study indicate that cryopreservation of otter sperm using an egg-yolk -TEST based extender containing 4% glycerol and 1% Equex, with the pellet freezing method, provided superior post-thaw sperm motility, longevity and acrosomal integrity compared to other combinations. Neither alterations in timing of glycerolated extender addition nor pre-freeze cooling rate had a discernable effect on post-thaw otter sperm parameters. These findings represent the first assessment of semen cryopreservation in any otter species and may be of value as a model for development of semen cryopreservation strategies in other endangered otter species. Copyright © 2017 Elsevier Inc. All rights reserved.

Investigation of the Active layer thickness and ground subsidence in Taimyr

NASA Astrophysics Data System (ADS)

Grebenets, V. I.; Tolmanov, V. A.; Streletskiy, D. A.

2017-12-01

The active layer of permafrost (ALT) is highly unstable and dynamic in space and time. Soil undergoes frost heave during the freezing process, and ground subsidence during the thawing. The problem of the development of soil sediments' deformations in ALT is relevant as for natural objects (influence on runoff, changing of landscape and vegetation, etc.), so for industrial infrastructure (pipelines, roads, buildings and structures). The observations in the frame of the CALM program in Taimyr were carried out since 2005 (site R-32) with the measurements of the geodetic level of soil surface since 2007. The results of these measurements were processed and the maps of thawing and changes in meso- and micro-relief were constructed. The differentiation of seasonally thawed layer and ground subsidence in different micro-landscape conditions was investigated. The depth of seasonal thawing and the changes of surface movements were found to be determined by three main systems: a) the weather conditions and the climate trends; b) the permafrost-lithological conditions and drainage; c) the micro-landscape characteristics. It was established that for the Norilsk region (Taimyr) the trend in increasing ALT was 0.3 cm / year (for the period of observations 2005-2016) with a certain slowdown in the last 3 to 4 years. Increase in the depth of the ALT was related to the rising Summer temperatures and reduction of the cold period. A strong high impact of the summer precipitation conditions was revealed: in rather cold summer of 2012, with large amount of precipitation mainly in the warmest month (July), the defrosting was the highest. In the year with the record-breaking number of positive degree days (from all the 85 years of regular meteorological observations) but anomalously dry year 2013 (in July - less than 10 mm atmospheric precipitation), the thawing was minimal at the R-32 site. It is interesting that the ground subsidence in 2012 was 30-40% less, than in 2013. This is due to the water saturation of the system, since water is practically incompressible. The maximum thawing and the ground subsidence are found in negative forms of relief. Differences in depths of the seasonally thawed layer and in change of the surface level within different landscapes can reach 50-70%.

Life in Ice: Microbial Growth Dynamics and Greenhouse Gas Production During Winter in a Thermokarst Bog Revealed by Stable Isotope Probing Targeted Metagenomics

NASA Astrophysics Data System (ADS)

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 pathways in growing organisms that likely drove C mineralization. Results indicate that winter microbial activities can play an important role in controlling seasonal C flux in recent thawed permafrost and characterization of growing organisms leads to stronger mechanistic linkages between the soil microbial community and ecosystem processes.

Seasonal Variability of Major Ions and δ13CDIC in Permafrost Watersheds of Arctic Alaska

NASA Astrophysics Data System (ADS)

Lehn, G. O.; Jacobson, A. D.; Douglas, T. A.; McClelland, J. W.; Khosh, M. S.; Barker, A. J.

2011-12-01

Models and observations predict that climate change will have more severe effects at higher latitudes. Many effects may already be underway. Increasing temperatures are expected to thaw permafrost soils, changing the hydrology and biogeochemistry of Arctic watersheds. These changes are particularly important because permafrost thaw could destabilize a large carbon reservoir, potentially leading to sizable greenhouse gas emissions. Tracking soil thaw and concomitant changes in carbon export are therefore critical to predicting feedbacks between Arctic climate change and global warming. As the climate warms, the seasonally thawed active layer will extend into deeper, previously frozen, mineral-rich soils, increasing the signal of chemical weathering in streams. Historical methods of monitoring active layer thaw depth are labor intensive and may not capture the heterogeneity of Arctic soils, whereas stream geochemistry provides a unique opportunity to integrate signals across vast spatial distances. We present major ion geochemistry and δ13C of dissolved inorganic carbon (DIC) variations that relate to seasonal changes in permafrost thaw depths. Samples were collected from six watersheds on the North Slope of Alaska. All rivers drain continuous permafrost but three drain tussock tundra-dominated watersheds and three drain bare bedrock catchments with minor tundra influences. Water samples were collected from April until October in 2009 and 2010. The major ion and δ13CDIC trends of tundra streams suggest that silicate weathering dominates during the spring melt while carbonate weathering dominates as the active layer deepens in the summer. In tundra streams, early season δ13CDIC values indicate carbonic acid-silicate weathering. Summer δ13CDIC values indicate carbonic acid-carbonate weathering. In both cases, carbonic acid forms from CO2 produced by the microbial decomposition of C3 organic matter. Bedrock streams have nearly constant δ13CDIC values and high dissolved sulfate concentrations through the year, indicating sulfuric acid-carbonate weathering. In late fall of 2010, δ13CDIC decreases in all streams suggest increased CO2 from a source with relatively negative δ13C values, possibly methane oxidation in soils. The difference between the tundra and bedrock streams allows us to clearly isolate the effect of soil thaw on stream geochemistry. Our initial findings illustrate how seasonal changes in mineral weathering have potential for tracking active layer dynamics.

Response of Carbon Fluxes to Soil Moisture Variability across an Alaskan Tundra Landscape

NASA Astrophysics Data System (ADS)

Melton, S.; Natali, S.; Schade, J. D.; Holmes, R. M.; Mann, P. J.; Fiske, G. J.

2017-12-01

Soils in arctic and sub-arctic permafrost regions store large amounts of carbon (C), which is becoming more biologically available as soils warm and permafrost thaws. Microbial decay of organic forms of C can result in the production and emission of carbon dioxide (CO2) and methane (CH4), and the amount and form of C released into the atmosphere depend on organic matter composition and soil conditions. Soil moisture, which is a strong driver of microbial processes, varies spatially and temporally across tundra landscapes and may change dramatically as a result of permafrost thaw. The Yukon-Kuskokwim Delta (YKD) of Alaska is underlain by discontinuous permafrost and is particularly vulnerable to permafrost thaw and soil moisture changes associated with thaw. As permafrost thaws, some areas may dry as drainage increases with increasing thaw depth. Alternatively, permafrost thaw may lead to ground subsidence and saturation of previously dry soils. Our objective was to investigate patterns in C storage and processing across the landscape and in response to changes in soil moisture in the YKD. We analyzed soil C pools (0-30 cm) and CO2 and CH4 concentrations in soils from sites of different land cover and landscape position, including moist and dry peat plateaus, high and low intensity burned plateaus, fens, and drained lakes. We also conducted aerobic and anaerobic soil incubations to determine changes in CO2 and CH4 production under a range of soil moisture conditions. Soils from burned plateaus, which were drier and had lower C content than unburned soils, had higher CO2 production (per g soil) than unburned soils during aerobic incubations. Both increased and decreased moisture reduced CO2 production from soils. Soil drying increased net CH4 uptake in all aerobically-incubated burned soils, and wetting resulted in CH4 emissions from low intensity burn soils. CO2 and CH4 production from fen soils were higher than from the other landscape positions analyzed here. Our results suggest that soil drying could lead to decreased microbial respiration, whereas subsidence may result in increased methanogenesis. Additionally, amplified CH4 release from burned soils after rainfall events or subsidence may accompany the increased fire frequency projected in tundra regions.

Unprecedented permafrost thaw dynamics on a decadal time scale: Batagay mega thaw slump development, Yana Uplands, Yakutia, Russia

NASA Astrophysics Data System (ADS)

Günther, F.; Grosse, G.; Jones, B. M.; Schirrmeister, L.; Romanovsky, V. E.; Kunitsky, V.

2016-12-01

Ice-rich permafrost that formed in glacial periods of the Quaternary is highly vulnerable to thaw under ongoing climate warming and anthropogenic disturbance. The mega thaw slump near the village of Batagay (Yakutia, Russia) is an outstanding example of permafrost degradation and demonstrates that thermo-erosion processes may occur in unexpected locations, develop very rapidly in particular after disturbances, and leave behind deep rutted badlands. Retrogressive thaw slumps are particularly frequent along riverbanks and coastlines of regions where buried glacier ice or ice-rich glacial till have been mapped. In East Siberia, syngenetic Late Pleistocene Ice Complex (Yedoma) permafrost deposits accumulate volumetric ground ice contents of up to 80-90% % and extend tens of meters below the ground surface. Beyond the Yedoma main distributional range in the coastal lowlands of the Laptev and East Siberian seas, these deposits are also found on slopes of the Verkhoyan Mountain Range and in valleys of surrounding foothills, providing favorable preconditions for rapid thaw development. The Batagay mega slump exposes a profile of 30m thick Yedoma deposits underlain by ice saturated alluvial sand of around 60 m thickness and another very ice-rich layer at the base. We present data from a multi-sensor remote sensing time series investigation of the mega slump in order to assess the planimetric and volumetric dimensions and its decadal and interannual expansion rates. For ortho-rectification purposes and for volumetric analyses, we photogrammetrically derived highly detailed digital elevation models. The height difference between the headwall and the slump outflow is 145 m along a distance of 2300 m, while the maximum slump width is 840 m. Our analysis does not show any signs of stabilization after several decades (since 1980s) of slump growth, with the headwall retreating with observed rates of generally >10 m and more recently up to 30m per year. Reconstruction of a paleo-surface revealed that the slump has carved into the rolling topography to a depth of up to 73 m. The current size of the slump is >69 ha, while it had thawed >25 × 106 m³ of ice-rich permafrost through 2016. The majority of sediment released from the slump episodically dams up the Batagay River, forming a large temporary lake which then empties catastrophically.