Sperm cryopreservation of African catfish, Clarias gariepinus: cryoprotectants, freezing rates and sperm:egg dilution ratio.

PubMed

Viveiros, A T; So, N; Komen, J

2000-12-01

Methods for cryopreserving spermatozoa and optimizing sperm:egg dilution ratio in African catfish Clarias gariepinus were developed. Five percent to 25% DMSO and methanol were tested as cryoprotectants, by diluting semen in Ginzburg fish ringer and freezing in 1-milliliter cryovials in a programmable freezer. To avoid an excess of spermatozoa per egg, post-thaw semen was diluted 1:20, 1:200 or 1:2,000 before fertilization. Highest hatching rates were obtained by spermatozoa frozen in 10% methanol and post-thaw diluted to 1:200. Then, slow freezing rates (-2, -5 or -10 degrees C/min) to various endpoint temperatures (range -25 to -70 degrees C) before fast freezing in liquid nitrogen (LN2) were evaluated. Hatching rates equal to control (P > 0.05) were obtained by spermatozoa frozen at -5 degrees C/min to -45 to -50 degrees C and at -10 degrees C/min to -55 degrees C. In 3-step freezing programs, at -5 degrees C/min, the effect of holding spermatozoa for 0, 2 or 5 min at -30, -35 or -40 degrees C before fast freezing in LN2 was analyzed. Hatching rates equal to control (P > 0.05) were produced by spermatozoa frozen to, and held at, -35 degrees C for 5 min and at -40 degrees C for 2 or 5 min. Finally, frozen spermatozoa (10% methanol, -5 degrees C/min, 5-min hold at -40 degrees C, LN2, post-thaw diluted to 1:200) were tested in on-farm fertilization conditions. Again, no difference (P > 0.05) in hatching rate was observed between frozen and fresh spermatozoa. Cryopreservation offers utility as a routine method of sperm storage and management for catfish.

Physiological Mechanisms Only Tell Half Story: Multiple Biological Processes are involved in Regulating Freezing Tolerance of Imbibed Lactuca sativa Seeds

PubMed Central

Jaganathan, Ganesh K.; Han, Yingying; Li, Weijie; Song, Danping; Song, Xiaoyan; Shen, Mengqi; Zhou, Qiang; Zhang, Chenxue; Liu, Baolin

2017-01-01

The physiological mechanisms by which imbibed seeds survive freezing temperatures in their natural environment have been categorized as freezing avoidance by supercooling and freezing tolerance by extracellular freeze-desiccation, but the biochemical and molecular mechanisms conferring seed freezing tolerance is unexplored. In this study, using imbibed Lactuca sativa seeds we show that fast cooled seeds (60 °C h−1) suffered significantly higher membrane damage at temperature between −20 °C and −10 °C than slow cooled (3 °Ch−1) seeds (P < 0.05), presumably explaining viability loss during fast cooling when temperature approaches −20 °C. Total soluble sugars increase in low temperature environment, but did not differ significantly between two cooling rates (P > 0.05). However, both SOD activity and accumulation of free proline were induced significantly after slow cooling to −20 °C compared with fast cooling. RNA-seq demonstrated that multiple pathways were differentially regulated between slow and fast cooling. Real-time verification of some differentially expressed genes (DEGs) revealed that fast cooling caused mRNA level changes of plant hormone and ubiquitionation pathways at higher sub-zero temperature, whilst slow cooling caused mRNA level change of those pathways at lower sub-zero ttemperatures. Thus, we conclude that imbibed seed tolerate low temperature not only by physiological mechanisms but also by biochemical and molecular changes. PMID:28287125

Physiological Mechanisms Only Tell Half Story: Multiple Biological Processes are involved in Regulating Freezing Tolerance of Imbibed Lactuca sativa Seeds.

PubMed

Jaganathan, Ganesh K; Han, Yingying; Li, Weijie; Song, Danping; Song, Xiaoyan; Shen, Mengqi; Zhou, Qiang; Zhang, Chenxue; Liu, Baolin

2017-03-13

The physiological mechanisms by which imbibed seeds survive freezing temperatures in their natural environment have been categorized as freezing avoidance by supercooling and freezing tolerance by extracellular freeze-desiccation, but the biochemical and molecular mechanisms conferring seed freezing tolerance is unexplored. In this study, using imbibed Lactuca sativa seeds we show that fast cooled seeds (60 °C h -1 ) suffered significantly higher membrane damage at temperature between -20 °C and -10 °C than slow cooled (3 °Ch -1 ) seeds (P < 0.05), presumably explaining viability loss during fast cooling when temperature approaches -20 °C. Total soluble sugars increase in low temperature environment, but did not differ significantly between two cooling rates (P > 0.05). However, both SOD activity and accumulation of free proline were induced significantly after slow cooling to -20 °C compared with fast cooling. RNA-seq demonstrated that multiple pathways were differentially regulated between slow and fast cooling. Real-time verification of some differentially expressed genes (DEGs) revealed that fast cooling caused mRNA level changes of plant hormone and ubiquitionation pathways at higher sub-zero temperature, whilst slow cooling caused mRNA level change of those pathways at lower sub-zero ttemperatures. Thus, we conclude that imbibed seed tolerate low temperature not only by physiological mechanisms but also by biochemical and molecular changes.

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.

Oocyte, embryo and blastocyst cryopreservation in ART: systematic review and meta-analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance.

PubMed

Rienzi, Laura; Gracia, Clarisa; Maggiulli, Roberta; LaBarbera, Andrew R; Kaser, Daniel J; Ubaldi, Filippo M; Vanderpoel, Sheryl; Racowsky, Catherine

2017-03-01

Successful cryopreservation of oocytes and embryos is essential not only to maximize the safety and efficacy of ovarian stimulation cycles in an IVF treatment, but also to enable fertility preservation. Two cryopreservation methods are routinely used: slow-freezing or vitrification. Slow-freezing allows for freezing to occur at a sufficiently slow rate to permit adequate cellular dehydration while minimizing intracellular ice formation. Vitrification allows the solidification of the cell(s) and of the extracellular milieu into a glass-like state without the formation of ice. The objective of our study was to provide a systematic review and meta-analysis of clinical outcomes following slow-freezing/thawing versus vitrification/warming of oocytes and embryos and to inform the development of World Health Organization guidance on the most effective cryopreservation method. A Medline search was performed from 1966 to 1 August 2016 using the following search terms: (Oocyte(s) [tiab] OR (Pronuclear[tiab] OR Embryo[tiab] OR Blastocyst[tiab]) AND (vitrification[tiab] OR freezing[tiab] OR freeze[tiab]) AND (pregnancy[tiab] OR birth[tiab] OR clinical[tiab]). Queries were limited to those involving humans. RCTs and cohort studies that were published in full-length were considered eligible. Each reference was reviewed for relevance and only primary evidence and relevant articles from the bibliographies of included articles were considered. References were included if they reported cryosurvival rate, clinical pregnancy rate (CPR), live-birth rate (LBR) or delivery rate for slow-frozen or vitrified human oocytes or embryos. A meta-analysis was performed using a random effects model to calculate relative risk ratios (RR) and 95% CI. One RCT study comparing slow-freezing versus vitrification of oocytes was included. Vitrification was associated with increased ongoing CPR per cycle (RR = 2.81, 95% CI: 1.05-7.51; P = 0.039; 48 and 30 cycles, respectively, per transfer (RR = 1.81, 95% CI 0.71-4.67; P = 0.214; 47 and 19 transfers) and per warmed/thawed oocyte (RR = 1.14, 95% CI: 1.02-1.28; P = 0.018; 260 and 238 oocytes). One RCT comparing vitrification versus fresh oocytes was analysed. In vitrification and fresh cycles, respectively, no evidence for a difference in ongoing CPR per randomized woman (RR = 1.03, 95% CI: 0.87-1.21; P = 0.744, 300 women in each group), per cycle (RR = 1.01, 95% CI: 0.86-1.18; P = 0.934; 267 versus 259 cycles) and per oocyte utilized (RR = 1.02, 95% CI: 0.82-1.26; P = 0.873; 3286 versus 3185 oocytes) was reported. Findings were consistent with relevant cohort studies. Of the seven RCTs on embryo cryopreservation identified, three met the inclusion criteria (638 warming/thawing cycles at cleavage and blastocyst stage), none of which involved pronuclear-stage embryos. A higher CPR per cycle was noted with embryo vitrification compared with slow-freezing, though this was of borderline statistical significance (RR = 1.89, 95% CI: 1.00-3.59; P = 0.051; three RCTs; I2 = 71.9%). LBR per cycle was reported by one RCT performed with cleavage-stage embryos and was higher for vitrification (RR = 2.28; 95% CI: 1.17-4.44; P =  0.016; 216 cycles; one RCT). A secondary analysis was performed focusing on embryo cryosurvival rate. Pooled data from seven RCTs (3615 embryos) revealed a significant improvement in embryo cryosurvival following vitrification as compared with slow-freezing (RR = 1.59, 95% CI: 1.30-1.93; P < 0.001; I2 = 93%). Data from available RCTs suggest that vitrification/warming is superior to slow-freezing/thawing with regard to clinical outcomes (low quality of the evidence) and cryosurvival rates (moderate quality of the evidence) for oocytes, cleavage-stage embryos and blastocysts. The results were confirmed by cohort studies. The improvements obtained with the introduction of vitrification have several important clinical implications in ART. Based on this evidence, in particular regarding cryosurvival rates, laboratories that continue to use slow-freezing should consider transitioning to the use of vitrification for cryopreservation. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology.m

Oocyte, embryo and blastocyst cryopreservation in ART: systematic review and meta-analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance

PubMed Central

Rienzi, Laura; Gracia, Clarisa; Maggiulli, Roberta; LaBarbera, Andrew R.; Kaser, Daniel J.; Ubaldi, Filippo M.; Vanderpoel, Sheryl; Racowsky, Catherine

2017-01-01

Abstract BACKGROUND Successful cryopreservation of oocytes and embryos is essential not only to maximize the safety and efficacy of ovarian stimulation cycles in an IVF treatment, but also to enable fertility preservation. Two cryopreservation methods are routinely used: slow-freezing or vitrification. Slow-freezing allows for freezing to occur at a sufficiently slow rate to permit adequate cellular dehydration while minimizing intracellular ice formation. Vitrification allows the solidification of the cell(s) and of the extracellular milieu into a glass-like state without the formation of ice. OBJECTIVE AND RATIONALE The objective of our study was to provide a systematic review and meta-analysis of clinical outcomes following slow-freezing/thawing versus vitrification/warming of oocytes and embryos and to inform the development of World Health Organization guidance on the most effective cryopreservation method. SEARCH METHODS A Medline search was performed from 1966 to 1 August 2016 using the following search terms: (Oocyte(s) [tiab] OR (Pronuclear[tiab] OR Embryo[tiab] OR Blastocyst[tiab]) AND (vitrification[tiab] OR freezing[tiab] OR freeze[tiab]) AND (pregnancy[tiab] OR birth[tiab] OR clinical[tiab]). Queries were limited to those involving humans. RCTs and cohort studies that were published in full-length were considered eligible. Each reference was reviewed for relevance and only primary evidence and relevant articles from the bibliographies of included articles were considered. References were included if they reported cryosurvival rate, clinical pregnancy rate (CPR), live-birth rate (LBR) or delivery rate for slow-frozen or vitrified human oocytes or embryos. A meta-analysis was performed using a random effects model to calculate relative risk ratios (RR) and 95% CI. OUTCOMES One RCT study comparing slow-freezing versus vitrification of oocytes was included. Vitrification was associated with increased ongoing CPR per cycle (RR = 2.81, 95% CI: 1.05–7.51; P = 0.039; 48 and 30 cycles, respectively, per transfer (RR = 1.81, 95% CI 0.71–4.67; P = 0.214; 47 and 19 transfers) and per warmed/thawed oocyte (RR = 1.14, 95% CI: 1.02–1.28; P = 0.018; 260 and 238 oocytes). One RCT comparing vitrification versus fresh oocytes was analysed. In vitrification and fresh cycles, respectively, no evidence for a difference in ongoing CPR per randomized woman (RR = 1.03, 95% CI: 0.87–1.21; P = 0.744, 300 women in each group), per cycle (RR = 1.01, 95% CI: 0.86–1.18; P = 0.934; 267 versus 259 cycles) and per oocyte utilized (RR = 1.02, 95% CI: 0.82–1.26; P = 0.873; 3286 versus 3185 oocytes) was reported. Findings were consistent with relevant cohort studies. Of the seven RCTs on embryo cryopreservation identified, three met the inclusion criteria (638 warming/thawing cycles at cleavage and blastocyst stage), none of which involved pronuclear-stage embryos. A higher CPR per cycle was noted with embryo vitrification compared with slow-freezing, though this was of borderline statistical significance (RR = 1.89, 95% CI: 1.00–3.59; P = 0.051; three RCTs; I2 = 71.9%). LBR per cycle was reported by one RCT performed with cleavage-stage embryos and was higher for vitrification (RR = 2.28; 95% CI: 1.17–4.44; P =  0.016; 216 cycles; one RCT). A secondary analysis was performed focusing on embryo cryosurvival rate. Pooled data from seven RCTs (3615 embryos) revealed a significant improvement in embryo cryosurvival following vitrification as compared with slow-freezing (RR = 1.59, 95% CI: 1.30–1.93; P < 0.001; I2 = 93%). WIDER IMPLICATIONS Data from available RCTs suggest that vitrification/warming is superior to slow-freezing/thawing with regard to clinical outcomes (low quality of the evidence) and cryosurvival rates (moderate quality of the evidence) for oocytes, cleavage-stage embryos and blastocysts. The results were confirmed by cohort studies. The improvements obtained with the introduction of vitrification have several important clinical implications in ART. Based on this evidence, in particular regarding cryosurvival rates, laboratories that continue to use slow-freezing should consider transitioning to the use of vitrification for cryopreservation. PMID:27827818

Equine ovarian tissue viability after cryopreservation and in vitro culture.

PubMed

Gastal, G D A; Aguiar, F L N; Alves, B G; Alves, K A; de Tarso, S G S; Ishak, G M; Cavinder, C A; Feugang, J M; Gastal, E L

2017-07-15

Ovarian tissue cryopreservation allows the preservation of the female fertility potential for an undetermined period. The objectives of this study were to compare the efficiency of cryoprotective agents (CPAs; dimethyl sulfoxide, DMSO; ethylene glycol, EG; and propylene glycol, PROH) using slow-freezing and vitrification methods, and evaluate the viability of cryopreserved equine ovarian tissue after 7 days of culture. Fresh and cryopreserved ovarian fragments were evaluated for preantral follicle morphology, stromal cell density, EGFR, Ki-67, Bax, and Bcl-2 protein expression, and DNA fragmentation. Vitrification with EG had the highest rate of morphologically normal preantral follicles, while DMSO had the lowest (76.1 ± 6.1% and 40.9 ± 14.8%, respectively; P < 0.05). In slow-freezing, despite that DMSO had the highest percentage of morphologically normal follicles (77.7 ± 5.8%), no difference among the CPAs was observed. Fluorescence intensity of EGFR and Ki-67 was greater when vitrification with EG was used. Regardless of the cryopreservation treatment, DMSO had the highest (P < 0.05) Bax/Bcl-2 ratio; however, DNA fragmentation was similar (P > 0.05) among treatments after thawing. After in vitro culture, the percentage of normal follicles was similar (P > 0.05) between slow-freezing and vitrification methods; however, vitrification had greater (P < 0.05) stromal cell density than slow-freezing. In summary, equine ovarian tissue was successfully cryopreserved, increasing the viability of the cells in the ovarian tissue after thawing when using DMSO and EG for slow-freezing and vitrification methods, respectively. Therefore, these results are relevant for fertility preservation programs. Copyright © 2017 Elsevier Inc. All rights reserved.

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

PubMed

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

2014-12-01

Adding dimethyl sulfoxide (DMSO) to hematopoietic progenitor cells (HPCs) causes an exothermic reaction, potentially affecting their viability. The freezing method might also influence this. The aim was to investigate the effect of 1) precooling of DMSO and plasma (D/P) and white blood cell (WBC)-enriched product, 2) DMSO concentration of D/P, 3) freezing program, and 4) storage method on WBC quality. WBC-enriched product without CD34+ cells was used instead of HPCs. This was divided into six or eight portions. D/P (20 or 50%; precooled or room temperature [RT]) was added to the WBC-enriched product (precooled or RT), resulting in 10% DMSO, while monitoring temperature. The product was frozen using controlled-rate freezing ("fast-rate" or "slow-rate") and placed in vapor-phase or liquid nitrogen. After thawing, WBC recovery and viability were determined. Temperature increased most for precooled D/P to precooled WBC-enriched product, without influence of 20 or 50% D/P, but remained for all variations below 30°C. WBC recovery for both freezing programs was more than 95%. Recovery of WBC viability was higher for slow-rate freezing compared to fast-rate freezing (74% vs. 61%; p < 0.05) and also for 50% compared to 20% D/P (two test variations). Effect of precooling D/P or WBC-enriched product and of storage in vapor-phase or liquid nitrogen was marginal. Based on these results, precooling is not necessary. Fifty percent D/P is preferred over 20% D/P. Slow-rate freezing is preferred over fast-rate freezing. For safety reasons storage in vapor-phase nitrogen is preferred over storage in liquid nitrogen. Additional testing using real HPCs might be necessary. © 2014 AABB.

Current results with slow freezing and vitrification of the human oocyte.

PubMed

Boldt, Jeffrey

2011-09-01

The past decade has witnessed renewed interest in human oocyte cryopreservation (OCP). This article reviews the two general methods used for OCP, slow freezing and vitrification, compares the outcomes associated with each technique and discusses the factors that might influence success with OCP (such as oocyte selection or day of transfer). Based on available data, OCP offers a reliable, reproducible method for preservation of the female gamete and will find increasing application in assisted reproductive technology. Oocyte cryopreservation can provide a number of advantages to couples undergoing assisted reproduction or to women interested in fertility preservation. Two methods, slow freezing and vitrification, have been used successfully for oocyte cryopreservation. This article reviews and compares these methods, and discusses various factors that can impact upon success of oocyte cryopreservation. Copyright © 2011 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Ice nucleating agents allow embryo freezing without manual seeding.

PubMed

Teixeira, Magda; Buff, Samuel; Desnos, Hugo; Loiseau, Céline; Bruyère, Pierre; Joly, Thierry; Commin, Loris

2017-12-01

Embryo slow freezing protocols include a nucleation induction step called manual seeding. This step is time consuming, manipulator dependent and hard to standardize. It requires access to samples, which is not always possible within the configuration of systems, such as differential scanning calorimeters or cryomicroscopes. Ice nucleation can be induced by other methods, e.g., by the use of ice nucleating agents. Snomax is a commercial preparation of inactivated proteins extracted from Pseudomonas syringae. The aim of our study was to investigate if Snomax can be an alternative to manual seeding in the slow freezing of mouse embryos. The influence of Snomax on the pH and osmolality of the freezing medium was evaluated. In vitro development (blastocyst formation and hatching rates) of fresh embryos exposed to Snomax and embryo cryopreserved with and without Snomax was assessed. The mitochondrial activity of frozen-thawed blastocysts was assessed by JC-1 fluorescent staining. Snomax didn't alter the physicochemical properties of the freezing medium, and did not affect embryo development of fresh embryos. After cryopreservation, the substitution of manual seeding by the ice nucleating agent (INA) Snomax did not affect embryo development or embryo mitochondrial activity. In conclusion, Snomax seems to be an effective ice nucleating agent for the slow freezing of mouse embryos. Snomax can also be a valuable alternative to manual seeding in research protocols in which manual seeding cannot be performed (i.e., differential scanning calorimetry and cryomicroscopy). Copyright © 2017 Elsevier Inc. All rights reserved.

Short-term and practice effects of metronome pacing in Parkinson's disease patients with gait freezing while in the 'on' state: randomized single blind evaluation.

PubMed

Cubo, Esther; Leurgans, Sue; Goetz, Christopher G

2004-12-01

In a randomized single blind parallel study, we tested the efficacy of an auditory metronome on walking speed and freezing in Parkinson's disease (PD) patients with freezing gait impairment during their 'on' function. No pharmacological treatment is effective in managing 'on' freezing in PD. Like visual cues that can help overcome freezing, rhythmic auditory pacing may provide cues that help normalize walking pace and overcome freezing. Non-demented PD patients with freezing during their 'on' state walked under two conditions, in randomized order: unassisted walking and walking with the use of an audiocassette with a metronome recording. The walking trials were randomized and gait variables were rated from videotapes by a blinded evaluator. Outcome measures were total walking time (total trial time-total freezing time), which was considered the time over a course of specified length, freezing time, average freeze duration and number of freezes. All outcomes were averaged across trials for each person and then compared across conditions using Signed Rank tests. Twelve non-demented PD patients with a mean age of 65.8 +/- 11.2 years, and mean PD duration of 12.4 +/- 7.3 years were included. The use of the metronome slowed ambulation and increased the total walking time (P < 0.0005) only during the first visit, without affecting any freezing variable. In the nine patients who took the metronome recording home and used it daily for 1 week while walking, freezing remained unimproved. Though advocated in prior publications as a walking aid for PD patients, auditory metronome pacing slows walking and is not a beneficial intervention for freezing during their 'on' periods.

Cryopreservation of animal oocytes and embryos: Current progress and future prospects.

PubMed

Mandawala, A A; Harvey, S C; Roy, T K; Fowler, K E

2016-10-15

Cryopreservation describes techniques that permit freezing and subsequent warming of biological samples without loss of viability. The application of cryopreservation in assisted reproductive technology encompasses the freezing of gametes, embryos, and primordial germ cells. Whilst some protocols still rely on slow-freezing techniques, most now use vitrification, or ultra-rapid freezing, for both oocytes and embryos due to an associated decreased risk of damage caused by the lack of ice crystal formation, unlike in slow-freezing techniques. Vitrification has demonstrated its use in many applications, not only following IVF procedures in human embryology clinics but also following in vitro production of embryos in agriculturally important, or endangered animal species, before embryo transfer. Here, we review the various cryopreservation and vitrification technologies that are used in both humans and other animals and discuss the most recent innovations in vitrification with a particular emphasis on their applicability to animal embryology. Copyright © 2016 Elsevier Inc. All rights reserved.

Contribution of extracellular ice formation and the solution effects to the freezing injury of PC-3 cells suspended in NaCl solutions.

PubMed

Takamatsu, Hiroshi; Zawlodzka, Sylwia

2006-08-01

The mechanism of cell injury during slow freezing was examined using PC-3 human prostate adenocarcinoma cells suspended in NaCl solutions. The objective was to evaluate contribution of extracellular ice and the 'solution effects' to freezing injury separately. The solution effects that designate the influence of elevated concentration were evaluated from a pseudo-freezing experiment, where cells were subjected to the milieu that simulated a freeze-thaw process by changing the NaCl concentration and the temperature at the same time. The effect of extracellular ice formation on cell injury was then estimated from the difference in cell survival between the pseudo-freezing experiment and a corresponding freezing experiment. When cells were frozen to a relatively higher freezing temperature at -10 degrees C, about 30% of cells were damaged mostly due to extracellular ice formation, because the concentration increase without ice formation to 2.5-M NaCl, i.e., the equilibrium concentration at -10 degrees C, had no effect on cell survival. In contrast, in the case of the lower freezing temperature at -20 degrees C, about 90% of cells were injured by both effects, particularly 60-80% by the solution effects among them. The present results suggested that the solution effects become more crucial to cell damage during slow freezing at lower temperatures, while the effect of ice is limited to some extent.

Slow and ultra-rapid freezing protocols for cryopreserving mouflon (Ovis musimon) and fallow deer (Dama dama) epididymal sperm.

PubMed

Bóveda, P; Esteso, M C; Castaño, C; Toledano-Díaz, A; López-Sebastián, A; Muñiz, A; Prieto, P; Mejía, O; Ungerfeld, R; Santiago-Moreno, J

2018-05-01

This study examines the effectiveness of two methods for cryopreserving post-mortem epididymal sperm - conventional slow freezing employing a short equilibration time with glycerol, and ultra-rapid freezing - from the wild ruminant species Ovis musimon (mouflon) and Dama dama (fallow deer). A Tris-citric acid-glucose (TCG) + 12% egg yolk-based medium was used for the conventional slow freezing of the fallow deer sperm, whereas a Tes-Tris-glucose (TEST) + 6% egg yolk-based medium was used for the mouflon sperm. Glycerol was added to a final concentration of 5% to both media. The same diluents were used for ultra-rapid freezing but replacing the glycerol with 100 mM of sucrose. Sperm variables (motility, viability, acrosome integrity, membrane integrity, and morphological abnormalities) were analyzed before and after cryopreservation. Although values were generally better after the thawing of the conventionally cryopreserved sperm, total sperm motility (38.40 ± 4.44% in mouflon and 31.25 ± 3.37% in fallow deer) and total live sperm (47.19 ± 5.18% in mouflon and 43.13 ± 2.43% in fallow deer) were acceptable for the ultra-rapidly cooled sperm. Independent of the cryopreservation method, membrane integrity, acrosome integrity and the percentages of dead sperm and sperms with a damaged acrosome were better for the cryopreserved mouflon sperm than the fallow deer sperm (P < 0.05). Despite exerting a more harmful effect on sperm variables than conventional freezing, ultra-rapid freezing may be a useful alternative for the cryopreservation of these species' epididymal sperm in the field, as this simple technique does not require sophisticated equipment and expertise. Copyright © 2018 Elsevier B.V. All rights reserved.

Conventional slow freezing cryopreserves mouflon spermatozoa better than vitrification.

PubMed

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

2017-04-01

This work examines the effectiveness of a TCG (Tris, citric acid, glucose, 6% egg yolk and 5% glycerol) and a TEST (TES, Tris, glucose, 6% egg yolk and 5% glycerol) sperm extender in the freezing of mouflon spermatozoa at slow cooling rates, using different pre-freezing equilibration times (2-3 hr). It also examines the tolerance of mouflon spermatozoa to different concentrations of cryoprotectants (5, 10, 20% glycerol; 5%, 10%, 20% dimethyl sulfoxide; 6% polyvinylpyrrolidone) and/or sucrose (100, 300, 500 mm). The highest quality (p < .01) thawed spermatozoa were obtained when using the TEST extender and an equilibration time of 3 hr. Sperm motility and membrane integrity were strongly reduced when using rapid freezing rates (60-85°C min -1 ), independent of the concentration of cryoprotectants. The lowest sucrose concentration (100 mm) provided the highest (p < .05) percentage of motile spermatozoa and live spermatozoa with an intact acrosome. Vitrified-warmed sperm variables were at their best when the spermatozoa was diluted in TCG-6% egg yolk + 100 mm sucrose and warmed at 60°C. Slow warming at 37°C strongly reduced (p < .05) sperm motility and viability. However, sperm vitrification returned lower fertility, sperm motility and sperm viability values than conventional sperm freezing. © 2016 Blackwell Verlag GmbH.

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.

PLEASE NOTE CHANGE OF ROOM! Particle Dark Matter (2/4)

ScienceCinema

Hooper, Dan

2018-04-16

Academic Training Lecture Regular Programme. Previously discussed the process of freeze-out of dark matter in the early universe, now discussing how other particles might affect the process of freeze-out.

The immunogenicity of thin-film freeze-dried, aluminum salt-adjuvanted vaccine when exposed to different temperatures.

PubMed

Thakkar, Sachin G; Ruwona, Tinashe B; Williams, Robert O; Cui, Zhengrong

2017-04-03

Insoluble aluminum salts such as aluminum oxyhydroxide have been used for decades as adjuvants in human vaccines, and many vaccines contain aluminum salts as adjuvants. Aluminum salt-adjuvanted vaccines must be managed in cold-chain (2-8° C) during transport and storage, as vaccine antigens in general are too fragile to be stable in ambient temperatures, and unintentional slowing freezing causes irreversible aggregation and permanent damage to the vaccines. Previously, we reported that thin-film freeze-drying can be used to convert vaccines adjuvanted with an aluminum salt from liquid suspension into dry powder without causing particle aggregation or decreasing in immunogenicity following reconstitution. In the present study, using ovalbumin (OVA)-adsorbed Alhydrogel® (i.e. aluminum oxyhydroxide, 2% w/v) as a model vaccine, we showed that the immunogenicity of thin-film freeze-dried OVA-adsorbed Alhydrogel® vaccine powder was not significantly changed after it was exposed for an extended period of time in temperatures as high as 40° C or subjected to repeated slow freezing-and-thawing. It is expected that immunization programs can potentially benefit by integrating thin-film freeze-drying into vaccine preparations.

The immunogenicity of thin-film freeze-dried, aluminum salt-adjuvanted vaccine when exposed to different temperatures

PubMed Central

Thakkar, Sachin G.; Ruwona, Tinashe B.; Williams, Robert O.; Cui, Zhengrong

2017-01-01

ABSTRACT Insoluble aluminum salts such as aluminum oxyhydroxide have been used for decades as adjuvants in human vaccines, and many vaccines contain aluminum salts as adjuvants. Aluminum salt-adjuvanted vaccines must be managed in cold-chain (2–8° C) during transport and storage, as vaccine antigens in general are too fragile to be stable in ambient temperatures, and unintentional slowing freezing causes irreversible aggregation and permanent damage to the vaccines. Previously, we reported that thin-film freeze-drying can be used to convert vaccines adjuvanted with an aluminum salt from liquid suspension into dry powder without causing particle aggregation or decreasing in immunogenicity following reconstitution. In the present study, using ovalbumin (OVA)-adsorbed Alhydrogel® (i.e. aluminum oxyhydroxide, 2% w/v) as a model vaccine, we showed that the immunogenicity of thin-film freeze-dried OVA-adsorbed Alhydrogel® vaccine powder was not significantly changed after it was exposed for an extended period of time in temperatures as high as 40° C or subjected to repeated slow freezing-and-thawing. It is expected that immunization programs can potentially benefit by integrating thin-film freeze-drying into vaccine preparations. PMID:28051903

Optimum Parameters for Freeze-Drying Decellularized Arterial Scaffolds

PubMed Central

Sheridan, William S.; Duffy, Garry P.

2013-01-01

Decellularized arterial scaffolds have achieved success in advancing toward clinical use as vascular grafts. However, concerns remain regarding long-term preservation and sterilization of these scaffolds. Freeze drying offers a means of overcoming these concerns. In this study, we investigated the effects of various freeze-drying protocols on decellularized porcine carotid arteries and consequently, determined the optimum parameters to fabricate a stable, preserved scaffold with unaltered mechanical properties. Freeze drying by constant slow cooling to two final temperatures ((Tf), −10°C and −40°C) versus instant freezing was investigated by histological examination and mechanical testing. Slow cooling to Tf= −10°C produced a stiffer and less distensible response than the non freeze-dried scaffolds and resulted in disruption to the collagen fibers. The mechanical response of Tf= −40°C scaffolds demonstrated disruption to the elastin network, which was confirmed with histology. Snap freezing scaffolds in liquid nitrogen and freeze drying to Tf= −40°C with a precooled shelf at −60°C produced scaffolds with unaltered mechanical properties and a histology resembling non-freeze-dried scaffolds. The results of this study demonstrate the importance of optimizing the nucleation and ice crystal growth/size to ensure homogenous drying, preventing extracellular matrix disruption and subsequent inferior mechanical properties. This new manufacturing protocol creates the means for the preservation and sterilization of decellularized arterial scaffolds while simultaneously maintaining the mechanical properties of the tissue. PMID:23614758

Vacuum Freeze-Drying, a Method Used To Salvage Water-Damaged Archival and Library Materials: A RAMP Study with Guidelines.

ERIC Educational Resources Information Center

McCleary, John M.

This Records and Archives Management Programme (RAMP) study covers the conservation of archival documents and the application of freeze-drying to the salvage of documents damaged by flood. Following an introductory discussion of the hazards of water, the study presents a broad summary of data on freeze-drying, including the behavior of…

Preservation of beluga (Delphinapterus leucas) spermatozoa using a trehalose-based cryodiluent and directional freezing technology.

PubMed

O'Brien, J K; Robeck, T R

2010-01-01

A beluga (Delphinapterus leucas) sperm preservation method was developed for use in genome banking and AI. In Study 1, glycerol-based cryodiluents (modified BF5F and modified Platz Diluent Variant (PDV)) were unable to maintain adequate progressive motility using straws (fast and slow freezing rate (FR)) or pellets (slow FR). Neither freezing method nor FR affected in vitro sperm characteristics (P > 0.05), but retention of prefreeze progressive motility following thawing was greater (P < 0.05) for BF5F (21%) than PDV (15%). In Study 2, examining the effects of straw freeze-thawing using BF5F with glycerol (1 and 3%, v/v) or trehalose (46 and 91 mM) on sperm characteristics, samples cryopreserved in trehalose exhibited superior (P < 0.05) in vitro parameters compared with their glycerol-treated counterparts. In Study 3, compared with a straw method, directional freezing using 91 mM trehalose enhanced (P < 0.05) sperm characteristics, with samples retaining 38%, 75% and 61% of their prefreeze progressive motility, curvilinear velocity and viability, respectively. A higher (P < 0.05) proportion of motile spermatozoa displayed rapid velocity after directional (21 +/- 1%) compared with straw (12 +/- 3%) freezing. Systematic development of a cryodiluent and the use of directional freezing resulted in beluga spermatozoa exhibiting adequate post-thaw quality for genome banking and use in AI.

Critical Slowing Down in Zn-Mg-Ho Quasicrystal

NASA Astrophysics Data System (ADS)

Sugiyama, Jun; Nozaki, Hiroshi; Ansaldo, Eduardo J.; Morris, Gerald D.; Brewer, Jess H.; Sato, Taku J.

By means of longitudinal field muon-spin spectroscopy, we have found a clear critical slowing down caused by spin fluctuation of Ho moments in the icosahedral quasicrystal (QC), i-ZnMgHo, with freezing temperature (Tf =1.95 K), for which the susceptibility showed an anomaly at5K. The difference is attributed to crystalline elec-tric field (CEF) effects. The muons experience a broad, fluctuating, field distribution, of width Δ ∼6.3Taround Tf . The effect of the CEF is also apparent in zero field and weak applied transverse field measurements, with an onset around 60 K. For the Cd-based QCs (CdMgHo and CdMgGd), which exhibited two freezing temperatures in the susceptibility, the change in fluctuation rate, i.e. freezing, occurs at the lower Tf .

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.

Cryopreservation of putative pre-pubertal bovine spermatogonial stem cells by slow freezing.

PubMed

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

2015-04-01

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

Influence of cell loss after vitrification or slow-freezing on further in vitro development and implantation of human Day 3 embryos.

PubMed

Van Landuyt, L; Van de Velde, H; De Vos, A; Haentjens, P; Blockeel, C; Tournaye, H; Verheyen, G

2013-11-01

Is the effect of cell loss on further cleavage and implantation different for vitrified than for slowly frozen Day 3 embryos? Vitrified embryos develop better overnight than slowly frozen embryos, regardless of the number of cells lost, but have similar implantation potential if further cleavage occurs overnight. After slow-freezing, similar implantation rates have been obtained for intact 4-cell embryos or 4-cell embryos with 1 cell damaged. For slowly frozen Day 3 embryos, lower implantation rates have been observed when at least 25% of cells were lost. Other studies reported similar implantation potential for 7- to 8-cell embryos with 0, 1 or 2 cells damaged. No data are available on further development of vitrified embryos in relation to cell damage. Survival and overnight cleavage were retrospectively assessed for 7664 slowly frozen Day 3 embryos (study period: January 2004-December 2008) and 1827 vitrified embryos (study period: April 2010-September 2011). Overnight cleavage was assessed according to cell stage at cryopreservation and post-thaw cell loss for both protocols. The relationship between cell loss and implantation rate was analysed in a subgroup of single-embryo transfers (SETs) with 780 slowly frozen and 294 vitrified embryos. Embryos with ≥6 blastomeres and ≤20% fragmentation were cryopreserved using slow controlled freezing [with dimethyl sulphoxide (DMSO) as cryoprotectant] or closed vitrification [with DMSO-ethylene glycol (EG)-sucrose (S) as cryoprotectants]. Only embryos with ≥50% of cells intact after thawing were cultured overnight and were only transferred if further cleaved. For each outcome, logistic regression analysis was performed. Survival was 94 and 64% after vitrification and slow-freezing respectively. Logistic regression analysis showed that overnight cleavage of surviving embryos was higher after vitrification than after slow-freezing (P < 0.001) and decreased according to the degree of cell damage (P < 0.001). If the embryo continued to cleave after thawing, there was no effect of the number of cells lost or the cryopreservation method on its implantation potential. The implantation rates of embryos with 0, 1 or 2 cells damaged were, respectively, 21% (n = 114), 21% (n = 28) and 20% (n = 12) after slow-freezing and 20% (n = 50), 21% (n = 5) and 27% (n = 4) after vitrification. This analysis is retrospective and study periods for vitrification and slow-freezing are different. The number of SETs with vitrified embryos is limited. However, a large number of vitrified embryos were available to analyse the further cleavage of surviving embryos. Although it is not proved that vitrified embryos are more viable than slowly frozen embryos in terms of pregnancy outcome, vitrification yields higher survival rates, better overnight development and higher transfer rates per embryo warmed. This increases the number of frozen transfer cycles originating from a single treatment and might result in a better cumulative clinical outcome. Based on the present data, the policy to warm an extra embryo before overnight culture depends on the cell stage at cryopreservation and the cell damage after warming. For 8-cell embryos, up to two cells may be damaged compared with only one cell for 6- to 7-cell embryos, before an additional embryo is warmed. none.

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.

Human single follicle growth in vitro from cryopreserved ovarian tissue after slow freezing or vitrification.

PubMed

Wang, Tian-ren; Yan, Jie; Lu, Cui-ling; Xia, Xi; Yin, Tai-lang; Zhi, Xu; Zhu, Xiao-hui; Ding, Ting; Hu, Wei-hong; Guo, Hong-yan; Li, Rong; Yan, Li-ying; Qiao, Jie

2016-04-01

What is the effect of human ovarian tissue cryopreservation on single follicular development in vitro? Vitrification had a greater negative effect on growth and gene expression of human ovarian follicles when compared with fresh follicles. For human ovarian cortex cryopreservation, the conventional option is slow freezing while more recently vitrification has been demonstrated to maintain good quality and function of ovarian tissues. Ovarian tissues were collected from 11 patients. For every patient, the ovarian cortex was divided into three samples: Fresh, slow-rate freezing (Slow) and vitrification (Vit). Tissue histology was performed and follicles were isolated for single-cell mRNA analysis and in vitro culture (IVC) in 1% alginate for 8 days. Follicle morphology was assessed with hematoxylin-eosin analysis. Follicles were individually embedded in alginate (1% w/v) and cultured in vitro for 8 days. Follicle survival and growth were assessed by microscopy. Follicle viability was observed after Calcein-AM and ethidium homodimer-I (Ca-AM/EthD-I) staining. Expression of genes, including GDF9 (growth differentiation factor 9), BMP15 (bone morphogenetic protein 15) and ZP3 (zona pellucida glycoprotein 3) in oocytes and AMH (anti-Mullerian hormone), FSHR (FSH receptor), CYP11A (cholesterol side-chain cleavage cytochrome P450) and STAR (steroidogenic acute regulatory protein) in GCs, was evaluated by single-cell mRNA analysis. A total of 129 follicles were separated from ovarian cortex (Fresh n = 44; Slow n = 40; Vit n = 45). The percentage of damaged oocytes and granulosa cells was significantly higher in both the Slow and Vit groups, as compared with Fresh control (P< 0.05). The growth of follicles in vitro was significantly delayed in the Vit group compared with the Fresh group (P< 0.05). Both slow freezing (P< 0.05) and vitrification (P< 0.05) down-regulated the mRNA levels of ZP3 and CYP11A compared with Fresh group, while there was no significant difference between the Slow and Vit groups (P> 0.05). Vitrification also down-regulates AMH mRNA levels compared with Fresh group (P< 0.05). Only short-term IVC studies (8 days) are reported. Further study should be performed to examine and improve follicular development in a long-term culture system after cryopreservation. This is the first comparison of gene expression and growth of single human ovarian follicles in vitro after either slow freezing or vitrification. With the decreased gene expression and growth during IVC, damage by cryopreservation still exists and needs to be minimized during the long-term IVC of follicles in the future for eventual clinical application. This work was supported by the National Natural Science Foundation of China (31230047, 81571386, 81471508, 31429004 and 81501247), National Natural Science Foundation of Beijing (7142166) and Mega-projects of Science Research for the 12th five-year plan (2012ba132b05). There are no conflicts of interest to declare. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Evaluation of combined effects of ageing period and freezing rate on quality attributes of beef loins.

PubMed

Kim, Yuan H Brad; Liesse, Charlotte; Kemp, Robert; Balan, Prabhu

2015-12-01

The objective of our study was to evaluate the combined effects of ageing period and different freezing rates on meat quality attributes of beef loins. Pairs of loins (M. longissimus at 1 day post mortem) from 12 carcasses were divided into four equal portions and randomly assigned to four ageing/freezing treatments (aged only, frozen only, and 3 or 4 weeks ageing at -1.5°C then frozen). Two freezing methods (fast freezing by calcium chloride immersion or slow freezing by air freezer at -18°C) were applied to the loin sections. Fast freezing had no effect on shear force (P>0.05), but significantly improved the water-holding capacity of the aged/frozen loins by reducing purge and drip losses. Ageing-then-freezing significantly improved shear force values of loins compared to both the aged only and frozen only loins. These observations suggest that fast freezing will add more value to the aged/frozen/thawed meat by minimising the amount of water-loss due to the freezing/thawing process. Copyright © 2015 Elsevier Ltd. All rights reserved.

Influence of 2 cryopreservation methods to induce CCL-13 from dental pulp cells.

PubMed

Ahn, Su-Jin; Jang, Ji-Hyun; Seo, Ji-Sung; Cho, Kyu Min; Jung, Su-Hee; Lee, Hyeon-Woo; Kim, Eun-Cheol; Park, Sang Hyuk

2013-12-01

Cryopreservation preserves periodontal ligament cells but has a lower success rate with dental pulp cells (DPCs) because it causes inflammation. There are 2 well-known cryopreservation methods that reduce inflammation, slow freezing and rapid freezing, but the effects of the 2 methods on inflammation are not well-established. The purpose of this study was to compare the effects of the 2 different cryopreservation methods on CCL-13 induction from DPCs by using microarrays, real-time polymerase chain reaction (PCR), Western blotting, enzyme-linked immunosorbent assay, and confocal laser scanning microscopy (CLSM). In this study, the concentration of cryoprotectant was fixed, and the methods compared differed with respect to freezing speed. Initially we screened the DPCs of cryopreserved teeth with expression microarrays, and CCL-13 was identified as a differentially expressed gene involved in generalized inflammation. We then compared the expression of CCL-13 after exposing teeth to the 2 cryopreservation methods by using real-time PCR, Western blot, enzyme-linked immunosorbent assay, and CLSM. Expression of CCL-13 was up-regulated significantly only in the rapid freezing group, except in measurements made by real-time PCR. CLSM analysis also confirmed this up-regulation visually. Rapid freezing increased the expression of CCL-13 in DPCs compared with slow freezing. Understanding the inflammatory effect of cryopreservation should help to establish an optimal cryoprofile to minimize inflammation of DPCs and reduce the need for endodontic treatment. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Comparison of different cryopreservation methods for horse and donkey embryos.

PubMed

Pérez-Marín, C C; Vizuete, G; Vazquez-Martinez, R; Galisteo, J J

2018-05-01

Few studies have been published about cryopreservation and embryo assessment in horses and donkeys. To evaluate the viability of embryos collected from mares and jennies that were cryopreserved by slow freezing or by vitrification. Randomised controlled experiment. Horse (n=19) and donkey (n=16) embryos (≤300 μm) were recovered on days 6.5-7.5 post-ovulation and assigned to control or cryopreservation protocols of slow freezing or vitrification. For slow freezing, 1.5 mol/L ethylene glycol (EG) was used. For vitrification, horse embryos were exposed to 1.4 mol/L glycerol, 1.4 mol/L glycerol + 3.6 mol/L EG and 3.4 mol/L glycerol + 4.6 mol/L EG, using Fibreplug or a 0.25 mL straw; donkey embryos were vitrified using Fibreplug with similar EG-glycerol solutions to above or 7.0 mol/L EG. Dead cells, apoptotic and fragmented nuclei, and cytoskeleton quality were assessed on thawed/warmed embryos. A significant decrease in embryo quality was observed after cryopreservation (P<0.05). Although the percentage of dead cells was lower (P<0.05) in control than in cryopreserved embryos, no differences were observed between freezing protocols used for horse or donkey embryos. While no differences were detected in the number of apoptotic cells in warmed horse embryos, in donkey embryos a higher incidence of apoptosis was measured after vitrification with EG-glycerol in Fibreplug (P<0.05). Vitrified horse embryos had a significantly (P<0.05) higher percentage of nonviable cells than donkey embryo. Actin cytoskeleton quality did not differ between treatments. Difficulties in obtaining a large number of embryos meant that the number of embryos per group was low. Vitrified horse and donkey embryos did not show higher susceptibility to cell damage than those preserved by slow freezing, whether using straws or Fibreplug. However, Fibreplug with EG 7 mol/L resulted in fewer nonviable and apoptotic cells in donkey embryos. Donkey embryos showed lower susceptibility to vitrification than horse embryos. THE SUMMARY IS AVAILABLE IN SPANISH - SEE SUPPORTING INFORMATION. © 2017 EVJ Ltd.

Amygdala kindling-resistant (SLOW) or -prone (FAST) rat strains show differential fear responses.

PubMed

Mohapel, P; McIntyre, D C

1998-12-01

The authors compared two rat strains, selectively bred for their susceptibility to amygdala kindling, with respect to their performance on various behavioral and learning tasks that are associated with fear and anxiety. The two rat strains differed significantly in measurements of exploration of novel and familiar environments, as well as in reactivity to footshock and fear-based learning. The kindling-resistant (SLOW) strain exhibited a lower ratio of open- to closed-arm entries in the elevated plus-maze, less activity over days in the open field, greater behavioral suppression in the open-field if previously footshocked, greater freezing in the inhibitory avoidance task, and slower acquisition and poorer retention in the one-way avoidance task than did the kindling-prone (FAST) strain. These experiments suggest that the SLOW rats are more expressively fearful than the FAST rats, particularly with respect to environmentally triggered freezing or immobility. Further, these observations imply that the relatively constrained excitability of the amygdala network in the SLOW rats might mediate their relatively greater expression of fear and anxiety compared with the FAST rats.

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.

Pharmaceutical patent applications in freeze-drying.

PubMed

Ekenlebie, Edmond; Einfalt, Tomaž; Karytinos, Arianna Irò; Ingham, Andrew

2016-09-01

Injectable products are often the formulation of choice for new therapeutics; however, formulation in liquids often enhances degradation through hydrolysis. Thus, freeze-drying (lyophilization) is regularly used in pharmaceutical manufacture to reduce water activity. Here we examine its contribution to 'state of the art' and look at its future potential uses. A comprehensive search of patent databases was conducted to characterize the international patent landscape and trends in the use of freeze-drying. A total of 914 disclosures related to freeze-drying, lyophilization or drying of solid systems in pressures and temperatures equivalent to those of freeze-drying were considered over the period of 1992-2014. Current applications of sublimation technology were contrasted across two periods those with patents due to expire (1992-1993) and those currently filed. The number of freeze-drying technology patents has stabilized after initial activity across the biotechnology sector in 2011 and 2012. Alongside an increasing trend for patent submissions, freeze-drying submissions have slowed since 2002 and is indicative of a level of maturity.

Effect of Freezing Conditions on Distances and Their Distributions Derived from Double Electron Electron Resonance (DEER): A Study of Doubly-Spin-Labeled T4 Lysozyme

PubMed Central

Georgieva, Elka R.; Roy, Aritro S.; Grigoryants, Vladimir M.; Borbat, Petr P.; Earle, Keith A.; Scholes, Charles P.; Freed, Jack H.

2012-01-01

Pulsed dipolar ESR spectroscopy, DEER and DQC, require frozen samples. An important issue in the biological application of this technique is how the freezing rate and concentration of cryoprotectant could possibly affect the conformation of biomacromolecule and/or spin-label. We studied in detail the effect of these experimental variables on the distance distributions obtained by DEER from a series of doubly spin-labeled T4 lysozyme mutants. We found that the rate of sample freezing affects mainly the ensemble of spin-label rotamers, but the distance maxima remain essentially unchanged. This suggests that proteins frozen in a regular manner in liquid nitrogen faithfully maintain the distance-dependent structural properties in solution. We compared the results from rapidly freeze-quenched (≤100 μs) samples to those from commonly shock-frozen (slow freeze, 1s or longer) samples. For all the mutants studied we obtained inter-spin distance distributions, which were broader for rapidly frozen samples than for slowly frozen ones. We infer that rapid freezing trapped a larger ensemble of spin label rotamers; whereas, on the time-scale of slower freezing the protein and spin-label achieve a population showing fewer low-energy conformers. We used glycerol as a cryoprotectant in concentrations of 10% and 30% by weight. With 10% glycerol and slow freezing, we observed an increased slope of background signals, which in DEER is related to increased local spin concentration, in this case due to insufficient solvent vitrification, and therefore protein aggregation. This effect was considerably suppressed in slowly frozen samples containing 30% glycerol and rapidly frozen samples containing 10% glycerol. The assignment of bimodal distributions to tether rotamers as opposed to protein conformations is aided by comparing results using MTSL and 4-Bromo MTSL spin-labels. The latter usually produce narrower distance distributions. PMID:22341208

Clinical outcomes following cryopreservation of blastocysts by vitrification or slow freezing: a population-based cohort study.

PubMed

Li, Z; Wang, Y A; Ledger, W; Edgar, D H; Sullivan, E A

2014-12-01

What are the clinical efficacy and perinatal outcomes following transfer of vitrified blastocysts compared with transfer of fresh or of slow frozen blastocysts? Compared with slow frozen blastocysts, vitrified blastocysts resulted in significantly higher clinical pregnancy and live delivery rates with similar perinatal outcomes at population level. Although vitrification has been reported to be associated with significantly increased post-thaw survival rates compared with slow freezing, there has been a lack of general consensus over which method of cryopreservation (vitrification versus slow freezing) is most appropriate for blastocysts. A population-based cohort of autologous fresh and initiated thaw cycles (a cycle where embryos were thawed with intention to transfer) performed between January 2009 and December 2011 in Australia and New Zealand was evaluated retrospectively. A total of 46 890 fresh blastocyst transfer cycles, 12 852 initiated slow frozen blastocyst thaw cycles and 20 887 initiated vitrified blastocyst warming cycles were included in the data analysis. Pairwise comparisons were made between the vitrified blastocyst group and slow frozen or fresh blastocyst group. A Chi-square test was used for categorical variables and t-test was used for continuous variables. Cox regression was used to examine the pregnancy outcomes (clinical pregnancy rate, miscarriage rate and live delivery rate) and perinatal outcomes (preterm delivery, low birthweight births, small for gestational age (SGA) births, large for gestational age (LGA) births and perinatal mortality) following transfer of fresh, slow frozen and vitrified blastocysts. The 46 890 fresh blastocyst transfers, 11 644 slow frozen blastocyst transfers and 19 978 vitrified blastocyst transfers resulted in 16 845, 2766 and 6537 clinical pregnancies, which led to 13 049, 2065 and 4955 live deliveries, respectively. Compared with slow frozen blastocyst transfer cycles, vitrified blastocyst transfer cycles resulted in a significantly higher clinical pregnancy rate (adjusted relative risk (ARR): 1.47, 95% confidence intervals (CI): 1.39-1.55) and live delivery rate (ARR: 1.41, 95% CI: 1.34-1.49). Compared with singletons born after transfer of fresh blastocysts, singletons born after transfer of vitrified blastocysts were at 14% less risk of being born preterm (ARR: 0.86, 95% CI: 0.77-0.96), 33% less risk of being low birthweight (ARR: 0.67, 95% CI: 0.58-0.78) and 40% less risk of being SGA (ARR: 0.60, 95% CI: 0.53-0.68). A limitation of this population-based study is the lack of information available on clinic-specific cryopreservation protocols and processes for slow freezing-thaw and vitrification-warm of blastocysts and the potential impact on outcomes. This study presents population-based evidence on clinical efficacy and perinatal outcomes associated with transfer of fresh, slow frozen and vitrified blastocysts. Vitrified blastocyst transfer resulted in significantly higher clinical pregnancy and live delivery rates with similar perinatal outcomes compared with slow frozen blastocyst transfer. Comparably better perinatal outcomes were reported for singletons born after transfer of vitrified blastocysts than singletons born after transfer of fresh blastocysts. Elective vitrification could be considered as an alternative embryo transfer strategy to achieve better perinatal outcomes following Assisted Reproduction Technology (ART) treatment. No specific funding was obtained. The authors have no conflicts of interest to declare. © The Author 2014. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Brine rejection from freezing salt solutions: a molecular dynamics study.

PubMed

Vrbka, Lubos; Jungwirth, Pavel

2005-09-30

The atmospherically and technologically very important process of brine rejection from freezing salt solutions is investigated with atomic resolution using molecular dynamics simulations. The present calculations allow us to follow the motion of each water molecule and salt ion and to propose a microscopic mechanism of brine rejection, in which a fluctuation (reduction) of the ion density in the vicinity of the ice front is followed by the growth of a new ice layer. The presence of salt slows down the freezing process, which leads to the formation of an almost neat ice next to a disordered brine layer.

Vogel-Fulcher-Tammann freezing of a thermally fluctuating artificial spin ice probed by x-ray photon correlation spectroscopy

DOE PAGES

Morley, S. A.; Alba Venero, D.; Porro, J. M.; ...

2017-03-16

We report on the crossover from the thermal to the athermal regime of an artificial spin ice formed from a square array of magnetic islands whose lateral size, 30 nm × 70 nm, is small enough that they are dynamic at room temperature.We used resonant magnetic soft x-ray photon correlation spectroscopy as a method to observe the time-time correlations of the fluctuating magnetic configurations of spin ice during cooling, which are found to slow abruptly as a freezing temperature of T 0 = 178 ± 5 K is approached. This slowing is well described by a Vogel-Fulcher-Tammann law, implying thatmore » the frozen state is glassy, with the freezing temperature being commensurate with the strength of magnetostatic interaction energies in the array. The activation temperature, T A = 40 ± 10 K, is much less than that expected from a Stoner-Wohlfarth coherent rotation model. Zerofield- cooled/field-cooled magnetometry reveals a freeing up of fluctuations of states within islands above this temperature, caused by variation in the local anisotropy axes at the oxidised edges. This Vogel-Fulcher-Tammann behavior implies that the system enters a glassy state upon freezing, which is unexpected for a system with a well-defined ground state.« less

Vogel-Fulcher-Tammann freezing of a thermally fluctuating artificial spin ice probed by x-ray photon correlation spectroscopy

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

Morley, S. A.; Alba Venero, D.; Porro, J. M.

We report on the crossover from the thermal to the athermal regime of an artificial spin ice formed from a square array of magnetic islands whose lateral size, 30 nm × 70 nm, is small enough that they are dynamic at room temperature.We used resonant magnetic soft x-ray photon correlation spectroscopy as a method to observe the time-time correlations of the fluctuating magnetic configurations of spin ice during cooling, which are found to slow abruptly as a freezing temperature of T 0 = 178 ± 5 K is approached. This slowing is well described by a Vogel-Fulcher-Tammann law, implying thatmore » the frozen state is glassy, with the freezing temperature being commensurate with the strength of magnetostatic interaction energies in the array. The activation temperature, T A = 40 ± 10 K, is much less than that expected from a Stoner-Wohlfarth coherent rotation model. Zerofield- cooled/field-cooled magnetometry reveals a freeing up of fluctuations of states within islands above this temperature, caused by variation in the local anisotropy axes at the oxidised edges. This Vogel-Fulcher-Tammann behavior implies that the system enters a glassy state upon freezing, which is unexpected for a system with a well-defined ground state.« less

Comparison of Cryopreservation Protocols (Single and Two-steps) and Thawing (Fast and Slow) for Canine Sperm.

PubMed

Brito, Maíra M; Lúcio, Cristina F; Angrimani, Daniel S R; Losano, João Diego A; Dalmazzo, Andressa; Nichi, Marcílio; Vannucchi, Camila I

2017-01-02

In addition to the existence of several cryopreservation protocols, no systematic research has been carried out in order to confirm the suitable protocol for canine sperm. This study aims to assess the effect of adding 5% glycerol during cryopreservation at 37°C (one-step) and 5°C (two-steps), in addition of testing two thawing protocols (37°C for 30 seconds, and 70°C for 8 seconds). We used 12 sperm samples divided into four experimental groups: Single-Step - Slow Thawing Group; Two-Step - Slow Thawing Group; Single-Step - Fast Thawing Group; and Two-Step - Fast Thawing Group. Frozen-thawed samples were submitted to automated analysis of sperm motility, evaluation of plasmatic membrane integrity, acrosomal integrity, mitochondrial activity, sperm morphology, sperm susceptibility to oxidative stress, and sperm binding assay to perivitellinic membrane of chicken egg yolk. Considering the comparison between freezing protocols, no statistical differences were verified for any of the response variables. When comparison between thawing protocols was performed, slow thawing protocol presented higher sperm count bound to perivitelline membrane of chicken egg yolk, compared to fast thawing protocol. Regardless of the freezing process, the slow thawing protocol can be recommended for the large scale cryopreservation of canine semen, since it shows a consistent better functional result.

Antifreeze Proteins Modify the Freezing Process In Planta12

PubMed Central

Griffith, Marilyn; Lumb, Chelsey; Wiseman, Steven B.; Wisniewski, Michael; Johnson, Robert W.; Marangoni, Alejandro G.

2005-01-01

During cold acclimation, winter rye (Secale cereale L. cv Musketeer) plants accumulate antifreeze proteins (AFPs) in the apoplast of leaves and crowns. The goal of this study was to determine whether these AFPs influence survival at subzero temperatures by modifying the freezing process or by acting as cryoprotectants. In order to inhibit the growth of ice, AFPs must be mobile so that they can bind to specific sites on the ice crystal lattice. Guttate obtained from cold-acclimated winter rye leaves exhibited antifreeze activity, indicating that the AFPs are free in solution. Infrared video thermography was used to observe freezing in winter rye leaves. In the absence of an ice nucleator, AFPs had no effect on the supercooling temperature of the leaves. However, in the presence of an ice nucleator, AFPs lowered the temperature at which the leaves froze by 0.3°C to 1.2°C. In vitro studies showed that apoplastic proteins extracted from cold-acclimated winter rye leaves inhibited the recrystallization of ice and also slowed the rate of migration of ice through solution-saturated filter paper. When we examined the possible role of winter rye AFPs in cryoprotection, we found that lactate dehydrogenase activity was higher after freezing in the presence of AFPs compared with buffer, but the same effect was obtained by adding bovine serum albumin. AFPs had no effect on unstacked thylakoid volume after freezing, but did inhibit stacking of the thylakoids, thus indicating a loss of thylakoid function. We conclude that rye AFPs have no specific cryoprotective activity; rather, they interact directly with ice in planta and reduce freezing injury by slowing the growth and recrystallization of ice. PMID:15805474

'Banking time': egg freezing and the negotiation of future fertility.

PubMed

Waldby, Catherine

2015-01-01

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

Effect of ultrasound-assisted freezing on the physico-chemical properties and volatile compounds of red radish.

PubMed

Xu, Bao-Guo; Zhang, Min; Bhandari, Bhesh; Cheng, Xin-Feng; Islam, Md Nahidul

2015-11-01

Power ultrasound, which can enhance nucleation rate and crystal growth rate, can also affect the physico-chemical properties of immersion frozen products. In this study, the influence of slow freezing (SF), immersion freezing (IF) and ultrasound-assisted freezing (UAF) on physico-chemical properties and volatile compounds of red radish was investigated. Results showed that ultrasound application significantly improved the freezing rate; the freezing time of ultrasound application at 0.26 W/cm(2) was shorten by 14% and 90%, compared to IF and SF, respectively. UAF products showed significant (p<0.05) reduction in drip loss and phytonutrients (anthocyanins, vitamin C and phenolics) loss. Compared to SF products, IF and UAF products showed better textural preservation and higher calcium content. The radish tissues exhibited better cellular structures under ultrasonic power intensities of 0.17 and 0.26 W/cm(2) with less cell separation and disruption. Volatile compound data revealed that radish aromatic profile was also affected in the freezing process. Copyright © 2015 Elsevier B.V. All rights reserved.

A comparative study of sodium dodecyl sulfate and freezing/thawing treatment on wheat starch: The role of water absorption.

PubMed

Tao, Han; Wang, Pei; Zhang, Bao; Wu, Fengfeng; Jin, Zhengyu; Xu, Xueming

2016-06-05

The effect of freezing on functionality of native and sodium dodecyl sulfate (SDS)-treated wheat starches was investigated, with the aim of understanding the role of water absorption during freezing process. SDS is one of most efficient detergents to remove non-starch components (such as proteins and lipids) for starches but does not cause any apparent damage on granular structure. Slow swelling could be converted to rapid swelling by SDS washing, indicating higher water absorption. Freezing process induced slight roughness on starch granules but the non-starch components content was little affected. Combined SDS+freezing treatment significantly decreased both amylose and proteins non-starch components contents, which was accompanied with high gelatinization temperatures, melting enthalpy, and pasting viscosities. A smaller bread specific volume was obtained from SDS+freezing-treated starches while the crumb firmness significantly increased (p<0.05). SDS mainly extracted the surface components from starch granules, leading to high water absorption and making granules sensitive to the freezing treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

Human oocyte cryopreservation in infertility and oncology.

PubMed

Porcu, Eleonora; Bazzocchi, Antonia; Notarangelo, Leonardo; Paradisi, Roberto; Landolfo, Chiara; Venturoli, Stefano

2008-12-01

To evaluate the present state of research and clinical application of human oocyte cryopreservation in infertility and oncology. Recent literature documents have an increasing interest in cryopreserving human eggs. A number of studies report on different freezing protocols and various types of clinical application. Increasing attention is paid to vitrification as an alternative to slow cooling for oocyte cryopreservation. Several studies cover the modification of meiotic spindle during cryopreservation in order to assess the less damaging cryopreservation system. The first births with cryopreserved oocytes in cancer patients are reported. Egg freezing may circumvent the ethical and legal concerns regarding embryo cryopreservation, increase assisted reproduction flexibility and be a concrete option to save fertility in women with cancer. Recently, egg survival and pregnancy rates improved, with the birth of more than 500 children. The birth rate per thawed oocyte is around 5-6%. As regards safety, data on birth defects seems to be reassuring so far but must be monitored by an international registry. Comparative studies between slow freezing and vitrification in the same patient population are needed to elucidate pros and cons of each technique.

No difference in mitochondrial distribution is observed in human oocytes after cryopreservation.

PubMed

Stimpfel, Martin; Vrtacnik-Bokal, Eda; Virant-Klun, Irma

2017-08-01

The primary aim of this study was to determine if any difference in mitochondrial distribution can be observed between fresh and cryopreserved (slow-frozen/thawed and vitrified/warmed) oocytes when oocytes are stained with Mitotracker Red CMXRos and observed under a conventional fluorescent microscope. Additionally, the influence of cryopreservation procedure on the viable rates of oocytes at different maturation stages was evaluated. The germinal vesicle (GV) and MII oocytes were cryopreserved with slow-freezing and vitrification. After thawing/warming, oocytes were stained using Mitotracker Red CMXRos and observed under a conventional fluorescent microscope. Mitotracker staining revealed that in GV oocytes the pattern of mitochondrial distribution appeared as aggregated clusters around the whole oocyte. In mature MII oocytes, three different patterns of mitochondrial distribution were observed; a smooth pattern around the polar body with aggregated clusters at the opposite side of the polar body, a smooth pattern throughout the whole cell, and aggregated clusters as can be seen in GV oocytes. There were no significant differences in the observed patterns between fresh, vitrified/warmed and frozen/thawed oocytes. When comparing the viable rates of oocytes after two different cryopreservation procedures, the results showed no significant differences, although the trend of viable MII oocytes tends to be higher after vitrification/warming and for viable GV oocytes it tends to be higher after slow-freezing/thawing. Mitotracker Red CMXRos staining of mitochondria in oocytes did not reveal differences in mitochondrial distribution between fresh and cryopreserved oocytes at different maturity stages. Additionally, no difference was observed in the viable rates of GV and MII oocytes after slow-freezing/thawing and vitrification/warming.

Effect of estrous cow serum during bovine embryo culture on blastocyst development and cryotolerance after slow freezing or vitrification.

PubMed

Mucci, N; Aller, J; Kaiser, G G; Hozbor, F; Cabodevila, J; Alberio, R H

2006-05-01

The present study investigated the effect of estrous cow serum (ECS) during culture of bovine embryos on blastocyst development and survival after cryopreservation by slow freezing or vitrification. Embryos were derived from in vitro maturation (IVM) and in vitro fertilization (IVF) of abbatoir-derived oocytes. At Day 3, embryos were cultured in three different media: Charles Ronsenkrans medium + amino acids (CR1aa; without bovine serum albumin (BSA)) + 5% estrous cow serum (CR1-ECS), CR1aa + 3 mg/mL BSA (CR1-BSA) or CR1aa + 5% ECS + 3 mg/mL BSA (CR1-ECS-BSA). At 7.5 d post-insemination (PI), blastocyst yield and quality were evaluated; blastocysts and expanded blastocysts from each media were cryopreserved by Open Pulled Straw (OPS) vitrification method or slow freezing (1.5 M ethylene glycol, EM). Total blastocyst yield did not differ among CR1-ECS, CR1-BSA and CR1-ECS-BSA (30.9, 33.1 and 32.9%, respectively, P < 0.05). Embryo survival (hatching rate) was higher in vitrified versus slow-frozen embryos (43% versus 12%, respectively, P < 0.01), and in embryos cultured in CR1-BSA (40.3%) compared with those cultured in serum-containing media (CR1-ECS, 21.5% and CR1-ECS-BSA, 19.8%; P < 0.01). (a) it was possible to produce in vitro bovine embryos in serum-free culture medium without affecting blastocyst yield and quality; (b) serum-free medium produced the best quality embryos (in terms of post-cryopreservation survival); and (c) vitrification yielded the highest post-cryopreservation survival rates, regardless of the presence of serum in the culture medium.

Freezing temperatures as a limit to forest recruitment above tropical Andean treelines.

PubMed

Rehm, Evan M; Feeley, Kenneth J

2015-07-01

The elevation of altitudinal treelines is generally believed to occur where low mean temperatures during the growing season limit growth and prevent trees from establishing at higher elevations. Accordingly, treelines should move upslope with increasing global temperatures. Contrary to this prediction, tropical treelines have remained stable over the past several decades despite increasing mean temperatures. The observed stability of tropical treelines, coupled with the drastically different temperature profiles between temperate and tropical treelines, suggests that using mean measures of temperature to predict tropical treeline movements during climate change may be overly simplistic. We hypothesize that frost events at tropical treelines may slow climate driven treeline movement by preventing tree recruitment beyond the established forest canopy. To assess this hypothesis, we measured freezing resistance of four canopy-forming treeline species (Weinmannia fagaroides, Polylepis pauta, Clethra cuneata, and Gynoxys nitida) at two life stages (juvenile and adult) and during two seasons (warm-wet and cold-dry). Freezing resistances were then compared to microclimatic data to determine if freezing events in the grassland matrix above treeline are too harsh for these forest species. Freezing resistance varied among species and life stages from -5.7 degrees C for juveniles of P. pauta to -11.1 degrees C for juveniles of W. fagaroides. Over a four-year period, the lowest temperatures recorded at 10 cm above ground level in the grasslands above treeline and at treeline itself were -8.9 degrees C and -6.8 degrees C, respectively. Juveniles maintained freezing resistances similar to adults during the coldest parts of the year and ontogenetic differences in freezing resistance were only present during the warm season when temperatures did not represent a significant threat to active plant tissue. These findings support the hypothesis that rare extreme freezing events at and above tropical treelines can prevent recruitment outside of closed canopy forest for some tree species and may significantly slow treeline advancement despite warming mean temperatures. Predictions of treeline shifts under climate change should be reevaluated to include species-specific' climatic tolerances and measures of climatic variability.

Influence of the freezing method on the changes that occur in grape samples after frozen storage.

PubMed

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

2013-09-01

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

Kinetics of Water Loss from Cells at Subzero Temperatures and the Likelihood of Intracellular Freezing

PubMed Central

Mazur, Peter

1963-01-01

The survival of various cells subjected to low temperature exposure is higher when they are cooled slowly. This increase is consistent with the view that slow cooling decreases the probability of intracellular freezing by permitting water to leave the cell rapidly enough to keep the protoplasm at its freezing point. The present study derives a quantitative relation between the amount of water in a cell and temperature. The relation is a differential equation involving cooling rate, surface-volume ratio, membrane permeability to water, and the temperature coefficient of the permeability constant. Numerical solutions to this equation give calculated water contents which permit predictions as to the likelihood of intracellular ice formation. Both the calculated water contents and the predictions on internal freezing are consistent with the experimental observations of several investigators. PMID:14085017

The Alpha consensus meeting on cryopreservation key performance indicators and benchmarks: proceedings of an expert meeting.

PubMed

2012-08-01

This proceedings report presents the outcomes from an international workshop designed to establish consensus on: definitions for key performance indicators (KPIs) for oocyte and embryo cryopreservation, using either slow freezing or vitrification; minimum performance level values for each KPI, representing basic competency; and aspirational benchmark values for each KPI, representing best practice goals. This report includes general presentations about current practice and factors for consideration in the development of KPIs. A total of 14 KPIs were recommended and benchmarks for each are presented. No recommendations were made regarding specific cryopreservation techniques or devices, or whether vitrification is 'better' than slow freezing, or vice versa, for any particular stage or application, as this was considered to be outside the scope of this workshop. Copyright © 2012 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Preservation of primordial follicles from lions by slow freezing and xenotransplantation of ovarian cortex into an immunodeficient mouse.

PubMed

Wiedemann, C; Hribal, R; Ringleb, J; Bertelsen, M F; Rasmusen, K; Andersen, C Y; Kristensen, S G; Jewgenow, K

2012-12-01

Assisted reproductive technology (ART) is considered an important tool in the conservation of endangered species, but often the most limiting factor of ART is the availability of mature oocytes. The aim of the present study was to investigate the feasibility of preserving female germ cells from ovaries of female lions (Panthera leo). Good quality cumulus-oocyte complexes (COCs) were isolated and subjected to in vitro maturation (IVM). In addition, ovarian cortex was obtained and cut into pieces for culture and cryopreservation by slow freezing. The survival of ovarian follicles was assessed by histology. Frozen-thawed samples of ovarian cortex samples were xenotransplanted under the skin of ovariectomized immunodeficient mouse for 28 days. Overall, 178 intact COCs were obtained from 13 lions, but only 28.1% were matured in vitro indicating insufficient IVM conditions. In contrast, almost all follicles within the ovarian cortex survived culture when the original sample was from a young healthy lion collected immediately after euthanasia. Within the xenotransplants, the number of primordial follicles decreased after 28 days by 20%, but the relation between primordial and growing follicles changed in favour of follicular growth. Female gamete rescue from valuable felids may be performed by slow freeze cryopreservation of ovarian cortex. Although the IVM protocol for lions is not yet optimized, mature oocytes may be obtained after long-term xenotransplantation and IVM and could potentially represent one way of salvage of endangered felid species in the future. © 2012 Blackwell Verlag GmbH.

Programmable fast-freezing method improves the post-thaw motion dynamics, integrities of plasmalemma, mitochondrial transmembrane, DNA and, acrosome, and in vivo fertility of water buffalo (Bubalus bubalis) spermatozoa.

PubMed

Khalil Ur Rehman, H; Andrabi, S M H; Ahmed, H; Shah, S A H

2017-10-01

The effects of freezing methods (FR1, nonprogrammable/static, 5 cm above liquid nitrogen [LN 2 ] for 10 min, plunging in LN 2 ; FR2, programmable medium, +4°C to -15°C at 3°C min -1 , from -15 to -80°C at 10°C min -1 and final holding for 1 min at -80°C, plunging in LN 2 ; FR3, programmable fast, from initial holding at +4°C for 2 min, from +4°C to -20°C at 10°C min -1 , from -20°C to -100°C at 30°C min -1 , final holding for 1 min at -100°C and plunging in LN 2 ) were assessed on post-thaw in vitro quality and in vivo fertility of water buffalo spermatozoa. Mean sperm progressive motility (%), rapid velocity (%), average path velocity (μm s -1 ), straight line velocity (μm s -1 ), curved line velocity (μm s -1 ), integrities (%) of plasmalemma, mitochondrial transmembrane, DNA and acrosome were higher (p < .05) in samples cryopreserved with FR3 compared to FR1 and FR2. Similarly, in vivo fertility (%) of buffalo spermatozoa was higher (p < .05) with FR3 than FR1 (%; 68.0 versus 50.0). We concluded that programmable fast-freezing method (FR3) improves the post-thaw in vitro quality and in vivo fertility of water buffalo spermatozoa. © 2016 Blackwell Verlag GmbH.

Freezing Bubbles

NASA Astrophysics Data System (ADS)

Kingett, Christian; Ahmadi, Farzad; Nath, Saurabh; Boreyko, Jonathan

2017-11-01

The two-stage freezing process of a liquid droplet on a substrate is well known; however, how bubbles freeze has not yet been studied. We first deposited bubbles on a silicon substrate that was chilled at temperatures ranging from -10 °C to -40 °C, while the air was at room temperature. We observed that the freeze front moved very slowly up the bubble, and in some cases, even came to a complete halt at a critical height. This slow freezing front propagation can be explained by the low thermal conductivity of the thin soap film, and can be observed more clearly when the bubble size or the surface temperature is increased. This delayed freezing allows the frozen portion of the bubble to cool the air within the bubble while the top part is still liquid, which induces a vapor pressure mismatch that either collapses the top or causes the top to pop. In cases where the freeze front reaches the top of the bubble, a portion of the top may melt and slowly refreeze; this can happen more than just once for a single bubble. We also investigated freezing bubbles inside of a freezer where the air was held at -20 °C. In this case, the bubbles freeze quickly and the ice grows radially from nucleation sites instead of perpendicular to the surface, which provides a clear contrast with the conduction limited room temperature bubbles.

Neural control of behavioural choice in juvenile crayfish.

PubMed

Liden, William H; Phillips, Mary L; Herberholz, Jens

2010-11-22

Natural selection leads to behavioural choices that increase the animal's fitness. The neuronal mechanisms underlying behavioural choice are still elusive and empirical evidence connecting neural circuit activation to adaptive behavioural output is sparse. We exposed foraging juvenile crayfish to approaching shadows of different velocities and found that slow-moving shadows predominantly activated a pair of giant interneurons, which mediate tail-flips that thrust the animals backwards and away from the approaching threat. Tail-flips also moved the animals farther away from an expected food source, and crayfish defaulted to freezing behaviour when faced with fast-approaching shadows. Under these conditions, tail-flipping, an ineffective and costly escape strategy was suppressed in favour of freezing, a more beneficial choice. The decision to freeze also dominated in the presence of a more desirable resource; however, the increased incentive was less effective in suppressing tail-flipping when paired with slow-moving visual stimuli that reliably evoked tail-flips in most animals. Together this suggests that crayfish make value-based decisions by weighing the costs and benefits of different behavioural options, and they select adaptive behavioural output based on the activation patterns of identifiable neural circuits.

Cryopreservation of human sperm: efficacy and use of a new nitrogen-free controlled rate freezer versus liquid nitrogen vapour freezing.

PubMed

Creemers, E; Nijs, M; Vanheusden, E; Ombelet, W

2011-12-01

Preservation of spermatozoa is an important aspect of assisted reproductive medicine. The aim of this study was to investigate the efficacy and use of a recently developed liquid nitrogen and cryogen-free controlled rate freezer and this compared with the classical liquid nitrogen vapour freezing method for the cryopreservation of human spermatozoa. Ten patients entering the IVF programme donated semen samples for the study. Samples were analysed according to the World Health Organization guidelines. No significant difference in total sperm motility after freeze-thawing between the new technique and classical technique was demonstrated. The advantage of the new freezing technique is that it uses no liquid nitrogen during the freezing process, hence being safer to use and clean room compatible. Investment costs are higher for the apparatus but running costs are only 1% in comparison with classical liquid nitrogen freezing. In conclusion, post-thaw motility of samples frozen with the classical liquid nitrogen vapour technique was comparable with samples frozen with the new nitrogen-free freezing technique. This latter technique can thus be a very useful asset to the sperm cryopreservation laboratory. © 2011 Blackwell Verlag GmbH.

Abscisic acid induced freezing tolerance in chilling-sensitive suspension cultures and seedlings of rice

PubMed Central

2013-01-01

Background The role of abscisic acid (ABA) as a possible activator of cold acclimation process was postulated since endogenous levels of ABA increase temporarily or constitutively during cold-hardening. Exogenous application of ABA has been known to induce freezing tolerance at ambient temperatures in in vitro systems derived from cold hardy plants. Yet, some cell cultures acquired much greater freezing tolerance by ABA than by cold whilst maintaining active growth. This raises questions about the relationships among ABA, cold acclimation and growth cessation. To address this question, we attempted to 1) determine whether exogenous ABA can confer freezing tolerance in chilling-sensitive rice suspension cells and seedlings, which obviously lack the mechanisms to acquire freezing tolerance in response to cold; 2) characterize this phenomenon by optimizing the conditions and compare with the case of cold hardy bromegrass cells. Results Non-embryogenic suspension cells of rice suffered serious chilling injury when exposed to 4°C. When incubated with ABA at the optimal conditions (0.5-1 g cell inoculum, 75 μM ABA, 25-30°C, 7–10 days), they survived slow freezing (2°C/h) to −9.0 ~ −9.3°C (LT50: 50% killing temperature) while control cells were mostly injured at −3°C (LT50: -0.5 ~ −1.5°C). Ice-inoculation of the cell suspension at −3°C and survival determination by regrowth confirmed that ABA-treated rice cells survived extracellular freezing at −9°C. ABA-induced freezing tolerance did not require any exposure to cold and was best achieved at 25-30°C where the rice cells maintained high growth even in the presence of ABA. ABA treatment also increased tolerance to heat (43°C) as determined by regrowth. ABA-treated cells tended to have more augmented cytoplasm and/or reduced vacuole sizes compared to control cultures with a concomitant increase in osmolarity and a decrease in water content. ABA-treated (2–7 days) in vitro grown seedlings and their leaves survived slow freezing to −3°C with only marginal injury (LT50: -4°C) whereas untreated seedlings were killed at −3°C (LT50: -2°C). Conclusions The results indicate that exogenous ABA can induce some levels of freezing tolerance in chilling-sensitive rice cells and seedlings, probably by eliciting mechanisms different from low temperature-induced cold acclimation. PMID:24004611

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.

Transcriptomic difference in bovine blastocysts following vitrification and slow freezing at morula stage

PubMed Central

Gupta, Alisha; Singh, Jaswant; Dufort, Isabelle; Robert, Claude; Dias, Fernanda Caminha Faustino

2017-01-01

Cryopreservation is known for its marked deleterious effects on embryonic health. Bovine compact morulae were vitrified or slow-frozen, and post-warm morulae were cultured to the expanded blastocyst stage. Blastocysts developed from vitrified and slow-frozen morulae were subjected to microarray analysis and compared with blastocysts developed from unfrozen control morulae for differential gene expression. Morula to blastocyst conversion rate was higher (P < 0.05) in control (72%) and vitrified (77%) than in slow-frozen (34%) morulae. Total 20 genes were upregulated and 44 genes were downregulated in blastocysts developed from vitrified morulae (fold change ≥ ± 2, P < 0.05) in comparison with blastocysts developed from control morulae. In blastocysts developed from slow-frozen morulae, 102 genes were upregulated and 63 genes were downregulated (fold change ≥ ± 1.5, P < 0.05). Blastocysts developed from vitrified morulae exhibited significant changes in gene expression mainly involving embryo implantation (PTGS2, CALB1), lipid peroxidation and reactive oxygen species generation (HSD3B1, AKR1B1, APOA1) and cell differentiation (KRT19, CLDN23). However, blastocysts developed from slow-frozen morulae showed changes in the expression of genes related to cell signaling (SPP1), cell structure and differentiation (DCLK2, JAM2 and VIM), and lipid metabolism (PLA2R1 and SMPD3). In silico comparison between blastocysts developed form vitrified and slow-frozen morulae revealed similar changes in gene expression as between blastocysts developed from vitrified and control morulae. In conclusion, blastocysts developed form vitrified morulae demonstrated better post-warming survival than blastocysts developed from slow-frozen morulae but their gene expression related to lipid metabolism, steroidogenesis, cell differentiation and placentation changed significantly (≥ 2 fold). Slow freezing method killed more morulae than vitrification but those which survived up to blastocyst stage did not express ≥ 2 fold change in their gene expression as compared with blastocysts from control morulae. PMID:29095916

The effect of low temperature on Antarctic endolithic green algae

NASA Technical Reports Server (NTRS)

Meyer, M. A.; Morris, G. J.; Friedmann, E. I.

1988-01-01

Laboratory experiments show that undercooling to about -5 degrees C occurs in colonized Beacon sandstones of the Ross Desert, Antarctica. High-frequency temperature oscillations between 5 degrees C and -5 degrees C or -10 degrees C (which occur in nature on the rock surface) did not damage Hemichloris antarctica. In a cryomicroscope, H. antarctica appeared to be undamaged after slow or rapid cooling to -50 degrees C. 14CO2 incorporation after freezing to -20 degrees C was unaffected in H. antarctica or in Trebouxia sp. but slightly depressed in Stichococcus sp. (isolated from a less extreme Antarctic habitat). These results suggest that the freezing regime in the Antarctic desert is not injurious to endolithic algae. It is likely that the freezing-point depression inside the rock makes available liquid water for metabolic activity at subzero temperatures. Freezing may occur more frequently on the rock surface and contribute to the abiotic nature of the surface.

In vitro sperm characterization and development of a sperm cryopreservation method using directional solidification in the killer whale (Orcinus orca).

PubMed

Robeck, T R; Gearhart, S A; Steinman, K J; Katsumata, E; Loureiro, J D; O'Brien, J K

2011-07-15

Research was conducted to characterize seminal traits and to develop a sperm cryopreservation method using directional freezing (DF) for the killer whale (Orcinus orca). Experiments evaluated effects of: (i) freezing rate (SLOW, MED, FAST) by diluent (BF5F, Biladyl®, EYC) in 0.5 mL straws; and (ii) freezing method (straw or DF) by glycerol (3, 6, or 9% final concentration, v:v) on in vitro sperm quality. Fresh ejaculates (n = 161) were (mean ± SD) 7.8 ± 7.4 mL at 740 × 10(6) sperm/mL with 92.2 ± 6.3% total motility (TM), 85.4 ± 6.9% progressive motility (PM), 89.6 ± 9.0% viability and 89.8 ± 9.2% acrosome integrity. Samples frozen using straws by the MED or SLOW method were improved (P < 0.05) over FAST across all diluents. At 3 h post thaw (PT), TM, PM, Rapid motility (RM), VAP, VCL, ALH and viability for 3% and 6% glycerol were improved (P < 0.05) over 9% glycerol. Directional freezing samples at 0 h and 3 h PT, at all glycerol concentrations, displayed higher (P < 0.001) TM, PM, RM, VAP, VSL, VCL and viability /intact acrosomes (PI/FITC-PNA) than straw. These data provided the first information on ejaculate characteristics and the development of a semen cryopreservation method using DF in the killer whale. Copyright © 2011 Elsevier Inc. All rights reserved.

Wheeled and standard walkers in Parkinson's disease patients with gait freezing.

PubMed

Cubo, Esther; Moore, Charity G; Leurgans, Sue; Goetz, Christopher G

2003-10-01

Compare the efficacy of two walking assistance devices (wheeled walker and standard walker) to unassisted walking for patients with PD and gait freezing. Although numerous walking devices are used clinically, their relative effects on freezing and walking speed have never been systematically tested. Nineteen PD patients (14 non-demented) walked under three conditions in randomized order: unassisted walking, standard walker, and wheeled walker. Patients walked up to three times in each condition through a standard course that included rising from a chair, walking through a doorway, straightway walking, pivoting, and return. Total walking time, freezing time and number of freezes were compared for the three conditions using mixed models (walking time) and Friedman's test (freezing). The wheeled walker was further studied by comparing the effect of an attached laser that projected a bar of light on the floor as a visual walking cue. Use of either type of device significantly slowed walking compared to unassisted walking. Neither walker reduced any index of freezing, nor the laser attachment offered any advantage to the wheeled walker. The standard walker increased freezing, and the wheeled walker had no effect on freezing. Among the non-demented subjects (n=14), the same patterns occurred, although the walking speed was less impaired by the wheeled walker than the standard walker in this group. Though walkers may stabilize patients and increase confidence, PD patients walk more slowly when using them, without reducing freezing. Because the wheeled walker was intermediate for walking time and does not aggravate freezing, if walkers are used for these subjects, this type of walker should be favored.

Multiple Glass Transitions and Freezing Events of Aqueous Citric Acid

PubMed Central

2014-01-01

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

Effect of freezing on the rheological, chemical and colour properties of Serpa cheese.

PubMed

Alvarenga, Nuno; Canada, João; Sousa, Isabel

2011-02-01

The effect of freezing on the properties of a raw ewes'-milk semi-soft cheese (Serpa cheese) was studied using small amplitude oscillatory (SAOS) and texture measurements, colour and chemical parameters. The freezing was introduced at three different stages of the ripening process (28, 35 and 42 days), and the cheeses were maintained frozen for 12 months. Cheeses were submitted to a slow or fast freezing method, and to different storage temperatures: -10 and -20°C (three replicates for each set conditions). Chemical data showed that only the proteolysis indicators exhibited differences between frozen and non-frozen samples; frozen samples showed higher values of NPN than the non-frozen samples, indicating that the freezing process did not prevent the secondary proteolysis of cheese. Frozen samples showed a significantly (P<0·05) stronger structure than the non-frozen, as indicated by hardness. However, the differences between the frozen and non-frozen samples were not significantly for storage modulus (G' 1Hz) and loss tangent (tan δ 1Hz) (P>0·05). Freezing affected mainly colour parameters: frozen samples were more luminous, and more yellow-green. The results allowed us to conclude that the damages caused by freezing to cheese properties could be minimized if this type of storage is introduced at the end of ripening (42 d) using a freezing temperature of -20°C.

Proteomic analyses reveal differences in cold acclimation mechanisms in freezing-tolerant and freezing-sensitive cultivars of alfalfa

PubMed Central

Chen, Jing; Han, Guiqing; Shang, Chen; Li, Jikai; Zhang, Hailing; Liu, Fengqi; Wang, Jianli; Liu, Huiying; Zhang, Yuexue

2015-01-01

Cold acclimation in alfalfa (Medicago sativa L.) plays a crucial role in cold tolerance to harsh winters. To examine the cold acclimation mechanisms in freezing-tolerant alfalfa (ZD) and freezing-sensitive alfalfa (W5), holoproteins, and low-abundance proteins (after the removal of RuBisCO) from leaves were extracted to analyze differences at the protein level. A total of 84 spots were selected, and 67 spots were identified. Of these, the abundance of 49 spots and 24 spots in ZD and W5, respectively, were altered during adaptation to chilling stress. Proteomic results revealed that proteins involved in photosynthesis, protein metabolism, energy metabolism, stress and redox and other proteins were mobilized in adaptation to chilling stress. In ZD, a greater number of changes were observed in proteins, and autologous metabolism and biosynthesis were slowed in response to chilling stress, thereby reducing consumption, allowing for homeostasis. The capability for protein folding and protein biosynthesis in W5 was enhanced, which allows protection against chilling stress. The ability to perceive low temperatures was more sensitive in freezing-tolerant alfalfa compared to freezing-sensitive alfalfa. This proteomics study provides new insights into the cold acclimation mechanism in alfalfa. PMID:25774161

Comparative Study on Two Different Methods for Determination of Hydraulic Conductivity of HeLa Cells During Freezing.

PubMed

Li, Lei; Gao, Cai; Zhao, Gang; Shu, Zhiquan; Cao, Yunxia; Gao, Dayong

2016-12-01

The measurement of hydraulic conductivity of the cell membrane is very important for optimizing the protocol of cryopreservation and cryosurgery. There are two different methods using differential scanning calorimetry (DSC) to measure the freezing response of cells and tissues. Devireddy et al. presented the slow-fast-slow (SFS) cooling method, in which the difference of the heat release during the freezing process between the osmotically active and inactive cells is used to obtain the cell membrane hydraulic conductivity and activation energy. Luo et al. simplified the procedure and introduced the single-slow (SS) cooling protocol, which requires only one cooling process although different cytocrits are required for the determination of the membrane transport properties. To the best of our knowledge, there is still a lack of comparison of experimental processes and requirements for experimental conditions between these two methods. This study made a systematic comparison between these two methods from the aforementioned aspects in detail. The SFS and SS cooling methods mentioned earlier were utilized to obtain the reference hydraulic conductivity (L pg ) and activation energy (E Lp ) of HeLa cells by fitting the model to DSC data. With the SFS method, it was determined that L pg  = 0.10 μm/(min·atm) and E Lp  = 22.9 kcal/mol; whereas the results obtained by the SS cooling method showed that L pg  = 0.10 μm/(min·atm) and E Lp  = 23.6 kcal/mol. The results indicated that the values of the water transport parameters measured by two methods were comparable. In other words, the two parameters can be obtained by comparing the heat releases between two slow cooling processes of the same sample according to the SFS method. However, the SS method required analyzing heat releases of samples with different cytocrits. Thus, more experimental time was required.

Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei.

PubMed

Wen, Yan-Zi; Su, Bi-Xiu; Lyu, Shu-Shen; Hide, Geoff; Lun, Zhao-Rong; Lai, De-Hua

2016-12-01

Trehalose, a non-permeating cryoprotective agent (CPA), has been documented as less toxic and highly efficient at cryopreserving different kinds of cells or organisms. In the present study, trehalose was evaluated for its application in cryopreservation of both Trypanosoma brucei procyclic and bloodstream form cells. The cryopreservation efficiency was determined by the motility of trypanosomes after thawing, as well as a subsequent recovery and infectivity assessment. The viability of trypanosomes from cultivation that were frozen in a serial concentrations of trehalose showed similar results to classical CPAs of glycerol and DMSO. Nevertheless, trypanosomes cryopreserved in 0.2M trehalose showed the best growth characteristic during subsequent cultivation. In addition, CPA cocktails with trehalose and permeating CPA glycerol or DMSO were developed and evaluated. Interestingly, trypanosomes in host (mouse) blood cryopreserved in 0.4M trehalose plus 5% glycerol showed higher infectivity than those preserved in trehalose/DMSO cocktails as well as individually. Further investigations showed that, in comparison with slow freezing at -80°C, flash freezing in liquid nitrogen provided better cryopreservation for bloodstream form cells than slow freezing. In conclusion, trehalose is an easy, safe and efficient CPA for cryopreservation of T. brucei and potentially for other protozoan species and cells. Copyright © 2016 Elsevier B.V. All rights reserved.

Short-term culture of ovarian cortex pieces to assess the cryopreservation outcome in wild felids for genome conservation

PubMed Central

2013-01-01

Background Cryopreservation of ovarian tissue has the potential to preserve female germ cells of endangered mammals. In the present study, a freezing protocol successfully used for human tissue, was adapted for preserving ovarian tissue of domestic and non-domestic felids. Ovaries from non-domestic felid species were obtained from seven freshly euthanized and two recently deceased wild felids kept in different European Zoos. In addition, ovaries from domestic cats were obtained after ovariectomy from local veterinary clinics for methological adaptations. Ovarian cortex was dissected and uniform sized pieces of 2 mm diameter were obtained. Using a slow freezing protocol (-0.3°C per min) in 1.5 mol/L ethylene glycol, 0.1 mol/L sucrose, the pieces were cultured for up to 14 days both before and after cryopreservation. The integrity of primordial follicles was assessed by histology, and the impact of different protein sources (FCS or BSA) and Vitamin C was determined during two weeks of culture. Results and conclusion During culture the number of primordial follicles decreased within the ovarian pieces (p < 0.05). This effect was less pronounced when FCS was used as the protein source instead of BSA. Supplementation with Vitamin C had a detrimental effect on follicle survival. Since the procedure of cryopreservation had no effect on the follicle survival after one week of culture we conclude that the freezing protocol was suitable for felids. This is the first report of preserving a huge amount of follicles within ovarian tissue by slow freezing performed in several wild feline species. PMID:23433001

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

NASA Astrophysics Data System (ADS)

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

2018-05-01

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

Migration of Water in Litopenaeus Vannamei Muscle Following Freezing and Thawing.

PubMed

Deng, Qi; Wang, Yaling; Sun, Lijun; Li, Jianrong; Fang, Zhijia; Gooneratne, Ravi

2018-06-15

Water and protein are major constituents of shrimp, any changes in protein and the state of water influence the quality of shrimp. Therefore, a study to examine the law of moisture migration and protein denaturation under different freezing and thawing conditions is important. The proton density images of thawed frozen-shrimp revealed that the water loss during quick-freezing was much greater than that during slow freezing or microfreezing. At room temperature (25 °C), the water loss from brine-thawing was more than still-water thawing and still-water thawing was more than thawing spontaneously. Freezing-thawing resulted in uniform water redistribution in shrimp muscle. Nuclear magnetic resonance technology (low field magnetic imaging) was used to directly monitor the dynamic processes of fluidity state in shrimp and indirectly monitor protein denaturation and thereby determine the optimal method of freezing-thawing shrimp. Our research showed that microfreezing preservation minimized weight loss, juice leakage and protein denaturation in shrimp muscle during thawing. Water is one of the major components in most organs and is an important factor that influences the shrimp muscle quality. Water migration patterns and subsequent effects on the shrimp muscle under different freezing and thawing conditions were examined using low field nuclear magnetic resonance (NMR) technology. This research provides a theoretical foundation for shrimp processing plants to improve the freezing and thawing process to obtain optimal quality and flavor of shrimp products. © 2018 Institute of Food Technologists®.

Porcine uterus cryopreservation: an analysis of contractile function using different uterotonics.

PubMed

Schölch, Daniel; Schölch, Sebastian; Strahl, Olga; Hoffmann, Inge; Beckmann, Matthias W; Dittrich, Ralf

2012-10-01

Cryopreservation of whole organs has become increasingly successful in recent years, and establishing reliable methods for confirming the success of specific cryopreservation procedures has therefore become extremely important. On the assumption that methods such as histological evaluation do not provide definitive evidence of long-term cryopreservation and that clear signs of conserved function in an organ are good evidence of its viability, contractile function was analysed in porcine uteri (n=60), either after long-term (group A) or short-term (group B) cryopreservation and post-thaw treatment with three different uterotonics. A slow freezing protocol was used to preserve the organs. Fifteen fresh uteri were analysed similarly for contractile function, which was evaluated by measuring intrauterine pressure after administration of oxytocin, prostaglandin E(1) (PGE(1)), and carbachol. After cryopreservation, all but three uteri (95%) showed rhythmic contractions similar to those in fresh uteri except for differences in the heights of contraction peaks, with lower contractions in PGE(1) subgroup B (P<0.05). With the exception of three nonresponsive uteri in group A, there were no differences in contractility between uteri after long-term cryopreservation and fresh uteri. The results of this study thus contribute to the debate on whether slow freezing or vitrification techniques are best for whole-organ cryopreservation. In summary, (1) preservation of muscular function in porcine uteri is feasible with a slow freezing protocol; (2) measurement of contractile function following administration of uterotonics is a useful method of confirming functionality; and (3) long-term cryopreservation does not significantly impair post-thaw contractibility in comparison with fresh uteri. Copyright © 2012 Elsevier Inc. All rights reserved.

Threat interferes with response inhibition.

PubMed

Hartikainen, Kaisa M; Siiskonen, Anna R; Ogawa, Keith H

2012-05-09

A potential threat, such as a spider, captures attention and engages executive functions to adjust ongoing behavior and avoid danger. We and many others have reported slowed responses to neutral targets in the context of emotional distractors. This behavioral slowing has been explained in the framework of attentional competition for limited resources with emotional stimuli prioritized. Alternatively, slowed performance could reflect the activation of avoidance/freezing-type motor behaviors associated with threat. Although the interaction of attention and emotion has been widely studied, little is known on the interaction between emotion and executive functions. We studied how threat-related stimuli (spiders) interact with executive performance and whether the interaction profile fits with a resource competition model or avoidance/freezing-type motor behaviors. Twenty-one young healthy individuals performed a Go-NoGo visual discrimination reaction time (RT) task engaging several executive functions with threat-related and emotionally neutral distractors. The threat-related distractors had no effect on the RT or the error rate in the Go trials. The NoGo error rate, reflecting failure in response inhibition, increased significantly because of threat-related distractors in contrast to neutral distractors, P less than 0.05. Thus, threat-related distractors temporarily impaired response inhibition. Threat-related distractors associated with increased commission errors and no effect on RT does not suggest engagement of avoidance/freezing-type motor behaviors. The results fit in the framework of the resource competition model. A potential threat calls for evaluation of affective significance as well as inhibition of undue emotional reactivity. We suggest that these functions tax executive resources and may render other executive functions, such as response inhibition, temporarily compromised when the demands for resources exceed availability.

Carbon nanotube scaffolds with controlled porosity as electromagnetic absorbing materials in the gigahertz range

NASA Astrophysics Data System (ADS)

González, M.; Crespo, M.; Baselga, J.; Pozuelo, J.

2016-05-01

Control of the microscopic structure of CNT nanocomposites allows modulation of the electromagnetic shielding in the gigahertz range. The porosity of CNT scaffolds has been controlled by two freezing protocols and a subsequent lyophilization step: fast freezing in liquid nitrogen and slow freezing at -20 °C. Mercury porosimetry shows that slowly frozen specimens present a more open pore size (100-150 μm) with a narrow distribution whereas specimens frozen rapidly show a smaller pore size and a heterogeneous distribution. 3D-scaffolds containing 3, 4, 6 and 7% CNT were infiltrated with epoxy and specimens with 2, 5 and 8 mm thicknesses were characterized in the GHz range. Samples with the highest pore size and porosity presented the lowest reflected power (about 30%) and the highest absorbed power (about 70%), which allows considering them as electromagnetic radiation absorbing materials.Control of the microscopic structure of CNT nanocomposites allows modulation of the electromagnetic shielding in the gigahertz range. The porosity of CNT scaffolds has been controlled by two freezing protocols and a subsequent lyophilization step: fast freezing in liquid nitrogen and slow freezing at -20 °C. Mercury porosimetry shows that slowly frozen specimens present a more open pore size (100-150 μm) with a narrow distribution whereas specimens frozen rapidly show a smaller pore size and a heterogeneous distribution. 3D-scaffolds containing 3, 4, 6 and 7% CNT were infiltrated with epoxy and specimens with 2, 5 and 8 mm thicknesses were characterized in the GHz range. Samples with the highest pore size and porosity presented the lowest reflected power (about 30%) and the highest absorbed power (about 70%), which allows considering them as electromagnetic radiation absorbing materials. Electronic supplementary information (ESI) available: Scheme of hydrogenated derivative of diglycidyl ether of bisphenol-A (HDGEBA) and m-xylylenediamine; X-ray diffractograms of pristine CNT and oxidized CNT; glass transition temperatures of composites; electromagnetic shielding analysis in the 1-18 GHz frequency range. See DOI: 10.1039/c6nr02133f

Current status of human oocyte and embryo cryopreservation.

PubMed

Herrero, Leyre; Martínez, Mónica; Garcia-Velasco, Juan A

2011-08-01

To summarize recent advances in oocyte and embryo cryopreservation techniques and outcomes. Vitrification is gradually replacing slow freezing due to a better survival rate after thawing. Most units use vitrification for both oocyte and blastocyst cryopreservation, as these two biological structures did not perform very well with slow freezing technique. Basic experiments show that cellular damage seems lower after vitrification. Taken all together, this is helping vitirification to be expanding rapidly, and new clinical indications are being incorporated as well (i.e., fertility preservation). Cryopreservation has been used as a complement to IVF, and recent publications indicate that pregnancy rates achieved with frozen oocytes and embryos are comparable with those achieved in fresh cycles. Multiple publications studying oocyte and embryo physiology during cryopreservation have been published recently; however, larger studies are needed to verify the efficacy of new cryopreservation techniques. Vitrification is a simple and robust technique that is being incorporated into the majority of IVF units, mainly for oocyte and blastocyst cryopreservation.

Small Molecule Ice Recrystallization Inhibitors Enable Freezing of Human Red Blood Cells with Reduced Glycerol Concentrations

PubMed Central

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

2015-01-01

In North America, red blood cells (RBCs) are cryopreserved in a clinical setting using high glycerol concentrations (40% w/v) with slow cooling rates (~1°C/min) prior to storage at −80°C, while European protocols use reduced glycerol concentrations with rapid freezing rates. After thawing and prior to transfusion, glycerol must be removed to avoid intravascular hemolysis. This is a time consuming process requiring specialized equipment. Small molecule ice recrystallization inhibitors (IRIs) such as β-PMP-Glc and β-pBrPh-Glc have the ability to prevent ice recrystallization, a process that contributes to cellular injury and decreased cell viability after cryopreservation. Herein, we report that addition of 110 mM β-PMP-Glc or 30 mM β-pBrPh-Glc to a 15% glycerol solution increases post-thaw RBC integrity by 30-50% using slow cooling rates and emphasize the potential of small molecule IRIs for the preservation of cells. PMID:25851700

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

PubMed

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

2015-04-08

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

Hierarchical Freezing in a Lattice Model

NASA Astrophysics Data System (ADS)

Byington, Travis W.; Socolar, Joshua E. S.

2012-01-01

A certain two-dimensional lattice model with nearest and next-nearest neighbor interactions is known to have a limit-periodic ground state. We show that during a slow quench from the high temperature, disordered phase, the ground state emerges through an infinite sequence of phase transitions. We define appropriate order parameters and show that the transitions are related by renormalizations of the temperature scale. As the temperature is decreased, sublattices with increasingly large lattice constants become ordered. A rapid quench results in a glasslike state due to kinetic barriers created by simultaneous freezing on sublattices with different lattice constants.

Anomalously low C6+/C5+ ratio in solar wind: ACE/SWICS observation

NASA Astrophysics Data System (ADS)

Zhao, L.; Landi, E.; Kocher, M.; Lepri, S. T.; Fisk, L. A.; Zurbuchen, T. H.

2016-03-01

The Carbon and Oxygen ionization states in the solar wind plasma freeze-in within 2 solar radii (Rs) from the solar surface, and then they do not change as they propagate with the solar wind into the heliosphere. Therefore, the O7+/O6+ and C6+/C5+ charge state ratios measured in situ maintain a record of the thermal properties (electron temperature and density) of the inner corona where the solar wind originates. Since these two ratios freeze-in at very similar height, they are expected to be correlated. However, an investigation of the correlation between these two ratios as measured by ACE/SWICS instrument from 1998 to 201l shows that there is a subset of "Outliers" departing from the expected correlation. We find about 49.4% of these Outliers is related to the Interplanetary Coronal Mass Ejections (ICMEs), while 49.6% of them is slow speed wind (Vp < 500 km/s) and about 1.0% of them is fast solar wind (Vp > 500 km/s). We compare the outlier-slow-speed wind with the normal slow wind (defined as Vp < 500 km/s and O7+/O6+ > 0.2) and find that the reason that causes the Outliers to depart from the correlation is their extremely depleted C6+/C5+ ratio which is decreased by 80% compared to the normal slow wind. We discuss the implication of the Outlier solar wind for the solar wind acceleration mechanism.

Ovarian and oocyte cryopreservation.

PubMed

Lornage, Jacqueline; Salle, Bruno

2007-08-01

The present article is an update on progress in the two available techniques of oocyte and ovarian cryopreservation: slow cooling/rapid thawing and vitrification. A new line of research has opened in recent years: freezing the whole ovary with its vascular pedicle, so as to enable vascular grafts limiting ischemia-related follicle reserve loss. The technique of mature oocyte vitrification has advanced significantly, with improved oocyte physiology, increased safety, and higher clinical pregnancy rates. The number of studies on whole ovary freezing has grown, and there has been a large-mammal (sheep) live birth by orthotopic graft with vascular anastomosis of a cryopreserved ovary. Ovarian and oocyte cryopreservation is essential to conserving the fertility of young women. Results of mature oocyte freezing techniques have improved significantly over the past few years, but remain poorer than those with embryo freezing. Mature oocyte vitrification is progressing well, but requires safety validation in view of the high cryoprotectant concentrations used. Ovarian cortex fragment freezing is widely used in patients, with two live births after orthotopic graft, worldwide. The problem of rapid graft exhaustion has led to a focus on whole ovary cryopreservation which has resulted in one live birth in a ewe.

Slow Freezing Coupled Static Magnetic Field Exposure Enhances Cryopreservative Efficiency—A Study on Human Erythrocytes

PubMed Central

Lin, Chun-Yen; Wei, Po-Li; Chang, Wei-Jen; Huang, Yung-Kai; Feng, Sheng-Wei; Lin, Che-Tong; Lee, Sheng-Yang; Huang, Haw-Ming

2013-01-01

The aim of this study was to assess the cryoprotective effect of static magnetic fields (SMFs) on human erythrocytes during the slow cooling procedure. Human erythrocytes suspended in 20% glycerol were slowly frozen with a 0.4-T or 0.8-T SMF and then moved to a −80°C freezer for 24 hr. The changes in survival rate, morphology, and metabolites of the thawed erythrocytes were examined. To understand possible cryoprotective mechanisms of SMF, membrane fluidity and dehydration stability of SMF-exposed erythrocytes were tested. For each test, sham-exposed erythrocytes were used as controls. Our results showed that freezing coupled with 0.4-T or 0.8-T SMFs significantly increased the relative survival ratios of the frozen-thawed erythrocytes by 10% and 20% (p<0.001), respectively. The SMFs had no effect on erythrocyte morphology and metabolite levels. However, membrane fluidity of the samples exposed to 0.8-T SMF decreased significantly (p<0.05) in the hydrophobic regions. For the dehydration stability experiments, the samples exposed to 0.8-T SMF exhibited significantly lower (p<0.05) hemolysis. These results demonstrate that a 0.8-T SMF decreases membrane fluidity and enhances erythrocyte membrane stability to resist dehydration damage caused by slow cooling procedures. PMID:23520546

Freeze-drying of tert-butyl alcohol/water cosolvent systems: effects of formulation and process variables on residual solvents.

PubMed

Wittaya-Areekul, S; Nail, S L

1998-04-01

The objective of this study was to identify significant formulation and processing variables affecting levels of tert-butyl alcohol (TBA) and isopropyl alcohol (IPA) in freeze-dried solids prepared from TBA/water cosolvent systems. The variables examined were the physical state of the solute (crystalline vs amorphous), initial TBA concentration, freezing rate, cake thickness, and the temperature and duration of secondary drying. Sucrose and glycine were used as models for noncrystallizing and crystallizing solutes, respectively. The TBA concentration above which eutectic crystallization takes place was determined by differential scanning calorimetry. Model formulations were subjected to extremes of freezing rate by either dipping in liquid nitrogen or by slowly freezing on the shelf of a freeze-dryer. Dynamics of solvent loss during secondary drying was determined by withdrawing samples as a function of time at different shelf temperatures using a thief system. On the basis of these studies, the most important determinant of residual TBA level is the physical state of the solute. Freeze-dried glycine contained very low levels of residual TBA (0.01-0.03%) regardless of freezing rate or initial TBA concentration. For freeze-dried sucrose, residual TBA levels were approximately 2 orders of magnitude higher and were significantly affected by initial TBA concentration and freezing rate. For the sucrose/TBA/water system, relatively low residual TBA levels were obtained when the initial TBA level was above the threshold concentration for eutectic crystallization of TBA, whereas samples freeze-dried from solutions containing TBA concentrations below this threshold contained significantly higher levels of TBA. Residual IPA levels increased continuously with initial concentration of TBA in the sucrose/TBA/water system. Formulations of sucrose/TBA/water which were frozen rapidly contained residual TBA levels which were approximately twice those measured in the same formulation after slow freezing and drying under the same conditions. For the sucrose/TBA/water system, the temperature and time of secondary drying had only minimal influence on residual TBA in the freeze-dried solid. At low initial TBA concentrations (2%), residual TBA increases with increased cake thickness, perhaps because of the influence of depth of fill on effective freezing rate.

Developmental potential of murine germinal vesicle stage cumulus-oocyte complexes following exposure to dimethylsulphoxide or cryopreservation: loss of membrane integrity of cumulus cells after thawing.

PubMed

Ruppert-Lingham, C J; Paynter, S J; Godfrey, J; Fuller, B J; Shaw, R W

2003-02-01

Cumulus cells of the cumulus-oocyte complex (COC) are important in oocyte maturation. Thus, in preserving immature oocytes it is prudent to also preserve their associated cumulus cells. The survival and function of oocytes and their associated cumulus cells was assessed following cryopreservation or exposure to cryoprotectant without freezing. Immature COCs were collected from mice primed with pregnant mare's serum. COCs were either slow-cooled or exposed to 1.5 mol/l dimethylsulphoxide without freezing. Treated and fresh COCs were stained for membrane integrity or, after in-vitro maturation and IVF, were assessed for developmental capability. Development of cumulus-denuded fresh oocytes, as well as denuded and frozen-thawed oocytes co-cultured with fresh cumulus cells, was assessed. Slow-cooled oocytes had significantly reduced coverage by intact cumulus cells compared with fresh COCs. Cumulus cell association and developmental capability were not substantially affected by exposure to cryoprotectant without freezing. Denuded fresh oocytes and cryopreserved COCs had decreased developmental potential that was not overcome by co-culture with fresh cumulus cells. Loss of association between oocyte and cumulus cells was induced by cryopreservation, but not by treatment with cryoprotectant alone. The data indicate that direct physical contact between cumulus cells and the oocyte, throughout maturation, improves subsequent embryo development.

Effects of cryoprotectant agents and freezing protocol on motility of black-lip pearl oyster (Pinctada margaritifera L.) spermatozoa.

PubMed

Acosta-Salmón, Héctor; Jerry, Dean R; Southgate, Paul C

2007-02-01

Gamete cryopreservation techniques have been applied to several bivalve mollusc species. However, research activity in this area has primarily focused on cryopreserving gametes from edible oysters (Ostreiidae). Few studies have examined the effect of cryoprotectants and freezing protocols in the preservation of spermatozoa from cultured pearl oysters (Pteriidae). Pearl oyster producers are increasingly looking towards the development of improved family lines and, as a consequence, the ability to cryopreserve gametes would bring about significant benefits to the cultured pearl industry. In response to this need, we evaluated the effect of three cryoprotectant additives (CPA) on motility of spermatozoa from the black-lip pearl oyster, Pinctada margaritifera. These additives have previously been used to cryopreserve gametes of other bivalve species. The following CPA mixtures were evaluated: (1) 0.45M trehalose and 0, 0.64, 1.02 and 1.53 M dimethyl sulfoxide (Me(2)SO); (2) 0.2M glucose and 2M Me(2)SO and (3) 1.31 M propylene glycol (PG). The effects of four different freezing protocols on motility of P. margaritifera spermatozoa were also evaluated (slow, medium, medium-rapid and rapid cooling). This study showed that total motility was best retained when spermatozoa were cryopreserved in 0.45 M trehalose and 0, 0.64, 1.02 or 1.53 M Me(2)SO and frozen using slow to medium-rapid cooling rates (2.1-5.2 degrees Cmin(-1)). Rapid freezing through direct plunging of spermatozoa into liquid nitrogen resulted in the lowest overall retention of motility regardless of the CPA additive; however, CPA mixture also influenced retention of motility, with 0.2M glucose in 2M Me(2)SO and 1.31 M PG retaining the lowest levels of motility for the CPAs evaluated.

Critical viewpoints on the methods of realizing the metal freezing points of the ITS-90

NASA Astrophysics Data System (ADS)

Ma, C. K.

1995-08-01

The time-honored method for realizing the freezing point tf of a metal (in practice necessarily a dilute alloy) is that of continuous, slow freezing where the plateau temperature (which is the result of solidifying material's being so pure that its phase-transition temperature is observably constant) is measured. The freezing point being an equilibrium temperature, Ancsin considers this method to be inappropriate in principle: equilibrium between the solid and liquid phases cannot be achieved while the solid is being cooled to dispose of the releasing latent heat and while it is accreting at the expense of the liquid. In place of the continuous freezing method he has employed the pulse-heating method (in which the sample is allowed to approach equilibrium after each heat pulse) in his study of Ag; his measurements suggest that freezing can produce non-negligible errors. Here we examine both methods and conclude that the freezing method, employing an inside solid-liquid interface thermally isolated by an outside interface, can provide realizations of the highest accuracy; in either method, perturbation, by inducing solid-liquid phase transition continuously or intermittently, is essential for detecting equilibrium thermally. The respective merits and disadvantages of these two methods and also of the inner-melt method are discussed. We conclude that in a freezing-point measurement what is being measured is in effect the however minutely varying phase transition, and nonconstitutional equilibrium, temperature ti at the solid-liquid interface. The objective is then to measure the ti that is the best measure of tf, which is, normally, the plateau temperature.

Device and method for determining freezing points

NASA Technical Reports Server (NTRS)

Mathiprakasam, Balakrishnan (Inventor)

1986-01-01

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

Hiring Freeze

ERIC Educational Resources Information Center

Cooper, Kenneth J.

2010-01-01

Recession reduces job opportunities in the professoriate, particularly in areas where minorities are clustered. It's a tough time to want to be a professor or try to climb the faculty ranks, particularly for minority faculty, as Dr. Christopher Tudico, a Mexican-American, and others are finding. The recession and its go-slow recovery have cut the…

Cooperativity and Heterogeneity in Plastic Crystals Studied by Nonlinear Dielectric Spectroscopy

NASA Astrophysics Data System (ADS)

Michl, M.; Bauer, Th.; Lunkenheimer, P.; Loidl, A.

2015-02-01

The glassy dynamics of plastic-crystalline cyclo-octanol and ortho-carborane, where only the molecular reorientational degrees of freedom freeze without long-range order, is investigated by nonlinear dielectric spectroscopy. Marked differences to canonical glass formers show up: While molecular cooperativity governs the glassy freezing, it leads to a much weaker slowing down of molecular dynamics than in supercooled liquids. Moreover, the observed nonlinear effects cannot be explained with the same heterogeneity scenario recently applied to canonical glass formers. This supports ideas that molecular relaxation in plastic crystals may be intrinsically nonexponential. Finally, no nonlinear effects were detected for the secondary processes in cyclo-octanol.

Non-equilibrium freezing behaviour of aqueous systems.

PubMed

MacKenzie, A P

1977-03-29

The tendencies to non-equilibrium freezing behaviour commonly noted in representative aqueous systems derive from bulk and surface properties according to the circumstances. Supercooling and supersaturation are limited by heterogeneous nucleation in the presence of solid impurities. Homogeneous nucleation has been observed in aqueous systems freed from interfering solids. Once initiated, crystal growth is ofter slowed and, very frequently, terminated with increasing viscosity. Nor does ice first formed always succeed in assuming its most stable crystalline form. Many of the more significant measurements on a given systeatter permitting the simultaneous representation of thermodynamic and non-equilibrium properties. The diagram incorporated equilibrium melting points, heterogeneous nucleation temperatures, homogeneous nucleation temperatures, glass transition and devitrification temperatures, recrystallization temperatures, and, where appropriate, solute solubilities and eutectic temperatures. Taken together, the findings on modle systems aid the identification of the kinetic and thermodynamic factors responsible for the freezing-thawing survival of living cells.

Controlling condensation and frost growth with chemical micropatterns.

PubMed

Boreyko, Jonathan B; Hansen, Ryan R; Murphy, Kevin R; Nath, Saurabh; Retterer, Scott T; Collier, C Patrick

2016-01-22

In-plane frost growth on chilled hydrophobic surfaces is an inter-droplet phenomenon, where frozen droplets harvest water from neighboring supercooled liquid droplets to grow ice bridges that propagate across the surface in a chain reaction. To date, no surface has been able to passively prevent the in-plane growth of ice bridges across the population of supercooled condensate. Here, we demonstrate that when the separation between adjacent nucleation sites for supercooled condensate is properly controlled with chemical micropatterns prior to freezing, inter-droplet ice bridging can be slowed and even halted entirely. Since the edge-to-edge separation between adjacent supercooled droplets decreases with growth time, deliberately triggering an early freezing event to minimize the size of nascent condensation was also necessary. These findings reveal that inter-droplet frost growth can be passively suppressed by designing surfaces to spatially control nucleation sites and by temporally controlling the onset of freezing events.

Controlling condensation and frost growth with chemical micropatterns

DOE PAGES

Boreyko, Jonathan B.; Hansen, Ryan R.; Murphy, Kevin R.; ...

2016-01-22

Frost growth on chilled hydrophobic surfaces is an inter-droplet phenomenon, where frozen droplets harvest water from supercooled liquid droplets to grow ice bridges that propagate across the surface in a chain reaction. To date, no surface has been able to passively prevent the in-plane growth of frost across the population of supercooled condensate. Here, we demonstrate that when the nucleation sites for supercooled condensate are properly controlled with chemical micropatterns, the speed of frost growth can be slowed and even halted entirely. This stoppage of frost growth is attributed to the large interdroplet separation between condensate upon the onset ofmore » freezing, which was controlled by the pitch of the chemical patterns and by deliberately triggering an early freezing event. Lastly, these findings reveal that frost growth can be passively suppressed by designing surfaces to spatially control nucleation sites and/or temporally control the onset of freezing events.« less

Carbon nanotube scaffolds with controlled porosity as electromagnetic absorbing materials in the gigahertz range.

PubMed

González, M; Crespo, M; Baselga, J; Pozuelo, J

2016-05-19

Control of the microscopic structure of CNT nanocomposites allows modulation of the electromagnetic shielding in the gigahertz range. The porosity of CNT scaffolds has been controlled by two freezing protocols and a subsequent lyophilization step: fast freezing in liquid nitrogen and slow freezing at -20 °C. Mercury porosimetry shows that slowly frozen specimens present a more open pore size (100-150 μm) with a narrow distribution whereas specimens frozen rapidly show a smaller pore size and a heterogeneous distribution. 3D-scaffolds containing 3, 4, 6 and 7% CNT were infiltrated with epoxy and specimens with 2, 5 and 8 mm thicknesses were characterized in the GHz range. Samples with the highest pore size and porosity presented the lowest reflected power (about 30%) and the highest absorbed power (about 70%), which allows considering them as electromagnetic radiation absorbing materials.

Controlling condensation and frost growth with chemical micropatterns

PubMed Central

Boreyko, Jonathan B.; Hansen, Ryan R.; Murphy, Kevin R.; Nath, Saurabh; Retterer, Scott T.; Collier, C. Patrick

2016-01-01

In-plane frost growth on chilled hydrophobic surfaces is an inter-droplet phenomenon, where frozen droplets harvest water from neighboring supercooled liquid droplets to grow ice bridges that propagate across the surface in a chain reaction. To date, no surface has been able to passively prevent the in-plane growth of ice bridges across the population of supercooled condensate. Here, we demonstrate that when the separation between adjacent nucleation sites for supercooled condensate is properly controlled with chemical micropatterns prior to freezing, inter-droplet ice bridging can be slowed and even halted entirely. Since the edge-to-edge separation between adjacent supercooled droplets decreases with growth time, deliberately triggering an early freezing event to minimize the size of nascent condensation was also necessary. These findings reveal that inter-droplet frost growth can be passively suppressed by designing surfaces to spatially control nucleation sites and by temporally controlling the onset of freezing events. PMID:26796663

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

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.

Characterizing the freezing behavior of liposomes as a tool to understand the cryopreservation procedures.

PubMed

Siow, Lee Fong; Rades, Thomas; Lim, Miang Hoong

2007-12-01

Freezing behaviors of egg yolk l-alpha-phosphatidylcholine (EPC) and 1,2-dipalmitoyl-rac-glycero-3-phosphocholine (DPPC) large unilamellar vesicles (LUV) were quantitatively characterized in relation to freezing temperatures, cooling rates, holding time, presence of sodium chloride and phospholipid phase transition temperature. Cooling of the EPC LUV showed an abrupt increase in leakage of the encapsulated carboxyfluorescein (CF) between -5 degrees C and -10 degrees C, which corresponded with the temperatures of the extraliposomal ice formation at around -7 degrees C. For the DPPC LUV, CF leakage started at -10 degrees C, close to the temperature of the extraliposomal ice formation; followed by a subsequent rapid increase in leakage between -10 degrees C and -25 degrees C. Scanning electron microscopy showed that both of these LUV were freeze-concentrated and aggregated at sub-freezing temperatures. We suggest that the formation of the extraliposomal ice and the decrease of the unfrozen fraction causes freeze-injury and leakage of the CF. The degree of leakage, however, differs between EPC LUV and DPPC LUV that inherently vary in their phospholipid phase transition temperatures. With increasing holding time, the EPC LUV were observed to have higher leakage when they were held at -15 degrees C compared to at -30 degrees C whilst leakage of the DPPC LUV was higher when holding at -40 degrees C than at -15 degrees C and -50 degrees C. At slow cooling rates, osmotic pressure across the bilayers may cause an additional stress to the EPC LUV. The present work elucidates freeze-injury mechanisms of the phospholipid bilayers through the liposomal model membranes.

Modeling the cell-type dependence of diffusion-limited intracellular ice nucleation and growth during both vitrification and slow freezing

NASA Astrophysics Data System (ADS)

Yang, Geer; Zhang, Aili; Xu, Lisa X.; He, Xiaoming

2009-06-01

In this study, a set of models for predicting the diffusion-limited ice nucleation and growth inside biological cells were established. Both the heterogeneous and homogeneous nucleation mechanisms were considered in the models. Molecular mobility including viscosity and mutual diffusion coefficient of aqueous cryoprotectant (i.e., glycerol here) solutions was estimated using models derived from the free volume theory for glass transition, which makes it possible to predict the two most important physical properties (i.e., viscosity and mutual diffusion coefficient) over wide ranges of temperature and concentration as encountered in cryopreservation. After being verified using experimental data, the models were used to predict the critical cooling rate (defined as the cooling rate required so that the crystallized volume is less than 0.1% of the cell volume) as a function of the initial glycerol concentration in a number of cell types with different sizes. For slowing freezing, it was found that the required critical cooling rate is cell-type dependent with influences from cell size and the ice nucleation and water transport parameters. In general, the critical cooling rate does not change significantly with the initial glycerol concentration used and tends to be higher for smaller cells. For vitrification, the required critical cooling rate does change significantly with the initial glycerol concentration used and tends to decrease with the decrease in cell size. However, the required critical cooling rate can be similar for cells with very different sizes. It was further found that the thermodynamic and kinetic parameters for intracellular ice formation associated with different cells rather than the cell size per se significantly affect the critical cooling rates required for vitrification. For all cell types, it was found that homogeneous nucleation dominates at ultrafast cooling rates and/or high glycerol concentrations, whereas heterogeneous nucleation becomes important only during slow freezing with a low initial glycerol concentration (<1.5-2M), particularly for large cells such as mouse oocytes.

A novel particle engineering technology to enhance dissolution of poorly water soluble drugs: spray-freezing into liquid.

PubMed

Rogers, True L; Nelsen, Andrew C; Hu, Jiahui; Brown, Judith N; Sarkari, Marazban; Young, Timothy J; Johnston, Keith P; Williams, Robert O

2002-11-01

A novel cryogenic spray-freezing into liquid (SFL) process was developed to produce microparticulate powders consisting of an active pharmaceutical ingredient (API) molecularly embedded within a pharmaceutical excipient matrix. In the SFL process, a feed solution containing the API was atomized beneath the surface of a cryogenic liquid such that the liquid-liquid impingement between the feed and cryogenic liquids resulted in intense atomization into microdroplets, which were frozen instantaneously into microparticles. The SFL micronized powder was obtained following lyophilization of the frozen microparticles. The objective of this study was to develop a particle engineering technology to produce micronized powders of the hydrophobic drug, danazol, complexed with hydroxypropyl-beta-cyclodextrin (HPbetaCD) and to compare these SFL micronized powders to inclusion complex powders produced from other techniques, such as co-grinding of dry powder mixtures and lyophilization of bulk solutions. Danazol and HPbetaCD were dissolved in a water/tetrahydrofuran cosolvent mixture prior to SFL processing or slow freezing. Identical quantities of the API and HPbetaCD used in the solutions were co-ground in a mortar and pestle and blended to produce a co-ground physical mixture for comparison. The powder samples were characterized by differential scanning calorimetry (DSC), powder X-ray diffraction (XRD), Fourier transform infrared spectrometry (FTIR), scanning electron microscopy, surface area analysis, and dissolution testing. The results provided by DSC, XRD, and FTIR suggested the formation of inclusion complexes by both slow-freezing and SFL. However, the specific surface area was significantly higher for the latter. Dissolution results suggested that equilibration of the danazol/HPbetaCD solution prior to SFL processing was required to produce the most soluble conformation of the resulting inclusion complex following SFL. SFL micronized powders exhibited better dissolution profiles than the slowly frozen aggregate powder. Results indicated that micronized SFL inclusion complex powders dissolved faster in aqueous dissolution media than inclusion complexes formed by conventional techniques due to higher surface areas and stabilized inclusion complexes obtained by ultra-rapid freezing.

Earth's Fiercely Cooling Core - 24 TW

NASA Astrophysics Data System (ADS)

Morgan, Jason P.; Vannucchi, Paola

2014-05-01

Earth's mantle and core are convecting planetary heat engines. The mantle convects to lose heat from slow cooling, internal radioactivity, and core heatflow across its base. Its convection generates plate tectonics, volcanism, and the loss of ~35 TW of mantle heat through Earth's surface. The core convects to lose heat from slow cooling, small amounts of internal radioactivity, and the freezing-induced growth of a compositionally denser inner core. Core convection produces the geodynamo generating Earth's geomagnetic field. The geodynamo was thought to be powered by ~4 TW of heatloss across the core-mantle boundary, a rate sustainable (cf. Gubbins et al., 2003; Nimmo, 2007) by freezing a compositionally denser inner core over the ~3 Ga that Earth is known to have had a strong geomagnetic field (cf. Tarduno, 2007). However, recent determinations of the outer core's thermal conductivity(Pozzo et al., 2012; Gomi et al., 2013) indicate that >15 TW of power should conduct down its adiabat. Conducted power is unavailable to drive thermal convection, implying that the geodynamo needs a long-lived >17 TW power source. Core cooling was thought too weak for this, based on estimates for the Clapeyron Slope for high-pressure freezing of an idealized pure-iron core. Here we show that the ~500-1000 kg/m3 seismically-inferred jump in density between the liquid outer core and solid inner core allows us to directly infer the core-freezing Clapeyron Slope for the outer core's actual composition which contains ~8±2% lighter elements (S,Si,O,Al, H,…) mixed into a Fe-Ni alloy. A PREM-like 600 kg/m3 - based Clapeyron Slope implies there has been ~774K of core cooling during the freezing and growth of the inner core, releasing ~24 TW of power during the past ~3 Ga. If so, core cooling can easily power Earth's long-lived geodynamo. Another major implication of ~24 TW heatflow across the core-mantle boundary is that the present-day mantle is strongly 'bottom-heated', and diapiric mantle plumes should dominate deep mantle upwelling.

Maple sap uptake, exudation, and pressure changes correlated with freezing exotherms and thawing endotherms.

PubMed

Tyree, M T

1983-10-01

Sap flow rates and sap pressure changes were measured in dormant sugar maple trees (Acer saccharum Marsh.). In the forest, sap flow rates and pressure changes were measured from tap holes drilled into tree trunks in mature trees and sap flow rates were measured from the base of excised branches. Excised branches were also brought into the laboratory where air temperature could be carefully controlled in a refrigerated box and sap flow rates and sap pressures were measured from the cut base of the branches.Under both forest and laboratory conditions, sap uptake occurred as the wood temperature declined but much more rapid sap uptake correlated with the onset of the freezing exotherm. When sap pressures were measured under conditions of negligible volume displacement, the sap pressure rapidly fell to -60 to -80 kilopascals at the start of the freezing exotherm. The volume of water uptake and the rate of uptake depended on the rate of freezing. A slow freezing rate correlated with a large volume of water uptake, a fast freezing rate induced a smaller volume of water uptake. The volume of water uptake ranged from 0.02 to 0.055 grams water per gram dry weight of sapwood. The volume of water exuded after thawing was usually less than the volume of uptake so that after several freezing and thawing cycles the sapwood water content increased from 0.7 to 0.8 grams water per gram dry weight.These results are discussed in terms of a physical model of the mechanism of maple sap uptake and exudation first proposed by P. E. R. O'Malley. The proposed mechanism of sap uptake is by vapor distillation in air filled wood fiber lumina during the freezing of minor branches. Gravity and pressurized air bubbles (compressed during freezing) cause sap flow from the canopy down the tree after the thaw.

Maple Sap Uptake, Exudation, and Pressure Changes Correlated with Freezing Exotherms and Thawing Endotherms 1

PubMed Central

Tyree, Melvin T.

1983-01-01

Sap flow rates and sap pressure changes were measured in dormant sugar maple trees (Acer saccharum Marsh.). In the forest, sap flow rates and pressure changes were measured from tap holes drilled into tree trunks in mature trees and sap flow rates were measured from the base of excised branches. Excised branches were also brought into the laboratory where air temperature could be carefully controlled in a refrigerated box and sap flow rates and sap pressures were measured from the cut base of the branches. Under both forest and laboratory conditions, sap uptake occurred as the wood temperature declined but much more rapid sap uptake correlated with the onset of the freezing exotherm. When sap pressures were measured under conditions of negligible volume displacement, the sap pressure rapidly fell to −60 to −80 kilopascals at the start of the freezing exotherm. The volume of water uptake and the rate of uptake depended on the rate of freezing. A slow freezing rate correlated with a large volume of water uptake, a fast freezing rate induced a smaller volume of water uptake. The volume of water uptake ranged from 0.02 to 0.055 grams water per gram dry weight of sapwood. The volume of water exuded after thawing was usually less than the volume of uptake so that after several freezing and thawing cycles the sapwood water content increased from 0.7 to 0.8 grams water per gram dry weight. These results are discussed in terms of a physical model of the mechanism of maple sap uptake and exudation first proposed by P. E. R. O'Malley. The proposed mechanism of sap uptake is by vapor distillation in air filled wood fiber lumina during the freezing of minor branches. Gravity and pressurized air bubbles (compressed during freezing) cause sap flow from the canopy down the tree after the thaw. PMID:16663208

NON-RANDOM CELL KILLING IN CRYOPRESERVATION: IMPLICATIONS FOR PERFORMANCE OF THE BATTERY OF LEUKOCYTE TESTS (BLT) - I. TOXIC AND IMMUNOTOXIC EFFECTS

EPA Science Inventory

To eliminate between-tests error in longitudinal studies, for specimen sharing, convenient scheduling, etc., it is necessary to freeze freshly separated leukocytes as well as non-transformed, continuous T lymphocyte (CTL) lines. o test the efficacy of a programmable reezer (tempe...

Cryopreservation of boar semen in mini- and maxi-straws.

PubMed

Bwanga, C O; de Braganca, M M; Einarsson, S; Rodriguez-Martinez, H

1990-10-01

Split ejaculates from four boars were frozen with a programmable freezing machine, in mini- (0.25 ml) and maxi- (5 ml) plastic straws with an extender at either acidic (6.3) or alkaline (7.4) pH. Glycerol (3%) was used as cryoprotectant. The freezing of the semen was monitored by way of thermocouples placed in the straws. Post-thaw motility and acrosome integrity were evaluated; the latter using phase contrast microscopy, eosin-nigrosin stain and electron microscopy. Post-thaw sperm motility was significantly higher when semen was frozen in mini-straws than in maxi-straws. For the mini-straws, the motility was better when semen was exposed to an acidic environment during freezing, but this beneficial effect of the low extracellular pH was not evident when maxi-straws were thawed. The motility of the spermatozoa diminished significantly during the thermoresistance test (0 h and 2 h time) at 37 degrees C in a similar way for both straws and extracellular pH's. The freezing procedure, no matter the extracellular pH, did not cause major acrosomal damages, but significantly more normal apical ridges were present in the mini-straws than in the maxi-straws. This in vitro evaluation indicated that the freezing method employed was better for mini- than for maxi-straws since the freezing of the 5 ml volumes was not homogeneous, due to the large section area between the surface and the core of the straw.

The feasibility and positive effects of a customised videogame rehabilitation programme for freezing of gait and falls in Parkinson's disease patients: a pilot study.

PubMed

Nuic, Dijana; Vinti, Maria; Karachi, Carine; Foulon, Pierre; Van Hamme, Angèle; Welter, Marie-Laure

2018-04-10

Freezing of gait and falls represent a major burden in patients with advanced forms of Parkinson's disease (PD). These axial motor signs are not fully alleviated by drug treatment or deep-brain stimulation. Recently, virtual reality has emerged as a rehabilitation option for these patients. In this pilot study, we aim to determine the feasibility and acceptability of rehabilitation with a customised videogame to treat gait and balance disorders in PD patients, and assess its effects on these disabling motor signs. We developed a customised videogame displayed on a screen using the Kinect system. To play, the patient had to perform large amplitude and fast movements of all four limbs, pelvis and trunk, in response to visual and auditory cueing, to displace an avatar to collect coins and avoid obstacles to gain points. We tested ten patients with advanced forms of PD (median disease duration = 16.5 years) suffering from freezing of gait and/or falls (Hoehn&Yahr score ≥ 3) resistant to antiparkinsonian treatment and deep brain stimulation. Patients performed 18 training sessions during a 6-9 week period. We measured the feasibility and acceptability of our rehabilitation programme and its effects on parkinsonian disability, gait and balance disorders (with clinical scales and kinematics recordings), positive and negative affects, and quality of life, after the 9th and 18th training sessions and 3 months later. All patients completed the 18 training sessions with high feasibility, acceptability and satisfaction scores. After training, the freezing-of-gait questionnaire, gait-and-balance scale and axial score significantly decreased by 39, 38 and 41%, respectively, and the activity-balance confidence scale increased by 35%. Kinematic gait parameters also significantly improved with increased step length and gait velocity and decreased double-stance time. Three months after the final session, no significant change persisted except decreased axial score and increased step length and velocity. This study suggests that rehabilitation with a customised videogame to treat gait and balance disorders is feasible, well accepted, and effective in parkinsonian patients. These data serve as preliminary evidence for further larger and controlled studies to propose this customised videogame rehabilitation programme at home. ClinicalTrials.gov NCT02469350 .

Anomalously low C{sup 6+}/C{sup 5+} ratio in solar wind: ACE/SWICS observation

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

Zhao, L., E-mail: lzh@umich.edu; Landi, E.; Kocher, M.

The Carbon and Oxygen ionization states in the solar wind plasma freeze-in within 2 solar radii (R{sub s}) from the solar surface, and then they do not change as they propagate with the solar wind into the heliosphere. Therefore, the O{sup 7+}/O{sup 6+} and C{sup 6+}/C{sup 5+} charge state ratios measured in situ maintain a record of the thermal properties (electron temperature and density) of the inner corona where the solar wind originates. Since these two ratios freeze-in at very similar height, they are expected to be correlated. However, an investigation of the correlation between these two ratios as measuredmore » by ACE/SWICS instrument from 1998 to 201l shows that there is a subset of “Outliers” departing from the expected correlation. We find about 49.4% of these Outliers is related to the Interplanetary Coronal Mass Ejections (ICMEs), while 49.6% of them is slow speed wind (V{sub p} < 500 km/s) and about 1.0% of them is fast solar wind (V{sub p} > 500 km/s). We compare the outlier-slow-speed wind with the normal slow wind (defined as V{sub p} < 500 km/s and O{sup 7+}/O{sup 6+} > 0.2) and find that the reason that causes the Outliers to depart from the correlation is their extremely depleted C{sup 6+}/C{sup 5+} ratio which is decreased by 80% compared to the normal slow wind. We discuss the implication of the Outlier solar wind for the solar wind acceleration mechanism.« less

Origin of diverse time scales in the protein hydration layer solvation dynamics: A simulation study

NASA Astrophysics Data System (ADS)

Mondal, Sayantan; Mukherjee, Saumyak; Bagchi, Biman

2017-10-01

In order to inquire the microscopic origin of observed multiple time scales in solvation dynamics, we carry out several computer experiments. We perform atomistic molecular dynamics simulations on three protein-water systems, namely, lysozyme, myoglobin, and sweet protein monellin. In these experiments, we mutate the charges of the neighbouring amino acid side chains of certain natural probes (tryptophan) and also freeze the side chain motions. In order to distinguish between different contributions, we decompose the total solvation energy response in terms of various components present in the system. This allows us to capture the interplay among different self- and cross-energy correlation terms. Freezing the protein motions removes the slowest component that results from side chain fluctuations, but a part of slowness remains. This leads to the conclusion that the slow component approximately in the 20-80 ps range arises from slow water molecules present in the hydration layer. While the more than 100 ps component has multiple origins, namely, adjacent charges in amino acid side chains, hydrogen bonded water molecules and a dynamically coupled motion between side chain and water. In addition, the charges enforce a structural ordering of nearby water molecules and helps to form a local long-lived hydrogen bonded network. Further separation of the spatial and temporal responses in solvation dynamics reveals different roles of hydration and bulk water. We find that the hydration layer water molecules are largely responsible for the slow component, whereas the initial ultrafast decay arises predominantly (approximately 80%) due to the bulk. This agrees with earlier theoretical observations. We also attempt to rationalise our results with the help of a molecular hydrodynamic theory that was developed using classical time dependent density functional theory in a semi-quantitative manner.

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

Utschig, L. M.; Dalosto, S. D.; Thurnauer, M. C.

Metal ion binding to a surface site on photosynthetic reaction centers (RCs) modulates light-induced electron and proton transfer events in the RC. Whereas many studies have elucidated aspects of metal ion modulation events in Rhodobacter sphaeroides RCs, much less is understood about the surface site in Blastochloris viridis (Blc. viridis) RCs. Interestingly, electron paramagnetic resonance studies revealed two spectroscopically distinct Cu{sup 2+} surface site environments in Blc. viridis RCs. Herein, Cu{sup 2+} has been used to spectroscopically probe the structure of these Cu{sup 2+} site(s) in response to freezing conditions, temperature, and charge separation. One Cu{sup 2+} environment in Blc.more » viridis RCs, termed CuA, exhibits temperature-dependent conformational flexibility. Different conformation states of the CuA{sup 2+} site are trapped when the RC is frozen in the dark either by fast-freeze or slow-freeze procedure. The second Cu{sup 2+} environment, termed CuB, is structurally invariant to different freezing conditions and shows resolved hyperfine coupling to three nitrogen atoms. Cu{sup 2+} is most likely binding at the same location on the RC, but in different coordination environments which may reflect two distinct conformational states of the isolated Blc. viridis RC protein.« less

Cryopreservative effects of the recombinant ice-binding protein from the arctic yeast Leucosporidium sp. on red blood cells.

PubMed

Lee, Sung Gu; Koh, Hye Yeon; Lee, Jun Hyuck; Kang, Sung-Ho; Kim, Hak Jun

2012-06-01

Antifreeze proteins (AFPs) have important functions in many freeze-tolerant organisms. The proteins non-colligatively lower the freezing point and functionally inhibit ice recrystallization in frozen solutions. In our previous studies, we found that the Arctic yeast Leucosporidium sp. produces an AFP (LeIBP), and that the protein could be successfully produced in Pichia expression system. The present study showed that recombinant LeIBP possesses the ability to reduce the damage induced to red blood cells (RBCs) by freeze thawing. In addition to 40 % glycerol, both 0.4 and 0.8 mg/ml LeIBPs significantly reduced freeze-thaw-induced hemolysis at either rapid- (45 °C) or slow-warming (22 °C) temperatures. Post-thaw cell counts of the cryopreserved RBCs were dramatically enhanced, in particular, in 0.8 mg/ml LeIBP. Interestingly, the cryopreserved cells in the presence of LeIBP showed preserved cell size distribution. These results indicate that the ability of LeIBP to inhibit ice recrystallization helps the RBCs avoid critically damaging electrolyte concentrations, which are known as solution effects. Considering all these data, LeIBP can be thought of as a key component in improving RBC cryopreservation efficiency.

The Triaging and Treatment of Cold-Induced Injuries.

PubMed

Sachs, Christoph; Lehnhardt, Marcus; Daigeler, Adrien; Goertz, Ole

2015-10-30

In Central Europe, cold-induced injuries are much less common than burns. In a burn center in western Germany, the mean ratio of these two types of injury over the past 10 years was 1 to 35. Because cold-induced injuries are so rare, physicians often do not know how to deal with them. This article is based on a review of publications (up to December 2014) retrieved by a selective search in PubMed using the terms "freezing," "frostbite injury," "non-freezing cold injury," and "frostbite review," as well as on the authors' clinical experience. Freezing and cold-induced trauma are part of the treatment spectrum in burn centers. The treatment of cold-induced injuries is not standardized and is based largely on case reports and observations of use. distinction is drawn between non-freezing injuries, in which there is a slow temperature drop in tissue without freezing, and freezing injuries in which ice crystals form in tissue. In all cases of cold-induced injury, the patient should be slowly warmed to 22°-27°C to prevent reperfusion injury. Freezing injuries are treated with warming of the body's core temperature and with the bathing of the affected body parts in warm water with added antiseptic agents. Any large or open vesicles that are already apparent should be debrided. To inhibit prostaglandin-mediated thrombosis, ibuprofen is given (12 mg/kg body weight b.i.d.). The treatment of cold-induced injuries is based on their type, severity, and timing. The recommendations above are grade C recommendations. The current approach to reperfusion has yielded promising initial results and should be further investigated in prospective studies.

Sensitization of thermotolerant SCK cells to hyperthermia and freezing with reduction of intracellular pH: implications for cryosurgery.

PubMed

Burgher, Abram H; Swanlund, David J; Griffin, Robert J; Song, Chang W; Bischof, John C; Roberts, Kenneth P

2003-03-01

During cryosurgery, cells frozen slowly at the outer part of the ice ball undergo severe dehydration and are subject to solute effects injury, which may be caused in part by protein denaturation. This study was undertaken to determine whether heat shock proteins (HSPs), the molecular chaperones that stabilize proteins against denaturation, have a protective effect on cells during slow freezing. In addition, we aimed to determine whether acidic conditions, similar to those found in many solid tumors, would effect this protection. SCK cells were frozen at 5 degrees C/min to -10 degrees C or -20 degrees C before or after induction of thermotolerance, and at neutral or low pH conditions. Lethal damage was determined by clonogenics. Clonogenic survival was decreased by 50% in thermotolerant cells frozen to -10 degrees C after culture in acidic conditions (pH 6.6) compared with non-thermotolerant cells cultured at neutral pH. Induction of thermotolerance alone or low pH alone did not significantly sensitize SCK cells to freezing. All treatment groups were equally susceptible to killing when frozen to -20 degrees C. Our results show that induction of thermal tolerance does not protect SCK cells against subsequent freezing injury and that a low pH environment actually sensitizes these cells to freeze injury. Copyright 2003 Wiley-Liss, Inc.

Experiements with an inactivated hepatitis leptospirosis vaccine in vaccination programmes for dogs.

PubMed

Wilson, J H; Hermann-Dekkers, W M; Leemans-Dessy, S; Meijer, J W

1977-06-25

A fluid adjuvanted vaccine consisting of inactivated hepatitis virus (iH) and leptospirae antigens (L) was developed. The vaccine (Kavak iHL; Duphar) was tested in several vaccination programmes both alone and in combination with freeze dried measles (M) or distemper (D) vaccines. The results demonstrate that this new vaccine is also effective in pups with maternally derived antibodies, although a second vaccination at 14 weeks of age is recommended to boost the first vaccination. For the booster vaccination either the iHL-vaccine or the liver attenuated hepatitis vaccine (H) can be used.

Effects of buffer composition and processing conditions on aggregation of bovine IgG during freeze-drying.

PubMed

Sarciaux, J M; Mansour, S; Hageman, M J; Nail, S L

1999-12-01

The objective of this study was to identify critical formulation and processing variables affecting aggregation of bovine IgG during freeze-drying when no lyoprotective solute is used. Parameters examined were phosphate buffer concentration and counterion (Na versus K phosphate), added salts, cooling rate, IgG concentration, residual moisture level, and presence of a surfactant. No soluble aggregates were detected in any formulation after either freezing/thawing or freeze-drying. No insoluble aggregates were detected in any formulation after freezing, but insoluble aggregate levels were always detectable after freeze-drying. The data are consistent with a mechanism of aggregate formation involving denaturation of IgG at the ice/freeze-concentrate interface which is reversible upon freeze-thawing, but becomes irreversible after freeze-drying and reconstitution. Rapid cooling (by quenching in liquid nitrogen) results in more and larger aggregates than slow cooling on the shelf of the freeze-dryer. This observation is consistent with surface area measurements and environmental electron microscopic data showing a higher surface area of freeze-dried solids after fast cooling. Annealing of rapidly cooled solutions results in significantly less aggregation in reconstituted freeze-dried solids than in nonannealed controls, with a corresponding decrease in specific surface area of the freeze-dried, annealed system. Increasing the concentration of IgG significantly improves the stability of IgG against freeze-drying-induced aggregation, which may be explained by a smaller percentage of the protein residing at the ice/freeze-concentrate interface as IgG concentration is increased. A sodium phosphate buffer system consistently results in more turbid reconstituted solids than a potassium phosphate buffer system at the same concentration, but this effect is not attributable to a pH shift during freezing. Added salts such as NaCl or KCl contribute markedly to insoluble aggregate formation. Both sodium and potassium chloride contribute more to turbidity of the reconstituted solid than either sodium or potassium phosphate buffers at similar ionic strength, with sodium chloride resulting in a substantially higher level of aggregates than potassium chloride. At a given cooling rate, the specific surface area of dried solids is approximately a factor of 2 higher for the formulation containing sodium chloride than the formulation containing potassium chloride. Turbidity is also influenced by the extent of secondary drying, which underscores the importance of minimizing secondary drying of this system. Including a surfactant such as polysorbate 80, either in the formulation or in the water used for reconstitution, decreased, but did not eliminate, insoluble aggregates. There was no correlation between pharmaceutically acceptability of the freeze-dried cake and insoluble aggregate levels in the reconstituted product.

Influence of Emulsified Asphalt on the Mechanical Property and Microstructure of Cement-Stabilized Gravel under Freezing and Thawing Cycle Conditions.

PubMed

Wang, Yiqi; Tan, Yiqiu; Guo, Meng; Wang, Xinglong

2017-05-06

Properties of cement-stabilized gravel modified by emulsified asphalt under freezing and thawing cycle conditions were investigated by adjusting the dosage of cement. Mercury intrusion porosimetry (MIP) and Scanning electron microscopy (SEM) were introduced to analyze the influential mechanism. The results indicate that cement emulsified asphalt stabilized gravel with 5 wt % of cement performed well in both mechanics and frost-resistance. Although the addition of emulsified asphalt would lead to a partial decrease of strength, it can extend the process of strength loss and improve the freezing resistance. The main reason for this is that the permeability can be improved by the filling effects of emulsified asphalt. The frost-heave stress caused by the phase transition of water can also be remitted by emulsified asphalt, the elasticity modulus of which is much lower than the matrix. The generating speed of the micro crack can also be slowed down by emulsified asphalt.

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

PubMed

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

2016-01-01

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

Latent injury in frozen-thawed bacteriophage T4Bo.

PubMed Central

Steele, P. R.

1976-01-01

Two interesting new phenomena have been observed in suspensions of T4Bo bacteriophage which were frozen to temperatures below the eutectic temperature of the salt (sodium chloride) in the suspending medium. Approximately 10% of the phage appeared to survive such a phase change as determined by plaque titre. However, exposure of these survivors to ultrasonic vibration or repeated freezing showed them to be hypersensitive and thus latently injured. The hypersensitivity was lost on incubating the phage at 37 degrees C. for 3 hr. Furthermore, following a eutectic phase change, the surviving phage could be inactivated by rapid cooling to -90 degrees C. followed by slow rewarming. Such inactivation cannot be accounted for by accepted theories of freezing injury. PMID:1068188

Maintaining Long-Term Therapeutic Programmes: A Qualitative Case Study

ERIC Educational Resources Information Center

Brown, J. F.

2006-01-01

Maintaining therapeutic programmes for people with learning disabilities over long periods of time presents a major challenge to care systems as the limited and slow pace of client behavioural change is often insufficient to sustain staff behaviour. In this study, a 17-year-old young man with profound learning difficulties was successfully…

High-Throughput Cryopreservation of Plant Cell Cultures for Functional Genomics

PubMed Central

Ogawa, Yoichi; Sakurai, Nozomu; Oikawa, Akira; Kai, Kosuke; Morishita, Yoshihiko; Mori, Kumiko; Moriya, Kanami; Fujii, Fumiko; Aoki, Koh; Suzuki, Hideyuki; Ohta, Daisaku; Saito, Kazuki; Shibata, Daisuke

2012-01-01

Suspension-cultured cell lines from plant species are useful for genetic engineering. However, maintenance of these lines is laborious, involves routine subculturing and hampers wider use of transgenic lines, especially when many lines are required for a high-throughput functional genomics application. Cryopreservation of these lines may reduce the need for subculturing. Here, we established a simple protocol for cryopreservation of cell lines from five commonly used plant species, Arabidopsis thaliana, Daucus carota, Lotus japonicus, Nicotiana tabacum and Oryza sativa. The LSP solution (2 M glycerol, 0.4 M sucrose and 86.9 mM proline) protected cells from damage during freezing and was only mildly toxic to cells kept at room temperature for at least 2 h. More than 100 samples were processed for freezing simultaneously. Initially, we determined the conditions for cryopreservation using a programmable freezer; we then developed a modified simple protocol that did not require a programmable freezer. In the simple protocol, a thick expanded polystyrene (EPS) container containing the vials with the cell–LSP solution mixtures was kept at −30°C for 6 h to cool the cells slowly (pre-freezing); samples from the EPS containers were then plunged into liquid nitrogen before long-term storage. Transgenic Arabidopsis cells were subjected to cryopreservation, thawed and then re-grown in culture; transcriptome and metabolome analyses indicated that there was no significant difference in gene expression or metabolism between cryopreserved cells and control cells. The simplicity of the protocol will accelerate the pace of research in functional plant genomics. PMID:22437846

Chronic warming stimulates growth of marsh grasses more than mangroves in a coastal wetland ecotone.

PubMed

Coldren, G A; Barreto, C R; Wykoff, D D; Morrissey, E M; Langley, J A; Feller, I C; Chapman, S K

2016-11-01

Increasing temperatures and a reduction in the frequency and severity of freezing events have been linked to species distribution shifts. Across the globe, mangrove ranges are expanding toward higher latitudes, likely due to diminishing frequency of freezing events associated with climate change. Continued warming will alter coastal wetland plant dynamics both above- and belowground, potentially altering plant capacity to keep up with sea level rise. We conducted an in situ warming experiment, in northeast Florida, to determine how increased temperature (+2°C) influences co-occurring mangrove and salt marsh plants. Warming was achieved using passive warming with three treatment levels (ambient, shade control, warmed). Avicennia germinans, the black mangrove, exhibited no differences in growth or height due to experimental warming, but displayed a warming-induced increase in leaf production (48%). Surprisingly, Distichlis spicata, the dominant salt marsh grass, increased in biomass (53% in 2013 and 70% in 2014), density (41%) and height (18%) with warming during summer months. Warming decreased plant root mass at depth and changed abundances of anaerobic bacterial taxa. Even while the poleward shift of mangroves is clearly controlled by the occurrences of severe freezes, chronic warming between these freeze events may slow the progression of mangrove dominance within ecotones. © 2016 by the Ecological Society of America.

Cryopreservation: Vitrification and Controlled Rate Cooling.

PubMed

Hunt, Charles J

2017-01-01

Cryopreservation is the application of low temperatures to preserve the structural and functional integrity of cells and tissues. Conventional cooling protocols allow ice to form and solute concentrations to rise during the cryopreservation process. The damage caused by the rise in solute concentration can be mitigated by the use of compounds known as cryoprotectants. Such compounds protect cells from the consequences of slow cooling injury, allowing them to be cooled at cooling rates which avoid the lethal effects of intracellular ice. An alternative to conventional cooling is vitrification. Vitrification methods incorporate cryoprotectants at sufficiently high concentrations to prevent ice crystallization so that the system forms an amorphous glass thus avoiding the damaging effects caused by conventional slow cooling. However, vitrification too can impose damaging consequences on cells as the cryoprotectant concentrations required to vitrify cells at lower cooling rates are potentially, and often, harmful. While these concentrations can be lowered to nontoxic levels, if the cells are ultra-rapidly cooled, the resulting metastable system can lead to damage through devitrification and growth of ice during subsequent storage and rewarming if not appropriately handled.The commercial and clinical application of stem cells requires robust and reproducible cryopreservation protocols and appropriate long-term, low-temperature storage conditions to provide reliable master and working cell banks. Though current Good Manufacturing Practice (cGMP) compliant methods for the derivation and banking of clinical grade pluripotent stem cells exist and stem cell lines suitable for clinical applications are available, current cryopreservation protocols, whether for vitrification or conventional slow freezing, remain suboptimal. Apart from the resultant loss of valuable product that suboptimal cryopreservation engenders, there is a danger that such processes will impose a selective pressure on the cells selecting out a nonrepresentative, freeze-resistant subpopulation. Optimizing this process requires knowledge of the fundamental processes that occur during the freezing of cellular systems, the mechanisms of damage and methods for avoiding them. This chapter draws together the knowledge of cryopreservation gained in other systems with the current state-of-the-art for embryonic and induced pluripotent stem cell preservation in an attempt to provide the background for future attempts to optimize cryopreservation protocols.

Continuous measurement of drying rate of crystalline and amorphous systems during freeze-drying using an in situ microbalance technique.

PubMed

Roth, C; Winter, G; Lee, G

2001-09-01

The use of a novel microbalance (Christ) technique to monitor continuously the weight loss of a vial standing on a shelf of a freeze-dryer has been investigated. The drying rates of the following aqueous solutions were measured during the primary drying phase of a complete freeze-drying cycle: sucrose (75 mg/mL, 2.5-mL fill volume), sucrose and phenylalanine (1:0.2 by weight, 75 mg/mL, 2.5-mL fill volume), and mannitol (75mg/mL, 2.5-mL fill volume). The microbalance yields the cumulative water loss, m(cu) in grams, and the momentary drying rate, Deltam(cu)/Deltat in mg/10 min, of the frozen cake. The momentary drying rate curves were especially useful for examining how Deltam(cu)/Deltat changes with time during primary drying. Initially, Deltam(cu)/Deltat rises to a sharp maximum and then decreases in a fashion depending on shelf temperature, chamber pressure, and the nature of the substance being dried. Different drying behavior was observed for the sucrose and sucrose/phenylalanine systems, which was attributed to the presence of crystalline phenylalanine in the amorphous sucrose. At low shelf-temperature (-24 degrees C) the crystalline mannitol showed lower Deltam(cu)/Deltat than with either sucrose or sucrose/phenylalanine. The balance could also detect differences in Deltam(cu)/Deltat when using different freezing protocols. "Slow" and "moderate" freezing protocols gave similar drying behavior, but "rapid" freezing in liquid nitrogen produced greatly altered drying rate and internal cake morphology. The balance also could be used to detect the endpoint of primary drying. Different endpoint criteria and their influence on final dried cake properties were examined. Copyright 2001 Wiley-Liss, Inc. and the American Pharmaceutical Association

Effect of cooling rate on sperm quality of cryopreserved Andalusian donkey spermatozoa.

PubMed

Demyda-Peyrás, S; Bottrel, M; Acha, D; Ortiz, I; Hidalgo, M; Carrasco, J J; Gómez-Arrones, V; Gósalvez, J; Dorado, J

2018-06-01

The aim of this study was to evaluate the effect of different cooling rates on post-thaw quality of cryopreserved donkey spermatozoa. Eighteen ejaculates from six adult Andalusian donkeys (three ejaculates per donkey) were collected using an artificial vagina. Pooled semen samples (two ejaculates per pool) were divided into three aliquots, and frozen in Gent freezing extender using three different cryopreservation protocols (P): P1 (conventional slow freezing, as control): semen pre-cooled in an Equitainer for 2 h and frozen in liquid nitrogen (LN 2 ) vapour; P2 (controlled pre-freeze cooling rate): semen pre-cooled at a controlled rate for 73 min and frozen in LN 2 vapour; and P3 (rapid freezing) semen frozen immediately in LN 2 vapour. After thawing at 37 °C for 30 s, semen samples were assessed for motility, morphology, acrosome and plasma membrane integrity; spermatozoa were also tested for DNA integrity. Significant (P < 0.01) differences were found between the cryopreservation protocols for all sperm parameters evaluated, except for DNA integrity. Semen samples frozen using P2 showed significantly (P < 0.01) higher values for sperm motility, morphology, sperm membrane integrity, and acrosome integrity. On the contrary, P3 reduced sperm motility (P < 0.01) and increased the percentage of spermatozoa with damaged plasma membrane (P < 0.001). In our study, we demonstrated that the sperm of Andalusian donkey is particularly sensitive to the cooling rate used before freezing. Furthermore, Andalusian donkey semen can be successfully cryopreserved using controlled cooling rates combined with freezing in LN 2 vapour. Copyright © 2018 Elsevier B.V. All rights reserved.

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).

Marine Antifreeze Proteins: Structure, Function, and Application to Cryopreservation as a Potential Cryoprotectant

PubMed Central

Kim, Hak Jun; Lee, Jun Hyuck; Hur, Young Baek; Lee, Chang Woo; Park, Sun-Ha; Koo, Bon-Won

2017-01-01

Antifreeze proteins (AFPs) are biological antifreezes with unique properties, including thermal hysteresis (TH), ice recrystallization inhibition (IRI), and interaction with membranes and/or membrane proteins. These properties have been utilized in the preservation of biological samples at low temperatures. Here, we review the structure and function of marine-derived AFPs, including moderately active fish AFPs and hyperactive polar AFPs. We also survey previous and current reports of cryopreservation using AFPs. Cryopreserved biological samples are relatively diverse ranging from diatoms and reproductive cells to embryos and organs. Cryopreserved biological samples mainly originate from mammals. Most cryopreservation trials using marine-derived AFPs have demonstrated that addition of AFPs can improve post-thaw viability regardless of freezing method (slow-freezing or vitrification), storage temperature, and types of biological sample type. PMID:28134801

Development of a slow non-viral DNA release system from PDLLA scaffolds fabricated using a supercritical CO2 technique.

PubMed

Heyde, Mieke; Partridge, Kris A; Howdle, Steven M; Oreffo, Richard O C; Garnett, Martin C; Shakesheff, Kevin M

2007-10-15

Polyamidoamine polymers (PAA) comprising methylene-bisacrylamide/dimethylethylene-diamine monomers were synthesized, complexed with DNA and incorporated into porous P(DL)LA scaffolds by using a supercritical CO(2) (scCO(2)) technique. Scaffolds were made in a dry state consequently there was a need to lyophilize the complexes. A statistically significant reduction of the transfection efficiency was observed in the absence of trehalose when compared to the original complex after freeze-drying. Increasing concentrations (0-10% w/v) of trehalose were added to the complex prior to freeze-drying. Structure dependent differences in DNA binding were evaluated by gel electrophoresis and thermal transition analysis. TEM and PCS showed aggregate formation after freeze-drying without trehalose. Scaffolds were characterized by pore sizes of 173 +/- 73 microm and a porosity of 71%. The transfection potential of the released DNA was investigated by seeding scaffolds with A549 cells and following firefly luciferase as a marker gene after 48 h exposure. Low but continuous levels of transfection were observed for PAA complexes during a 60-day study. Complexes made with Lipofectaminetrade mark gave initially higher levels of DNA release but no further expression was seen after 40 days. Uncomplexed DNA showed background levels of transfection. Culturing cells on 3D scaffolds showed a benefit in retention of transfection activity with time compared to 2D controls. Transfection levels could be increased when cells were grown in OptiMEM. This study demonstrated that PAA/DNA complexes incorporated into a P(DL)LA scaffold made by using scCO(2) processing exhibited a slow release and extended gene expression profile. Copyright 2007 Wiley Periodicals, Inc.

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

Mamontov, Eugene; O'Neil, Hugh

In this paper, we have studied microscopic dynamics of a protein in carbon disulfide, a non-glass forming solvent, down to its freezing temperature of ca. 160 K. We have utilized quasielastic neutron scattering. A comparison of lysozyme hydrated with water and dissolved in carbon disulfide reveals a stark difference in the temperature dependence of the protein's microscopic relaxation dynamics induced by the solvent. In the case of hydration water, the common protein glass-forming solvent, the protein relaxation slows down in response to a large increase in the water viscosity on cooling down, exhibiting a well-known protein dynamical transition. The dynamicalmore » transition disappears in non-glass forming carbon disulfide, whose viscosity remains a weak function of temperature all the way down to freezing at just below 160 K. The microscopic relaxation dynamics of lysozyme dissolved in carbon disulfide is sustained down to the freezing temperature of its solvent at a rate similar to that measured at ambient temperature. Finally, our results demonstrate that protein dynamical transition is not merely solvent-assisted, but rather solvent-induced, or, more precisely, is a reflection of the temperature dependence of the solvent's glass-forming dynamics.« less

Studies on Rapidly Frozen Suspensions of Yeast Cells by Differential Thermal Analysis and Conductometry

PubMed Central

Mazur, Peter

1963-01-01

Few, if any, yeast cells survived rapid cooling to -196°C and subsequent slow warming. After rapid freezing, the suspensions absorbed latent heat of fusion between -15° and 0°C during warming, and the relation between the amount of heat absorbed and the concentration of cells was the same as that in equivalent KCl solutions, indicating that frozen suspensions behave thermally like frozen solutions. The amount of heat absorbed was such that more than 80 per cent of the intracellular solution had to be frozen. The conductometric behavior of frozen suspensions showed that cell solutes were still inside the cells and surrounded by an intact cell membrane at the time heat was being absorbed. Two models are consistent with these findings. The first assumes that intracellular freezing has taken place; the second that all freezable water has left the cells and frozen externally. The latter model is ruled out because rapidly cooled cells do not shrink by an amount equal to the volume of water that would have to be withdrawn to prevent internal freezing. PMID:13934216

Chemical consequences of compaction within the freezing front of a crystallizing magma ocean

NASA Astrophysics Data System (ADS)

Hier-Majumder, S.; Hirschmann, M. M.

2013-12-01

The thermal and compositional evolution of planetary magma oceans have profound influences on the early development and differentiation of terrestrial planets. During crystallization, rejection of elements incompatible in precipitating solids leads to petrologic and geochemical planetary differentiation, including potentially development of a compositionally stratified early mantle and evolution of thick overlying atmospheres. In cases of extremely efficient segregation of melt and crystals, solidified early mantles can be nearly devoid of key incompatible species including heat-producing (U, Th, K) and volatile (H,C,N,& noble gas) elements. A key structural component of a crystallizing magma ocean is the partially molten freezing front. The dynamics of this region influences the distribution of incompatible elements between the earliest mantle and the initial surficial reservoirs. It also can be the locus of heating owing to the dissipation of large amounts of tidal energy potentially available from the early Moon. The dynamics are influenced by the solidification rate, which is coupled to the liberation of volatiles owing to the modulating greenhouse effects in the overlying thick atmosphere. Compaction and melt retention in the freezing front of a magma ocean has received little previous attention. While the front advances during the course of crystallization, coupled conservation of mass, momentum, and energy within the front controls distribution and retention of melt within this layer. Due to compaction within this layer, melt distribution is far from uniform, and the fraction of melt trapped within this front depends on the rate of freezing of the magma ocean. During phases of rapid freezing, high amount of trapped melt within the freezing front retains a larger quantity of dissolved volatiles and the reverse is true during slow periods of crystallization. Similar effects are known from inferred trapped liquid fractions in layered mafic intrusions. Here we develop a simple 1-D model of melt retention in the freezing front of a crystallizing magma ocean, and apply it to the thermal and chemical evolution of the early Earth.

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.

The effect of water plasticization on the molecular mobility and crystallization tendency of amorphous disaccharides.

PubMed

Heljo, Ville Petteri; Nordberg, Antti; Tenho, Mikko; Virtanen, Tommi; Jouppila, Kirsi; Salonen, Jarno; Maunu, Sirkka Liisa; Juppo, Anne Mari

2012-10-01

To study how water plasticization affects the molecular mobility and crystallization tendency of freeze-dried trehalose, sucrose, melibiose and cellobiose. Freeze-dried disaccharides were subjected to different relative humidity atmospheres and their physical stabilities were evaluated. Lyophilizate water sorption tendencies and glass transition temperatures were modeled using Brunauer-Emmett-Teller (BET) and Gordon-Taylor (GT) equations, respectively. Sucrose and cellobiose crystallization tendencies were compared by using the concept of reduced crystallization temperature (RCT), and the molecular mobilities of trehalose and melibiose were compared by measuring their T(1)H relaxation time constants. Based on the BET and GT models, water sorption tendency and the resulting plasticizing effect were different in sucrose when compared to the other disaccharides. Trehalose and melibiose exhibited generally slower crystallization rates when compared to sucrose and cellobiose. Amorphous melibiose was shown to be particularly stable within the studied water content range, which may have partly been caused by its relatively slow molecular mobility. Slow amorphous-to-crystalline transition rate is known to be important for lyoprotecting excipients when formulating a robust drug product. The physical stabilities of amorphous trehalose and melibiose even with relatively high water contents might make their use advantageous in this respect compared to sucrose and cellobiose.

High post-thaw survival of ram sperm after partial freeze-drying.

PubMed

Arav, Amir; Idda, Antonella; Nieddu, Stefano Mario; Natan, Yehudit; Ledda, Sergio

2018-03-14

Recrystallization damages occur when a frozen sample is held at high subzero temperatures and when the warming process is too slow. In this work, ram semen diluted in two different concentrations of sugar solutions (Lyo A consisted of 0.4 M sorbitol and 0.25 M trehalose, and the second, Lyo B composed of 0.26 M sorbitol and 0.165 M trehalose) in egg yolk and Tris medium were compared after freezing 10 μL samples to: (1) - 10, - 25, and - 35 °C and thawing. (2) Freezing to - 10 and - 25 °C, holding for 1 h and then thawing, and (3) freezing to - 10 and - 25 °C and drying for 1 h at these temperatures at a vacuum of 80 mTorr, prior thawing. For drying, we used a new freeze-drying apparatus (Darya, FertileSafe, Israel) having a condensation temperature below - 110 °C and a vacuum pressure of 10-100 mTorr that is reached in less than 10s. Results showed that samples in Lyo B solution frozen at - 25 °C had significantly higher sperm motility in partially freeze-dried samples than frozen samples (46.6 ± 2.8% vs 1.2 ± 2.5%, P < 0.001). Moreover, partially dried samples in Lyo B showed higher motility than Lyo A at - 25 °C (46.6 ± 2.8% vs 35 ± 4%). Cryomicroscopy and low-temperature/low-pressure environmental scanning electronic microscope demonstrated that the amount of the ice crystals present in partially dried samples was lower than in the frozen samples. Holding the sperm at high subzero temperatures is necessary for the primary drying of cells during the freeze-drying process. Rapid freeze-drying can be achieved using this new device, which enables to reduce recrystallization damages.

Equilibrium, quasi-equilibrium, and nonequilibrium freezing of mammalian embryos.

PubMed

Mazur, P

1990-08-01

The first successful freezing of early embryos to -196 degrees C in 1972 required that they be cooled slowly at approximately 1 degree C/min to about -70 degrees C. Subsequent observations and physical/chemical analyses indicate that embryos cooled at that rate dehydrate sufficiently to maintain the chemical potential of their intracellular water close to that of the water in the partly frozen extracellular solution. Consequently, such slow freezing is referred to as equilibrium freezing. In 1972 and since, a number of investigators have studied the responses of embryos to departures from equilibrium freezing. When disequilibrium is achieved by the use of higher constant cooling rates to -70 degrees C, the results is usually intracellular ice formation and embryo death. That result is quantitatively in accord with the predictions of the physical/chemical analysis of the kinetics of water loss as a function of cooling rate. However, other procedures involving rapid nonequilibrium cooling do not result in high mortality. One common element in these other nonequilibrium procedures is that, before the temperature has dropped to a level that permits intracellular ice formation, the embryo water content is reduced to the point at which the subsequent rapid nonequilibrium cooling results in either the formation of small innocuous intracellular ice crystals or the conversion of the intracellular solution into a glass. In both cases, high survival requires that subsequent warming be rapid, to prevent recrystallization or devitrification. The physical/chemical analysis developed for initially nondehydrated cells appears generally applicable to these other nonequilibrium procedures as well.

Slow Cooling Cryopreservation Optimized to Human Pluripotent Stem Cells.

PubMed

Miyazaki, Takamichi; Suemori, Hirofumi

2016-01-01

Human pluripotent stem cells (hPSCs) have the potential for unlimited expansion and differentiation into cells that form all three germ layers. Cryopreservation is one of the key processes for successful applications of hPSCs, because it allows semi-permanent preservation of cells and their easy transportation. Most animal cell lines, including mouse embryonic stem cells, are standardly cryopreserved by slow cooling; however, hPSCs have been difficult to preserve and their cell viability has been extremely low whenever cryopreservation has been attempted.Here, we investigate the reasons for failure of slow cooling in hPSC cryopreservation. Cryopreservation involves a series of steps and is not a straightforward process. Cells may die due to various reasons during cryopreservation. Indeed, hPSCs preserved by traditional methods often suffer necrosis during the freeze-thawing stages, and the colony state of hPSCs prior to cryopreservation is a major factor contributing to cell death.It has now become possible to cryopreserve hPSCs using conventional cryopreservation methods without any specific equipment. This review summarizes the advances in this area and discusses the optimization of slow cooling cryopreservation for hPSC storage.

The Effect of Liquid Nitrogen on Bone Graft Survival.

PubMed

Sirinoglu, Hakan; Çilingir, Özlem Tuğçe; Çelebiler, Ozhan; Ercan, Feriha; Numanoglu, Ayhan

2015-08-01

Liquid nitrogen is used in medicine for cancer treatment and tissue preservation; however, bone viability after its application is controversial. This study aims to evaluate both the tissue viability and the clinical and histopathologic findings following liquid nitrogen application with different thawing techniques in rats. Mandibular bone grafts were taken from 45 Wistar rats and freezed in liquid nitrogen for 20 minutes. In the rapid-thawing technique (Rapid Thawing-1, Rapid Thawing-2), the grafts were held for 20 minutes in room temperature; in the slow-thawing technique (Slow Thawing-1, Slow Thawing-2), 20 minutes in -20°C, 20 minutes in +4°C, and 20 minutes in room temperature, respectively. In Rapid Thawing-2 and Slow Thawing-2 groups, autografts were implanted to their origin for 3 weeks and bone staining with India ink was performed and samples taken for histologic examination. The amount of cells and blood vessels and the density of bone canaliculi were significantly reduced in Rapid Thawing-1 and Slow Thawing-1 groups comparing to the Control group. However, the reduction rate was more significant in the Slow Thawing-1 group. Histomorphometric evaluation of the healing autografts after 3 weeks revealed that the decreased amounts of canaliculi were not changed in Slow Thawing-2 group. The study results demonstrated that bone tissue survives after liquid nitrogen treatment regardless of the performed thawing technique; however, slow thawing causes more tissue damage and metabolism impairment. Thieme Medical Publishers 333 Seventh Avenue, New York, NY 10001, USA.

FPGA implementation of bit controller in double-tick architecture

NASA Astrophysics Data System (ADS)

Kobylecki, Michał; Kania, Dariusz

2017-11-01

This paper presents a comparison of the two original architectures of programmable bit controllers built on FPGAs. Programmable Logic Controllers (which include, among other things programmable bit controllers) built on FPGAs provide a efficient alternative to the controllers based on microprocessors which are expensive and often too slow. The presented and compared methods allow for the efficient implementation of any bit control algorithm written in Ladder Diagram language into the programmable logic system in accordance with IEC61131-3. In both cases, we have compared the effect of the applied architecture on the performance of executing the same bit control program in relation to its own size.

Activation of Actuating Hydrogels with WS2 Nanosheets for Biomimetic Cellular Structures and Steerable Prompt Deformation.

PubMed

Zong, Lu; Li, Xiankai; Han, Xiangsheng; Lv, Lili; Li, Mingjie; You, Jun; Wu, Xiaochen; Li, Chaoxu

2017-09-20

Macroscopic soft actuation is intrinsic to living organisms in nature, including slow deformation (e.g., contraction, bending, twisting, and curling) of plants motivated by microscopic swelling and shrinking of cells, and rapid motion of animals (e.g., deformation of jellyfish) motivated by cooperative nanoscale movement of motor proteins. These actuation behaviors, with an exceptional combination of tunable speed and programmable deformation direction, inspire us to design artificial soft actuators for broad applications in artificial muscles, nanofabrication, chemical valves, microlenses, soft robotics, etc. However, so far artificial soft actuators have been typically produced on the basis of poly(N-isopropylacrylamide) (PNiPAM), whose deformation is motived by volumetric shrinkage and swelling in analogue to plant cells, and exhibits sluggish actuation kinetics. In this study, alginate-exfoliated WS 2 nanosheets were incorporated into ice-template-polymerized PNiPAM hydrogels with the cellular microstructures which mimic plant cells, yet the prompt steerable actuation of animals. Because of the nanosheet-reinforced pore walls formed in situ in freezing polymerization and reasonable hierarchical water channels, this cellular hybrid hydrogel achieves super deformation speed (on the order of magnitude of 10° s), controllable deformation direction, and high near-infrared light responsiveness, offering an unprecedented platform of artificial muscles for various soft robotics and devices (e.g., rotator, microvalve, aquatic swimmer, and water-lifting filter).

Experimental observation of the influence of furnace temperature profile on convection and segregation in the vertical Bridgman crystal growth technique

NASA Technical Reports Server (NTRS)

Neugebauer, G. T.; Wilcox, W. R.

1990-01-01

Azulene-doped naphtalene was directionally solidified using the vertical Bridgman-Stockbarger technique. Doping homogeneity and convection are determined as a function of the temperature profile in the furnace and the freezing rate. Convective velocities are two orders of magnitude lower when the temperature increases with height. The cross sectional variation in azulene concentration tends to be asymmetric. Neither rotation of the ampoule nor deliberate introduction of thermal asymmetries during solidification had a significant influence on cross sectional variations in doping. It is predicted that slow directional solidification under microgravity conditions can produce greater inhomogeneities than on earth. Thus when low freezing rates are necessary in order to avoid constitutional supercooling, it may be necessary to combine microgravity and magnetic fields in order to achieve homogeneous crystals.

Microscopic relaxations in a protein sustained down to 160 K in a non-glass forming organic solvent

DOE PAGES

Mamontov, Eugene; O'Neil, Hugh

2016-05-03

In this paper, we have studied microscopic dynamics of a protein in carbon disulfide, a non-glass forming solvent, down to its freezing temperature of ca. 160 K. We have utilized quasielastic neutron scattering. A comparison of lysozyme hydrated with water and dissolved in carbon disulfide reveals a stark difference in the temperature dependence of the protein's microscopic relaxation dynamics induced by the solvent. In the case of hydration water, the common protein glass-forming solvent, the protein relaxation slows down in response to a large increase in the water viscosity on cooling down, exhibiting a well-known protein dynamical transition. The dynamicalmore » transition disappears in non-glass forming carbon disulfide, whose viscosity remains a weak function of temperature all the way down to freezing at just below 160 K. The microscopic relaxation dynamics of lysozyme dissolved in carbon disulfide is sustained down to the freezing temperature of its solvent at a rate similar to that measured at ambient temperature. Finally, our results demonstrate that protein dynamical transition is not merely solvent-assisted, but rather solvent-induced, or, more precisely, is a reflection of the temperature dependence of the solvent's glass-forming dynamics.« less

Improved Cryopreservation of Human Umbilical Vein Endothelial Cells: A Systematic Approach

NASA Astrophysics Data System (ADS)

Sultani, A. Billal; Marquez-Curtis, Leah A.; Elliott, Janet A. W.; McGann, Locksley E.

2016-10-01

Cryopreservation of human umbilical vein endothelial cells (HUVECs) facilitated their commercial availability for use in vascular biology, tissue engineering and drug delivery research; however, the key variables in HUVEC cryopreservation have not been comprehensively studied. HUVECs are typically cryopreserved by cooling at 1 °C/min in the presence of 10% dimethyl sulfoxide (DMSO). We applied interrupted slow cooling (graded freezing) and interrupted rapid cooling with a hold time (two-step freezing) to identify where in the cooling process cryoinjury to HUVECs occurs. We found that linear cooling at 1 °C/min resulted in higher membrane integrities than linear cooling at 0.2 °C/min or nonlinear two-step freezing. DMSO addition procedures and compositions were also investigated. By combining hydroxyethyl starch with DMSO, HUVEC viability after cryopreservation was improved compared to measured viabilities of commercially available cryopreserved HUVECs and viabilities for HUVEC cryopreservation studies reported in the literature. Furthermore, HUVECs cryopreserved using our improved procedure showed high tube forming capability in a post-thaw angiogenesis assay, a standard indicator of endothelial cell function. As well as presenting superior cryopreservation procedures for HUVECs, the methods developed here can serve as a model to optimize the cryopreservation of other cells.

When a water drop freezes before it solidifies

NASA Astrophysics Data System (ADS)

Kavehpour, Pirouz; Davis, Stephen; Tavakoli, Faryar

2012-11-01

When a drop of liquid is placed on a substrate which temperature is below the melting point of the liquid, one would expect the drop to solidify instantaneously. However, many liquids, such as water, must be subcooled to solidify below its melting temperature due to homogeneous nucleation's high activation energy. Most of the drop solidification research, particularly for water, phase change is assumed to occur at equilibrium freezing temperature; however, this is not the case. We found that after a certain degree of supercooling, a kinetic based nucleation begins and latent heat of fusion is suddenly liberated, causing an increase in liquid temperature. At the end of this stage, approximately 20% of the drop is crystallized. This phenomenon is known among metallurgists as recalescence. This is followed by a slow solidification process at the melting point. As a water droplet spreads on a cold substrate, its contact line stops just prior to freezing inception from the liquid-solid interface. In this study, we assert that recalescence prior to solidification may be the cause of water's sudden immobility, which results in a fixed contact angle and droplet diameter. In our experiments, the nucleation front initiates from the trijunction point and propagates to the drop volume.

Toward understanding life under subzero conditions: the significance of exploring psychrophilic "cold-shock" proteins.

PubMed

Kuhn, Emanuele

2012-11-01

Understanding the behavior of proteins under freezing conditions is vital for detecting and locating extraterrestrial life in cold environments, such as those found on Mars and the icy moons of Jupiter and Saturn. This review highlights the importance of studying psychrophilic "cold-shock" proteins, a topic that has yet to be explored. A strategy for analyzing the psychrophilic RNA helicase protein CsdA (Psyc_1082) from Psychrobacter arcticus 273-4 as a key protein for life under freezing temperatures is proposed. The experimental model presented here was developed based on previous data from investigations of Escherichia coli, P. arcticus 273-4, and RNA helicases. P. arcticus 273-4 is considered a model for life in freezing environments. It is capable of growing in temperatures as cold as -10°C by using physiological strategies to survive not only in freezing temperatures but also under low-water-activity and limited-nutrient-availability conditions. The analyses of its genome, transcriptome, and proteome revealed specific adaptations that allow it to inhabit freezing environments by adopting a slow metabolic strategy rather than a cellular dormancy state. During growth at subzero temperatures, P. arcticus 273-4 genes related to energy metabolism and carbon substrate incorporation are downregulated, and genes for maintenance of membranes, cell walls, and nucleic acid motion are upregulated. At -6°C, P. arcticus 273-4 does not upregulate the expression of either RNA or protein chaperones; however, it upregulates the expression of its cold-shock induced DEAD-box RNA helicase protein A (CsdA - Psyc_1082). CsdA - Psyc_1082 was investigated as a key helper protein for sustaining life in subzero conditions. Proving CsdA - Psyc_1082 to be functional as a key protein for life under freezing temperatures may extend the known minimum growth temperature of a mesophilic cell and provide key information about the mechanisms that underlie cold-induced biological systems in icy worlds.

History of cryobiology, with special emphasis in evolution of mouse sperm cryopreservation.

PubMed

Sztein, Jorge M; Takeo, Toru; Nakagata, Naomi

2018-06-01

Confucius said study the past if you would define the future and a popular statement says that history depends on who writes it. To talk about history it is necessary to find and define a milestone where to start the narration. The intention of this quick review is to take the reader through moments and selected publications; part and pieces of memories showing how the concept of cryopreservation, specifically for mouse sperm, was conceived and sustained as we know it today. Beginning with the development of the microscope (1677) and continuing through the 17th century with the first documented observation by L. Spallanzani describing that sperm could maintain the motility under cold conditions. As J. Sherman suggested, we divide the cryopreservation evolution into two sequences, previous to and after 1949 when Polge, Smith and Parkes discovered the property of glycerol as cryoprotectant. Later, in 1972, D. Whittingham, S. Leibo, and P. Mazur applying a slow freezing process achieved the first embryo freezing (mouse). During that time many theories were scientifically confirmed. Among those, Peter Mazur demonstrated the relation between the speed of freezing and intracellular ice formation, and Stanley Leibo that each cell type has their unique freezing curve. In 1950, after the discovery of the protective aspect of glycerol, sperm from many mammals were frozen, except from the mouse. It was in the early 90's when the mouse sperm freezing becomes important and it was a real challenge for many groups, nevertheless, the technique using skim milk and raffinose modified by Dr Nakagata was the beginning of a different story …. Copyright © 2018 Elsevier Inc. All rights reserved.

PpCBF3 from Cold-Tolerant Kentucky Bluegrass Involved in Freezing Tolerance Associated with Up-Regulation of Cold-Related Genes in Transgenic Arabidopsis thaliana

PubMed Central

Chen, Yu; Xu, Bin; Yang, Zhimin; Huang, Bingru

2015-01-01

Dehydration-Responsive Element Binding proteins (DREB)/C-repeat (CRT) Binding Factors (CBF) have been identified as transcriptional activators during plant responses to cold stress. The objective of this study was to determine the physiological roles of a CBF gene isolated from a cold-tolerant perennial grass species, Kentucky bluegrass (Poa pratensis L.), which designated as PpCBF3, in regulating plant tolerance to freezing stress. Transient transformation of Arabidopsis thaliana mesophyll protoplast with PpCBF3-eGFP fused protein showed that PpCBF3 was localized to the nucleus. RT-PCR analysis showed that PpCBF3 was specifically induced by cold stress (4°C) but not by drought stress [induced by 20% polyethylene glycol 6000 solution (PEG-6000)] or salt stress (150 mM NaCl). Transgenic Arabidopsis overexpressing PpCBF3 showed significant improvement in freezing (-20°C) tolerance demonstrated by a lower percentage of chlorotic leaves, lower cellular electrolyte leakage (EL) and H2O2 and O2 .- content, and higher chlorophyll content and photochemical efficiency compared to the wild type. Relative mRNA expression level analysis by qRT-PCR indicated that the improved freezing tolerance of transgenic Arabidopsis plants overexpressing PpCBF3 was conferred by sustained activation of downstream cold responsive (COR) genes. Other interesting phenotypic changes in the PpCBF3-transgenic Arabidopsis plants included late flowering and slow growth or ‘dwarfism’, both of which are desirable phenotypic traits for perennial turfgrasses. Therefore, PpCBF3 has potential to be used in genetic engineering for improvement of turfgrass freezing tolerance and other desirable traits. PMID:26177510

Effect of cooling rate on the survival of cryopreserved rooster sperm: Comparison of different distances in the vapor above the surface of the liquid nitrogen.

PubMed

Madeddu, M; Mosca, F; Abdel Sayed, A; Zaniboni, L; Mangiagalli, M G; Colombo, E; Cerolini, S

2016-08-01

The aim of the present trial was to study the effect of different freezing rates on the survival of cryopreserved rooster semen packaged in straws. Slow and fast freezing rates were obtained keeping straws at different distances in the vapor above the surface of the nitrogen during freezing. Adult Lohmann roosters (n=27) were used. Two experiments were conducted. In Experiment 1, semen was packaged in straws and frozen comparing the distances of 1, 3 and 5cm in nitrogen vapor above the surface of the liquid nitrogen. In Experiment 2, the distances of 3, 7 and 10cm above the surfaces of the liquid nitrogen were compared. Sperm viability, motility and progressive motility and the kinetic variables were assessed in fresh and cryopreserved semen samples. The recovery rates after freezing/thawing were also calculated. In Experiment 1, there were no significant differences among treatments for all semen quality variables. In Experiment 2, the percentage of viable (46%) and motile (22%) sperm in cryopreserved semen was greater when semen was placed 3cm compared with 7 and 10cm in the vapor above the surface of the liquid nitrogen. The recovery rate of progressive motile sperm after thawing was also greater when semen was stored 3cm in the vapor above the surface of the liquid nitrogen. More rapid freezing rates are required to improve the survival of rooster sperm after cryopreservation and a range of distances from 1 to 5cm in nitrogen vapor above the surface of the liquid nitrogen is recommended for optimal sperm viability. Copyright © 2016 Elsevier B.V. All rights reserved.

Modelling Cryovolcanism Due to Subsurface Ocean Freezing on Pluto and Charon

NASA Astrophysics Data System (ADS)

Conrad, J. W.; Nimmo, F.; Singer, K. N.

2016-12-01

The New Horizons spacecraft identified various possible cryovolcanic features on the surfaces of both Pluto and Charon [1]. However, there are major differences between the cryovolcanism on Pluto and Charon. Pluto has two mound-flanked depressions which are possibly cryovolcanic [2], while Charon's putative cryovolcanism is more widespread within its smooth southern plains. If Pluto or Charon have (or had) subsurface oceans, slow refreezing of these oceans would lead to extensional surface tectonics [3,4] and pressurization of the ocean [5]. Sufficiently large pressurization can overcome the overburden pressure and cause an eruption. We applied thermal evolution models based on [3] to determine likely freezing scenarios. Eruptions on Charon are possible under most conditions, and occur after tens of kilometers of freezing of an ice shell initially 100 km thick. This would produce an areal extensional strain of 1%. The implied globally-averaged thickness of erupted material is a few hundred meters and the critical crack width for propagation through the entire ice shell [6] is about half a meter for all eruption scenarios. Eruptions on Pluto require probably unrealistic freezing scenarios, because of the larger body size and higher overburden pressure. We conclude that ocean freezing is a possible source of cryovolcanism on Charon and may explain the smooth plains in its southern hemisphere [1]. Pluto, on the other hand, requires more complex models to explain the putative cryovolcanic features on its surface. [1] Moore et al., Science 351 (2016): 1284-1293. [2] Singer et al., LPSC 47 (2016): 2276 [3] Robuchon and Nimmo, Icarus 216 (2011): 426-439. [4] Hammond et al., GRL 43 (2016). [5] Manga and Wang, GRL 34 (2007). [6] Porco et al., The Astronomical Journal 148 (2014): 45.

Should we isolate human preantral follicles before or after cryopreservation of ovarian tissue?

PubMed

Vanacker, Julie; Luyckx, Valérie; Amorim, Christiani; Dolmans, Marie-Madeleine; Van Langendonckt, Anne; Donnez, Jacques; Camboni, Alessandra

2013-04-01

To evaluate the survival and growth potential of human preantral follicles isolated before and after cryopreservation. Pilot study. Gynecology research unit in a university hospital. Six women aged 27 to 32 years. Six ovarian biopsy samples were cut into two equal parts, half subjected to slow-freezing followed by follicle isolation (cryo-iso group) and alginate-matrigel embedding, and half immediately processed for follicle isolation and alginate-matrigel embedding followed by slow-freezing (iso-cryo group) or used as fresh controls (fresh group). Follicle number, viability, diameter, and morphology. After 1,134 preantral follicles had been isolated from fresh biopsy samples and 1,132 from frozen specimens, the three groups were compared before and after 7 days of in vitro culture (IVC) in alginate-matrigel beads. No statistically significant differences in viability were found between the three groups before or after IVC, but follicle diameter increased in all three groups after IVC. Morphology analysis revealed well-preserved follicles in both the iso-cryo and cryo-iso groups after IVC. Human preantral follicles can be successfully cryopreserved before or after isolation without impairing their ability to survive and grow in vitro. This could lead to development of new protocols for follicle cryopreservation, IVC, and grafting in clinical and research settings for fertility preservation. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Assessment of imaging parameters correlated with the effects of cryopreservation on embryo development

NASA Astrophysics Data System (ADS)

Zarnescu, Livia; Abeyta, Mike; Baer, Thomas M.; Behr, Barry; Ellerbee, Audrey K.

2014-03-01

Embryo cryopreservation is an increasingly common technique that allows patients to undergo multiple cycles of in vitro fertilization (IVF) without being subjected to repeated ovarian stimulation and oocyte retrieval. There are two types of cryopreservation commonly used in IVF clinics today: slow freezing and vitrification. Because vitrification has been shown to result in higher rates of embryo survival post-thaw compared to slow freezing, it is rapidly gaining popularity in clinics worldwide. However, several studies have shown that vitrification can still cause damage to embryos in the form of DNA fragmentation, altered mitochondrial distribution and changes in transcriptional activity, all of which are impossible to assess noninvasively. In this paper we demonstrate a new method of quantitatively and noninvasively assessing changes in embryo appearance due to vitrification. Using full-field optical coherence tomography (FF-OCT), we show that vitrification causes striking changes in the appearance of the cytoplasm that are not visible under conventional brightfield microscopy. Using an automated algorithm that extracts parameters to describe these changes, we show that these parameters can also predict viability in embryos that have undergone vitrification. An automated, noninvasive assessment of embryo viability after vitrification and thawing could have significant clinical impact: allowing clinicians to more accurately choose the most viable embryos to transfer back to patients could reduce the average number of IVF cycles that patients must undergo to achieve pregnancy.

Freeze or flee? Negative stimuli elicit selective responding.

PubMed

Estes, Zachary; Verges, Michelle

2008-08-01

Humans preferentially attend to negative stimuli. A consequence of this automatic vigilance for negative valence is that negative words elicit slower responses than neutral or positive words on a host of cognitive tasks. Some researchers have speculated that negative stimuli elicit a general suppression of motor activity, akin to the freezing response exhibited by animals under threat. Alternatively, we suggest that negative stimuli only elicit slowed responding on tasks for which stimulus valence is irrelevant for responding. To discriminate between these motor suppression and response-relevance hypotheses, we elicited both lexical decisions and valence judgments of negative words and positive words. Relative to positive words (e.g., kitten), negative words (e.g., spider) elicited slower lexical decisions but faster valence judgments. Results therefore indicate that negative stimuli do not cause a generalized motor suppression. Rather, negative stimuli elicit selective responding, with faster responses on tasks for which stimulus valence is response-relevant.

Cryopreservation of embryos and oocytes in human assisted reproduction.

PubMed

Konc, János; Kanyó, Katalin; Kriston, Rita; Somoskői, Bence; Cseh, Sándor

2014-01-01

Both sperm and embryo cryopreservation have become routine procedures in human assisted reproduction and oocyte cryopreservation is being introduced into clinical practice and is getting more and more widely used. Embryo cryopreservation has decreased the number of fresh embryo transfers and maximized the effectiveness of the IVF cycle. The data shows that women who had transfers of fresh and frozen embryos obtained 8% additional births by using their cryopreserved embryos. Oocyte cryopreservation offers more advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation, and postponing childbirth, and eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In this review, the basic principles, methodology, and practical experiences as well as safety and other aspects concerning slow cooling and ultrarapid cooling (vitrification) of human embryos and oocytes are summarized.

Oocyte vitrification for elective fertility preservation: the past, present, and future.

PubMed

Gunnala, Vinay; Schattman, Glenn

2017-02-01

Oocyte cryopreservation is no longer experimental and one of its rapidly growing indications is elective fertility preservation. Currently there is no sufficient evidence to support its practice and therefore its place in IVF remains uncertain. Vitrification has superior post-thaw survival and fertilization outcomes compared with oocytes that were frozen with the slow-freeze technique. Oocyte vitrification produces similar IVF outcomes compared with fresh oocytes and is not associated with further obstetrical or perinatal morbidity. Undergoing elective oocyte cryopreservation between ages 35 and 37 will optimize live birth rates as well as cost effectiveness from mathematical models. In women who delay child bearing, elective oocyte cryopreservation in the mid 30s may be beneficial in terms of live birth rates and cost effectiveness. Prospective studies of women who have undergone oocyte cryopreservation and are now attempting conception are needed before official recommendations can be made regarding elective egg freezing.

Physiological limitation at alpine treeline: relationships of threshold responses of conifers to their establishment patterns

NASA Astrophysics Data System (ADS)

Germino, M. J.; Lazarus, B.; Castanha, C.; Moyes, A. B.; Kueppers, L. M.

2014-12-01

An understanding of physiological limitations to tree establishment at alpine treeline form the basis for predicting how this climate-driven boundary will respond to climate shifts. Most research on this topic has focused on limitations related to carbon balance and growth of trees. Carbon balance could limit survival and establishment primarily through slow-acting, chronic means. We asked whether tree survival and thus establishment patterns reflect control by chronic effects in comparison to acute, threshold responses, such as survival of frost events. Seedling survivorship patterns were compared to thresholds in freezing (temperature causing leaf freezing, or freezing point, FP; and physiological response to freezing) and water status (turgor loss point, TLP; and related physiological adjustments). Subject seedlings were from forest, treeline, and alpine sites in the Alpine Treeline Warming Experiment in Colorado, and included limber and lodgepole pine (a low-elevation species), and Engelmann Spruce. Preliminary results show survival increases with seedling age, but the only corresponding increase in stress acclimation was photosynthetic resistance to freezing and TLP, not FP. Differences in survivorship among the species were not consistent with variation in FP but they generally agreed with variation in photosynthetic resistance to deep freezing and to early-season drought avoidance. Mortality of limber pine increased 35% when minimum temperatures decreased below -9C, which compares with FPs of >-8.6C, and about 1/3 of its mortality occurred during cold/wet events, particularly in the alpine. The other major correlate of mortality is midsummer drying events, as previously reported. Also in limber pine, the TLP for year-old seedlings (-2.5 MPa) corresponded with seasonal-drought mortality. In summary, we show several examples of correspondence in physiological thresholds to mortality events within a species, although the relationships are not strong. Across species, photosynthetic resistance to freezing and early-season drought avoidance related well to mortality patterns. These results are generally more supportive of the role of chronic rather than acute climate effects in broad patterns of tree seedling establishment at treeline.

A Novel Approach to Identifying Physical Markers of Cryo-Damage in Bull Spermatozoa

PubMed Central

Yoon, Sung-Jae; Kwon, Woo-Sung; Rahman, Md Saidur; Lee, June-Sub; Pang, Myung-Geol

2015-01-01

Cryopreservation is an efficient way to store spermatozoa and plays a critical role in the livestock industry as well as in clinical practice. During cryopreservation, cryo-stress causes substantial damage to spermatozoa. In present study, the effects of cryo-stress at various cryopreservation steps, such as dilution / cooling, adding cryoprtectant, and freezing were studied in spermatozoa collected from 9 individual bull testes. The motility (%), motion kinematics, capacitation status, mitochondrial activity, and viability of bovine spermatozoa at each step of the cryopreservation process were assessed using computer-assisted sperm analysis, Hoechst 33258/chlortetracycline fluorescence, rhodamine 123 staining, and hypo-osmotic swelling test, respectively. The results demonstrate that the cryopreservation steps reduced motility (%), rapid speed (%), and mitochondrial activity, whereas medium/slow speed (%), and the acrosome reaction were increased (P < 0.05). Differences (Δ) of the acrosome reaction were higher in dilution/cooling step (P < 0.05), whereas differences (Δ) of motility, rapid speed, and non-progressive motility were higher in cryoprotectant and freezing as compared to dilution/cooling (P < 0.05). On the other hand, differences (Δ) of mitochondrial activity, viability, and progressive motility were higher in freezing step (P < 0.05) while the difference (Δ) of the acrosome reaction was higher in dilution/cooling (P < 0.05). Based on these results, we propose that freezing / thawing steps are the most critical in cryopreservation and may provide a logical ground of understanding on the cryo-damage. Moreover, these sperm parameters might be used as physical markers of sperm cryo-damage. PMID:25938413

[Testicular tissue vitrification: evolution or revolution?].

PubMed

Wyns, C; Abu-Ghannam, G; Poels, J

2013-09-01

Preservation of reproductive health is a major concern for patient long-term quality of life. While sperm freezing has proven to be effective to preserve fertility after puberty, cryopreservation of immature testicular tissue (ITT) is emerging as a promising approach for fertility preservation in young boys. Slow-freezing (SF) is the conventional method used to preserve ITT and has resulted in the birth of mice offspring. In humans, methods to preserve ITT are still at the research stage. Controlled SF using dimethyl sulfoxide showed preservation of proliferative spermatogonia after thawing in a xenotransplantation model used to evaluate the efficiency of freezing and thawing procedures. However, spermatogonial recovery was low and normal differentiation could not be achieved. Both freezing/thawing and the environment of the xenotransplantation model may be implicated. Indeed, with SF, ice crystal formation could damage tissue and cells. For this reason, vitrification, leading to solidification of a liquid without crystallization, may be a promising alternative. ITT vitrification has been investigated in different species and shown spermatogonial survival and differentiation to the round or elongated spermatids stage. Offspring were also recently obtained after vitrification and allotransplantation in avians, confirming the potential of vitrification for fertility preservation. In humans, vitrification appears to be as efficient as SF in terms of spermatogonial survival and initiation of differentiation after xenotransplantation. However, before validation of such fertility preservation methods, completion of normal spermatogenesis and the fertilization capacity of sperm retrieved from cryopreserved and transplanted tissue should be fully investigated. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Behavioural and physiological adaptations to low-temperature environments in the common frog, Rana temporaria.

PubMed

Muir, Anna P; Biek, Roman; Mable, Barbara K

2014-05-23

Extreme environments can impose strong ecological and evolutionary pressures at a local level. Ectotherms are particularly sensitive to low-temperature environments, which can result in a reduced activity period, slowed physiological processes and increased exposure to sub-zero temperatures. The aim of this study was to assess the behavioural and physiological responses that facilitate survival in low-temperature environments. In particular, we asked: 1) do high-altitude common frog (Rana temporaria) adults extend the time available for larval growth by breeding at lower temperatures than low-altitude individuals?; and 2) do tadpoles sampled from high-altitude sites differ physiologically from those from low-altitude sites, in terms of routine metabolic rate (RMR) and freeze tolerance? Breeding date was assessed as the first day of spawn observation and local temperature recorded for five, paired high- and low-altitude R. temporaria breeding sites in Scotland. Spawn was collected and tadpoles raised in a common laboratory environment, where RMR was measured as oxygen consumed using a closed respiratory tube system. Freeze tolerance was measured as survival following slow cooling to the point when all container water had frozen. We found that breeding did not occur below 5°C at any site and there was no significant relationship between breeding temperature and altitude, leading to a delay in spawning of five days for every 100 m increase in altitude. The relationship between altitude and RMR varied by mountain but was lower for individuals sampled from high- than low-altitude sites within the three mountains with the highest high-altitude sites (≥900 m). In contrast, individuals sampled from low-altitudes survived freezing significantly better than those from high-altitudes, across all mountains. Our results suggest that adults at high-altitude do not show behavioural adaptations in terms of breeding at lower temperatures. However, tadpoles appear to have the potential to adapt physiologically to surviving at high-altitude via reduced RMR but without an increase in freeze tolerance. Therefore, survival at high-altitude may be facilitated by physiological mechanisms that permit faster growth rates, allowing completion of larval development within a shorter time period, alleviating the need for adaptations that extend the time available for larval growth.

Workplace ART programmes: Why do companies invest in them and are they working?

PubMed

George, Gavin

2006-09-01

Prevalence data indicates that certain sectors within the private sector are particularly affected by HIV/AIDS. Companies in southern Africa began implementing treatment programmes in early 2002 as the corporate sector came to realise the financial imperative of offsetting employee morbidity and mortality. This article sets about to explain the rationale behind antiretroviral treatment (ART) programmes within the private sector while uncovering some of the obstacles businesses face when treating HIV-infected employees. Data suggest that in many cases employees' uptake of voluntary counselling and testing (VCT) and ART are slow. At this early stage of workplace treatment provision, data indicate that employers must seek ways to increase uptake of VCT and treatment in an attempt to make programmes more cost-effective.

Cryosurgery for treatment of trichiasis.

PubMed Central

Sullivan, J H; Beard, C; Bullock, J D

1976-01-01

We cryosurgically destroyed eyelashes in rabbits and applied the technique to treat 23 selected patients with trichiasis. Liquid nitrogen was sprayed on the eyelid margin by using a double, rapid-freeze, slow-thaw cycle monitored by a subcutaneous thermocouple to -30 degrees C. It was an improvement on electrolysis and a simple alternative to surgery. Images FIGURE 1 FIGURE 2 A FIGURE 2 B FIGURE 3 A FIGURE 3 B FIGURE 4 A FIGURE 4 B FIGURE 5 A FIGURE 5 B FIGURE 7 A FIGURE 7 B PMID:867626

Evaluating De-centralised and Distributional Options for the Distributed Electronic Warfare Situation Awareness and Response Test Bed

DTIC Science & Technology

2013-12-01

effectors (deployed on ground based or aerial platforms) to detect , identify, locate, track or suppress stationary or slow moving surface based RF...ground based or aerial platforms) to detect , identify, locate, track or suppress stationary or slow moving surface based RF emitting targets. In the...Electronic Support EO Electro-Optic FPGAs Field Programmable Gate Arrays IR Infra-red LADAR Laser Detection and Ranging OSX Mac OS X; the apple

Closed-loop carrier-envelope phase stabilization with an acousto-optic programmable dispersive filter.

PubMed

Forget, N; Canova, L; Chen, X; Jullien, A; Lopez-Martens, R

2009-12-01

We demonstrate arbitrary carrier-envelope (CE) phase control of femtosecond laser pulses by an acousto-optic programmable dispersive filter (AOPDF), with an accuracy better than pi/100 at a repetition rate of 1 kHz. We also demonstrate, for the first time to the best of our knowledge, 15 Hz closed-loop CE phase stabilization using an AOPDF inside a 1 kHz chirped pulse amplifier to correct for slow CE phase drifts.

Virtual reality walking and dopamine: opening new doorways to understanding freezing of gait in Parkinson's disease.

PubMed

Matar, E; Shine, J M; Naismith, S L; Lewis, S J G

2014-09-15

Freezing of gait (FOG) is a disabling form of gait disturbance that is common in the advanced stages of Parkinson's disease (PD). Despite its prevalence, methods of studying and assessing FOG are limited. We have previously shown that a virtual reality paradigm was able to distinguish between those who report FOG ("freezers") and those who do not report FOG ("non-freezers"). In this paradigm, 'freezers' were found to have prolonged footstep latency in response to known triggers of FOG including doorways, sliding doors and dual-tasking. In this study, we employed the same paradigm to assess performance of 27 freezers and 14 non-freezers in their clinical 'on' and 'off' medication states. In this study, only participants in the freezing group demonstrated statistically significant increases in latencies experienced in the 'off' state compared to the 'on' state in response to wide and narrow doorways and the opening of a sliding door. By contrast, these behavioral differences were not apparent in non-freezers. Furthermore the delay was specific to environmental cues and was not due to generalized slowing in the 'off' state. The findings suggest that this motor delay when processing environmentally salient cues is specific to freezers and is partially mediated by dopamine-dependent neurocircuitry. Copyright © 2014 Elsevier B.V. All rights reserved.

The protective effect of rapid cold-hardening develops more quickly in frozen versus supercooled larvae of the Antarctic midge, Belgica antarctica.

PubMed

Kawarasaki, Yuta; Teets, Nicholas M; Denlinger, David L; Lee, Richard E

2013-10-15

During the austral summer, larvae of the terrestrial midge Belgica antarctica (Diptera: Chironomidae) experience highly variable and often unpredictable thermal conditions. In addition to remaining freeze tolerant year-round, larvae are capable of swiftly increasing their cold tolerance through the rapid cold-hardening (RCH) response. The present study compared the induction of RCH in frozen versus supercooled larvae. At the same induction temperature, RCH occurred more rapidly and conferred a greater level of cryoprotection in frozen versus supercooled larvae. Furthermore, RCH in frozen larvae could be induced at temperatures as low as -12°C, which is the lowest temperature reported to induce RCH. Remarkably, as little as 15 min at -5°C significantly enhanced larval cold tolerance. Not only is protection from RCH acquired swiftly, but it is also quickly lost after thawing for 2 h at 2°C. Because the primary difference between frozen and supercooled larvae is cellular dehydration caused by freeze concentration of body fluids, we also compared the effects of acclimation in dehydrated versus frozen larvae. Because slow dehydration without chilling significantly increased larval survival to a subsequent cold exposure, we hypothesize that cellular dehydration caused by freeze concentration promotes the rapid acquisition of cold tolerance in frozen larvae.

Safe and efficient method for cryopreservation of human induced pluripotent stem cell-derived neural stem and progenitor cells by a programmed freezer with a magnetic field.

PubMed

Nishiyama, Yuichiro; Iwanami, Akio; Kohyama, Jun; Itakura, Go; Kawabata, Soya; Sugai, Keiko; Nishimura, Soraya; Kashiwagi, Rei; Yasutake, Kaori; Isoda, Miho; Matsumoto, Morio; Nakamura, Masaya; Okano, Hideyuki

2016-06-01

Stem cells represent a potential cellular resource in the development of regenerative medicine approaches to the treatment of pathologies in which specific cells are degenerated or damaged by genetic abnormality, disease, or injury. Securing sufficient supplies of cells suited to the demands of cell transplantation, however, remains challenging, and the establishment of safe and efficient cell banking procedures is an important goal. Cryopreservation allows the storage of stem cells for prolonged time periods while maintaining them in adequate condition for use in clinical settings. Conventional cryopreservation systems include slow-freezing and vitrification both have advantages and disadvantages in terms of cell viability and/or scalability. In the present study, we developed an advanced slow-freezing technique using a programmed freezer with a magnetic field called Cells Alive System (CAS) and examined its effectiveness on human induced pluripotent stem cell-derived neural stem/progenitor cells (hiPSC-NS/PCs). This system significantly increased cell viability after thawing and had less impact on cellular proliferation and differentiation. We further found that frozen-thawed hiPSC-NS/PCs were comparable with non-frozen ones at the transcriptome level. Given these findings, we suggest that the CAS is useful for hiPSC-NS/PCs banking for clinical uses involving neural disorders and may open new avenues for future regenerative medicine. Copyright © 2015 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

Freezing solution containing dimethylsulfoxide and fetal calf serum maintains survival and ultrastructure of goat preantral follicles after cryopreservation and in vitro culture of ovarian tissue.

PubMed

Castro, Simone Vieira; de Carvalho, Adeline Andrade; da Silva, Cleidson Manoel Gomes; Faustino, Luciana Rocha; Campello, Cláudio Cabral; Lucci, Carolina Madeira; Báo, Sônia Nair; de Figueiredo, José Ricardo; Rodrigues, Ana Paula Ribeiro

2011-11-01

Goat ovarian cortex fragments were subjected to slow freezing in the presence of various solutions containing intracellular cryoprotectants, including 1.0 M ethylene glycol (EG), propanediol (PROH), or dimethyl sulfoxide (DMSO), with or without sucrose and/or fetal calf serum (FCS). Histological examination revealed that only the DMSO-containing solutions were able to maintain a follicular ultrastructure similar to the morphology observed in the fresh control. Therefore, fragments previously cryopreserved in DMSO solutions (with and without sucrose and/or FCS) were cultured in vitro for 48 h and then subjected to viability, histological, and ultrastructural analysis. No significant differences were observed among the percentages of morphologically normal follicles in cryopreserved ovarian tissue before in vitro culture (DMSO: 62.5%; DMSO + sucrose: 68.3%; DMSO + FCS: 60.0%; DMSO + sucrose + FCS: 60.0%) and after culture (DMSO: 60.8%; DMSO + sucrose: 64.2%; DMSO + FCS: 70.8%; DMSO + sucrose + FCS: 55.0%). Following in vitro culture, the viability analysis showed that only the freezing solution containing DMSO and FCS (75.6%) maintained a percentage of viable follicles similar to that observed after culture without cryopreservation (89.3%). As determined by ultrastructural analysis, morphologically normal preantral follicles were detected in the fresh control and in fragments cultured before and after cryopreservation with DMSO and FCS. Thus, a freezing solution containing DMSO and FCS, under the experimental conditions tested here, guaranteed the maintenance of viability and follicular ultrastructure after short-term in vitro culture.

Effects of α-tocopherol and freezing rates on the quality and heterologous in vitro fertilization capacity of stallion sperm after cryopreservation.

PubMed

de Vasconcelos Franco, J S; Faheem, M; Chaveiro, A; Moreira da Silva, F

2016-09-01

The effects of supplementation of α-tocopherol and different freezing rates (FRs) on the ability of stallion sperm to fertilize bovine oocytes with intact zona pellucida were investigated, in an attempt to develop a model to assess cryopreserved sperm function. Semen was obtained from four purebred Lusitano stallions (n = 4). Each ejaculate was subjected to cryopreservation with a commercial extender (Ghent, Minitub Iberia, Spain), without any supplementation (control) or supplemented with 2-mM α-tocopherol. The semen was exposed to two different FRs between 5 °C and -15 °C: slow (5 °C/min) and moderate (10 °C/min). After thawing, the viability (SYBR®-14 and propidium iodide [PI]), mitochondrial membrane potential (JC-1, 5,5',6,6'-tetrachloro-1,1',3,3'tetraethylbenzimidazolyl carbocyanine iodine) and membrane lipid peroxidation (C11-BODIPY(581/591)) of each sample were determined by flow cytometry. Moreover, the heterologous IVF rate was measured to evaluate the fertilization capacity of postthaw semen in the four different treatments. For both extenders, the viability was higher for spermatozoa cooled slowly (39.40 ± 2.17 vs. 17.59 ± 2.25-control; 31.96 ± 2.19 vs. 11.46 ± 1.34-Tocopherol; P < 0.05). The α-tocopherol extender improved (P < 0.05) postthaw lipid peroxidation (10.28 ± 0.70 vs. 15.40 ± 0.95-slow FR; 10.14 ± 0.40 vs. 13.48 ± 0.34-moderate FR); however, it did not improve viability and mitochondrial membrane potential. Regarding the IVF rate, in the moderate FR, α-tocopherol supplementation reported a higher percentage of IVF (20.50 ± 2.11; P < 0.05), comparing with the control (14.00 ± 1.84). Regarding the slow FR, no significance differences were observed for percentage of IVF between the two extenders and the FRs. However, it seems that the α-tocopherol supplementation improved the IVF rate. In conclusion, this research reported that bovine oocytes intact zona pellucida can be used to evaluate the quality of postthaw stallion semen and α-tocopherol supplementation in the stallion freezing extender might exert a protective effect against oxidative damage during heterologous IVF. Copyright © 2016 Elsevier Inc. All rights reserved.

Feline spermatozoa from fresh and cryopreserved testicular tissues have comparable ability to fertilize matured oocytes and sustain the embryo development after intracytoplasmic sperm injection.

PubMed

Buarpung, S; Tharasanit, T; Comizzoli, P; Techakumphu, M

2013-01-01

Cryopreservation of testicular tissue associated with intracytoplasmic sperm injection (ICSI) is a critical tool that still needs to be explored for preserving the fertility of endangered species. Using the domestic cat as a model for wild felids, the study aimed at determining the effect of different cryoprotectants and freezing techniques (two-step freezing vs. controlled slow freezing) on the sperm quality (membrane and DNA integrity). Then, spermatozoa were extracted from frozen-thawed testicular tissues and used for ICSI to assess early gamete activation or developmental competence in vitro. The percentage of spermatozoa with intact plasma membrane was not different (P > 0.05) among nonfrozen control, glycerol-, and ethylene glycol-frozen tissues (63.2 ± 2%, 58.2 ± 2.6%, 53.3 ± 2.3%, respectively). However, these percentages were significantly lower (P < 0.05) in groups of dimethyl sulfoxide (46.3 ± 3.3%) and 1,2 propanediol (44.3 ± 2.9%) when compared with control. Conventional freezing combined with 5% (vol/vol) glycerol best preserved sperm membrane integrity (55.0 ± 2.7%) when compared with other freezing techniques. The incidence of DNA fragmentation was found to be low (0.2%-1.1%) in all freezing protocols. After ICSI with frozen testicular spermatozoa, male and female gametes were asynchronously activated and the percentages of normal fertilization at 6, 12, and 18 hours were 11.2%, 20.6%, and 22.1%, respectively. Metaphase II-arrested oocytes containing or not a decondensed sperm head were predominantly found after ICSI with frozen testicular spermatozoa. Although two-pronucleus formation could be observed as soon as 6 hours post ICSI (10%), the rate increased dramatically after 12 and 18 hours post ICSI (17.2% and 19.5%, respectively). ICSI using frozen-thawed testicular spermatozoa yielded cleavage (32.7%), morula (6.5%), and blastocyst (4.4%) percentages similar to nonfrozen control (P > 0.05). It is concluded that conventional freezing technique with glycerol as a principle cryoprotectant is simplified and applicable for cat testicular tissue cryopreservation. We also demonstrate for the first time that feline spermatozoa derived from frozen-thawed testicular tissues retain their fertilizing ability and can be used to produce ICSI-derived embryos. Copyright © 2013 Elsevier Inc. All rights reserved.

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

PubMed

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

2014-06-01

Specific protocols for the cryopreservation of endangered Cantabrian brown bear spermatozoa are critical to create a genetic resource bank. The aim of this study was to assess the effect of cooling rates and equilibration time before freezing on post-thawed brown bear spermatozoa quality. Electroejaculates from 11 mature bears were extended to 100 × 10(6) spermatozoa/mL in a TES-Tris-Fructose-based extender, cryopreserved following performance of the respective cooling/equilibration protocol each sample was assigned to, and stored at -196 °C for further assessment. Before freezing, after thawing, and after 1 hour's incubation post-thawing at 37 °C (thermal stress test), the quality of the samples was assessed for motility by computer-assisted semen analysis, and for viability (SYBR-14/propidium iodide), acrosomal status (peanut agglutinin-fluorescein isothiocyanate /propidium iodide), and sperm chromatin stability (SCSA) by flow cytometry. In experiment 1, three cooling rates (0.25 °C/min, 1 °C/min, and 4 °C/min) to 5 °C were assessed. After thawing, total motility (%TM) was higher and percentage of damaged acrosomes (%dACR) was lower (P < 0.05) for 0.25 °C/min than for 4 °C/min. The thermal stress test data indicated equally poor quality (P < 0.05) for the 4 °C/min cooled samples in viability (%VIAB), %dACR, %TM, and progressive motility (%PM). In experiment 2, the effect of a pre-freezing equilibration period at 5 °C for 1 hour (cooling at 0.25 °C/min) was evaluated. Samples kept at 5 °C for 1 hour showed higher (P < 0.05) values than the nonequilibrated ones for both thawing (%dACR) and thermal stress test (%VIAB, %TM, and %PM). In experiment 3, samples stored without cooling and equilibration (direct freezing) were compared with the samples cooled at 0.25 °C/min and equilibrated for 1 hour (control freezing). Using thermal stress test, we observed that direct freezing causes damage in viability, acrosomal status, and motility of spermatozoa compared with the control group (P < 0.05). In conclusion, our results suggest that slow cooling rates to 5 °C and at least 1 hour equilibration time are necessary for the effective cryopreservation of brown bear sperm. Copyright © 2014 Elsevier Inc. All rights reserved.

The development of cat testicular sperm cryopreservation protocols: Effects of tissue fragments or sperm cell suspension.

PubMed

Chatdarong, Kaywalee; Thuwanut, Paweena; Morrell, Jane M

2016-01-15

In endangered animals that have been found dead or sterilized for medical reasons, testis is the ultimate source of haploid DNA or sperm. Thus, preservation of testicular sperm may be performed to rescue their genetics. The aim of this study was to evaluate protocols for testicular sperm freezing: as tissue fragments or cell suspension in domestic cats as a model. A pair of testes from each cat (n = 9) were cut into eight equal pieces. Four randomly selected pieces were cryopreserved as: (1) tissue pieces using two-step freezing; (2) tissue pieces using a slow passive cooling device (CoolCell); (3) sperm suspension after single-layer centrifugation (SLC) through colloids; and (4) sperm suspension without being processed through SLC. A testicular piece from each cat served as fresh control. Testicular sperm membrane and DNA integrity were evaluated before, and after, the cryopreservation process. In addition, spermatogenic cell types (testicular sperm, spermatogonia, spermatocyte, and spermatid) present in the suspension samples were counted before and after SLC. The results found that testicular sperm membrane integrity in the suspension after SLC process was higher than that in the fragment form neither using the two-step nor CoolCell freezing, both before and after freezing (before freezing: 92.3 ± 3.4 vs. 81 ± 4.5 and 80.0 ± 7.0; after freezing: 84.5 ± 4.6 vs. 71.2 ± 12 and 76.2 ± 4.6; P ≤ 0.05). Testicular sperm DNA integrity was, however, not different among groups. Furthermore, the samples processed through the SLC had higher ration of sperm cells: other spermatogenic cells than those were not processed through the SLC (88.9 ± 3.8 vs. 30 ± 7.9; P ≤ 0.05). In summary, testicular sperm cryopreserved as a minced suspension is considered suitable in terms of preventing sperm membrane integrity, and SLC is considered a selection tool for enriching haploid sperm cells from castrated or postmortem cats. Copyright © 2016 Elsevier Inc. All rights reserved.

A comparison of the low temperature transcriptomes and CBF regulons of three plant species that differ in freezing tolerance: Solanum commersonii, Solanum tuberosum, and Arabidopsis thaliana

PubMed Central

Pino, María-Teresa; Jeknić, Zoran; Zou, Cheng; Shiu, Shin-Han; Chen, Tony H. H.; Thomashow, Michael F.

2011-01-01

Solanum commersonii and Solanum tuberosum are closely related plant species that differ in their abilities to cold acclimate; whereas S. commersonii increases in freezing tolerance in response to low temperature, S. tuberosum does not. In Arabidopsis thaliana, cold-regulated genes have been shown to contribute to freezing tolerance, including those that comprise the CBF regulon, genes that are controlled by the CBF transcription factors. The low temperature transcriptomes and CBF regulons of S. commersonii and S. tuberosum were therefore compared to determine whether there might be differences that contribute to their differences in ability to cold acclimate. The results indicated that both plants alter gene expression in response to low temperature to similar degrees with similar kinetics and that both plants have CBF regulons composed of hundreds of genes. However, there were considerable differences in the sets of genes that comprised the low temperature transcriptomes and CBF regulons of the two species. Thus differences in cold regulatory programmes may contribute to the differences in freezing tolerance of these two species. However, 53 groups of putative orthologous genes that are cold-regulated in S. commersonii, S. tuberosum, and A. thaliana were identified. Given that the evolutionary distance between the two Solanum species and A. thaliana is 112–156 million years, it seems likely that these conserved cold-regulated genes—many of which encode transcription factors and proteins of unknown function—have fundamental roles in plant growth and development at low temperature. PMID:21511909

A comparison of the low temperature transcriptomes and CBF regulons of three plant species that differ in freezing tolerance: Solanum commersonii, Solanum tuberosum, and Arabidopsis thaliana.

PubMed

Carvallo, Marcela A; Pino, María-Teresa; Jeknic, Zoran; Zou, Cheng; Doherty, Colleen J; Shiu, Shin-Han; Chen, Tony H H; Thomashow, Michael F

2011-07-01

Solanum commersonii and Solanum tuberosum are closely related plant species that differ in their abilities to cold acclimate; whereas S. commersonii increases in freezing tolerance in response to low temperature, S. tuberosum does not. In Arabidopsis thaliana, cold-regulated genes have been shown to contribute to freezing tolerance, including those that comprise the CBF regulon, genes that are controlled by the CBF transcription factors. The low temperature transcriptomes and CBF regulons of S. commersonii and S. tuberosum were therefore compared to determine whether there might be differences that contribute to their differences in ability to cold acclimate. The results indicated that both plants alter gene expression in response to low temperature to similar degrees with similar kinetics and that both plants have CBF regulons composed of hundreds of genes. However, there were considerable differences in the sets of genes that comprised the low temperature transcriptomes and CBF regulons of the two species. Thus differences in cold regulatory programmes may contribute to the differences in freezing tolerance of these two species. However, 53 groups of putative orthologous genes that are cold-regulated in S. commersonii, S. tuberosum, and A. thaliana were identified. Given that the evolutionary distance between the two Solanum species and A. thaliana is 112-156 million years, it seems likely that these conserved cold-regulated genes-many of which encode transcription factors and proteins of unknown function-have fundamental roles in plant growth and development at low temperature.

Solitosynthesis: Cosmological evolution of non-topological solitons

NASA Technical Reports Server (NTRS)

Griest, Kim; Kolb, Edward W.

1989-01-01

The thermal creation, fusion, evaporation, and destruction of non-topological solitons (NTS) after a phase transition in the early universe is considered. By defining and following NTS statistical equilibrium and departures from it, and depending on particle physics parameters, one of three possible scenarios occurs. If reaction rates are high enough, a period of equilibrium occurs and relic abundances are determined by the freeze-out temperature. Equilibrium first drives most NTS's into their constituents (free phi particles) and then causes rapid fusion into large NTS's. If freeze-out occurs during the first phase, the NTS's are almost entirely destroyed, while if it occurs during the second phase, solitosynthesis occurs and NTS's may be cosmically relevant. For slow reaction rates the NTS's are born frozen out and have the abundance determined by the phase transition. Analytic approximations for determining the abundances are developed, and tested by numerically integrating a reaction network in an expanding universe. Unfortunately, for most of the parameter space considered, solito-destruction/evaporation occurs.

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.

Structure, phonon properties, and order-disorder transition in the metal formate framework of [NH4][Mg(HCOO)3].

PubMed

Mączka, Mirosław; Pietraszko, Adam; Macalik, Bogusław; Hermanowicz, Krzysztof

2014-01-21

We report the synthesis, crystal structure, thermal, dielectric, IR, and Raman studies of [NH4][Mg(HCOO)3] formate. Single-crystal X-ray diffraction shows that it crystallizes in the hexagonal space group P6322, with orientationally disordered NH4(+) ions located in the cages of the network. Upon cooling, [NH4][Mg(HCOO)3] undergoes a phase transition at around 255 K to the ferroelectric P63 structure. Raman and IR spectra show a strong increase in intensity of the N-H stretching bands as well as narrowing of the bands related to the NH4(+) ions upon cooling. These changes indicate that the phase transition is due to orientational ordering of the NH4(+) ions. Analysis of the Raman data show, however, that the rotational and translational motions of NH4(+) do not freeze completely at the phase transition but exhibit further slowing down below 255 K, and the motional freezing becomes nearly complete below 140 K.

Embryo survival and birth rate after minimum volume vitrification or slow freezing of in vivo and in vitro produced ovine embryos.

PubMed

Dos Santos-Neto, P C; Cuadro, F; Barrera, N; Crispo, M; Menchaca, A

2017-10-01

The objective was to evaluate pregnancy outcomes and birth rate of in vivo derived vs. in vitro produced ovine embryos submitted to different cryopreservation methods. A total of 197 in vivo and 240 in vitro produced embryos were cryopreserved either by conventional freezing, or by vitrification with Cryotop or Spatula MVD methods on Day 6 after insemination/fertilization. After thawing/warming and transfer, embryo survival rate on Day 30 of gestation was affected by the source of the embryos (in vivo 53.3%, in vitro 20.8%; P < 0.05) and by the method of cryopreservation (conventional freezing 26.5%, Cryotop 52.0%, Spatula MVD 22.2%; P < 0.05). For in vivo derived embryos, survival rate after embryo transfer was 45.6% for conventional freezing, 67.1% for Cryotop, and 40.4% for Spatula MVD. For in vitro produced embryos, survival rate was 7.3% for conventional freezing, 38.7% for Cryotop, and 11.4% for Spatula MVD. Fetal loss from Day 30 to birth showed a tendency to be greater for in vitro (15.0%) rather than for in vivo produced embryos (5.7%), and was not affected by the cryopreservation method. Gestation length, weight at birth and lamb survival rate after birth were not affected by the source of the embryo, the cryopreservation method or stage of development (average: 150.5 ± 1.8 days; 4232.8 ± 102.8 g; 85.4%; respectively). This study demonstrates that embryo survival and birth rate of both in vivo and in vitro produced ovine embryos are improved by vitrification with the minimum volume Cryotop method. Copyright © 2017 Elsevier Inc. All rights reserved.

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.

Chapter 10 Human Oocyte Vitrification.

PubMed

Rienzi, Laura; Cobo, Ana; Ubaldi, Filippo Maria

2017-01-01

Discovery and widespread application of successful cryopreservation methods for MII-phase oocytes was one of the greatest successes in human reproduction during the past decade. Although considerable improvements in traditional slow-rate freezing were also achieved, the real breakthrough was the result of introduction of vitrification. Here we describe the method that is most commonly applied for this purpose, provides consistent survival and in vitro developmental rates, results in pregnancy and birth rates comparable to those achievable with fresh oocytes, and does not result in higher incidence of gynecological or postnatal complications.

Microscopic relaxations in a protein sustained down to 160K in a non-glass forming organic solvent.

PubMed

Mamontov, E; O'Neill, H

2017-01-01

We have studied microscopic dynamics of a protein in carbon disulfide, a non-glass forming solvent, down to its freezing temperature of ca. 160K. We have utilized quasielastic neutron scattering. A comparison of lysozyme hydrated with water and dissolved in carbon disulfide reveals a stark difference in the temperature dependence of the protein's microscopic relaxation dynamics induced by the solvent. In the case of hydration water, the common protein glass-forming solvent, the protein relaxation slows down in response to a large increase in the water viscosity on cooling down, exhibiting a well-known protein dynamical transition. The dynamical transition disappears in non-glass forming carbon disulfide, whose viscosity remains a weak function of temperature all the way down to freezing at just below 160K. The microscopic relaxation dynamics of lysozyme dissolved in carbon disulfide is sustained down to the freezing temperature of its solvent at a rate similar to that measured at ambient temperature. Our results demonstrate that protein dynamical transition is not merely solvent-assisted, but rather solvent-induced, or, more precisely, is a reflection of the temperature dependence of the solvent's glass-forming dynamics. We hypothesize that, if the long debated idea regarding the direct link between the microscopic relaxations and the biological activity in proteins is correct, then not only the microscopic relaxations, but also the activity, could be sustained in proteins all the way down to the freezing temperature of a non-glass forming solvent with a weak temperature dependence of its viscosity. This article is part of a Special Issue entitled "Science for Life" Guest Editor: Dr. Austen Angell, Dr. Salvatore Magazù and Dr. Federica Migliardo. Copyright © 2016 Elsevier B.V. All rights reserved.

The effect of Me2SO overexposure during cryopreservation on HOS TE85 and hMSC viability, growth and quality.

PubMed

Morris, Timothy J; Picken, Andrew; Sharp, Duncan M C; Slater, Nigel K H; Hewitt, Christopher J; Coopman, Karen

2016-12-01

With the cell therapy industry continuing to grow, the ability to preserve clinical grade cells, including mesenchymal stem cells (MSCs), whilst retaining cell viability and function remains critical for the generation of off-the-shelf therapies. Cryopreservation of MSCs, using slow freezing, is an established process at lab scale. However, the cytotoxicity of cryoprotectants, like Me 2 SO, raises questions about the impact of prolonged cell exposure to cryoprotectant at temperatures >0 °C during processing of large cell batches for allogenic therapies prior to rapid cooling in a controlled rate freezer or in the clinic prior to administration. Here we show that exposure of human bone marrow derived MSCs to Me 2 SO for ≥1 h before freezing, or after thawing, degrades membrane integrity, short-term cell attachment efficiency and alters cell immunophenotype. After 2 h's exposure to Me 2 SO at 37 °C post-thaw, membrane integrity dropped to ∼70% and only ∼50% of cells retained the ability to adhere to tissue culture plastic. Furthermore, only 70% of the recovered MSCs retained an immunophenotype consistent with the ISCT minimal criteria after exposure. We also saw a similar loss of membrane integrity and attachment efficiency after exposing osteoblast (HOS TE85) cells to Me 2 SO before, and after, cryopreservation. Overall, these results show that freezing medium exposure is a critical determinant of product quality as process scale increases. Defining and reporting cell sensitivity to freezing medium exposure, both before and after cryopreservation, enables a fair judgement of how scalable a particular cryopreservation process can be, and consequently whether the therapy has commercial feasibility. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Bacterial Ice Nucleation in Monodisperse D2O and H2O-in-Oil Emulsions.

PubMed

Weng, Lindong; Tessier, Shannon N; Smith, Kyle; Edd, Jon F; Stott, Shannon L; Toner, Mehmet

2016-09-13

Ice nucleation is of fundamental significance in many areas, including atmospheric science, food technology, and cryobiology. In this study, we investigated the ice-nucleation characteristics of picoliter-sized drops consisting of different D2O and H2O mixtures with and without the ice-nucleating bacteria Pseudomonas syringae. We also studied the effects of commonly used cryoprotectants such as ethylene glycol, propylene glycol, and trehalose on the nucleation characteristics of D2O and H2O mixtures. The results show that the median freezing temperature of the suspension containing 1 mg/mL of a lyophilized preparation of P. syringae is as high as -4.6 °C for 100% D2O, compared to -8.9 °C for 100% H2O. As the D2O concentration increases every 25% (v/v), the profile of the ice-nucleation kinetics of D2O + H2O mixtures containing 1 mg/mL Snomax shifts by about 1 °C, suggesting an ideal mixing behavior of D2O and H2O. Furthermore, all of the cryoprotectants investigated in this study are found to depress the freezing phenomenon. Both the homogeneous and heterogeneous freezing temperatures of these aqueous solutions depend on the water activity and are independent of the nature of the solute. These findings enrich our fundamental knowledge of D2O-related ice nucleation and suggest that the combination of D2O and ice-nucleating agents could be a potential self-ice-nucleating formulation. The implications of self-nucleation include a higher, precisely controlled ice seeding temperature for slow freezing that would significantly improve the viability of many ice-assisted cryopreservation protocols.

Fundamental aspects of the freezing of cells, with emphasis on mammalian ova and embryos. (Aspectos fundamentales de la congelacion de celulas, especialmente ovulos y embriones de mamiferos

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

Mazur, P.

1980-01-01

For most cells there exists an optimum cooling rate. Both supraoptimal rates and suboptimal rates can be very damaging. The optimal rate varies enormously from less than or equal to 1/sup 0/C/min for mammalian preimplantation embryos to greater than or equal to 800/sup 0/C/min for the human red cell. Death at supraoptimal rates is the result of the formation of intracellular ice and its recrystallization during warming. Intracellular ice occurs when cells are cooled too rapidly to allow them to equilibrate by the osmotic withdrawal of intracellular water. The definition of too rapid depends chiefly on the size of themore » cell and its permeability to water. Death at suboptimal rates is a consequence of the major alterations in aqueous solutions produced by ice formation. The chief effects are a major reduction in the fraction of the solution remaining unfrozen at a given temperature and a major increase in the solute concentration of that fraction. Presumably, slow freezing injury is a consequence of one or both of these solution effects. The introduction of molar concentrations of protective solutes (additives) greatly reduces both the fraction frozen and the concentration of electrolytes in the unfrozen channels and in the cell interior. Usually, freezing either kills cells outright or it yields survivors that retain full capacity to function. Although there is the possibility that in some cases survivors may in fact be impaired genetically, all evidence indicates that genetic damage does not occur. But there are clear examples in which freezing does induce nonlethal physiological damage. Particularly striking examples are found in certain mammalian sperm.« less

Population policies, programmes and the environment

PubMed Central

Speidel, J. Joseph; Weiss, Deborah C.; Ethelston, Sally A.; Gilbert, Sarah M.

2009-01-01

Human consumption is depleting the Earth's natural resources and impairing the capacity of life-supporting ecosystems. Humans have changed ecosystems more rapidly and extensively over the past 50 years than during any other period, primarily to meet increasing demands for food, fresh water, timber, fibre and fuel. Such consumption, together with world population increasing from 2.6 billion in 1950 to 6.8 billion in 2009, are major contributors to environmental damage. Strengthening family-planning services is crucial to slowing population growth, now 78 million annually, and limiting population size to 9.2 billion by 2050. Otherwise, birth rates could remain unchanged, and world population would grow to 11 billion. Of particular concern are the 80 million annual pregnancies (38% of all pregnancies) that are unintended. More than 200 million women in developing countries prefer to delay their pregnancy, or stop bearing children altogether, but rely on traditional, less-effective methods of contraception or use no method because they lack access or face other barriers to using contraception. Family-planning programmes have a successful track record of reducing unintended pregnancies, thereby slowing population growth. An estimated $15 billion per year is needed for family-planning programmes in developing countries and donors should provide at least $5 billion of the total, however, current donor assistance is less than a quarter of this funding target. PMID:19770155

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

PubMed

Zhang, Jing; Zhou, Aimei; Deng, Aipeng; Yang, Yang; Gao, Lihu; Zhong, Zhaocai; Yang, Shulin

2015-04-01

Pore architecture of 3D scaffolds used in tissue engineering plays a critical role in the maintenance of cell survival, proliferation and further promotion of tissue regeneration. We investigated the pore size and structure, porosity, swelling as well as cell viability of a series of recombinant human collagen-peptide-chitosan (RHCC) scaffolds fabricated by lyophilization. In this paper, freezing regime containing a final temperature of freezing (Tf) and cooling rates was applied to obtain scaffolds with pore size ranging from 100μm to 120μm. Other protocols of RHC/chitosan suspension concentration and ratio modification were studied to produce more homogenous and appropriate structural scaffolds. The mean pore size decreased along with the decline of Tf at a slow cooling rate of 0.7°C/min; a more rapid cooling rate under 5°C/min resulted to a smaller pore size and more homogenous microstructure. High concentration could reduce pore size and lead to thick well of scaffold, while improved the ratio of RHC, lamellar and fiber structure coexisted with cellular pores. Human umbilical vein endothelial cells (HUVECs) were seeded on these manufactured scaffolds, the cell viability represented a negative correlation to the pore size. This study provides an alternative method to fabricate 3D RHC-chitosan scaffolds with appropriate pores for potential tissue engineering. Copyright © 2014 Elsevier B.V. All rights reserved.

Water at protein surfaces studied with femtosecond nonlinear spectroscopy

NASA Astrophysics Data System (ADS)

Bakker, Huib J.

We report on an investigation of the structure and dynamics of water molecules near protein surfaces with femtosecond nonlinear spectroscopic techniques. We measured the reorientation dynamics of water molecules near the surface of several globular protein surfaces, using polarization-resolved femtosecond infrared spectroscopy. We found that water molecules near the protein surface have a much slower reorientation than water molecules in bulk liquid water. The number of slow water molecules scales scales with the size of the hydrophobic surface of the protein. When we denature the proteins by adding an increasing amount of urea to the protein solution, we observe that the water-exposed surface increases by 50% before the secondary structure of the proteins changes. This finding indicates that protein unfolding starts with the protein structure becoming less tight, thereby allowing water to enter. With surface vibrational sum frequency generation (VSFG) spectroscopy, we studied the structure of water at the surface of antifreeze protein III. The measured VSFG spectra showed the presence of ice-like water layers at the ice-binding site of the protein in aqueous solution, at temperatures well above the freezing point. This ordered ice-like hydration layers at the protein surface likely plays an important role in the specific recognition and binding of anti-freeze protein III to nascent ice crystallites, and thus in its anti-freeze mechanism. This research is supported by the ''Nederlandse organisatie voor Wetenschappelijk Onderzoek (NWO).

Individual differences in bodily freezing predict emotional biases in decision making

PubMed Central

Ly, Verena; Huys, Quentin J. M.; Stins, John F.; Roelofs, Karin; Cools, Roshan

2014-01-01

Instrumental decision making has long been argued to be vulnerable to emotional responses. Literature on multiple decision making systems suggests that this emotional biasing might reflect effects of a system that regulates innately specified, evolutionarily preprogrammed responses. To test this hypothesis directly, we investigated whether effects of emotional faces on instrumental action can be predicted by effects of emotional faces on bodily freezing, an innately specified response to aversive relative to appetitive cues. We tested 43 women using a novel emotional decision making task combined with posturography, which involves a force platform to detect small oscillations of the body to accurately quantify postural control in upright stance. On the platform, participants learned whole body approach-avoidance actions based on monetary feedback, while being primed by emotional faces (angry/happy). Our data evidence an emotional biasing of instrumental action. Thus, angry relative to happy faces slowed instrumental approach relative to avoidance responses. Critically, individual differences in this emotional biasing effect were predicted by individual differences in bodily freezing. This result suggests that emotional biasing of instrumental action involves interaction with a system that controls innately specified responses. Furthermore, our findings help bridge (animal and human) decision making and emotion research to advance our mechanistic understanding of decision making anomalies in daily encounters as well as in a wide range of psychopathology. PMID:25071491

Tracking surface and subsurface lakes on the Greenland Ice Sheet using Sentinel-1 SAR and Landsat-8 OLI imagery

NASA Astrophysics Data System (ADS)

Miles, Katie; Willis, Ian; Benedek, Corinne; Williamson, Andrew; Tedesco, Marco

2017-04-01

Supraglacial lakes (SGLs) on the Greenland Ice Sheet (GrIS) are an important component of the ice sheet's mass balance and hydrology, with their drainage affecting ice dynamics. This study uses imagery from the recently launched Sentinel-1A Synthetic Aperture Radar (SAR) to investigate SGLs in West Greenland. SAR can image through cloud and in darkness, overcoming some of the limitations of commonly used optical sensors. A semi automated algorithm is developed to detect surface lakes from Sentinel images during the 2015 summer. It generally detects water in all locations where a Landsat-8 NDWI classification (with a relatively high threshold value) detects water. A combined set of images from Landsat-8 and Sentinel-1 is used to track lake behaviour at a comparable temporal resolution to that which is possible with MODIS, but at a higher spatial resolution. A fully automated lake drainage detection algorithm is used to investigate both rapid and slow drainages for both small and large lakes through the summer. Our combined Landsat-Sentinel dataset, with a temporal resolution of three days, could track smaller lakes (mean 0.089 km2) than are resolvable in MODIS (minimum 0.125 km2). Small lake drainage events (lakes smaller than can be detected using MODIS) were found to occur at lower elevations ( 200 m) and slightly earlier in the melt season than larger events, as were slow lake drainage events compared to rapid events. The Sentinel imagery allows the analysis to be extended manually into the early winter to calculate the dates and elevations of lake freeze-through more precisely than is possible with optical imagery (mean 30 August, 1270 m mean elevation). Finally, the Sentinel imagery allows subsurface lakes (which are invisible to optical sensors) to be detected, and, for the first time, their dates of appearance and freeze-through to be calculated (mean 9 August and 7 October, respectively). These subsurface lakes occur at higher elevations than the surface lakes detected in this study (1593 m mean elevation). Sentinel imagery therefore provides great potential for tracking melting, water movement and freezing within the firn zone of the GrIS.

Developing asynchronous online interprofessional education.

PubMed

Sanborn, Heidi

2016-09-01

For many health programmes, developing interprofessional education (IPE) has been a challenge. Evidence on the best method for design and implementation of IPE has been slow to emerge, with little research on how to best incorporate IPE in the asynchronous online learning environment. This leaves online programmes with no clear guidance when embarking upon an initiative to integrate IPE into the curriculum. One tool that can be effective at guiding the incorporation of IPE across all learning platforms is the Interprofessional Education Collaborative (IPEC) competencies. A project was designed to integrate the nationally defined IPEC competencies throughout an asynchronous, online baccalaureate nursing completion programme. A programme-wide review led to targeted revision of course and unit-level objectives, learning experiences, and assessments based on the IPEC framework. As a result of this effort, the programme curriculum now provides interprofessional learning activities across all courses. This report provides a method for using the IPEC competencies to incorporate IPE within various asynchronous learning assessments, assuring students learn about, with, and from other professions.

Improving ovarian tissue cryopreservation for oncologic patients: slow freezing versus vitrification, effect of different procedures and devices.

PubMed

Herraiz, Sonia; Novella-Maestre, Edurne; Rodríguez, Beatriz; Díaz, César; Sánchez-Serrano, María; Mirabet, Vicente; Pellicer, Antonio

2014-03-01

To compare slow freezing (SF) with four vitrification techniques (VT) for cryopreservation of ovarian tissue (OT) and to evaluate the best protocol for human OT in a xenograft model. Experimental study. University hospital. Patients undergoing fertility preservation. Ovariectomized nude mice. Cryopreservation of bovine OT after SF and four VTs (VT1, VT2, VT3, and VT4) by combining two cryoprotectant vitrification solutions (VS1 and VS2) and two devices (metallic grid and ethyl vinyl acetate bag), after which the cryopreservation of human OT by SF and VT1 and xenograft into nude mice. Follicular densities, proliferation, vascularization, fibrosis, apoptosis, tissue viability. The in vitro study in bovine OT showed a lower percentage of quiescent follicles in the SF group but not in the vitrification groups (VT1-VT4). Apoptosis increased and cell proliferation decreased in all the experimental groups except VT1 (20% ethylene glycol, 20% dimethyl sulfoxide, 0.5 M sucrose, and 20% synthetic serum substitute in HEPES-buffered M199 culture media with Cryotissue metallic grids). Tissue viability was diminished in VT3, and the SF-xenografted human samples showed reduced primordial and secondary densities and unbalanced follicular populations when compared with fresh and VT1 tissue. VT1 offers similar conditions to fresh tissue for follicular density, proliferation, viability, and cell death and preserves a larger population of quiescent follicles than SF after transplantation, thus ensuring the maintenance of graft potential fertility. Copyright © 2014 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Dynamics of water in LiCl and CaCl 2 aqueous solutions confined in silica matrices: A backscattering neutron spectroscopy study

NASA Astrophysics Data System (ADS)

Mamontov, E.; Cole, D. R.; Dai, S.; Pawel, M. D.; Liang, C. D.; Jenkins, T.; Gasparovic, G.; Kintzel, E.

2008-09-01

Backscattering neutron spectroscopy was used to probe the dynamics of water molecules in LiCl and CaCl 2 aqueous solutions confined in 2.7, 1.9, and 1.4 nm diameter pores of various silica matrices. The pore size of 2.7 nm was found to be sufficiently large for the confined liquids to exhibit characteristic traits of bulk behavior, such as a freezing-melting transition and a phase separation. On the other hand, none of the fluids in the 1.4 nm pores exhibited a clear freezing-melting transition; instead, their dynamics at low temperatures gradually became too slow for the nanosecond resolution of the experiment. The greatest suppression of water mobility was observed in the CaCl 2 solutions, which suggests that cation charge and perhaps the cation hydration environment have a profound influence on the dynamics of the water molecules. Quasielastic neutron scattering measurements of pure H 2O and 1 m LiCl-H 2O solution confined in 1.9 nm pores revealed a dynamic transition in both liquids at practically the same temperature of 225-226 K, even though the dynamics of the solution at room temperature appeared to slow down by more than an order of magnitude compared to the pure water. The observation of the dynamic transition in the solution suggests that this transition may be a universal feature of water governed by processes acting on the local scale, such as a change in the hydrogen bonding.

[Disease management programs from a health insurer's point of view].

PubMed

Szymkowiak, Christof; Walkenhorst, Karen; Straub, Christoph

2003-06-01

Disease Management Programmes represent a great challenge to the German statutory health insurance system. According to politicians, disease management programmes are an appropriate tool for increasing the level of care for chronically ill patients significantly, while at the same time they can slow down the cost explosion in health care. The statutory health insurers' point of view yields a more refined picture of the chances and risks involved. The chances are that a medical guideline-based, evidence-based, co-operative care of the chronically ill could be established. But also, there are the risks of misuse of disease management programmes and of misallocation of funds due to the ill-advised linkage with the so-called risk compensation scheme (RSA) balancing the sickness funds' structural deficits through redistribution. The nation-wide introduction of disease management programmes appears to be a gigantic experiment whose aim is to change the care of chronically ill patients and whose outcome is unpredictable.

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.

Oocyte cryopreservation and in vitro culture affect calcium signalling during human fertilization.

PubMed

Nikiforaki, D; Vanden Meerschaut, F; Qian, C; De Croo, I; Lu, Y; Deroo, T; Van den Abbeel, E; Heindryckx, B; De Sutter, P

2014-01-01

What are the precise patterns of calcium oscillations during the fertilization of human oocytes matured either in vivo or in vitro or aged in vitro and what is the effect of cryopreservation? Human oocytes matured in vivo exhibit a specific pattern of calcium oscillations, which is affected by in vitro maturation, in vitro ageing and cryopreservation. Oscillations in cytoplasmic calcium concentration are crucial for oocyte activation and further embryonic development. While several studies have described in detail the calcium oscillation pattern during fertilization in animal models, studies with human oocytes are scarce. This was a laboratory-based study using human MII oocytes matured in vivo or in vitro either fresh or after cryopreservation with slow freezing or vitrification. Altogether, 205 human oocytes were included in the analysis. In vivo and in vitro matured human oocytes were used for this research either fresh or following vitrification/warming (V/W) and slow freezing/thawing (F/T). Human oocytes were obtained following written informed consent from patients undergoing ovarian hyperstimulation. For the calcium pattern analysis, oocytes were loaded with the ratiometric calcium indicator fluorescent dye Fura-2. Following ICSI using sperm from a single donor, intracellular calcium was measured for 16 h at 37°C under 6% CO(2). The calcium oscillation parameters were calculated for all intact oocytes that showed calcium oscillations and were analyzed using the Mann-Whitney U-test. Human in vivo MII oocytes display a specific pattern of calcium oscillations following ICSI. This pattern is significantly affected by in vitro ageing, with the calcium oscillations occurring over a longer period of time and with a lower frequency, shorter duration and higher amplitude (P < 0.05). In vitro matured oocytes from the GV and MI stage exhibit a different pattern of calcium oscillations with calcium transients being of lower frequency and shorter duration compared with in vivo matured MII. In MI oocytes that reached the MII stage within 3 h the calcium oscillations additionally appear over a longer period of time (P < 0.05). In vivo MII oocytes show a different calcium oscillation pattern following V/W with calcium oscillations occurring over a longer period of time, with a higher amplitude and a lower frequency (P < 0.05). In vitro matured oocytes, either from the GV or the MI stage, also display an altered pattern of calcium oscillations after V/W and the parameters that were similarly affected in all these oocyte groups are the frequency and the amplitude of the calcium transients. Slow freezing/thawing differentially affects the calcium oscillation pattern of in vitro matured and in vitro aged oocytes. The relationship between a specific pattern of calcium oscillations and subsequent human embryonic development could not be evaluated since the calcium indicator used and the high-intensity excitation light impair development. Furthermore, all oocytes were derived from stimulated cycles and immature oocytes were denuded prior to in vitro maturation. Our data show for the first time how calcium signalling during human fertilization is affected by oocyte in vitro maturation, in vitro ageing as well as V/W and slow freezing/thawing. The analysis of calcium oscillations could be used as an oocyte quality indicator to evaluate in vitro culture and cryopreservation techniques of human oocytes. This work was supported by a clinical research mandate from the Flemish Foundation of Scientific Research (FWO-Vlaanderen, FWO09/ASP/063) to F.V.M, a fundamental clinical research mandate from the FWO-Vlaanderen (FWO05/FKM/001) to P.D.S and a Ghent University grant (KAN-BOF E/01321/01) to B.H. The authors have no conflict of interest to declare.

Generalized slow roll in the unified effective field theory of inflation

NASA Astrophysics Data System (ADS)

Motohashi, Hayato; Hu, Wayne

2017-07-01

We provide a compact and unified treatment of power spectrum observables for the effective field theory (EFT) of inflation with the complete set of operators that lead to second-order equations of motion in metric perturbations in both space and time derivatives, including Horndeski and Gleyzes-Langlois-Piazza-Vernizzi theories. We relate the EFT operators in ADM form to the four additional free functions of time in the scalar and tensor equations. Using the generalized slow-roll formalism, we show that each power spectrum can be described by an integral over a single source that is a function of its respective sound horizon. With this correspondence, existing model independent constraints on the source function can be simply reinterpreted in the more general inflationary context. By expanding these sources around an optimized freeze-out epoch, we also provide characterizations of these spectra in terms of five slow-roll hierarchies whose leading-order forms are compact and accurate as long as EFT coefficients vary only on time scales greater than an e -fold. We also clarify the relationship between the unitary gauge observables employed in the EFT and the comoving gauge observables of the postinflationary universe.

Relation of behaviour and macrophage function to life span in a murine model of premature immunosenescence.

PubMed

Guayerbas, Noelia; Catalán, Marina; Víctor, Víctor M; Miquel, Jaime; De la Fuente, Mónica

2002-08-21

According to our previous work, mice of the same strain and age show striking inter-individual differences in behaviour when exposed to a T-maze test. Further, the animals exploring the maze slowly (slow mice) or staying at the starting point (freezing behaviour), which show high levels of emotionality/anxiety in other standard behavioural tests, have a less competent immune system (earlier immunosenescence) than those which explore it quickly (fast mice). The present longitudinal study on OF-1 Swiss female mice confirms and extends the above findings. Thus, the animals showing a lower performance in the T-test (slow mice) which is accompanied by a poor neuromuscular coordination in a tightrope test, have a shorter life span than the good performers (fast mice). Moreover, the slow mice have a less competent immune system as regards the following functions of peritoneal macrophages: adherence to substrate, chemotaxis, ingestion of particles and superoxide anion production. This suggests that, at the same chronological age and as regards their immune competence, the slow mice are biologically older than the fast mice. This agrees with current ideas on the close functional relationship between the nervous and the immune system in the physiological adaptation to stress, and supports the concept that an optimum level of performance of these two systems is needed to attain a long life span. Copyright 2002 Elsevier Science B.V.

GMP Cryopreservation of Large Volumes of Cells for Regenerative Medicine: Active Control of the Freezing Process

PubMed Central

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

2014-01-01

Cryopreservation protocols are increasingly required in regenerative medicine applications but must deliver functional products at clinical scale and comply with Good Manufacturing Process (GMP). While GMP cryopreservation is achievable on a small scale using a Stirling cryocooler-based controlled rate freezer (CRF) (EF600), successful large-scale GMP cryopreservation is more challenging due to heat transfer issues and control of ice nucleation, both complex events that impact success. We have developed a large-scale cryocooler-based CRF (VIA Freeze) that can process larger volumes and have evaluated it using alginate-encapsulated liver cell (HepG2) spheroids (ELS). It is anticipated that ELS will comprise the cellular component of a bioartificial liver and will be required in volumes of ∼2 L for clinical use. Sample temperatures and Stirling cryocooler power consumption was recorded throughout cooling runs for both small (500 μL) and large (200 mL) volume samples. ELS recoveries were assessed using viability (FDA/PI staining with image analysis), cell number (nuclei count), and function (protein secretion), along with cryoscanning electron microscopy and freeze substitution techniques to identify possible injury mechanisms. Slow cooling profiles were successfully applied to samples in both the EF600 and the VIA Freeze, and a number of cooling and warming profiles were evaluated. An optimized cooling protocol with a nonlinear cooling profile from ice nucleation to −60°C was implemented in both the EF600 and VIA Freeze. In the VIA Freeze the nucleation of ice is detected by the control software, allowing both noninvasive detection of the nucleation event for quality control purposes and the potential to modify the cooling profile following ice nucleation in an active manner. When processing 200 mL of ELS in the VIA Freeze—viabilities at 93.4%±7.4%, viable cell numbers at 14.3±1.7 million nuclei/mL alginate, and protein secretion at 10.5±1.7 μg/mL/24 h were obtained which, compared well with control ELS (viability −98.1%±0.9%; viable cell numbers −18.3±1.0 million nuclei/mL alginate; and protein secretion −18.7±1.8 μg/mL/24 h). Large volume GMP cryopreservation of ELS is possible with good functional recovery using the VIA Freeze and may also be applied to other regenerative medicine applications. PMID:24410575

Effect of equilibration times, freezing, and thawing rates on post-thaw quality of buffalo (Bubalus bubalis) bull spermatozoa.

PubMed

Shah, S A H; Andrabi, S M H; Qureshi, I Z

2016-09-01

The effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manual, 5 cm above liquid nitrogen (LN2 ) for 10 min, plunging in LN2 ; FR2, programmable ultra-fast, holding at +4 °C for 2 min, from 4 to -10 °C at -10 °C/min, from -10 to -20 °C at -15 °C/min, from -20 to -120 °C at -60 °C/min, holding at -120 °C for 30 sec, plunging in LN2 ), and thawing rates (T1, 37 °C for 30 sec; T2, 50 °C for 15 sec; T3, 70 °C for 7 sec) were evaluated on quality of buffalo bull spermatozoa. Progressive motility (%), rapid velocity (%), average path velocity (VAP, μm/s), straight line velocity (VSL, μm/s), and mitochondrial transmembrane potential (%) were higher (p < 0.05) with E2, FR2, and T3 compared to other groups. Sperm curved line velocity (VCL, μm/s) was higher (p < 0.05) with E2 and FR2 compared to other groups. Sperm straightness (%) and linearity (LIN, %) were higher (p < 0.05) with E2 compared to other groups. Sperm LIN was affected (p < 0.05) with T3 compared to other groups. Supravital-plasma membrane integrity (%), viability and acrosome integrity (%) of spermatozoa were higher (p < 0.05) with E2 and FR2 compared to other groups. Sperm DNA integrity (%) was higher (p < 0.05) with FR2 and T1 compared to other groups. We concluded that inclusion of 4 h-equilibration time, programmable ultra-fast freezing rate, and rapid thawing at 70 °C for 7 sec in cryopreservation protocol improves the post-thaw quality of buffalo bull spermatozoa. © 2016 American Society of Andrology and European Academy of Andrology.

Digital Library Education: Global Trends and Issues

ERIC Educational Resources Information Center

Shem, Magaji

2015-01-01

The paper examines trends and issues in digital education programmes globally, drawing examples of developmental growth of Library Information Science (LIS), schools and digital education courses in North America, Britain, and Southern Asia, the slow growth of LIS schools and digital education in Nigeria and some countries in Africa and India. The…

Impact of Seasonal Heat Accumulation on Operation of Geothermal Heat Pump System with Vertical Ground Heat Exchanger

NASA Astrophysics Data System (ADS)

Timofeev, D. V.; Malyavina, E. G.

2017-11-01

The subject of the investigation was to find out the influence of heat pump operation in summer on its function in winter. For this purpose a mathematical model of a ground coupled heat pump system has been developed and programmed. The mathematical model of a system ground heat exchanger uses the finite difference method to describe the heat transfer in soil and the analytical method to specify the heat transfer in the U-tubes heat exchanger. The thermal diffusivity by the heat transfer in the soil changes during gradual freezing of the pore moisture and thus slows soil freezing. The mathematical model of a heat pump includes the description of a scroll compressor and the simplified descriptions of the evaporator and condenser. The analysis showed that heating during the cold season and cooling in the warm season affect the average heat transfer medium temperature in the soil loop in the winter season. It has been also showed that the degree of this effect depends on the clay content in the soil.

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.

Osmotolerance and leavening ability in sweet and frozen sweet dough. Comparative analysis between Torulaspora delbrueckii and Saccharomyces cerevisiae baker's yeast strains.

PubMed

Hernandez-Lopez, M J; Prieto, J A; Randez-Gil, F

2003-01-01

The response of Saccharomyces cerevisiae and freeze-tolerant Torulaspora delbrueckii strains to osmotic stress and their CO2 production capacity in sweet and frozen-sweet dough has been examined. T. delbrueckii strains, IGC5321 and IGC5323 showed higher leavening ability than Saccharomyces, specially after exposure to hyperosmotic stress of bread dough containing 20% sucrose and 2% salt added. In addition, Torulaspora and especially T. delbrueckii IGC5321 exhibited no loss of CO2 production capacity during freeze-thaw stress. Overall, these results appeared to indicate that Torulaspora cells are more tolerant than Saccharomyces to osmotic stress of bread dough. This trait correlated with a low invertase activity, a slow rate of trehalose mobilisation and the ability to respond rapidly to osmotic stress. Growth behaviour on high osmotic synthetic media was also examined. Cells of the IGC5321 strain showed intrinsic osmotolerance and ion toxicity resistance. However, T. delbrueckii IGC5323 exhibited a clear phenotype of osmosensitivity. Hence, this characteristic may not be essential or the only determinant for leavening ability in salted high-sugar dough.

Study on the feasibility of provision of distance learning programmes in surgery to Malawi.

PubMed

Mains, Edward A A; Blackmur, James P; Dewhurst, David; Ward, Ross M; Garden, O James; Wigmore, Stephen J

2011-12-01

Medical educational opportunities and resources are considerably limited in the developing world. The expansion of computing and Internet access means that there exists a potential to provide education to students through distance learning programmes. This study investigated the feasibility of providing distance learning course in surgery in Malawi. The study investigated the user requirements, technical requirements and Internet connections in two teaching hospitals in Malawi. In addition the appropriateness of current course material from the Edinburgh Surgical Sciences Qualification to Malawi trainees was assessed. The study found a high degree of interest from Malawian trainees in distance learning. The provision of basic science modules such as anatomy and physiology and the ability to access journals were considered highly desirable. The current ESSQ course would require extensive re-modelling to make it suitable to an African trainee's requirements. Internet speeds remain slow and access is currently expensive. There is considerable interest in distance learning programmes in Malawi but access to them is limited partly because of slow and expensive Internet access. Understanding the needs of trainees in countries such as Malawi will allow better direction of educational aid and resources to support surgical training. Copyright © 2010 Royal College of Surgeons of Edinburgh (Scottish charity number SC005317) and Royal College of Surgeons in Ireland. Published by Elsevier Ltd. All rights reserved.

Why are IPTp coverage targets so elusive in sub-Saharan Africa? A systematic review of health system barriers.

PubMed

Thiam, Sylla; Kimotho, Victoria; Gatonga, Patrick

2013-10-03

Use of intermittent preventive treatment (IPTp) is a proven cost-effective intervention for preventing malaria in pregnancy. However, despite the roll-out of IPTp policies across Africa more than ten years ago, utilization levels remain low. This review sought to consolidate scattered evidence as to the health system barriers for IPTp coverage in the continent. Relevant literature from Africa was systematically searched, reviewed and synthesized. Only studies containing primary data were considered. Studies reveal that: (i) poor leadership and governance contribute to slow decentralization of programme management, lack of harmonized guidelines, poor accountability mechanisms, such as robust monitoring and evaluation systems; (ii) low budgetary allocation towards policy implementation slows scale-up, while out-of-pocket expenditure deters women from seeking antenatal services that include IPTp; (iii) there are rampant human resource challenges including low staff motivation levels attributed to such factors as incorrect knowledge of IPTp recommendations and inadequate staffing; (iv) implementation of IPTp policies is hampered by prevailing service delivery barriers, such as long waiting time, long distances to health facilities and poor service provider/client relations; and (v) drug stock-outs and poor management of information and supply chains impair sustained availability of drugs for IPTp. For successful IPTp policy implementation, it is imperative that malaria control programmes target health system barriers that result in low coverage and hence programme ineffectiveness.

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 frozen to −4 and −11 C and kept frozen for up to 12 days. After slow thawing, a 2.5-cm square from a bulb scale was transferred to 25 ml deionized H2O. After shaking for standard times, measurements were made on the effusate and on the effused cells. The results obtained were as follows. Even when the scale tissue was completely infiltrated, and when up to 85% of the ions had diffused out, all of the cells were still alive, as revealed by cytoplasmic streaming and ability to plasmolyze. The osmotic concentration of the cell sap, as measured plasmolytically, decreased in parallel to the rise in conductivity of the effusate. The K+ content of the effusate, plus its assumed counterion, accounted for only 20% of the total solutes, but for 100% of the conductivity. A large part of the nonelectrolytes in the remaining 80% of the solutes was sugars. The increased cell injury and infiltration in the −11 C treatment, relative to the −4 C and control (unfrozen) treatments, were paralleled by increases in conductivity, K+ content, sugar content, and pH of the effusate. In spite of the 100% infiltration of the tissue and the large increase in conductivity of the effusate following freezing, no increase in permeability of the cells to water could be detected. The above observations may indicate that freezing or thawing involves a disruption of the active transport system before the cells reveal any injury microscopically. PMID:16660100

Temporal evolution of solar wind ion composition and their source coronal holes during the declining phase of cycle 23. I. Low-latitude extension of polar coronal holes

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

Ko, Yuan-Kuen; Wang, Yi-Ming; Muglach, Karin

2014-06-01

We analyzed 27 solar wind (SW) intervals during the declining phase of cycle 23, whose source coronal holes (CHs) can be unambiguously identified and are associated with one of the polar CHs. We found that the SW ions have a temporal trend of decreasing ionization state, and such a trend is different between the slow and fast SW. The photospheric magnetic field, both inside and at the outside boundary of the CH, also exhibits a trend of decrease with time. However, EUV line emissions from different layers of the atmosphere exhibit different temporal trends. The coronal emission inside the CHmore » generally increases toward the CH boundary as the underlying field increases in strength and becomes less unipolar. In contrast, this relationship is not seen in the coronal emission averaged over the entire CH. For C and O SW ions that freeze-in at lower altitude, stronger correlation between their ionization states and field strength (both signed and unsigned) appears in the slow SW, while for Fe ions that freeze-in at higher altitude, stronger correlation appears in the fast SW. Such correlations are seen both inside the CH and at its boundary region. On the other hand, the coronal electron temperature correlates well with the SW ion composition only in the boundary region. Our analyses, although not able to determine the likely footpoint locations of the SW of different speeds, raise many outstanding questions for how the SW is heated and accelerated in response to the long-term evolution of the solar magnetic field.« less

An All-Freeze-Casting Strategy to Design Typographical Supercapacitors with Integrated Architectures.

PubMed

Wang, Qingrong; Wang, Xinyu; Wan, Fang; Chen, Kena; Niu, Zhiqiang; Chen, Jun

2018-06-01

The emergence of flexible and wearable electronics has raised the demand for flexible supercapacitors with accurate sizes and aesthetic shapes. Here, a strategy is developed to prepare flexible all-in-one integrated supercapacitors by combining all-freeze-casting with typography technique. The continuous seamless connection of all-in-one supercapacitor devices enhances the load and/or electron transfer capacity and avoids displacing and detaching between their neighboring components at bending status. Therefore, such a unique structure of all-in-one integrated devices is beneficial for retaining stable electrochemical performance at different bending levels. More importantly, the sizes and aesthetic shapes of integrated supercapacitors could be controlled by the designed molds, like type matrices of typography. The molds could be assembled together and typeset randomly, achieving the controllable construction and series and/or parallel connection of several supercapacitor devices. The preparation of flexible integrated supercapacitors will pave the way for assembling programmable all-in-one energy storage devices into highly flexible electronics. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Vagotonicity of Violence: Biochemical and Cardiac Responses to Violent Films and Television Programmes

PubMed Central

Carruthers, Malcolm; Taggart, Peter

1973-01-01

In a search for a reproducible means of evoking different types of emotional stress it was found that in spite of increased adrenaline secretion slowing of the heart occurred when watching violent television programmes. Further evidence of increased vagal tone was provided by the “sinus arrhythmia” effect, a widening of the gap between the maximum and minimum heart rates during the respiratory cycle in parts of the humour, violence, and suspense sections of the television programme. Groups of people taken to see two particularly violent films showed similar evidence suggesting vagal overactivity, together with increases in plasma free fatty acids and decreases in triglycerides. As these changes occurred even with β-blockade it is suggested that they might be caused by non-sympathetically mediated changes in the levels of hormones, such as growth hormone, producing lipolysis. The ability to assess objectively an individual's reaction to viewing violence might make it possible to judge the likely social impact of violent films and television programmes. PMID:4730188

Fresh versus frozen embryo transfers in assisted reproduction.

PubMed

Wong, Kai Mee; van Wely, Madelon; Mol, Femke; Repping, Sjoerd; Mastenbroek, Sebastiaan

2017-03-28

In general, in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) implies a single fresh and one or more frozen-thawed embryo transfers. Alternatively, the 'freeze-all' strategy implies transfer of frozen-thawed embryos only, with no fresh embryo transfers. In practice, both strategies can vary technically including differences in freezing techniques and timing of transfer of cryopreservation, that is vitrification versus slow freezing, freezing of two pro-nucleate (2pn) versus cleavage-stage embryos versus blastocysts, and transfer of cleavage-stage embryos versus blastocysts.In the freeze-all strategy, embryo transfers are disengaged from ovarian stimulation in the initial treatment cycle. This could avoid a negative effect of ovarian hyperstimulation on the endometrium and thereby improve embryo implantation. It could also reduce the risk of ovarian hyperstimulation syndrome (OHSS) in the ovarian stimulation cycle by avoiding a pregnancy.We compared the benefits and risks of the two treatment strategies. To evaluate the effectiveness and safety of the freeze-all strategy compared to the conventional IVF/ICSI strategy in women undergoing assisted reproductive technology. We searched the Cochrane Gynaecology and Fertility Group Trials Register, the Cochrane Central Register of Studies (CRSO), MEDLINE, Embase, PsycINFO, CINAHL, and two registers of ongoing trials in November 2016 together with reference checking and contact with study authors and experts in the field to identify additional studies. We included randomised clinical trials comparing a freeze-all strategy with a conventional IVF/ICSI strategy which includes fresh transfer of embryos in women undergoing IVF or ICSI treatment. We used standard methodological procedures recommended by Cochrane. The primary review outcomes were cumulative live birth and OHSS. Secondary outcomes included other adverse effects (miscarriage rate). We included four randomised clinical trials analysing a total of 1892 women comparing a freeze-all strategy with a conventional IVF/ICSI strategy. The evidence was of moderate to low quality due to serious risk of bias and (for some outcomes) serious imprecision. Risk of bias was associated with unclear blinding of investigators for preliminary outcomes of the study, unit of analysis error, and absence of adequate study termination rules.There was no clear evidence of a difference in cumulative live birth rate between the freeze-all strategy and the conventional IVF/ICSI strategy (odds ratio (OR) 1.09, 95% confidence interval (CI) 0.91 to 1.31; 4 trials; 1892 women; I 2 = 0%; moderate-quality evidence). This suggests that if the cumulative live birth rate is 58% following a conventional IVF/ICSI strategy, the rate following a freeze-all strategy would be between 56% and 65%.The prevalence of OHSS was lower after the freeze-all strategy compared to the conventional IVF/ICSI strategy (OR 0.24, 95% CI 0.15 to 0.38; 2 trials; 1633 women; I 2 = 0%; low-quality evidence). This suggests that if the OHSS rate is 7% following a conventional IVF/ICSI strategy, the rate following a freeze-all strategy would be between 1% and 3%.The freeze-all strategy was associated with fewer miscarriages (OR 0.67, 95% CI 0.52 to 0.86; 4 trials; 1892 women; I 2 = 0%; low-quality evidence) and a higher rate of pregnancy complications (OR 1.44, 95% CI 1.08 to 1.92; 2 trials; 1633 women; low-quality evidence). There was no difference in multiple pregnancies per woman after the first transfer (OR 1.11, 95% CI 0.85 to 1.44; 2 trials; 1630 women; low-quality evidence), and no data were reported for time to pregnancy. We found moderate-quality evidence showing that one strategy is not superior to the other in terms of cumulative live birth rates. Time to pregnancy was not reported, but it can be assumed to be shorter using a conventional IVF/ICSI strategy in the case of similar cumulative live birth rates, as embryo transfer is delayed in a freeze-all strategy. Low-quality evidence suggests that not performing a fresh transfer lowers the OHSS risk for women at risk of OHSS.

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.

Has Earth's Plate Tectonics Led to Rapid Core Cooling?

NASA Astrophysics Data System (ADS)

de Montserrat Navarro, A.; Morgan, J. P.; Vannucchi, P.; Connolly, J. A.

2016-12-01

Earth's mantle and core are convecting planetary heat engines. The mantle convects to lose heat from secular cooling, internal radioactivity, and core heatflow across its base. Its convection generates plate tectonics, volcanism, and the loss of 35 TW of mantle heat through Earth's surface. The core convects to lose heat from secular cooling, small amounts of internal radioactivity, and the freezing-induced growth of a compositionally denser inner core. Until recently, the geodynamo was thought to be powered by 4 TW of heatloss across the core-mantle boundary. More recent determinations of the outer core's thermal conductivity (Pozzo et al., 2012; Gomi et al., 2013) would imply that >15 TW of power should conduct down its adiabat. Secular core cooling has been previously thought to be too slow for this, based on estimates for the Clapeyron Slope for high-pressure freezing of an idealized pure-iron core (cf. Nimmo, 2007). The 500-1000 kg m-3 seismically-inferred jump in density between the liquid outer core and solid inner core allows a direct estimate of the Clapeyron Slope for the outer core's actual composition which contains 0.08±0.02 lighter elements (S,Si,O,Al, H,…) mixed into a Fe-Ni alloy. A PREM-like 600 kg m-3 density jump yields a Clapeyron Slope for which there has been 774K of core cooling during the freezing and growth of the inner core, cooling that has been releasing an average of 21 TW of power during the past 3 Ga. If so, core cooling could easily have powered Earth's long-lived geodynamo. Another implication is that the present-day mantle is strongly `bottom-heated', and diapiric mantle plumes should dominate deep mantle upwelling. This mode of core and mantle convection is consistent with slow, 37.5K/Ga secular cooling of Earth's mantle linked to more rapid secular cooling of the core (cf. Morgan, Rüpke, and White, 2016). Efficient plate subduction, hence plate tectonics, is a key ingredient for such rapid secular core cooling.We also show how a more complete thermodynamic version of Birch's accretional energy calculation predicts that accretion with FeNi-sinking-linked differentiation between an Earth-like mantle and core would naturally generate a core that, post-accretion, was both hotter than overlying mantle and 1000K hotter than today.

The formation of supercooled brines, viscous liquids, and low-temperature perchlorate glasses in aqueous solutions relevant to Mars

NASA Astrophysics Data System (ADS)

Toner, J. D.; Catling, D. C.; Light, B.

2014-05-01

Salt solutions on Mars can stabilize liquid water at low temperatures by lowering the freezing point of water. The maximum equilibrium freezing-point depression possible, known as the eutectic temperature, suggests a lower temperature limit for liquid water on Mars; however, salt solutions can supercool below their eutectic before crystallization occurs. To investigate the magnitude of supercooling and its variation with salt composition and concentration, we performed slow cooling and warming experiments on pure salt solutions and saturated soil-solutions of MgSO4, MgCl2, NaCl, NaClO4, Mg(ClO4)2, and Ca(ClO4)2. By monitoring solution temperatures, we identified exothermic crystallization events and determined the composition of precipitated phases from the eutectic melting temperature. Our results indicate that supercooling is pervasive. In general, supercooling is greater in more concentrated solutions and with salts of Ca and Mg. Slowly cooled MgSO4, MgCl2, NaCl, and NaClO4 solutions investigated in this study typically supercool 5-15 °C below their eutectic temperature before crystallizing. The addition of soil to these salt solutions has a variable effect on supercooling. Relative to the pure salt solutions, supercooling decreases in MgSO4 soil-solutions, increases in MgCl2 soil-solutions, and is similar in NaCl and NaClO4 soil-solutions. Supercooling in MgSO4, MgCl2, NaCl, and NaClO4 solutions could marginally extend the duration of liquid water during relatively warm daytime temperatures in the martian summer. In contrast, we find that Mg(ClO4)2 and Ca(ClO4)2 solutions do not crystallize during slow cooling, but remain in a supercooled, liquid state until forming an amorphous glass near -120 °C. Even if soil is added to the solutions, a glass still forms during cooling. The large supercooling effect in Mg(ClO4)2 and Ca(ClO4)2 solutions has the potential to prevent water from freezing over diurnal and possibly annual cycles on Mars. Glasses are also potentially important for astrobiology because of their ability to preserve pristine cellular structures intact compared to solutions that crystallize.

Relative contributions of the fraction of unfrozen water and of salt concentration to the survival of slowly frozen human erythrocytes.

PubMed Central

Mazur, P; Rall, W F; Rigopoulos, N

1981-01-01

As suspensions of cells freeze, the electrolytes and other solutes in the external solution concentrate progressively, and the cells undergo osmotic dehydration if cooling is slow. The progressive concentration of solute comes about as increasing amounts of pure ice precipitate out of solution and cause the liquid-filled channels in which the cells are sequestered to dwindle in size. The consensus has been that slow freezing injury is related to the composition of the solution in these channels and not to the amount of residual liquid. The purpose of the research reported here was to test this assumption on human erythrocytes. Ordinarily, solute concentration and the amount of liquid in the unfrozen channels are inversely coupled. To vary them independently, one must vary the initial solute concentration. Two solutes were used here: NaCl and the permeating protective additive glycerol. To vary the total initial solute concentration while holding the mass ratio of glycerol to NaCl constant, we had to allow the NaCl tonicity to depart from isotonic. Specifically, human red cells were suspended in solutions with weight ratios of glycerol to NaCl of either 5.42 or 11.26, where the concentrations of NaCl were 0.6, 0.75, 1.0, 2.0, 3.0, or 4.0 times isotonic. Samples were then frozen to various subzero temperatures, which were chosen to produce various molalities of NaCl (0.24-3.30) while holding the fraction of unfrozen water constant, or conversely to produce various unfrozen fractions (0.03-0.5) while holding the molality of salt constant. (Not all combinations of these values were possible). The following general findings emerged: (a) few cells survived the freezing of greater than 90% of the extracellular water regardless of the salt concentration in the residual unfrozen portion. (b) When the fraction of frozen water was less than 75% the majority of the cells survived even when the salt concentration in the unfrozen portion exceeded 2 molal. (c) Salt concentration affected survival significantly only when the frozen fraction lay between 75 and 90%. To find a major effect on survival of the fraction of water that remains unfrozen was unexpected. It may require major modifications in how cryobiologists view solution-effect injury and its prevention. PMID:7326328

Observation of a brine layer on an ice surface with an environmental scanning electron microscope at higher pressures and temperatures.

PubMed

Krausko, Ján; Runštuk, Jiří; Neděla, Vilém; Klán, Petr; Heger, Dominik

2014-05-20

Observation of a uranyl-salt brine layer on an ice surface using backscattered electron detection and ice surface morphology using secondary-electron detection under equilibrium conditions was facilitated using an environmental scanning electron microscope (ESEM) at temperatures above 250 K and pressures of hundreds of Pa. The micrographs of a brine layer over ice grains prepared by either slow or shock freezing provided a complementary picture of the contaminated ice grain boundaries. Fluorescence spectroscopy of the uranyl ions in the brine layer confirmed that the species exists predominately in the solvated state under experimental conditions of ESEM.

Cosmological history in York time: inflation and perturbations

NASA Astrophysics Data System (ADS)

Roser, Philipp; Valentini, Antony

2017-02-01

The constant mean extrinsic curvature on a spacelike slice may constitute a physically preferred time coordinate, `York time'. One line of enquiry to probe this idea is to understand processes in our cosmological history in terms of York time. Following a review of the theoretical motivations, we focus on slow-roll inflation and the freezing and Hubble re-entry of cosmological perturbations. While the physics is, of course, observationally equivalent, we show how the mathematical account of these processes is distinct from the conventional account in terms of standard cosmological or conformal time. We also consider the cosmological York-timeline more broadly and contrast it with the conventional cosmological timeline.

Cryopreservation of umbilical cord blood with a novel freezing solution that mimics intracellular ionic composition.

PubMed

Nicoud, Ian B; Clarke, Dominic M; Taber, Greta; Stolowski, Kristin M; Roberge, Sarah E; Song, Melissa K; Mathew, Aby J; Reems, Jo-Anna

2012-09-01

Cryopreservation protocols have remained relatively unchanged since the first umbilical cord blood banking program was established. This study evaluated the preservation efficacy of a novel intracellular-like cryopreservation solution (CryoStor, BioLife Solutions, Inc.), the rate of addition of two cryopreservation solutions to cord blood units (CBUs), and reduced final dimethyl sulfoxide (DMSO) concentration of 5%. Split-sample CBUs were cryopreserved with either an in-house 20% DMSO-based cryopreservation solution or CryoStor CS10 at a rate of 1 mL/min (n = 10; i.e., slow addition) or as a bolus injection (n = 6; i.e., fast addition). Infrared images of exothermic effects of the cryopreservation solutions were monitored relative to the rate of addition. Prefreeze and postthaw colony-forming unit assays, total nucleated cells, and CD34+ cell counts were compared. Maximum temperature excursions observed were less than 6°C, regardless of the rate of solution addition. Fast addition resulted in peak excursions approximately twice that of slow addition but the magnitude and duration were minimal and transient. Slow addition of CryoStor CS10 (i.e., final concentration ≤ 5% DMSO) resulted in significantly better postthaw CD34+ cell recoveries; no other metrics were significantly different. Fast addition of CryoStor resulted in similar postthaw metrics compared to slow addition of the in-house solution. Slow and fast addition of cryopreservation solutions result in mean temperature changes of approximately 3.3 to 4.45°C. Postthaw recoveries with CryoStor were equivalent to or slightly better than with the in-house cryopreservation solution. CryoStor also provides several advantages including reduced processing time, formulation consistency, and reduced DMSO in the frozen product (≤ 5%). © 2012 American Association of Blood Banks.

Frozen and fresh ovarian tissue require different culture media to promote in vitro development of bovine preantral follicles.

PubMed

Castro, Simone Vieira; Carvalho, Adeline Andrade; Silva, Cleidson Manoel Gomes; Santos, Francielli Weber; Campello, Cláudio Cabral; de Figueiredo, José Ricardo; Rodrigues, Ana Paula Ribeiro

2014-10-01

The aim of this study was to evaluate the efficiency of different media in the in vitro culture of bovine preantral follicles that were used either fresh or following slow freezing treatment. Frozen and fresh noncultured or cultured ovarian fragments were processed for histological, viability, and cell proliferation analyses. For cryopreservation, a solution containing 1.5 M ethylene glycol was frozen in a programmable biological freezer. After thawing, a portion of the samples was destined for frozen controls. The remainder were cultured in vitro for 5 days in three media: α-MEM, McCoy, or M199. Samples from these culture media were collected on days 1 and 5 for quantification of reactive oxygen species (ROS) and for hormonal assays. In fresh-cultured tissues, the percentage of morphologically normal follicles was significantly higher when cultured in M199 compared to that in the other media. In frozen-cultured tissues, McCoy medium was significantly superior to the other media, and was the only treatment that helped in maintaining the viability similar to fresh and frozen controls. Upon quantification of the nucleolus organizer region, we observed greater proliferation of granulosa cells in the frozen-cultured tissues with McCoy medium, and lesser proliferation in fresh-cultured tissues only with α-MEM. In frozen-cultured tissues, ROS levels were highest at day 1 and progressively reduced during culture, independent of the media used. In conclusion, under the conditions used in this study, the M199 and McCoy media are recommended for the culture of follicles derived from fresh and frozen ovarian tissues, respectively.

Estuarine conservation and restoration: the Somme and the Seine case studies (English Channel, France).

PubMed

Ducrotoy, Jean-Paul; Dauvin, Jean-Claude

2008-01-01

Megatidal estuaries such as the Seine and the Somme (North-Western France) are rather well delimited and human impacts on them are well understood. Since the middle of the 19th Century, there has been a slow but irreversible degradation of the state of these English Channel estuaries. However, current conservation and restoration strategies tend to freeze habitats in a particular state, their status being defined, most often, through a patrimonial or utilitarian approach. Connectedness between biotopes (sensu habitat+community) has a tendency to be neglected, especially with regard to main ecological gradients, i.e., salinity. In this paper, evaluation methodologies are proposed with the intention of assessing changes to ecosystem functions, under anthropogenic disturbance, controlled or otherwise. The Seine (a heavily industrialised ecosystem) is compared to the Somme (considered here for its pseudo-natural features) in order to discriminate between oceanic processes (siltation and plugging of estuaries) and anthropogenic influences. Preservation and restoration of habitats rely on a robust scientific methodology. The multi-scale approach adopted in the projects presented here relies on sensitive socio-ecological assessment procedures, tools for evaluating ecological quality, and well-built monitoring programmes based upon pertinent indicators. Such managerial tools were used to refine strategies and make them compatible with the sustainable co-development of resources in a European context. This paper demonstrates how scientists were able to acquire and apply knowledge in the field of rehabilitation and restoration. Jointly with managers and policy-makers, they have brought scientific information and socio-economics together in order to answer questions about the restoration of sites or habitats and to anticipate future propositions in the spirit of Integrated Coastal Zone Management (ICZM).

Measurement of Total Calcium in Neurons by Electron Probe X-ray Microanalysis

PubMed Central

Pivovarova, Natalia B.; Andrews, S. Brian

2013-01-01

In this article the tools, techniques, and instruments appropriate for quantitative measurements of intracellular elemental content using the technique known as electron probe microanalysis (EPMA) are described. Intramitochondrial calcium is a particular focus because of the critical role that mitochondrial calcium overload plays in neurodegenerative diseases. The method is based on the analysis of X-rays generated in an electron microscope (EM) by interaction of an electron beam with the specimen. In order to maintain the native distribution of diffusible elements in electron microscopy specimens, EPMA requires "cryofixation" of tissue followed by the preparation of ultrathin cryosections. Rapid freezing of cultured cells or organotypic slice cultures is carried out by plunge freezing in liquid ethane or by slam freezing against a cold metal block, respectively. Cryosections nominally 80 nm thick are cut dry with a diamond knife at ca. -160 °C, mounted on carbon/pioloform-coated copper grids, and cryotransferred into a cryo-EM using a specialized cryospecimen holder. After visual survey and location mapping at ≤-160 °C and low electron dose, frozen-hydrated cryosections are freeze-dried at -100 °C for ~30 min. Organelle-level images of dried cryosections are recorded, also at low dose, by means of a slow-scan CCD camera and subcellular regions of interest selected for analysis. X-rays emitted from ROIs by a stationary, focused, high-intensity electron probe are collected by an energy-dispersive X-ray (EDX) spectrometer, processed by associated electronics, and presented as an X-ray spectrum, that is, a plot of X-ray intensity vs. energy. Additional software facilitates: 1) identification of elemental components by their "characteristic" peak energies and fingerprint; and 2) quantitative analysis by extraction of peak areas/background. This paper concludes with two examples that illustrate typical EPMA applications, one in which mitochondrial calcium analysis provided critical insight into mechanisms of excitotoxic injury and another that revealed the basis of ischemia resistance. PMID:24300079

A pilot treatment study for mild traumatic brain injury: Neuroimaging changes detected by MEG after low-intensity pulse-based transcranial electrical stimulation.

PubMed

Huang, Ming-Xiong; Swan, Ashley Robb; Quinto, Annemarie Angeles; Matthews, Scott; Harrington, Deborah L; Nichols, Sharon; Bruder, Barry J; Snook, Corey C; Huang, Charles W; Baker, Dewleen G; Lee, Roland R

2017-01-01

Mild traumatic brain injury (mTBI) is a leading cause of sustained impairments in military service members, Veterans, and civilians. However, few treatments are available for mTBI, partially because the mechanism of persistent mTBI deficits is not fully understood. We used magnetoencephalography (MEG) to investigate neuronal changes in individuals with mTBI following a passive neurofeedback-based treatment programme called IASIS. This programme involved applying low-intensity pulses using transcranial electrical stimulation (LIP-tES) with electroencephalography monitoring. Study participants included six individuals with mTBI and persistent post-concussive symptoms (PCS). MEG exams were performed at baseline and follow-up to evaluate the effect of IASIS on brain functioning. At the baseline MEG exam, all participants had abnormal slow-waves. In the follow-up MEG exam, the participants showed significantly reduced abnormal slow-waves with an average reduction of 53.6 ± 24.6% in slow-wave total score. The participants also showed significant reduction of PCS scores after IASIS treatment, with an average reduction of 52.76 ± 26.4% in PCS total score. The present study demonstrates, for the first time, the neuroimaging-based documentation of the effect of LIP-tES treatment on brain functioning in mTBI. The mechanisms of LIP-tES treatment are discussed, with an emphasis on LIP-tES's potentiation of the mTBI healing process.

"Working Together": An Intercultural Academic Leadership Programme to Build Health Science Educators' Capacity to Teach Indigenous Health and Culture

ERIC Educational Resources Information Center

Durey, Angela; Taylor, Kate; Bessarab, Dawn; Kickett, Marion; Jones, Sue; Hoffman, Julie; Flavell, Helen; Scott, Kim

2017-01-01

Progress has been slow in improving health disparities between Aboriginal and Torres Strait Islander (Indigenous) Australians and other Australians. While reasons for this are complex, delivering healthcare respectful of cultural differences is one approach to improving Indigenous health outcomes. This paper presents and evaluates an intercultural…

Slow Controls Using the Axiom M5235BCC

NASA Astrophysics Data System (ADS)

Hague, Tyler

2008-10-01

The Forward Vertex Detector group at PHENIX plans to adopt the Axiom M5235 Business Card Controller for use as slow controls. It is also being evaluated for slow controls on FermiLab e906. This controller features the Freescale MCF5235 microprocessor. It also has three parallel buses, these being the MCU port, BUS port, and enhanced Time Processing Unit (eTPU) port. The BUS port uses a chip select module with three external chip selects to communicate with peripherals. This will be used to communicate with and configure Field Programmable Gate Arrays (FPGAs). The controller also has an Ethernet port which can use several different protocols such as TCP and UDP. This will be used to transfer files with computers on a network. The M5235 Business Card Controller will be placed in a VME crate along with VME card and a Spartan-3 FPGA.

Can thermostable vaccines help address cold-chain challenges? Results from stakeholder interviews in six low- and middle-income countries

PubMed Central

Kristensen, Debra D.; Lorenson, Tina; Bartholomew, Kate; Villadiego, Shirley

2016-01-01

Introduction This study captures the perspectives of stakeholders at multiple levels of the vaccine supply chain regarding their assessment of challenges with storing vaccines within recommended temperature ranges and their perceptions on the benefits of having vaccines with improved stability, including the potential short-term storage and transport of vaccines in a controlled-temperature chain. Methods Semi-structured interviews were undertaken with 158 immunization stakeholders in six countries. Interviewees included national decision-makers and advisors involved in vaccine purchasing decisions, national Expanded Programme on Immunization managers, and health and logistics personnel at national, subnational, and health facility levels. Results Challenges with both heat and freeze-exposure of vaccines were recognized in all countries, with heat-exposure being a greater concern. Conditions leading to freeze-exposure including ice build-up due to poor refrigerator performance and improper icepack conditioning were reported by 53% and 28% of participants, respectively. Respondents were interested in vaccine products with improved heat/freeze-stability characteristics. The majority of those involved in vaccine purchasing indicated they would be willing to pay a US$0.05 premium per dose for a freeze-stable pentavalent vaccine (68%) or a heat-stable rotavirus vaccine (59%), although most (53%) preferred not to pay the premium for a heat-stable pentavalent vaccine if the increased stability required changing from a liquid to a lyophilized product. Most respondents (73%) were also interested in vaccines labeled for short-term use in a controlled-temperature chain. The majority (115/158) recognized the flexibility this would provide during outreach or should cold-chain breaks occur. Respondents were also aware that possible confusion might arise and additional training would be required if handling conditions were changed for some, but not all vaccines. Conclusion Participating immunization stakeholders recognized the benefits of vaccine products with improved stability characteristics and of labeling vaccines for controlled-temperature chain use as a means to help address cold-chain issues in their immunization programs. PMID:26778422

Aluminum Fixed Point: Impact of the Time Spent in the Liquid Phase on the Liquid-Solid Transition and Obviousness of the Pollution of the Ingot

NASA Astrophysics Data System (ADS)

Renaot, E.; Martin, C.

2011-08-01

In order to improve the uncertainty on the aluminum fixed point, a study was launched by Laboratoire Commun de Métrologie LNE-CNAM in the frame of the EURAMET Project 732 "Toward more accurate temperature fixed points" (coordinating laboratory: France, 17 partner countries). An earlier study completed in this laboratory showed that in regular realization of the melting-freezing plateaus, there is no diffusion of impurities in the thickness of the ingot, or the diffusion is excessively slow and cannot allow a uniform distribution of the impurities. On the other hand, it is frequently noticed that the experimental conditions before the freezing plateau have an impact on its characteristics (value, slope,…). Up to now, no systematic study was performed on the influence of this parameter. So, the objective of the task started recently in this laboratory is to investigate the influence of the time spent in the liquid phase on the phase transition. As a final result, it is demonstrated that in order to reach the equilibrium of the concentration of impurities, it is necessary to ensure that the metal remains in the liquid phase at least 24 h before initiating the freeze. At the end of the process, the aluminum ingot was chemically analyzed. The analyses reveal large contaminations of the surface of the ingot (sodium, sulfur, and phosphorus). One of the important outputs of this study is that the conditions of usage of the cells should be given important attention since large contaminations can be brought by the furnace.

Criteria to assess human oocyte quality after cryopreservation.

PubMed

Coticchio, G; Bonu, M A; Bianchi, V; Flamigni, C; Borini, A

2005-10-01

Oocyte cryopreservation certainly represents one of the most attractive developments in the field of assisted reproduction, with the aim of preserving female fertility and circumventing the ethical and legal drawbacks associated with embryo freezing. Despite the achievement of the first pregnancy from frozen oocytes dating back as early as 1987, since then fewer than 150 pregnancies have been reported. Over a long period of time, application of oocyte storage on a large scale has been prevented by various factors, namely poor post-thaw survival. Fertilization rates remained low even after the introduction of intracytoplasmic sperm injection. Modifications of slow-freezing protocols, mainly based on the increase of the concentration of sucrose used as non-penetrating cryoprotectant (CPA) and the replacement of sodium with choline, appear to have decisively improved survival rates to over 80%. Investigations at the cellular level on thawed oocytes are largely lacking. Fertilization rates have also benefited from protocol modifications, reaching values indistinguishable from those normally obtained with fresh material. Vitrification protocols have also been tested, giving rise to improvements whose reproducibility is still uncertain. Data on the dynamics of fertilization and preimplantation development of embryos derived from frozen oocytes are extremely scarce. At the moment, clinical efficiency of oocyte cryopreservation cannot be precisely assessed because of the lack of controlled studies, although it appears to be considerably lower than that achieved with embryo freezing. In summary, encouraging advances have been made in the field of oocyte cryopreservation, but presently no protocol can ensure standards of success and safety comparable to those guaranteed by embryo storage.

Community dynamics drive punctuated engraftment of the fecal microbiome following transplantation using freeze-dried, encapsulated fecal microbiota.

PubMed

Staley, Christopher; Vaughn, Byron P; Graiziger, Carolyn T; Singroy, Stephanie; Hamilton, Matthew J; Yao, Dan; Chen, Chi; Khoruts, Alexander; Sadowsky, Michael J

2017-05-04

Fecal microbiota transplantation (FMT) is a highly effective treatment of recurrent and recalcitrant Clostridium difficile infection (rCDI). In a recent study oral-delivery of encapsulated, freeze-dried donor material, resulted in comparable rates of cure to colonoscopic approaches. Here we characterize shifts in the fecal bacterial community structure of patients treated for rCDI using encapsulated donor material. Prior to FMT, patient fecal samples showed declines in diversity and abundance of Firmicutes and Bacteroidetes, with concurrent increases in members of the Proteobacteria, specifically Enterobacteriaceae. Moreover, patients who experienced recurrence of CDI within the 2-month clinical follow-up had greater abundances of Enterobacteriaceae and did not show resolution of dysbioses. Despite resolution of rCDI following oral-administration of encapsulated fecal microbiota, community composition was slow to return to a normal donor-like assemblage. Post-FMT taxa within the Firmicutes showed rapid increases in relative abundance and did not vary significantly over time. Conversely, Bacteroidetes taxa only showed significant increases in abundance after one month post-FMT, corresponding to significant increases in the community attributable to the donors. Changes in the associations among dominant OTUs were observed at days, weeks, and months post-FMT, suggesting shifts in community dynamics may be related to the timing of increases in abundance of specific taxa. Administration of encapsulated, freeze-dried, fecal microbiota to rCDI patients resulted in restoration of bacterial diversity and resolution of dysbiosis. However, shifts in the fecal microbiome were incremental rather than immediate, and may be driven by changes in community dynamics reflecting changes in the host environment.

Vitrification as an alternative means of cryopreserving ovarian tissue.

PubMed

Amorim, Christiani A; Curaba, Mara; Van Langendonckt, Anne; Dolmans, Marie-Madeleine; Donnez, Jacques

2011-08-01

Because of the simplicity of vitrification, many authors have investigated it as an alternative to slow freezing for cryopreserving ovarian tissue. In the last decade, numerous studies have evaluated vitrification of ovarian tissue from both humans and animals.Different vitrification solutions and protocols, mostly adapted from embryo and oocyte vitrification, have been applied. The results have been discrepant from species to species and even within the same species, but lately they appear to indicate that vitrification can achieve similar or even superior results to conventional freezing. Despite the encouraging results obtained with vitrification of ovarian tissue from humans and different animal species, it is necessary to understand how vitrification solutions and protocols can affect ovarian tissue, notably preantral follicles. In addition, it is important to bear in mind that the utilization of different approaches to assess tissue functionality and oocyte quality is essential in order to validate the promising results already obtained with vitrification procedures. This review summarizes the principles of vitrification, discusses the advantages of vitrification protocols for ovarian tissue cryopreservation and describes different studies conducted on the vitrification of ovarian tissue in humans and animal species. Copyright © 2011 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

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

NASA Astrophysics Data System (ADS)

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

2016-03-01

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

Vitreous Cryopreservation of Human Umbilical Vein Endothelial Cells with Low Concentration of Cryoprotective Agents for Vascular Tissue Engineering

PubMed Central

Zheng, Yuanyuan; Panhwar, Fazil

2016-01-01

Cryopreservation of human umbilical vein endothelial cells (HUVECs) is important to tissue engineering applications and the study of the role of endothelial cells in cardiovascular and cerebrovascular diseases. The traditional methods for cryopreservation by vitrification (cooling samples to a cryogenic temperature without apparent freezing) using high concentration of cryoprotective agents (CPAs) and slow freezing are suboptimal due to the severe toxicity of high concentration of CPAs and ice formation-induced cryoinjuries, respectively. In this study, we developed a method to cryopreserve HUVECs by vitrification with low concentration of CPAs. This is achieved by optimizing the CPAs and using highly thermally conductive quartz capillary (QC) to contain samples for vitrification. The latter minimizes the thermal mass to create ultra-fast cooling/warming rates. Our data demonstrate that HUVECs can be vitrified in the QC using 1.4 mol/L ethylene glycol and 1.1 mol/L dimethyl sulfoxide with more than 90% viability. Moreover, this method significantly improves the attachment efficiency of the cryopreserved HUVECs. The attached cells post-cryopreservation proliferate similarly to fresh cells. Therefore, this study may provide an effective vitrification technique to bank HUVECs for vascular tissue engineering and other applications. PMID:27673413

The current challenges to efficient immature oocyte cryopreservation.

PubMed

Brambillasca, Fausta; Guglielmo, Maria Cristina; Coticchio, Giovanni; Mignini Renzini, Mario; Dal Canto, Mariabeatrice; Fadini, Rubens

2013-12-01

Oocyte cryopreservation represents an important tool for assisted reproductive technology. It offers the opportunity to preserve fertility in women at risk of loss of the ovarian function for various pathologies. It also represents a treatment alternative for couples that cannot benefit from embryo cryopreservation because of moral, religious, or legal constrains. On the other hand, in vitro oocyte maturation has a range of applications. It can be applied in patients with a contraindication to ovarian stimulation to prevent ovarian hyperstimulation syndrome or to eliminate the risk of stimulation of hormone-sensitive tumours in cancer patients. However, while mature oocyte cryopreservation has found wide-spread application and oocyte in vitro maturation has a place for the treatment of specific clinical conditions, data on the efficiency of freezing of immature or in vitro matured oocytes are poorer. In this review we will focus on the combination of oocyte in vitro maturation with oocyte cryopreservation with particular emphasis on the biological implications of the cryopreservation of immature or in vitro matured oocytes. The two cryopreservation approaches, slow freezing and vitrification, will be discussed in relation to possible cryodamage occurring to subcellular structures of the oocyte and the functional interaction between oocyte and cumulus cells.

Progressive freezing of interacting spins in isolated finite magnetic ensembles

NASA Astrophysics Data System (ADS)

Bhattacharya, Kakoli; Dupuis, Veronique; Le-Roy, Damien; Deb, Pritam

2017-02-01

Self-organization of magnetic nanoparticles into secondary nanostructures provides an innovative way for designing functional nanomaterials with novel properties, different from the constituent primary nanoparticles as well as their bulk counterparts. Collective magnetic properties of such complex closed packing of magnetic nanoparticles makes them more appealing than the individual magnetic nanoparticles in many technological applications. This work reports the collective magnetic behaviour of magnetic ensembles comprising of single domain Fe3O4 nanoparticles. The present work reveals that the ensemble formation is based on the re-orientation and attachment of the nanoparticles in an iso-oriented fashion at the mesoscale regime. Comprehensive dc magnetic measurements show the prevalence of strong interparticle interactions in the ensembles. Due to the close range organization of primary Fe3O4 nanoparticles in the ensemble, the spins of the individual nanoparticles interact through dipolar interactions as realized from remnant magnetization measurements. Signature of super spin glass like behaviour in the ensembles is observed in the memory studies carried out in field cooled conditions. Progressive freezing of spins in the ensembles is corroborated from the Vogel-Fulcher fit of the susceptibility data. Dynamic scaling of relaxation reasserted slow spin dynamics substantiating cluster spin glass like behaviour in the ensembles.

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

PubMed Central

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

2016-01-01

During cryopreservation, ice recrystallization is a major cause of cellular damage. Conventional cryoprotectants such as dimethyl sulfoxide (DMSO) and glycerol function by a number of different mechanisms but do not mitigate or control ice recrystallization at concentrations utilized in cryopreservation procedures. In North America, cryopreservation of human red blood cells (RBCs) utilizes high concentrations of glycerol. RBC units frozen under these conditions must be subjected to a time-consuming deglycerolization process after thawing in order to remove the glycerol to <1% prior to transfusion thus limiting the use of frozen RBC units in emergency situations. We have identified several low molecular mass ice recrystallization inhibitors (IRIs) that are effective cryoprotectants for human RBCs, resulting in 70–80% intact RBCs using only 15% glycerol and slow freezing rates. These compounds are capable of reducing the average ice crystal size of extracellular ice relative to a 15% glycerol control validating the positive correlation between a reduction in ice crystal size and increased post-thaw recovery of RBCs. The most potent IRI from this study is also capable of protecting frozen RBCs against the large temperature fluctuations associated with transient warming. PMID:27021850

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

PubMed

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

2016-03-29

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

Reducing child mortality in India in the new millennium.

PubMed Central

Claeson, M.; Bos, E. R.; Mawji, T.; Pathmanathan, I.

2000-01-01

Globally, child mortality rates have been halved over the last few decades, a developmental success story. Nevertheless, progress has been uneven and in recent years mortality rates have increased in some countries. The present study documents the slowing decline in infant mortality rates in india; a departure from the longer-term trends. The major causes of childhood mortality are also reviewed and strategic options for the different states of India are proposed that take into account current mortality rates and the level of progress in individual states. The slowing decline in childhood mortality rates in India calls for new approaches that go beyond disease-, programme- and sector-specific approaches. PMID:11100614

[Corifollitropin alfa in women stimulated for the first time in in vitro fertilization programme].

PubMed

Vraná-Mardešićová, N; Vobořil, J; Melicharová, L; Jelínková, L; Vilímová, Š; Mardešić, T

2017-01-01

To compare results after stimulation with corifollitropin alfa (Elonva) in unselected group of women entering for the first time in in vitro fertilization programme (IVF) with results from Phase III randomized trials with selected groups of women. Prospective study. Sanatorium Pronatal, Praha. 40 unselected women with adequat ovarian reserve entering for the first time in IVF programme were stimulated with corifollitropin alfa and GnRH antagonists. Avarage age in the study group was 32,8 years (29-42 years), women younger then 36 and less then 60 kg received Elonva 100 µg , all others (age > 36 let, weight > 60 kg) Elonva 150 µg. Five days after egg retrieval one blastocyst was transferred (single embryo transfer - eSET). Our results were compared with the resuls in higly selected groups of women from Phase III randomized trials. After stimulation with corifollitropin alfa and GnRH antagonists on average 10,6 (9,2 ± 4,2) eggs could be retrieved, among them 7,3 (6,6 ± 3,9) were M II oocytes (68,9%) and fertilisation rate was 84,6%. After first embryo transfer ("fresh" embryos and embryos from "freeze all" cycles) 14 pregnancies were achieved (37,8%), three pregnancies were achieved later from transfer of frozen-thawed embryos (cumulative pregnancy rate 45,9%). There were three abortions. No severe hyperstimulation syndrom occured. Our results in unselected group of women stimulated for the first in an IVF programme with corifollitropin alfa are fully comparable with results published in randomized trials with selected group of patiens. Corifollitropin alfa in combination with daily GnRH antagonist can be successfully used in normal-responder patients stimulated for the first time in an IVF programmeKeywords: corifollitropin alfa, GnRH antagonists, ovarian stimulation, pregnancy.

Associations between Expressive and Receptive Language and Internalizing and Externalizing Behaviours in a Community-Based Prospective Study of Slow-to-Talk Toddlers

ERIC Educational Resources Information Center

Conway, Laura J.; Levickis, Penny A.; Mensah, Fiona; McKean, Cristina; Smith, Kylie; Reilly, Sheena

2017-01-01

Background: Evidence suggests that language and social, emotional and behavioural (SEB) difficulties are associated in children and adolescents. When these associations emerge and whether they differ by language or SEB difficulty profile is unclear. This knowledge is crucial to guide prevention and intervention programmes for children with…

Perceived Benefits and Barriers to the Use of High-Speed Broadband in Ireland's Second-Level Schools

ERIC Educational Resources Information Center

Coyne, Bryan; Devitt, Niamh; Lyons, Seán; McCoy, Selina

2015-01-01

As part of Ireland's National Digital Strategy, high-speed broadband is being rolled out to all second-level schools to support greater use of information and communication technology (ICT) in education. This programme signals a move from slow and unreliable broadband connections for many schools to a guaranteed high-speed connection with…

Cooling rate dependence of the glass transition at free surfaces

NASA Astrophysics Data System (ADS)

Streit-Nierobisch, S.; Gutt, C.; Paulus, M.; Tolan, M.

2008-01-01

In situ x-ray reflectivity measurements are used to determine the cooling rate dependent freezing of capillary waves on the oligomer poly(propylene glycol). Only above the glass transition temperature TG can the surface roughness σ be described by the capillary wave model for simple liquids, whereas the surface fluctuations are frozen-in at temperatures below TG . As the state of a glass forming liquid strongly depends on its thermal history, this effect occurs for fast cooling rates already at a higher temperature than for slow cooling. For the fastest cooling rates a very large shift of TG up to 240K compared to the bulk value of 196K was observed.

The Tethered Moon

NASA Technical Reports Server (NTRS)

Zahnle, Kevin; Lupu, Roxana Elena; Dubrovolskis, A. R.

2014-01-01

A reasonable initial condition on Earth after the Moonforming impact is that it begins as a hot global magma ocean1,2. We therefore begin our study with the mantle as a liquid ocean with a surface temperature on the order of 3000- 4000 K at a time some 100-1000 years after the impact, by which point we can hope that early transients have settled down. A 2nd initial condition is a substantial atmosphere, 100-1000 bars of H2O and CO2, supplemented by smaller amounts of CO, H2, N2, various sulfur-containing gases, and a suite of geochemical volatiles evaporated from the magma. Third, we start the Moon with its current mass at the relevant Roche limit. The 4th initial condition is the angular momentum of the Earth-Moon system. Canonical models hold this constant, whilst some recent models begin with considerably more angular momentum than is present today. Here we present a ruthlessly simplified model of Earth's cooling magmasphere based on a full-featured atmosphere and including tidal heating by the newborn Moon. Thermal blanketing by H2O-CO2 atmospheres slows cooling of a magma ocean. Geochemical volatiles - chiefly S, Na, and Cl - raise the opacity of the magma ocean's atmosphere and slow cooling still more. We assume a uniform mantle with a single internal (potential) temperature and a global viscosity. The important "freezing point" is the sharp rheological transition between a fluid carrying suspended crystals and a solid matrix through which fluids percolate. Most tidal heating takes place at this "freezing point" in a gel that is both pliable and viscous. Parameterized convection links the cooling rate to the temperature and heat generation inside the Earth. Tidal heating is a major effect. Tidal dissipation in the magma ocean is described by viscosity. The Moon is entwined with Earth by the negative feedback between thermal blanketing and tidal heating that comes from the temperature-dependent viscosity of the magma ocean. Because of this feedback, the rate that the Moon's orbit evolves is limited by the modest radiative cooling rate of Earth's atmosphere, which in effect tethers the Moon to the Earth. Consequently the Moon's orbit evolves orders of magnitude more slowly than in conventional models. Slow orbital evolution promotes capture by orbital resonances that may have been important in the Earth-Moon system

Men's perspectives on fall risk and fall prevention following participation in a group-based programme conducted at Men's Sheds, Australia.

PubMed

Liddle, Jeannine L M; Lovarini, Meryl; Clemson, Lindy M; Jang, Haeyoung; Willis, Karen; Lord, Stephen R; Sherrington, Catherine

2017-05-01

Research on older men's views regarding fall prevention is limited. The purpose of this qualitative study was to explore the experiences and perspectives of older men regarding fall risk and prevention so that fall prevention programmes can better engage older men. Eleven men who had taken part in a group-based fall prevention programme called Stepping On conducted at Men's Sheds in Sydney, Australia, participated in semi-structured interviews during June and July 2015 which were audio-recorded and transcribed. Data were coded and analysed using constant comparative methods. Over-arching theoretical categories were developed into a conceptual framework linking programme context and content with effects of programme participation on men. Men's Sheds facilitated participation in the programme by being inclusive, male-friendly places, where Stepping On was programmed into regular activities and was conducted in an enjoyable, supportive atmosphere. Programme content challenged participants to think differently about themselves and their personal fall risk, and provided practical options to address fall risk. Two major themes were identified: adjusting the mindset where men adopted a more cautious mindset paying greater attention to potential fall risks, being careful, concentrating and slowing down; and changing the ways where men acted purposefully on environmental hazards at home and incorporated fall prevention exercises into their routine schedules. Practitioners can engage and support older men to address falls by better understanding men's perspectives on personal fall risk and motivations for action. © 2016 John Wiley & Sons Ltd.

Identifying and Evaluating Teachers' Knowledge in Relation to Child Abuse and Neglect: A Qualitative Study with Australian Early Childhood Teachers

ERIC Educational Resources Information Center

Walsh, Kerryann; Farrell, Ann

2008-01-01

Child abuse and neglect are serious social problems that make extraordinary demands on teachers' knowledge and professionalism. Yet the field of education has been slow to develop a discipline-specific knowledge base about child abuse and neglect for teachers and teacher education programmes and there is a paucity empirical research into teachers'…

Towards monitoring surface and subsurface lakes on the Greenland Ice Sheet using Sentinel-1 SAR and Landsat-8 OLI imagery

NASA Astrophysics Data System (ADS)

Miles, Katie E.; Willis, Ian C.; Benedek, Corinne L.; Williamson, Andrew G.; Tedesco, Marco

2017-07-01

Supraglacial lakes are an important component of the Greenland Ice Sheet’s mass balance and hydrology, with their drainage affecting ice dynamics. This study uses imagery from the recently launched Sentinel-1A Synthetic Aperture Radar (SAR) satellite to investigate supraglacial lakes in West Greenland. A semi-automated algorithm is developed to detect surface lakes from Sentinel-1 images during the 2015 summer. A combined Landsat-8 and Sentinel-1 dataset, which has a comparable temporal resolution to MODIS (3 days versus daily) but a higher spatial resolution (25-40 m versus 250-500 m), is then used together with a fully-automated lake drainage detection algorithm. Rapid (< 4 days) and slow (> 4 days) drainages are investigated for both small (< 0.125 km2, the minimum size detectable by MODIS) and large (≥ 0.125 km2) lakes through the summer. Drainage events of small lakes occur at lower elevations (mean 159 m), and slightly earlier (mean 4.5 days) in the melt season than those of large lakes. The analysis is extended manually into the early winter to calculate the dates and elevations of lake freeze-through more precisely than is possible with optical imagery (mean 30 August; 1270 m mean elevation). Finally, the Sentinel-1 imagery is used to detect subsurface lakes and, for the first time, their dates of appearance and freeze-through (mean 9 August and 7 October, respectively). These subsurface lakes occur at higher elevations than the surface lakes detected in this study (mean 1593 m and 1185 m, respectively). Sentinel-1 imagery therefore provides great potential for tracking melting, water movement and freezing within both the firn zone and ablation area of the Greenland Ice Sheet.

[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.

Ovarian tissue cryopreservation and transplantation among alternatives for fertility preservation in the Nordic countries - compilation of 20 years of multicenter experience.

PubMed

Rodriguez-Wallberg, Kenny A; Tanbo, Tom; Tinkanen, Helena; Thurin-Kjellberg, Ann; Nedstrand, Elizabeth; Kitlinski, Margareta Laczna; Macklon, Kirsten T; Ernst, Erik; Fedder, Jens; Tiitinen, Aila; Morin-Papunen, Laure; Einarsson, Snorri; Jokimaa, Varpu; Hippeläinen, Maritta; Lood, Mikael; Gudmundsson, Johannes; Olofsson, Jan I; Andersen, Claus Yding

2016-09-01

The aim of this study is to report the current status of ovarian tissue cryopreservation among alternatives for fertility preservation in the Nordic countries. A questionnaire was sent to 14 Nordic academic reproductive centers with established fertility preservation programs. It covered fertility preservation cases performed up to December 2014, standard procedures for ovarian tissue cryopreservation and oocyte cryopreservation and reproductive outcomes following ovarian tissue transplantation. Among the Nordic countries, Denmark and Norway practice ovarian tissue cryopreservation as a clinical treatment (822 and 164 cases, respectively) and their programs are centralized. In Sweden (457 cases), ovarian tissue cryopreservation is practiced at five of six centers and in Finland at all five centers (145 cases). Nearly all considered ovarian tissue cryopreservation to be experimental. In Iceland, embryo cryopreservation is the only option for fertility preservation. Most centers use slow-freezing methods for ovarian tissue cryopreservation. Most patients selected for ovarian tissue cryopreservation were newly diagnosed with cancer and the tissue was predominantly retrieved laparoscopically by unilateral oophorectomy. Only minor complications were reported. In total, 46 women have undergone ovarian tissue transplantation aiming at recovering fertility, 17 healthy children have been born and several additional pregnancies are currently ongoing. Whenever patients' clinical condition is permissive, oocyte cryopreservation after hormonal stimulation is preferred for fertility preservation. Between 2012 and 2014, a smaller proportion of females have undergone fertility preservation in the Nordic centers, in comparison to males (1:3). Overall, ovarian tissue cryopreservation was reported to be safe. Slow freezing methods are still preferred. Promising results of recovery of fertility have been reported in Nordic countries that have initiated ovarian tissue transplantation procedures. © 2016 The Authors. Acta Obstetricia et Gynecologica Scandinavica published by John Wiley & Sons Ltd on behalf of Nordic Federation of Societies of Obstetrics and Gynecology (NFOG).

VizieR Online Data Catalog: Herschel FIR observations of NGC3603 (Di Cecco+, 2015)

NASA Astrophysics Data System (ADS)

di Cecco, A.; Faustini, F.; Paresce, F.; Correnti, M.; Calzoletti, L.

2015-06-01

The cloud complex surrounding NGC 3603 YC was observed by Herschel SPIRE/PACS during the Hi-GAL Survey (Molinari et al. 2010A&A...518L.100M, 2010PASP..122..314M), a Key Programme that mapped the Galactic plane in five photometric bands (70, 160, 250, 350, and 500um). The MSX (Egan et al. 2003AAS...203.5708E) images were taken at (wavelengths) 8.3, 12.1, 14.7, and 21.3um using the scan observing mode, and the WISE (Wright et al. 2010AJ....140.1868W) images were acquired in freeze-frame scan mode at 3.4, 4.6, 12, and 22um. (2 data files).

Novel ultrasonic real-time scanner featuring servo controlled transducers displaying a sector image.

PubMed

Matzuk, T; Skolnick, M L

1978-07-01

This paper describes a new real-time servo controlled sector scanner that produces high resolution images and has functionally programmable features similar to phased array systems, but possesses the simplicity of design and low cost best achievable in a mechanical sector scanner. The unique feature is the transducer head which contains a single moving part--the transducer--enclosed within a light-weight, hand held, and vibration free case. The frame rate, sector width, stop action angle, are all operator programmable. The frame rate can be varied from 12 to 30 frames s-1 and the sector width from 0 degrees to 60 degrees. Conversion from sector to time motion (T/M) modes are instant and two options are available, a freeze position high density T/M and a low density T/M obtainable simultaneously during sector visualization. Unusual electronic features are: automatic gain control, electronic recording of images on video tape in rf format, and ability to post-process images during video playback to extract T/M display and to change time gain control (tgc) and image size.

Electronic health information system at an opioid treatment programme: roadblocks to implementation

PubMed Central

Louie, Ben; Kritz, Steven; Brown, Lawrence S.; Chu, Melissa; Madray, Charles; Zavala, Roberto

2012-01-01

Rationale Electronic health systems are commonly included in health care reform discussions. However, their embrace by the health care community has been slow. Methods At Addiction Research and Treatment Corporation, a methadone maintenance programme that also provides primary medical care, HIV medical care and case management, substance abuse counselling and vocational services, we describe our experience in implementing an electronic health information system that encompasses all of these areas. Results We describe the challenges and opportunities of this process in terms of change management, hierarchy of corporate objectives, process mastering, training issues, information technology governance, electronic security, and communication and collaboration. Conclusion This description may provide practical insights to other institutions seeking to pursue this technology. PMID:21414111

Electronic health information system at an opioid treatment programme: roadblocks to implementation.

PubMed

Louie, Ben; Kritz, Steven; Brown, Lawrence S; Chu, Melissa; Madray, Charles; Zavala, Roberto

2012-08-01

Electronic health systems are commonly included in health care reform discussions. However, their embrace by the health care community has been slow. At Addiction Research and Treatment Corporation, a methadone maintenance programme that also provides primary medical care, HIV medical care and case management, substance abuse counselling and vocational services, we describe our experience in implementing an electronic health information system that encompasses all of these areas. We describe the challenges and opportunities of this process in terms of change management, hierarchy of corporate objectives, process mastering, training issues, information technology governance, electronic security, and communication and collaboration. This description may provide practical insights to other institutions seeking to pursue this technology. © 2011 Blackwell Publishing Ltd.

[Impact of heavy snow storm and freezing rain disasters on soil fauna in Chinese fir plantation in southern China].

PubMed

Yan, Shao-kui; Zhang, Wei-dong; Liu, Yan-xin; Fu, Sheng-lei; Li, Yuan-liang; Wang, Si-long

2009-01-01

In January 2008, southern China suffered an unusual heavy snowstorm and freezing rain over a large area for almost a month long. This catastrophic event was the worst one in past 50 years, which brought the area a serious impact on the infrastructure, ecology, and environment. To understand the long-term impact of this catastrophic event on the forest ecosystems in this area, a field investigation was conducted on the soil fauna in a pure Chinese fir plantation and a mixed Chinese fir plantation-alder plantation in Huitong County of Hunan Province on March 23, 2008, the date 40 days after the heavy snowstorm and freezing rain. With the abundance and community composition as the main parameters and the monitoring data from the two plantations on March 23, 2007 as the reference, the flexibility and resistance of soil fauna to the disturbances of the catastrophic event was preliminarily evaluated. The results showed that there was a significant deviation of soil fauna communities in the two plantations from the reference. An outbreak increase in microfauna nematode abundance was found from 12216.9 ind x m(-2) to 118343.9 ind x m(-2) in pure Chinese fir plantation and from 25435.9 ind x m(-2) to 84573.0 ind x m(-2) in mixed Chinese fir plantation-alder plantation, while a 27.0% and 85.6% decrease of macrofauna abundance was found in the two plantations, respectively, compared with the reference. Mesofauna abundance also had a significant decrease in litter layer but not in soil. The abundance recovery displayed a trend from quick rate for microfauna to slow rate for macrofauna, which indicated that the soil fauna functional groups, in terms of body size, could be used as a vulnerable indicator in evaluating disturbance event and post-disturbance recovery. By using community ordinations, no shift in soil fauna community composition was detected 40 days after the catastrophic event, suggesting that the community composition of soil invertebrate had a high resistance to catastrophic snowstorm and freezing rain disturbances.

Volatile generation in bell peppers during frozen storage and thawing using selected ion flow tube mass spectrometry (SIFT-MS).

PubMed

Wampler, Brendan; Barringer, Sheryl A

2012-06-01

To determine volatile formation during storage and thawing, whole, pureed, blanched, and raw green and red bell peppers (Capsicum annuum) were frozen quickly or slowly then stored at -18 °C for up to 7 mo, with and without SnCl(2) addition during thawing. Headspace analysis was performed by a Selected Ion Flow Tube Mass Spectrometer (SIFT-MS). After blanching, (Z)-3-hexenal had a large significant decrease in concentration since it is a heat labile compound while most other volatiles did not change in concentration. The freezing process increased volatile levels in the puree only. Slow freeze peppers had higher levels of some LOX generated volatiles during storage than quick freeze. During frozen storage of blanched samples (E)-2-hexenal, (Z and E)-hexen-1-ol, and (E)-2-pentenal increased likely because of nonenzymatic autoxidation of fatty acids while other volatiles remained constant. In Raw Whole peppers, (Z)-3-hexenal, hexanal, and 2-pentylfuran were generated during storage likely because the LOX enzyme is still active during frozen storage. However, blanched samples had higher concentrations of (E)-2-hexenal, (Z and E)-hexen-1-ol, 1-penten-3-one, and (E)-2-heptenal because of enzymatic destruction of these volatiles in the raw samples. The levels of many of the volatiles in the raw samples, including (Z)-3-hexenal, (E)-2-hexenal, (Z and E)-hexen-1-ol, hexanal, (E)-2-pentenal, and 2-pentylfuran, appeared to peak around 34 d after freezing. Pureed samples had significantly higher levels of volatiles than the whole samples, and volatiles peaked earlier. Green bell pepper volatile levels were always higher than red bell pepper. Significantly higher volatile formation occurred during thawing than it did during frozen storage. Studying and monitoring the headspace volatiles with a SIFT-MS can give information that will help manufacturers better understand how the volatiles in bell peppers change during frozen storage. This will give valuable information to processors on how to minimize volatile changes during storage of frozen peppers. © 2012 Institute of Food Technologists®

Prevention of Osmotic Injury to Human Umbilical Vein Endothelial Cells for Biopreservation: A First Step Toward Biobanking of Endothelial Cells for Vascular Tissue Engineering.

PubMed

Niu, Dan; Zhao, Gang; Liu, Xiaoli; Zhou, Ping; Cao, Yunxia

2016-03-01

High-survival-rate cryopreservation of endothelial cells plays a critical role in vascular tissue engineering, while optimization of osmotic injuries is the first step toward successful cryopreservation. We designed a low-cost, easy-to-use, microfluidics-based microperfusion chamber to investigate the osmotic responses of human umbilical vein endothelial cells (HUVECs) at different temperatures, and then optimized the protocols for using cryoprotective agents (CPAs) to minimize osmotic injuries and improve processes before freezing and after thawing. The fundamental cryobiological parameters were measured using the microperfusion chamber, and then, the optimized protocols using these parameters were confirmed by survival evaluation and cell proliferation experiments. It was revealed for the first time that HUVECs have an unusually small permeability coefficient for Me2SO. Even at the concentrations well established for slow freezing of cells (1.5 M), one-step removal of CPAs for HUVECs might result in inevitable osmotic injuries, indicating that multiple-step removal is essential. Further experiments revealed that multistep removal of 1.5 M Me2SO at 25°C was the best protocol investigated, in good agreement with theory. These results should prove invaluable for optimization of cryopreservation protocols of HUVECs.

The effect of solution nonideality on modeling transmembrane water transport and diffusion-limited intracellular ice formation during cryopreservation

NASA Astrophysics Data System (ADS)

Zhao, Gang; Takamatsu, Hiroshi; He, Xiaoming

2014-04-01

A new model was developed to predict transmembrane water transport and diffusion-limited ice formation in cells during freezing without the ideal-solution assumption that has been used in previous models. The model was applied to predict cell dehydration and intracellular ice formation (IIF) during cryopreservation of mouse oocytes and bovine carotid artery endothelial cells in aqueous sodium chloride (NaCl) solution with glycerol as the cryoprotectant or cryoprotective agent. A comparison of the predictions between the present model and the previously reported models indicated that the ideal-solution assumption results in under-prediction of the amount of intracellular ice at slow cooling rates (<50 K/min). In addition, the lower critical cooling rates for IIF that is lethal to cells predicted by the present model were much lower than those estimated with the ideal-solution assumption. This study represents the first investigation on how accounting for solution nonideality in modeling water transport across the cell membrane could affect the prediction of diffusion-limited ice formation in biological cells during freezing. Future studies are warranted to look at other assumptions alongside nonideality to further develop the model as a useful tool for optimizing the protocol of cell cryopreservation for practical applications.

Human oocyte cryopreservation.

PubMed

Tao, Tao; Zhang, Wenling; Del Valle, Alfonso

2009-06-01

This review summarized the clinical breakthroughs in the human oocyte cryopreservation field in the past 2 years and gave special emphasis on the role of vitrification method. Human oocyte cryopreservation is an attractive strategy to preserve female fertility, as it offers more opportunities to the future destination of the female gametes and also raises fewer legal and ethical questions compared with embryo cryopreservation. It became promising in recent years because of dramatic improvement in cryopreservation technologies. Human oocyte cryopreservation would not become a clinical routine until the availability of reliable cryopreservation methods and long-term follow-up results of the babies born by this technique. Oocyte cryopreservation produced very exciting results with pregnancy and implantation rates comparable to embryo cryopreservation and in some cases comparable to fresh in-vitro fertilization cycles with both modified slow-freezing and vitrification methods. A cancer patient conceived and delivered her own babies by this technology after recovery from the disease. Oocyte cryopreservation became a new focus in assisted reproductive technology. We witnessed the advanced development of human oocyte cryopreservation in the past years because of increasing demand, medically, legally and ethically, and also because of the dramatic improvement of the freezing technique. There is still a long way to go to integrate it into a routine clinical procedure to benefit more patients and encourage clinicians to follow the standard protocols.

Development and testing of a human collagen graft material.

PubMed

Quteish, D; Singh, G; Dolby, A E

1990-06-01

Human Type I collagen was extracted from placenta using pepsin and salt fractionation. The collagen was characterized by SDS-PAG electrophoresis dispersed in acidic medium, freeze-dried, and cross-linked in an 0.25% glutaraldehyde solution pH 4.5 for 2 days. After washing for 7 days and freeze drying the resultant collagen sponge was tested with regard to mechanical, physical, enzymatic degradation properties and biological responses. The modulus of elasticity was found to be 289 +/- 10 g/mm2 and the sponge was insoluble in water, buffered saline, or tissue culture medium over a period of 6 weeks with swelling occurring at less than 5% of volume. The sponge had a high fluid binding capacity, amounting to 56 +/- 5 mL tissue culture medium per gram of dry weight. Bacterial collagenase produced slow degradation of the sponge with complete disappearance by 24 h only when high concentrations (200 units enzyme per mg of the collagen sponge) were used. Cytotoxicity studies using human gingival and periodontal ligament fibroblasts revealed less than 5% apparent cytotoxicity or proliferation. Subcutaneous implantation was followed by resorption and vascularization over a period of 6-8 weeks. It was concluded that the collagen sponge prepared from human Type I collagen has potential as a graft material in oral surgical procedures.

The effect of solution nonideality on modeling transmembrane water transport and diffusion-limited intracellular ice formation during cryopreservation.

PubMed

Zhao, Gang; Takamatsu, Hiroshi; He, Xiaoming

2014-04-14

A new model was developed to predict transmembrane water transport and diffusion-limited ice formation in cells during freezing without the ideal-solution assumption that has been used in previous models. The model was applied to predict cell dehydration and intracellular ice formation (IIF) during cryopreservation of mouse oocytes and bovine carotid artery endothelial cells in aqueous sodium chloride (NaCl) solution with glycerol as the cryoprotectant or cryoprotective agent. A comparison of the predictions between the present model and the previously reported models indicated that the ideal-solution assumption results in under-prediction of the amount of intracellular ice at slow cooling rates (<50 K/min). In addition, the lower critical cooling rates for IIF that is lethal to cells predicted by the present model were much lower than those estimated with the ideal-solution assumption. This study represents the first investigation on how accounting for solution nonideality in modeling water transport across the cell membrane could affect the prediction of diffusion-limited ice formation in biological cells during freezing. Future studies are warranted to look at other assumptions alongside nonideality to further develop the model as a useful tool for optimizing the protocol of cell cryopreservation for practical applications.

SEED: a tool for disseminating systematic review data into Wikipedia.

PubMed

Schmidt, Lena; Friedel, Johannes; Adams, Clive E

2017-10-17

Wikipedia, the free-content online encyclopaedia, contains many heavily accessed pages relating to healthcare. Cochrane systematic reviews contain much high-grade evidence but dissemination into Wikipedia has been slow. New skills are needed to both translate and relocate data from Cochrane reviews to implant into Wikipedia pages. This letter introduces a programme to greatly simplify the process of disseminating the summary of findings of Cochrane reviews into Wikipedia pages.

Laser scanning confocal microscope with programmable amplitude, phase, and polarization of the illumination beam.

PubMed

Boruah, B R; Neil, M A A

2009-01-01

We describe the design and construction of a laser scanning confocal microscope with programmable beam forming optics. The amplitude, phase, and polarization of the laser beam used in the microscope can be controlled in real time with the help of a liquid crystal spatial light modulator, acting as a computer generated hologram, in conjunction with a polarizing beam splitter and two right angled prisms assembly. Two scan mirrors, comprising an on-axis fast moving scan mirror for line scanning and an off-axis slow moving scan mirror for frame scanning, configured in a way to minimize the movement of the scanned beam over the pupil plane of the microscope objective, form the XY scan unit. The confocal system, that incorporates the programmable beam forming unit and the scan unit, has been implemented to image in both reflected and fluorescence light from the specimen. Efficiency of the system to programmably generate custom defined vector beams has been demonstrated by generating a bottle structured focal volume, which in fact is the overlap of two cross polarized beams, that can simultaneously improve both the lateral and axial resolutions if used as the de-excitation beam in a stimulated emission depletion confocal microscope.

Impact of a narrative medicine programme on healthcare providers' empathy scores over time.

PubMed

Chen, Po-Jui; Huang, Chien-Da; Yeh, San-Jou

2017-07-05

The cultivation of empathy for healthcare providers is an important issue in medical education. Narrative medicine (NM) has been shown to foster empathy. To our knowledge, there has been no research that examines whether a NM programme affects multi-professional healthcare providers' empathy. Our study aims to fill this gap by investigating whether a NM programme effects multi-professional healthcare providers' empathy. A pre-post questionnaire method was used.142 participants (n = 122 females) who attended the NM programme were divided into single (n = 58) and team groups (n = 84) on the basis of inter-professional education during a period of 2 months. Perceptions of the NM programme were collected using our developed questionnaire. Empathy levels were measured using the Chinese version of Jefferson Scale of Empathy - Healthcare Providers Version (JSE-HP) - at three time points: prior to (Time 1), immediately after (T2), and 1.5 years (T3) after the programme. Participants' perceptions about the NM programme (n = 116; n = 96 females) suggested an in enhancement of empathy (90.5%). Empathy scores via the JSE-HP increased after the NM programme (T1 mean 111.05, T2 mean 116.19) and were sustainable for 1.5 years (T3 mean 116.04) for all participants (F(2297) = 3.74, p < .025). A main effect of gender on empathy scores was found (F(1298) = 5.33, p < .022). No significant effect of gender over time was found but there was a trend that showed females increasing empathy scores at T2, sustaining at T3, but males demonstrating a slow rise in empathy scores over time. NM programme as an educational tool for empathy is feasible. However, further research is needed to examine gender difference as it might be that males and females respond differently to a NM programme intervention.

Impact of conditional cash transfers on maternal and newborn health.

PubMed

Glassman, Amanda; Duran, Denizhan; Fleisher, Lisa; Singer, Daniel; Sturke, Rachel; Angeles, Gustavo; Charles, Jodi; Emrey, Bob; Gleason, Joanne; Mwebsa, Winnie; Saldana, Kelly; Yarrow, Kristina; Koblinsky, Marge

2013-12-01

Maternal and newborn health (MNH) is a high priority for global health and is included among the Millennium Development Goals (MDGs). However, the slow decline in maternal and newborn mortality jeopardizes achievements of the targets of MDGs. According to UNICEF, 60 million women give birth outside of health facilities, and family planning needs are satisfied for only 50%. Further, skilled birth attendance and the use of antenatal care are most inequitably distributed in maternal and newborn health interventions in low- and middle-income countries. Conditional cash transfer (CCT) programmes have been shown to increase health service utilization among the poorest but little is written on the effects of such programmes on maternal and newborn health. We carried out a systematic review of studies on CCT that report maternal and newborn health outcomes, including studies from 8 countries. The CCT programmes have increased antenatal visits, skilled attendance at birth, delivery at a health facility, and tetanus toxoid vaccination for mothers and reduced the incidence of low birthweight. The programmes have not had a significant impact on fertility while the impact on maternal and newborn mortality has not been well-documented thus far. Given these positive effects, we make the case for further investment in CCT programmes for maternal and newborn health, noting gaps in knowledge and providing recommendations for better design and evaluation of such programmes. We recommend more rigorous impact evaluations that document impact pathways and take factors, such as cost-effectiveness, into account.

Ferroelectric hydration shells around proteins: electrostatics of the protein-water interface.

PubMed

LeBard, David N; Matyushov, Dmitry V

2010-07-22

Numerical simulations of hydrated proteins show that protein hydration shells are polarized into a ferroelectric layer with large values of the average dipole moment magnitude and the dipole moment variance. The emergence of the new polarized mesophase dramatically alters the statistics of electrostatic fluctuations at the protein-water interface. The linear response relation between the average electrostatic potential and its variance breaks down, with the breadth of the electrostatic fluctuations far exceeding the expectations of the linear response theories. The dynamics of these non-Gaussian electrostatic fluctuations are dominated by a slow (approximately = 1 ns) component that freezes in at the temperature of the dynamical transition of proteins. The ferroelectric shell propagates 3-5 water diameters into the bulk.

Live birth of twins after IVF of oocytes that were cryopreserved almost 12 years before.

PubMed

Quintans, Carlos J; Donaldson, Monica J; Urquiza, M Fernanda; Carretero, Inés; Pasqualini, R Agustín; Horton, Marcos; Pasqualini, R Sergio

2012-12-01

A 45-year-old woman received embryos from IVF by intracytoplasmic sperm injection (ICSI) with her own oocytes that were cryopreserved (slow freezing in a low-sodium medium) 11 years and 7 and a half months before, when she was 33 years old. From seven metaphase-II oocytes thawed, five survived, four were fertilized after ICSI and two cleaving embryos were transferred on day 3. A diamniotic dichorionic term pregnancy was achieved, ending with the delivery of two healthy girls. As far as is known, this case represents, to date, the longest storage period of cryopreserved human oocytes resulting in a live birth. Copyright © 2012 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Can thermostable vaccines help address cold-chain challenges? Results from stakeholder interviews in six low- and middle-income countries.

PubMed

Kristensen, Debra D; Lorenson, Tina; Bartholomew, Kate; Villadiego, Shirley

2016-02-10

This study captures the perspectives of stakeholders at multiple levels of the vaccine supply chain regarding their assessment of challenges with storing vaccines within recommended temperature ranges and their perceptions on the benefits of having vaccines with improved stability, including the potential short-term storage and transport of vaccines in a controlled-temperature chain. Semi-structured interviews were undertaken with 158 immunization stakeholders in six countries. Interviewees included national decision-makers and advisors involved in vaccine purchasing decisions, national Expanded Programme on Immunization managers, and health and logistics personnel at national, subnational, and health facility levels. Challenges with both heat and freeze-exposure of vaccines were recognized in all countries, with heat-exposure being a greater concern. Conditions leading to freeze-exposure including ice build-up due to poor refrigerator performance and improper icepack conditioning were reported by 53% and 28% of participants, respectively. Respondents were interested in vaccine products with improved heat/freeze-stability characteristics. The majority of those involved in vaccine purchasing indicated they would be willing to pay a US$0.05 premium per dose for a freeze-stable pentavalent vaccine (68%) or a heat-stable rotavirus vaccine (59%), although most (53%) preferred not to pay the premium for a heat-stable pentavalent vaccine if the increased stability required changing from a liquid to a lyophilized product. Most respondents (73%) were also interested in vaccines labeled for short-term use in a controlled-temperature chain. The majority (115/158) recognized the flexibility this would provide during outreach or should cold-chain breaks occur. Respondents were also aware that possible confusion might arise and additional training would be required if handling conditions were changed for some, but not all vaccines. Participating immunization stakeholders recognized the benefits of vaccine products with improved stability characteristics and of labeling vaccines for controlled-temperature chain use as a means to help address cold-chain issues in their immunization programs. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Observations and modeling of northern mid-latitude recurring slope lineae (RSL) suggest recharge by a present-day martian briny aquifer

NASA Astrophysics Data System (ADS)

Stillman, David E.; Michaels, Timothy I.; Grimm, Robert E.; Hanley, Jennifer

2016-02-01

Recurring slope lineae (RSL) are narrow (0.5-5 m) dark features on Mars that incrementally lengthen down steep slopes, fade in colder seasons, and recur annually. These features have been identified from the northern to southern mid-latitudes. Here, we describe how observations of northern mid-latitude RSL in northern Chryse Planitia and southwestern Acidalia Planitia (CAP) suggest that brines start flowing before northern spring equinox and continue for more than half a Mars-year (490 ± 40 sols, spanning solar longitude 337° ± 11°-224° ± 20°). All CAP RSL are found on the steep slopes of craters and their source zones are at or below the elevation of the surrounding plains. Spacecraft-derived surface temperature observations cannot resolve individual RSL, so thermal modeling was used to determine that CAP RSL have a freezing temperature of 238-252 K, freeze and melt diurnally, and flow only occurs within the top ∼8 cm of the regolith. Furthermore, we calculate that a typical CAP RSL has a water budget of 1.5-5.6 m3/m of headwall. Therefore, such a large water budget makes annual recharge via atmospheric or subsurface diffusion sources unlikely. Alternatively, we hypothesize that the most plausible RSL source is a briny aquifer with a freezing temperature less than or equal to the mean annual CAP surface temperature (220-225 K). The annual cycle is as follows: in late autumn, the shallowest part of the brine feeding the source zone freezes, forming an ice dam. As spring approaches, temperatures rise and the dam is breached. Brine is discharged and the RSL initially lengthens rapidly (>1.86 m/sol), the lengthening rate then slows considerably, to ∼0.25 m/sol. Eventually, the losses equal the discharge rate and the RSL reaches its equilibrium phase. As brine flows in the RSL some of the water is lost to the atmosphere, therefore the freezing temperature of the brine within the RSL is higher (238-252 K) as the brine transitions to a super-eutectic salt concentration. In the late autumn, falling temperatures restore the ice dam and the H2O in the RSL slowly sublimates away. Overall, CAP RSL possess a significantly different seasonality and much longer duration than typical southern mid-latitude RSL, suggesting that RSL at different latitude bands have different source types. Lastly, CAP RSL are the best evidence that shallow groundwater may still exist on Mars.

Slow dynamics in the geometrically frustrated magnet ZnFe2O4: Universal features of aging phenomena in spin glasses

NASA Astrophysics Data System (ADS)

Mamiya, H.; Tsujii, N.; Terada, N.; Nimori, S.; Kitazawa, H.; Hoshikawa, A.; Ishigaki, T.

2014-07-01

To clarify the universal features of spin glasses, we carefully studied slow dynamics in a geometrically frustrated magnet ZnFe2O4 with slight disorders, regarded as an "unconventional" Heisenberg spin glass, using time-resolved neutron diffractometry and magnetometry. The results indicate that "aging" can be attributed not to growth of the short-range order detected by a diffuse scattering but to aging of a hidden aperiodic correlation, as expected from theories for spin glasses. Concerning aging, peculiar behavior was found; the decay of thermoremanent magnetization is extremely accelerated if the sample is heated/cooled briefly midway through the isothermal slow relaxation. Conversely, magnetization surprisingly increases despite the absence of a magnetic field when the temperature returns after the brief heating/cooling. The behavior can be explained as a destabilization of the aged spin configuration due to the thermal perturbations and subsequent spontaneous restoration of the original spin configuration after the destabilization. Whereas such destabilization and restoration do not occur during freezing into numerous metastable states in a fixed energy landscape, these are possible in an energy landscape with a temperature-sensitive funnel-like structure. These features, consistent with the ghost domain scenario of the droplet picture, are the same as for conventional Heisenberg spin glasses such as dilute magnetic alloys and dilute magnetic semiconductors. In other words, they are universal features in Heisenberg spin glasses including unconventional ones.

Effect of rapid addition and dilution of dimethyl sulfoxide and 37 degrees C equilibration on viability of rabbit morulae thawed rapidly.

PubMed

Kojima, T; Soma, T; Oguri, N

1987-06-01

The aim of the present study was to examine the effects of various conditions of addition and dilution of dimethyl sulfoxide (Me2SO) and 37 degrees C equilibration, and also the effects of freezing in the solution which was prepared in advance and stored in plastic straws at -20 degrees C on the viability of rabbit morulae thawed rapidly. The embryos were cooled from room temperature to -30 degrees C at 1 degree C/min in the presence of 1.5 M Me2SO using a programmable liquid nitrogen vapor freezing machine with an automatic seeding device, then cooled rapidly, and stored in liquid nitrogen. The frozen straws were thawed rapidly (greater than 1000 degrees C/min). When Me2SO was added in a single step, equilibrated with embryos at 37 degrees C for 15 min and diluted out in a single step, a very high survival was obtained: transferable/recovered, 90%: developed/recovered, 96%. When embryos were pipetted into 1.5 M Me2SO that was prepared in advance, stocked in straws at -20 degrees C, and cooled, the proportions of transferable and developed embryos were equivalent to those of embryos frozen in the solution that was prepared immediately before use.

Development of a method for long-term preservation of Bombyx mori silkworm strains using frozen ovaries.

PubMed

Banno, Yutaka; Nagasaki, Kiyomi; Tsukada, Marino; Minohara, Yuko; Banno, Junko; Nishikawa, Kazuhiro; Yamamoto, Kazunori; Tamura, Kei; Fujii, Tsuguru

2013-06-01

Development of long-term preservation is essential for conservation of stocks of silkworm genetic resources. Thus far, a few methods have been reported, but more improvement is required for practical use. We have developed two effective modifications of a method for long-term preservation using frozen ovaries. One was slow cooling (1 °C per min) until -80 °C of the donor ovaries made possible by use of a BICELL freezing vessel. Using donor ovaries of 4th instar larvae, the average number of eggs laid per moth increased significantly from 110.7 ± 53.4 eggs per moth by slow cooling with the BICELL vessel vs 12.3 ± 10.3 eggs per moth by direct cooling in liquid nitrogen. A second improvement was connecting the thread bodies of the donor ovaries with those of the host in the transplantation step. Females operated on with the new method yielded a significantly higher percentage of moths that laid fertilized eggs than those transplanted with the standard procedure (70.4 ± 21.6% vs 22.9 ± 9.3%). Copyright © 2013 Elsevier Inc. All rights reserved.

NASA Tech Briefs, January 2004

NASA Technical Reports Server (NTRS)

2004-01-01

Topics covered include: Multisensor Instrument for Real-Time Biological Monitoring; Sensor for Monitoring Nanodevice-Fabrication Plasmas; Backed Bending Actuator; Compact Optoelectronic Compass; Micro Sun Sensor for Spacecraft; Passive IFF: Autonomous Nonintrusive Rapid Identification of Friendly Assets; Finned-Ladder Slow-Wave Circuit for a TWT; Directional Radio-Frequency Identification Tag Reader; Integrated Solar-Energy-Harvesting and -Storage Device; Event-Driven Random-Access-Windowing CCD Imaging System; Stroboscope Controller for Imaging Helicopter Rotors; Software for Checking State-charts; Program Predicts Broadband Noise from a Turbofan Engine; Protocol for a Delay-Tolerant Data-Communication Network; Software Implements a Space-Mission File-Transfer Protocol; Making Carbon-Nanotube Arrays Using Block Copolymers: Part 2; Modular Rake of Pitot Probes; Preloading To Accelerate Slow-Crack-Growth Testing; Miniature Blimps for Surveillance and Collection of Samples; Hybrid Automotive Engine Using Ethanol-Burning Miller Cycle; Fabricating Blazed Diffraction Gratings by X-Ray Lithography; Freeze-Tolerant Condensers; The StarLight Space Interferometer; Champagne Heat Pump; Controllable Sonar Lenses and Prisms Based on ERFs; Measuring Gravitation Using Polarization Spectroscopy; Serial-Turbo-Trellis-Coded Modulation with Rate-1 Inner Code; Enhanced Software for Scheduling Space-Shuttle Processing; Bayesian-Augmented Identification of Stars in a Narrow View; Spacecraft Orbits for Earth/Mars-Lander Radio Relay; and Self-Inflatable/Self-Rigidizable Reflectarray Antenna.

Programmable logic controller performance enhancement by field programmable gate array based design.

PubMed

Patel, Dhruv; Bhatt, Jignesh; Trivedi, Sanjay

2015-01-01

PLC, the core element of modern automation systems, due to serial execution, exhibits limitations like slow speed and poor scan time. Improved PLC design using FPGA has been proposed based on parallel execution mechanism for enhancement of performance and flexibility. Modelsim as simulation platform and VHDL used to translate, integrate and implement the logic circuit in FPGA. Xilinx's Spartan kit for implementation-testing and VB has been used for GUI development. Salient merits of the design include cost-effectiveness, miniaturization, user-friendliness, simplicity, along with lower power consumption, smaller scan time and higher speed. Various functionalities and applications like typical PLC and industrial alarm annunciator have been developed and successfully tested. Results of simulation, design and implementation have been reported. Copyright © 2014 ISA. Published by Elsevier Ltd. All rights reserved.

D0 Superconducting Solenoid Quench Data and Slow Dump Data Acquisition

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

Markley, D.; /Fermilab

1998-06-09

This Dzero Engineering note describes the method for which the 2 Tesla Superconducting Solenoid Fast Dump and Slow Dump data are accumulated, tracked and stored. The 2 Tesla Solenoid has eleven data points that need to be tracked and then stored when a fast dump or a slow dump occur. The TI555(Texas Instruments) PLC(Programmable Logic Controller) which controls the DC power circuit that powers the Solenoid, also has access to all the voltage taps and other equipment in the circuit. The TI555 constantly logs these eleven points in a rotating memory buffer. When either a fast dump(dump switch opens) ormore » a slow dump (power supply turns off) occurs, the TI555 organizes the respective data and will down load the data to a file on DO-CCRS2. This data in this file is moved over ethernet and is stored in a CSV (comma separated format) file which can easily be examined by Microsoft Excel or any other spreadsheet. The 2 Tesla solenoid control system also locks in first fault information. The TI555 decodes the first fault and passes it along to the program collecting the data and storing it on DO-CCRS2. This first fault information is then part of the file.« less

Analytical solution and numerical simulation of the liquid nitrogen freezing-temperature field of a single pipe

NASA Astrophysics Data System (ADS)

Cai, Haibing; Xu, Liuxun; Yang, Yugui; Li, Longqi

2018-05-01

Artificial liquid nitrogen freezing technology is widely used in urban underground engineering due to its technical advantages, such as simple freezing system, high freezing speed, low freezing temperature, high strength of frozen soil, and absence of pollution. However, technical difficulties such as undefined range of liquid nitrogen freezing and thickness of frozen wall gradually emerge during the application process. Thus, the analytical solution of the freezing-temperature field of a single pipe is established considering the freezing temperature of soil and the constant temperature of freezing pipe wall. This solution is then applied in a liquid nitrogen freezing project. Calculation results show that the radius of freezing front of liquid nitrogen is proportional to the square root of freezing time. The radius of the freezing front also decreases with decreased the freezing temperature, and the temperature gradient of soil decreases with increased distance from the freezing pipe. The radius of cooling zone in the unfrozen area is approximately four times the radius of the freezing front. Meanwhile, the numerical simulation of the liquid nitrogen freezing-temperature field of a single pipe is conducted using the Abaqus finite-element program. Results show that the numerical simulation of soil temperature distribution law well agrees with the analytical solution, further verifies the reliability of the established analytical solution of the liquid nitrogen freezing-temperature field of a single pipe.

Slow and fast solar wind - data selection and statistical analysis

NASA Astrophysics Data System (ADS)

Wawrzaszek, Anna; Macek, Wiesław M.; Bruno, Roberto; Echim, Marius

2014-05-01

In this work we consider the important problem of selection of slow and fast solar wind data measured in-situ by the Ulysses spacecraft during two solar minima (1995-1997, 2007-2008) and solar maximum (1999-2001). To recognise different types of solar wind we use a set of following parameters: radial velocity, proton density, proton temperature, the distribution of charge states of oxygen ions, and compressibility of magnetic field. We present how this idea of the data selection works on Ulysses data. In the next step we consider the chosen intervals for fast and slow solar wind and perform statistical analysis of the fluctuating magnetic field components. In particular, we check the possibility of identification of inertial range by considering the scale dependence of the third and fourth orders scaling exponents of structure function. We try to verify the size of inertial range depending on the heliographic latitudes, heliocentric distance and phase of the solar cycle. Research supported by the European Community's Seventh Framework Programme (FP7/2007 - 2013) under grant agreement no 313038/STORM.

[Update rehabilitation therapy for Parkinson disease].

PubMed

Hayashi, Akito

2013-01-01

Rehabilitation is essential for treatment of Parkinson's disease. New rehabilitation therapy is updated, in addition to evidence shown with "Parkinson's disease treatment guidelines 2011". Furthermore, a portable gait rhythmogram (acceleration sensor) is presented (not publication). Parkinsonian gait was significantly slow and the steps were small, but the cadence was not different compared as that of normal control. The strength of parkinsonian gait was apparently week compared as normal control. We also could examine consecutive changes of gait rhythm and detect freezing gait in patients. In this study, we could extract the characteristic of the parkinsonian gait and evaluate especially freeing events more objectively. This method may bring us to evaluate severity of parkinsonian gait not only in a consulting room but also daily profile even not to see directly, using the portable gait rhythmogram.

Pressure ramp programmer; IMBLMS Phase B4 Additional Tasks: Task 3.0 pressure ramp programmer

NASA Technical Reports Server (NTRS)

Fogal, G. L.; Reinhardt, C. G.

1972-01-01

A pressure ramp programmer model was designed, fabricated and tested. This model, in conjunction with an automatic blood pressure monitor, automatically controls the pressure in the blood pressure monitor arterial cuff. The cuff pressurization cycle is designed to maximize accuracy and repeatability of blood pressure measurements. The key feature of this automatic cycle is rapid blood pressure cuff bleed down from an initial setting until systolic (diastolic) pressure is encountered followed by a short repressurization and slow bleed, long enough to permit accurate systolic (diastolic) pressure determination. The system includes a pressure reservoir which bleeds the cuff through a precision needle valve; a solenoid valve which permits rapid pressurization from the reservoir; and a pressure sensor which provides information for bleed rate and set point controls. Korotkoff sound signals from a microphone in the blood pressure cuff (not part of the system) provide decision information to the digital control system. The system completed a series of engineering tests using simulated Korotkoff sound inputs. The system performed successfully in all cases and was stable over an extended period of time.

Programmable snapping composites with bio-inspired architecture.

PubMed

Schmied, Jascha U; Le Ferrand, Hortense; Ermanni, Paolo; Studart, André R; Arrieta, Andres F

2017-03-13

The development of programmable self-shaping materials enables the onset of new and innovative functionalities in many application fields. Commonly, shape adaptation is achieved by exploiting diffusion-driven swelling or nano-scale phase transition, limiting the change of shape to slow motion predominantly determined by the environmental conditions and/or the materials specificity. To address these shortcomings, we report shape adaptable programmable shells that undergo morphing via a snap-through mechanism inspired by the Dionaea muscipula leaf, known as the Venus fly trap. The presented shells are composite materials made of epoxy reinforced by stiff anisotropic alumina micro-platelets oriented in specific directions. By tailoring the microstructure via magnetically-driven alignment of the platelets, we locally tune the pre-strain and stiffness anisotropy of the composite. This novel approach enables the fabrication of complex shapes showing non-orthotropic curvatures and stiffness gradients, radically extending the design space when compared to conventional long-fibre reinforced multi-stable composites. The rare combination of large stresses, short actuation times and complex shapes, results in hinge-free artificial shape adaptable systems with large design freedom for a variety of morphing applications.

Spin-glass-like freezing of inner and outer surface layers in hollow γ-Fe 2O 3 nanoparticles

DOE PAGES

Khurshid, Hafsa; Lampen-Kelley, Paula; Iglesias, Òscar; ...

2015-10-27

Disorder among surface spins largely dominates the magnetic response of ultrafine magnetic particle systems. In this work, we examine time-dependent magnetization in high-quality, monodisperse hollow maghemite nanoparticles (NPs) with a 14.8±0.5 nm outer diameter and enhanced surface-to-volume ratio. The nanoparticle ensemble exhibits spin-glass-like signatures in dc magnetic aging and memory protocols and ac magnetic susceptibility. The dynamics of the system slow near 50 K, and becomes frozen on experimental time scales below 20 K. Remanence curves indicate the development of magnetic irreversibility concurrent with the freezing of the spin dynamics. A strong exchange-bias effect and its training behavior point tomore » highly frustrated surface spins that rearrange much more slowly than interior spins with bulk coordination. Monte Carlo simulations of a hollow particle reproducing the experimental morphology corroborate strongly disordered surface layers with complex energy landscapes that underlie both glass-like dynamics and magnetic irreversibility. Calculated hysteresis loops reveal that magnetic behavior is not identical at the inner and outer surfaces, with spins at the outer surface layer of the 15 nm hollow particles exhibiting a higher degree of frustration. Lastly, our combined experimental and simulated results shed light on the origin of spin-glass-like phenomena and the important role played by the surface spins in magnetic hollow nanostructures.« less

Spin-glass-like freezing of inner and outer surface layers in hollow γ-Fe2O3 nanoparticles

PubMed Central

Khurshid, Hafsa; Lampen-Kelley, Paula; Iglesias, Òscar; Alonso, Javier; Phan, Manh-Huong; Sun, Cheng-Jun; Saboungi, Marie-Louise; Srikanth, Hariharan

2015-01-01

Disorder among surface spins is a dominant factor in the magnetic response of magnetic nanoparticle systems. In this work, we examine time-dependent magnetization in high-quality, monodisperse hollow maghemite nanoparticles (NPs) with a 14.8 ± 0.5 nm outer diameter and enhanced surface-to-volume ratio. The nanoparticle ensemble exhibits spin-glass-like signatures in dc magnetic aging and memory protocols and ac magnetic susceptibility. The dynamics of the system slow near 50 K, and become frozen on experimental time scales below 20 K. Remanence curves indicate the development of magnetic irreversibility concurrent with the freezing of the spin dynamics. A strong exchange-bias effect and its training behavior point to highly frustrated surface spins that rearrange much more slowly than interior spins. Monte Carlo simulations of a hollow particle corroborate strongly disordered surface layers with complex energy landscapes that underlie both glass-like dynamics and magnetic irreversibility. Calculated hysteresis loops reveal that magnetic behavior is not identical at the inner and outer surfaces, with spins at the outer surface layer of the 15 nm hollow particles exhibiting a higher degree of frustration. Our combined experimental and simulated results shed light on the origin of spin-glass-like phenomena and the important role played by the surface spins in magnetic hollow nanostructures. PMID:26503506

Freezing of Dynamics of a Methyl Group in a Protein Hydrophobic Core at Cryogenic Temperatures by Deuteron NMR Spectroscopy

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

Vugmeyster, Liliya; Ostrovsky, Dmitry; Ford, Joseph J.

2010-03-31

Proteins undergo a number of changes when their temperature is dropped from the physiological range to much lower values. One of the most well-known dynamical changes undergone by proteins in a solid state is a so-called protein glass-transition, which is a dynamic transition occurring at about 200-230K leading to a loss of biological activity.1,2 X-ray diffraction, neutron scattering studies, and dielectric spectroscopy, as well as evidence from NMR relaxation measurements, indicate freezing of slow collective modes of motion below the glass transition temperature.3-8 Various arguments have been presented that connect the transition to solvent participation.1,4,8-10 In addition to the solvent-relatedmore » modes that are frozen below the glass-transition temperature, there are anharmonic motions at temperatures below 200K which are likely to be dominated by methyl group dynamics down to about 100K.2,5,7 Recent neutron-scattering and NMR studies emphasize the role of these modes in low temperature dynamics. 2,5,7,11,12 One of the latest works on the subject by Bajaj et al.11 has reported a structural transition associated with dynamic processes in a solvent-free polypeptide. Thus, protein dynamics at low temperatures are complex and more studies are required to discern their pattern.« less

Vitrification and xenografting of human ovarian tissue.

PubMed

Amorim, Christiani Andrade; Dolmans, Marie-Madeleine; David, Anu; Jaeger, Jonathan; Vanacker, Julie; Camboni, Alessandra; Donnez, Jacques; Van Langendonckt, Anne

2012-11-01

To assess the efficiency of two vitrification protocols to cryopreserve human preantral follicles with the use of a xenografting model. Pilot study. Gynecology research unit in a university hospital. Ovarian biopsies were obtained from seven women aged 30-41 years. Ovarian tissue fragments were subjected to one of three cryopreservation protocols (slow freezing, vitrification protocol 1, and vitrification protocol 2) and xenografted for 1 week to nude mice. The number of morphologically normal follicles after cryopreservation and grafting and fibrotic surface area were determined by histologic analysis. Apoptosis was assessed by the TUNEL method. Morphometric analysis of TUNEL-positive surface area also was performed. Follicle proliferation was evaluated by immunohistochemistry. After xenografting, a difference was observed between the cryopreservation procedures applied. According to TUNEL analysis, both vitrification protocols showed better preservation of preantral follicles than the conventional freezing method. Moreover, histologic evaluation showed a significantly higher proportion of primordial follicles in vitrified (protocol 2)-warmed ovarian tissue than in frozen-thawed tissue. The proportion of growing follicles and fibrotic surface area was similar in all groups. Vitrification procedures appeared to preserve not only the morphology and survival of preantral follicles after 1 week of xenografting, but also their ability to resume folliculogenesis. In addition, vitrification protocol 2 had a positive impact on the quiescent state of primordial follicles after xenografting. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Valorization of postharvest sweet cherry discard for the development of dehydrated fruit ingredients: compositional, physical, and mechanical properties.

PubMed

Franceschinis, Lorena; Sette, Paula; Salvatori, Daniela; Schebor, Carolina

2018-04-20

Sweet cherries are an excellent source of phenolic compounds, which may contribute to a healthy diet. The objective of this work was to generate dehydrated ingredients from postharvest discard of sweet cherries. Four dried ingredients were obtained from fresh sweet cherry discard (Lapins var.) using an osmotic dehydration pretreatment and freeze drying or air drying. The ingredients showed an important phenolic contribution (2.8-6.6 g gallic acid kg -1 of product) and preserved the natural color of the fruit to a great extent. Freeze-dried ingredients were less hygroscopic than air-dried ones, and presented with a softer texture. All the ingredients were in a supercooled state at room temperature (T g range: -23.0 to -18.8 °C). Sugar infusion pretreatment caused a decrease in water sorption capacity and molecular mobility; it also reduced the initial rehydration rate. Relevant differences in nutritional and structural characteristics of the ingredients were observed depending on the processing method used. These ingredients could be incorporated into different processed foods, such as snacks, cereal mixtures, cereal bars, and bakery and confectionery products. Air-dried control ingredients presented better nutritional qualities and air-dried sweet cherries with sugar infusion pretreatment could be appropriate ingredients for applications where sweet flavor and slow rehydration rate are required. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

Development of gelatin/carboxymethyl chitosan/nano-hydroxyapatite composite 3D macroporous scaffold for bone tissue engineering applications.

PubMed

Maji, Somnath; Agarwal, Tarun; Das, Joyjyoti; Maiti, Tapas Kumar

2018-06-01

The present study delineates a relatively simpler approach for fabrication of a macroporous three-dimensional scaffold for bone tissue engineering. The novelty of the work is to obtain a scaffold with macroporosity (interconnected networks) through a combined approach of high stirring induced foaming of the gelatin/carboxymethyl chitosan (CMC)/nano-hydroxyapatite (nHAp) matrix followed by freeze drying. The fabricated macroporous (SGC) scaffold had a greater pore size, higher porosity, higher water retention capacity, slow and sustained enzymatic degradation rate along with higher compressive strength compared to that of non-macroporous (NGC, prepared by conventional freeze drying methodology) scaffold. The biological studies revealed the increased percentage of viability, proliferation, and differentiation as well as higher mineralization of differentiated human Wharton's jelly MSC microtissue (wjhMSC-MT) on SGC as compared to NGC scaffold. RT-PCR also showed enhanced expression level of collagen type I, osteocalcin and Runx2 when seeded on SGC. μCT and histological analysis further revealed a penetration of cellular spheroid to a greater depth in SGC scaffold than NGC scaffold. Furthermore, the effect of cryopreservation on microtissue survival on the three-dimensional construct revealed significant higher viability upon revival in macroporous SGC scaffolds. These results together suggest that high stirring based macroporous scaffolds could have a potential application in bone tissue engineering. Copyright © 2018 Elsevier Ltd. All rights reserved.

Molecular dynamics studies of the size and temperature dependence of the kinetics of freezing of Fe nanoparticles

NASA Astrophysics Data System (ADS)

Zhao, Bo; Huang, Jinfan; Bartell, Lawrence S.

2013-11-01

Molecular dynamics (MD) computer simulations have been carried out and a novel modified technique of Voronoi polyhedra has been performed to identify solid-like particles in a molten nanoparticle. This technique works quite well in analyzing the effects of particle size on nucleation rates of iron nanoparticles in the temperature range of 750-1160 K. Nanoparticles with 1436 and 2133 Fe atoms have been examined and the results are compared with those obtained earlier with Fe331 nanoparticles. Nucleation rates for freezing obtained from MD simulations for Fe2133 vary from 8.8×1034 m3/s to 4.1×1035 m3/s at over a temperature range from 1160 K to 900 K, Rates for. Fe1436 and Fe331 are somewhat higher. Nucleation rates increase as supercooling deepens until the viscosity of the liquid increases sharply enough to slow down the rate. Bt applying classical nucleation theory, the interfacial free energy between solid and liquid cab be estimated From this and other thermodynamic information can be derived a theoretical expression for the size-dependence of the heat of fusion of nanoparticles. Results agreed quite well with those observed in our MD observations. An earlier expression in the literature for this size-dependence was shown to be incorrect. The size dependence of melting point is discussed.

Freezing induces a loss of freeze tolerance in an overwintering insect.

PubMed

Brown, C L; Bale, J S; Walters, K F A

2004-07-22

Cold-hardy insects overwinter by one of two main strategies: freeze tolerance and freeze avoidance by supercooling. As a general model, many freeze-tolerant species overwinter in extreme climates, freeze above -10 degrees C via induction by ice-nucleating agents, and once frozen, can survive at temperatures of up to 40 degrees C or more below the initial freezing temperature or supercooling point (SCP). It has been assumed that the SCP of freeze-tolerant insects is unaffected by the freezing process and that the freeze-tolerant state is therefore retained in winter though successive freeze-thaw cycles of the body tissues and fluids. Studies on the freeze-tolerant larva of the hoverfly Syrphus ribesii reveal this assumption to be untrue. When a sample with a mean 'first freeze' SCP of -7.6 degrees C (range of -5 degrees C to -9.5 degrees C) were cooled, either to -10 degrees C or to their individual SCP, on five occasions, the mean SCP was significantly depressed, with some larvae subsequently freezing as low as -28 degrees C. Only larvae that froze at the same consistently high temperature above -10 degrees C were alive after being frozen five times. The wider occurrence of this phenomenon would require a fundamental reassessment of the dynamics and distinctions of the freeze-tolerant and freeze-avoiding strategies of insect overwintering. Copyright 2004 The Royal Society

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.

Critical interactions between Global Fund-supported programmes and health systems: a case study in Papua New Guinea.

PubMed

Rudge, James W; Phuanakoonon, Suparat; Nema, K Henry; Mounier-Jack, Sandra; Coker, Richard

2010-11-01

In Papua New Guinea, investment by the Global Fund to Fight AIDS, Tuberculosis and Malaria (the Global Fund) has played an important role in scaling up the response to HIV and tuberculosis (TB). As part of a series of case studies on how Global Fund-supported programmes interact with national health systems, we assessed the nature and extent of integration of the Global Fund portfolios within the national HIV and TB programmes, the integration of the HIV and TB programmes within the general health system, and system-wide effects of Global Fund support in Papua New Guinea. The study relied on a literature review and 30 interviews with key stakeholders using the Systemic Rapid Assessment Toolkit and thematic analysis. Global Fund-supported activities were found to be largely integrated, or at least coordinated, with the national HIV and TB programmes. However, this has reinforced the vertical nature of these programmes with respect to the general health system, with parallel systems established to meet the demands of programme scale-up and the performance-based nature of Global Fund investment in the weak health system context of Papua New Guinea. The more parallel functions include monitoring and evaluation, and procurement and supply chain systems, while human resources and infrastructure for service delivery are increasingly integrated at more local levels. Positive synergies of Global Fund support include engagement of civil-society partners, and a reliable supply of high-quality drugs which may have increased patient confidence in the health system. However, the severely limited and overburdened pool of human resources has been skewed towards the three diseases, both at management and service delivery levels. There is also concern surrounding the sustainability of the disease programmes, given their dependence on donors. Increasing Global Fund attention towards health system strengthening was viewed positively, but should acknowledge that system changes are slow, difficult to measure and require long-term support.

The ability of the Antarctic nematode Panagrolaimus davidi to survive intracellular freezing is dependent upon nutritional status.

PubMed

Raymond, Mélianie R; Wharton, David A

2013-02-01

The Antarctic nematode Panagrolaimus davidi is the best documented example of an animal surviving intracellular freezing and the only animal so far shown to survive such freezing throughout its tissues. However, a recent study found that after exposure to a freezing stress that produced intracellular freezing in a proportion of nematodes, the resulting survival levels could be explained if those nematodes that froze intracellularly had died. We have thus re-examined the survival of intracellular freezing in this nematode. The ability to survive a freezing exposure that is likely to produce intracellular freezing (freezing at -10 °C) declines with culture age. In cultures that are fed regularly, the ability to survive freezing at -10 °C increases, but in starved cultures freezing survival declines. Survival of intracellular freezing in fed cultures was confirmed using cryomicroscopy, staining of cells with vital dyes and by freeze substitution and transmission electron microscopy. We have thus confirmed that P. davidi can survive intracellular freezing and shown that this ability is dependent upon them being well fed. The effect of culture conditions on the nutrient status of the nematodes should thus be an important factor in the design of experiments.

Emerging techniques for assisting and accelerating food freezing processes: A review of recent research progresses.

PubMed

Cheng, Lina; Sun, Da-Wen; Zhu, Zhiwei; Zhang, Zi

2017-03-04

Freezing plays an important role in food preservation and the emergence of rapid freezing technologies can be highly beneficial to the food industry. This paper reviews some novel food freezing technologies, including high-pressure freezing (HPF), ultrasound-assisted freezing (UAF), electrically disturbed freezing (EF) and magnetically disturbed freezing (MF), microwave-assisted freezing (MWF), and osmo-dehydro-freezing (ODF). HPF and UAF can initiate ice nucleation rapidly, leading to uniform distribution of ice crystals and the control of their size and shape. Specifically, the former is focused on increasing the degree of supercooling, whereas the latter aims to decrease it. Direct current electric freezing (DC-EF) and alternating current electric freezing (AC-EF) exhibit different effects on ice nucleation. DC-EF can promote ice nucleation and AC-EF has the opposite effect. Furthermore, ODF has been successfully used for freezing various vegetables and fruit. MWF cannot control the nucleation temperature, but can decrease supercooling degree, thus decreasing the size of ice crystals. The heat and mass transfer processes during ODF have been investigated experimentally and modeled mathematically. More studies should be carried out to understand the effects of these technologies on food freezing process.

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.

Complex-formation between reduced xanthine oxidase and purine substrates demonstrated by electron paramagnetic resonance

PubMed Central

Pick, Frances M.; Bray, R. C.

1969-01-01

The origin of the Rapid molybdenum electron-paramagnetic-resonance signals, which are obtained on reducing xanthine oxidase with purine or with xanthine, and whose parameters were measured by Bray & Vänngård (1969), was studied. It is concluded that these signals represent complexes of reduced enzyme with substrate molecules. Xanthine forms one complex at high concentrations and a different one at low concentrations. Purine forms a complex indistinguishable from the low-concentration xanthine complex. There are indications that some other substrates also form complexes, but uric acid, a reaction product, does not appear to do so. The possible significance of the complexes in the catalytic cycle of the enzyme is discussed and it is suggested that they represent substrate molecules bound at the reduced active site, waiting their turn to react there, when the enzyme has been reoxidized. Support for this role for the complexes was deduced from experiments in which frozen samples of enzyme–xanthine mixtures, prepared by the rapid-freezing method, were warmed until the signals began to change. Under these conditions an increase in amplitude of the Very Rapid signal took place. Data bearing on the origin of the Slow molybdenum signal are also discussed. This signal disappears only slowly in the presence of oxygen, and its appearance rate is unaffected by change in the concentration of dithionite. It is concluded that, like other signals from the enzyme, it is due to Mov but that a slow change of ligand takes place before it is seen. The Slow species, like the Rapid, seems capable of forming complexes with purines. PMID:4310056

A Low Temperature Limit for Life on Earth

PubMed Central

Clarke, Andrew; Morris, G. John; Fonseca, Fernanda; Murray, Benjamin J.; Price, Hannah C.

2013-01-01

There is no generally accepted value for the lower temperature limit for life on Earth. We present empirical evidence that free-living microbial cells cooling in the presence of external ice will undergo freeze-induced desiccation and a glass transition (vitrification) at a temperature between −10°C and −26°C. In contrast to intracellular freezing, vitrification does not result in death and cells may survive very low temperatures once vitrified. The high internal viscosity following vitrification means that diffusion of oxygen and metabolites is slowed to such an extent that cellular metabolism ceases. The temperature range for intracellular vitrification makes this a process of fundamental ecological significance for free-living microbes. It is only where extracellular ice is not present that cells can continue to metabolise below these temperatures, and water droplets in clouds provide an important example of such a habitat. In multicellular organisms the cells are isolated from ice in the environment, and the major factor dictating how they respond to low temperature is the physical state of the extracellular fluid. Where this fluid freezes, then the cells will dehydrate and vitrify in a manner analogous to free-living microbes. Where the extracellular fluid undercools then cells can continue to metabolise, albeit slowly, to temperatures below the vitrification temperature of free-living microbes. Evidence suggests that these cells do also eventually vitrify, but at lower temperatures that may be below −50°C. Since cells must return to a fluid state to resume metabolism and complete their life cycle, and ice is almost universally present in environments at sub-zero temperatures, we propose that the vitrification temperature represents a general lower thermal limit to life on Earth, though its precise value differs between unicellular (typically above −20°C) and multicellular organisms (typically below −20°C). Few multicellular organisms can, however, complete their life cycle at temperatures below ∼−2°C. PMID:23840425

A Low Temperature Limit for Life on Earth.

PubMed

Clarke, Andrew; Morris, G John; Fonseca, Fernanda; Murray, Benjamin J; Acton, Elizabeth; Price, Hannah C

2013-01-01

There is no generally accepted value for the lower temperature limit for life on Earth. We present empirical evidence that free-living microbial cells cooling in the presence of external ice will undergo freeze-induced desiccation and a glass transition (vitrification) at a temperature between -10°C and -26°C. In contrast to intracellular freezing, vitrification does not result in death and cells may survive very low temperatures once vitrified. The high internal viscosity following vitrification means that diffusion of oxygen and metabolites is slowed to such an extent that cellular metabolism ceases. The temperature range for intracellular vitrification makes this a process of fundamental ecological significance for free-living microbes. It is only where extracellular ice is not present that cells can continue to metabolise below these temperatures, and water droplets in clouds provide an important example of such a habitat. In multicellular organisms the cells are isolated from ice in the environment, and the major factor dictating how they respond to low temperature is the physical state of the extracellular fluid. Where this fluid freezes, then the cells will dehydrate and vitrify in a manner analogous to free-living microbes. Where the extracellular fluid undercools then cells can continue to metabolise, albeit slowly, to temperatures below the vitrification temperature of free-living microbes. Evidence suggests that these cells do also eventually vitrify, but at lower temperatures that may be below -50°C. Since cells must return to a fluid state to resume metabolism and complete their life cycle, and ice is almost universally present in environments at sub-zero temperatures, we propose that the vitrification temperature represents a general lower thermal limit to life on Earth, though its precise value differs between unicellular (typically above -20°C) and multicellular organisms (typically below -20°C). Few multicellular organisms can, however, complete their life cycle at temperatures below ∼-2°C.

Cognitive Factors Affecting Freeze-like Behavior in Humans.

PubMed

Alban, Michael W; Pocknell, Victoria

2017-01-01

Contemporary research on survival-related defensive behaviors has identified physiological markers of freeze/flight/fight. Our research focused on cognitive factors associated with freeze-like behavior in humans. Study 1 tested if an explicit decision to freeze is associated with the psychophysiological state of freezing. Heart rate deceleration occurred when participants chose to freeze. Study 2 varied the efficacy of freezing relative to other defense options and found "freeze" was responsive to variations in the perceived effectiveness of alternative actions. Study 3 tested if individual differences in motivational orientation affect preference for a "freeze" option when the efficacy of options is held constant. A trend in the predicted direction suggested that naturally occurring cognitions led loss-avoiders to select "freeze" more often than reward-seekers. In combination, our attention to the cognitive factors affecting freeze-like behavior in humans represents a preliminary step in addressing an important but neglected research area.

Spin dynamics in the single-ion magnet [Er(W5O18) 2 ] 9 -

NASA Astrophysics Data System (ADS)

Mariani, M.; Borsa, F.; Graf, M. J.; Sanna, S.; Filibian, M.; Orlando, T.; Sabareesh, K. P. V.; Cardona-Serra, S.; Coronado, E.; Lascialfari, A.

2018-04-01

In this work we present a detailed NMR and μ+SR investigation of the spin dynamics in the new hydrated sodium salt containing the single-ion magnet [Er(W5O18) 2 ] 9 -. The 1HNMR absorption spectra at various applied magnetic fields present a line broadening on decreasing temperature which indicates a progressive spin freezing of the single-molecule magnetic moments. The onset of quasistatic local magnetic fields, due to spin freezing, is observed also in the muon relaxation curves at low temperature. Both techniques yield a local field distribution of the order of 0.1-0.2 T, which appears to be of dipolar origin. On decreasing the temperature, a gradual loss of the 1HNMR signal intensity is observed, a phenomenon known as wipe-out effect. The effect is analyzed quantitatively on the basis of a simple model which relies on the enhancement of the NMR spin-spin, T2-1, relaxation rate due to the slowing down of the magnetic fluctuations. Measurements of spin-lattice relaxation rate T1-1 for 1HNMR and of the muon longitudinal relaxation rate λ show an increase as the temperature is lowered. However, while for the NMR case the signal is lost before reaching the very slow fluctuation region, the muon spin-lattice relaxation λ can be followed until very low temperatures and the characteristic maximum, reached when the electronic spin fluctuation frequency becomes of the order of the muon Larmor frequency, can be observed. At high temperatures, the data can be well reproduced with a simple model based on a single correlation time τ =τ0exp (Δ /T ) for the magnetic fluctuations. However, to fit the relaxation data for both NMR and μ+SR over the whole temperature and magnetic field range, one has to use a more detailed model that takes into account spin-phonon transitions among the E r3 + magnetic sublevels. A good agreement for both proton NMR and μ+SR relaxation is obtained, which confirms the validity of the energy level scheme previously calculated from an effective crystal field Hamiltonian.

A Chemically Defined Medium for Rabbit Embryo Cryopreservation

PubMed Central

Bruyère, Pierre; Baudot, Anne; Joly, Thierry; Commin, Loris; Pillet, Elodie; Guérin, Pierre; Louis, Gérard; Josson-Schramme, Anne; Buff, Samuel

2013-01-01

This study evaluates a new synthetic substitute (CRYO3, Ref. 5617, Stem Alpha, France) for animal-based products in rabbit embryo cryopreservation solutions. This evaluation was performed using two approaches: a thermodynamic approach using differential scanning calorimetry and a biological approach using rabbit embryo slow-freezing. During the experiment, foetal calf serum (FCS) was used as a reference. Because FCS varies widely by supplier, three different FCS were selected for the thermodynamic approach. The rabbit embryo slow-freezing solutions were made from Dulbecco's phosphate buffer saline containing 1.5 M Dimethyl Sulfoxide and 18% (v.v−1) of CRYO3 or 18% (v.v−1) of FCS. These solutions were evaluated using four characteristics: the end of melting temperature, the enthalpy of crystallisation (thermodynamic approach) and the embryo survival rates after culture and embryo transfer (biological approach). In the thermodynamic approach, the solutions containing one of the three different FCS had similar mean thermodynamic characteristics but had different variabilities in the overall data with aberrant values. The solution containing CRYO3 had similar thermodynamic properties when compared to those containing FCS. Moreover, no aberrant value was measured in the solution containing CRYO3. This solution appears to be more stable than the solutions containing a FCS. In the biological approach, the in vitro embryo survival rates obtained with the solution containing CRYO3 (73.7% and 81.3%) and with the solution containing a FCS (77.6% and 71.9%) were similar (p = 0.7). Nevertheless, during the in vivo evaluation, the implantation rate (21.8%) and the live-foetuses rate (18.8%) of the CRYO3 group were significantly higher than the implantation rate (7.1%, p = 0.0002) and the live-foetuses rate (5.3%, p = 0.0002) of the FCS group. The pregnancy rate was also higher in the CRYO3 group compared to the FCS group (81.3% and 43.8%, respectively, p = 0.066). We conclude that CRYO3 can be used as a chemically defined substitute for animal-based products in rabbit embryo cryopreservation solutions. PMID:23977074

The role of ice dynamics in shaping vegetation in flowing waters.

PubMed

Lind, Lovisa; Nilsson, Christer; Polvi, Lina E; Weber, Christine

2014-11-01

Ice dynamics is an important factor affecting vegetation in high-altitude and high-latitude streams and rivers. During the last few decades, knowledge about ice in streams and rivers has increased significantly and a respectable body of literature is now available. Here we review the literature on how ice dynamics influence riparian and aquatic vegetation. Traditionally, plant ecologists have focused their studies on the summer period, largely ignoring the fact that processes during winter also impact vegetation dynamics. For example, the freeze-up period in early winter may result in extensive formation of underwater ice that can restructure the channel, obstruct flow, and cause flooding and thus formation of more ice. In midwinter, slow-flowing reaches develop a surface-ice cover that accumulates snow, protecting habitats under the ice from formation of underwater ice but also reducing underwater light, thus suppressing photosynthesis. Towards the end of winter, ice breaks up and moves downstream. During this transport, ice floes can jam up and cause floods and major erosion. The magnitudes of the floods and their erosive power mainly depend on the size of the watercourse, also resulting in different degrees of disturbance to the vegetation. Vegetation responds both physically and physiologically to ice dynamics. Physical action involves the erosive force of moving ice and damage caused by ground frost, whereas physiological effects - mostly cell damage - happen as a result of plants freezing into the ice. On a community level, large magnitudes of ice dynamics seem to favour species richness, but can be detrimental for individual plants. Human impacts, such as flow regulation, channelisation, agriculturalisation and water pollution have modified ice dynamics; further changes are expected as a result of current and predicted future climate change. Human impacts and climate change can both favour and disfavour riverine vegetation dynamics. Restoration of streams and rivers may mitigate some effects of anticipated climate change on ice and vegetation dynamics by, for example, slowing down flows and increasing water depth, thus reducing the potential for massive formation of underwater ice. © 2014 The Authors. Biological Reviews © 2014 Cambridge Philosophical Society.

Response of seasonal soil freeze depth to climate change across China

NASA Astrophysics Data System (ADS)

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

2017-05-01

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

Automated Software Vulnerability Analysis

NASA Astrophysics Data System (ADS)

Sezer, Emre C.; Kil, Chongkyung; Ning, Peng

Despite decades of research, software continues to have vulnerabilities. Successful exploitations of these vulnerabilities by attackers cost millions of dollars to businesses and individuals. Unfortunately, most effective defensive measures, such as patching and intrusion prevention systems, require an intimate knowledge of the vulnerabilities. Many systems for detecting attacks have been proposed. However, the analysis of the exploited vulnerabilities is left to security experts and programmers. Both the human effortinvolved and the slow analysis process are unfavorable for timely defensive measure to be deployed. The problem is exacerbated by zero-day attacks.

[Principles of fast track surgery. Multimodal perioperative therapy programme].

PubMed

Kehlet, H

2009-08-01

Recent evidence has documented that a combination of single-modality evidence-based care principles into a multimodal effort to enhance postoperative recovery (the fast track methodology) has led to enhanced recovery with reduced medical morbidity, need for hospitalisation and convalescence. Nevertheless, general implementation of fast track surgery has been relatively slow despite concomitant economic benefits. Further improvement in postoperative outcome may be obtained by developments within each care principle with a specific focus on minimally invasive surgery, effective multimodal, non-opioid analgesia and pharmacological stress reduction.

Field programmable chemistry: integrated chemical and electronic processing of informational molecules towards electronic chemical cells.

PubMed

Wagler, Patrick F; Tangen, Uwe; Maeke, Thomas; McCaskill, John S

2012-07-01

The topic addressed is that of combining self-constructing chemical systems with electronic computation to form unconventional embedded computation systems performing complex nano-scale chemical tasks autonomously. The hybrid route to complex programmable chemistry, and ultimately to artificial cells based on novel chemistry, requires a solution of the two-way massively parallel coupling problem between digital electronics and chemical systems. We present a chemical microprocessor technology and show how it can provide a generic programmable platform for complex molecular processing tasks in Field Programmable Chemistry, including steps towards the grand challenge of constructing the first electronic chemical cells. Field programmable chemistry employs a massively parallel field of electrodes, under the control of latched voltages, which are used to modulate chemical activity. We implement such a field programmable chemistry which links to chemistry in rather generic, two-phase microfluidic channel networks that are separated into weakly coupled domains. Electric fields, produced by the high-density array of electrodes embedded in the channel floors, are used to control the transport of chemicals across the hydrodynamic barriers separating domains. In the absence of electric fields, separate microfluidic domains are essentially independent with only slow diffusional interchange of chemicals. Electronic chemical cells, based on chemical microprocessors, exploit a spatially resolved sandwich structure in which the electronic and chemical systems are locally coupled through homogeneous fine-grained actuation and sensor networks and play symmetric and complementary roles. We describe how these systems are fabricated, experimentally test their basic functionality, simulate their potential (e.g. for feed forward digital electrophoretic (FFDE) separation) and outline the application to building electronic chemical cells. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

Clinical SAS programming in India: A study of industry needs versus wants.

PubMed

Ananthakrishnan, Nithiyanandhan

2014-07-01

The clinical SAS (www.sas.com) programming industry, in India, has seen a rapid growth in the last decade and the trend seems set to continue, for the next couple of years, due to cost advantage and the availability of skilled labor. On one side the industry needs are focused on less execution time, high margins, segmented tasks and the delivery of high quality output with minimal oversight. On the other side, due to the increased demand for skilled resources, the wants of the programmers have taken a different shift toward diversifying exposure, unsustainable wage inflation due to multiple opportunities and generally high expectations around career progression. If the industry needs are not going to match with programmers want, or vice versa, then there is the possibility that the current year on year growth may start to slow or even go into decline. This paper is intended to identify the gap between wants and need and puts forwards some suggestions, for both sides, in ways to change the equation to benefit all. Questionnaire on similar themes created to survey managers and programmers working in clinical SAS programming industry and was surveyed online to collect their perspectives. Their views are compared for each theme and presented as results. Two surveys were created in www.surveymonkey.com. https://www.surveymonkey.com/s/SAS_India_managment_needvswant_survey. Programmer: https://www.surveymonkey.com/s/SAS_India_programmer_needvswant_survey. Bar chart and pie chart used on data collect to show segmentation of data. In conclusion, it seeks to highlight the future industry direction and the skillset that existing programmers need to have, in order to sustain the momentum and remain competitive, to contribute to the future pipeline and the development of the profession in India.

Mechanisms of deterioration of nutrients. [of freeze dried foods

NASA Technical Reports Server (NTRS)

Karel, M.; Flink, J. M.

1976-01-01

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

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

PubMed

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

2015-04-01

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

Hedgehog regulation of superficial slow muscle fibres in Xenopus and the evolution of tetrapod trunk myogenesis.

PubMed

Grimaldi, Annalisa; Tettamanti, Gianluca; Martin, Benjamin L; Gaffield, William; Pownall, Mary E; Hughes, Simon M

2004-07-01

In tetrapod phylogeny, the dramatic modifications of the trunk have received less attention than the more obvious evolution of limbs. In somites, several waves of muscle precursors are induced by signals from nearby tissues. In both amniotes and fish, the earliest myogenesis requires secreted signals from the ventral midline carried by Hedgehog (Hh) proteins. To determine if this similarity represents evolutionary homology, we have examined myogenesis in Xenopus laevis, the major species from which insight into vertebrate mesoderm patterning has been derived. Xenopus embryos form two distinct kinds of muscle cells analogous to the superficial slow and medial fast muscle fibres of zebrafish. As in zebrafish, Hh signalling is required for XMyf5 expression and generation of a first wave of early superficial slow muscle fibres in tail somites. Thus, Hh-dependent adaxial myogenesis is the likely ancestral condition of teleosts, amphibia and amniotes. Our evidence suggests that midline-derived cells migrate to the lateral somite surface and generate superficial slow muscle. This cell re-orientation contributes to the apparent rotation of Xenopus somites. Xenopus myogenesis in the trunk differs from that in the tail. In the trunk, the first wave of superficial slow fibres is missing, suggesting that significant adaptation of the ancestral myogenic programme occurred during tetrapod trunk evolution. Although notochord is required for early medial XMyf5 expression, Hh signalling fails to drive these cells to slow myogenesis. Later, both trunk and tail somites develop a second wave of Hh-independent slow fibres. These fibres probably derive from an outer cell layer expressing the myogenic determination genes XMyf5, XMyoD and Pax3 in a pattern reminiscent of amniote dermomyotome. Thus, Xenopus somites have characteristics in common with both fish and amniotes that shed light on the evolution of somite differentiation. We propose a model for the evolutionary adaptation of myogenesis in the transition from fish to tetrapod trunk.

Assessing storage of stability and mercury reduction of freeze-dried Pseudomonas putida within different types of lyoprotectant

NASA Astrophysics Data System (ADS)

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

2017-12-01

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

Synchrotron X-Ray Visualisation of Ice Formation in Insects during Lethal and Non-Lethal Freezing

PubMed Central

Sinclair, Brent J.; Gibbs, Allen G.; Lee, Wah-Keat; Rajamohan, Arun; Roberts, Stephen P.; Socha, John J.

2009-01-01

Although the biochemical correlates of freeze tolerance in insects are becoming well-known, the process of ice formation in vivo is subject to speculation. We used synchrotron x-rays to directly visualise real-time ice formation at 3.3 Hz in intact insects. We observed freezing in diapausing 3rd instar larvae of Chymomyza amoena (Diptera: Drosophilidae), which survive freezing if it occurs above −14°C, and non-diapausing 3rd instar larvae of C. amoena and Drosophila melanogaster (Diptera: Drosophilidae), neither of which survive freezing. Freezing was readily observed in all larvae, and on one occasion the gut was seen to freeze separately from the haemocoel. There were no apparent qualitative differences in ice formation between freeze tolerant and non-freeze tolerant larvae. The time to complete freezing was positively related to temperature of nucleation (supercooling point, SCP), and SCP declined with decreasing body size, although this relationship was less strong in diapausing C. amoena. Nucleation generally occurred at a contact point with the thermocouple or chamber wall in non-diapausing larvae, but at random in diapausing larvae, suggesting that the latter have some control over ice nucleation. There were no apparent differences between freeze tolerant and non-freeze tolerant larvae in tracheal displacement or distension of the body during freezing, although there was markedly more distension in D. melanogaster than in C. amoena regardless of diapause state. We conclude that although control of ice nucleation appears to be important in freeze tolerant individuals, the physical ice formation process itself does not differ among larvae that can and cannot survive freezing. This suggests that a focus on cellular and biochemical mechanisms is appropriate and may reveal the primary adaptations allowing freeze tolerance in insects. PMID:20011523

Interfacial Free Energy as the Key to the Pressure-Induced Deceleration of Ice Nucleation

NASA Astrophysics Data System (ADS)

Espinosa, Jorge R.; Zaragoza, Alberto; Rosales-Pelaez, Pablo; Navarro, Caridad; Valeriani, Chantal; Vega, Carlos; Sanz, Eduardo

2016-09-01

The avoidance of water freezing is the holy grail in the cryopreservation of biological samples, food, and organs. Fast cooling rates are used to beat ice nucleation and avoid cell damage. This strategy can be enhanced by applying high pressures to decrease the nucleation rate, but the physics behind this procedure has not been fully understood yet. We perform computer experiments to investigate ice nucleation at high pressures consisting in embedding ice seeds in supercooled water. We find that the slowing down of the nucleation rate is mainly due to an increase of the ice I -water interfacial free energy with pressure. Our work also clarifies the molecular mechanism of ice nucleation for a wide pressure range. This study is not only relevant to cryopreservation, but also to water amorphization and climate change modeling.

Using Power Ultrasound to Accelerate Food Freezing Processes: Effects on Freezing Efficiency and Food Microstructure.

PubMed

Zhang, Peizhi; Zhu, Zhiwei; Sun, Da-Wen

2018-05-31

Freezing is an effective way of food preservation. However, traditional freezing methods have the disadvantages of low freezing efficiency and generation of large ice crystals, leading to possible damage of food quality. Power ultrasound assisted freezing as a novel technique can effectively reduce the adverse effects during freezing process. This paper gives an overview on recent researches of power ultrasound technique to accelerate the food freezing processes and illustrates the main principles of power ultrasound assisted freezing. The effects of power ultrasound on liquid food, model solid food as well as fruit and vegetables are discussed, respectively, from the aspects of increasing freezing rate and improving microstructure. It is shown that ultrasound assisted freezing can effectively improve the freezing efficiency and promote the formation of small and evenly distributed ice crystals, resulting in better food quality. Different inherent properties of food samples affect the effectiveness of ultrasound application and optimum ultrasound parameters depend on the nature of the samples. The application of ultrasound to the food industry is more likely on certain types of food products and more efforts are still needed to realize the industrial translation of laboratory results.

An FPGA-Based Test-Bed for Reliability and Endurance Characterization of Non-Volatile Memory

NASA Technical Reports Server (NTRS)

Rao, Vikram; Patel, Jagdish; Patel, Janak; Namkung, Jeffrey

2001-01-01

Memory technologies are divided into two categories. The first category, nonvolatile memories, are traditionally used in read-only or read-mostly applications because of limited write endurance and slow write speed. These memories are derivatives of read only memory (ROM) technology, which includes erasable programmable ROM (EPROM), electrically-erasable programmable ROM (EEPROM), Flash, and more recent ferroelectric non-volatile memory technology. Nonvolatile memories are able to retain data in the absence of power. The second category, volatile memories, are random access memory (RAM) devices including SRAM and DRAM. Writing to these memories is fast and write endurance is unlimited, so they are most often used to store data that change frequently, but they cannot store data in the absence of power. Nonvolatile memory technologies with better future potential are FRAM, Chalcogenide, GMRAM, Tunneling MRAM, and Silicon-Oxide-Nitride-Oxide-Silicon (SONOS) EEPROM.

Weight management in obese pets: the tailoring concept and how it can improve results.

PubMed

German, Alexander J

2016-10-20

Obesity is now recognised as the most important medical disease in pets worldwide. All current strategies for weight management involve dietary energy restriction with a purpose-formulated diet. Whilst current weight management regimes can be successful, outcomes are often disappointing with the rate of weight loss progressively slowing down as time goes on. Success is most challenging for the most obese dogs and cats that are more likely to discontinue the programme before reaching target weight. To improve outcomes, clinicians must focus carefully on better tailoring programmes, paying particular to setting an appropriate target weight so as to maximise the benefits for the individual. In this opinionated review, the author will discuss findings from recent clinical research studies examining weight management in obese dogs and cats. A strategy for tailoring weight management targets will then be discussed, illustrated with case examples.

N-acetyltransferase gene polymorphisms & plasma isoniazid concentrations in patients with tuberculosis.

PubMed

Hemanth Kumar, A K; Ramesh, K; Kannan, T; Sudha, V; Haribabu, Hemalatha; Lavanya, J; Swaminathan, Soumya; Ramachandran, Geetha

2017-01-01

Variations in the N-acetyltransferase (NAT2) gene among different populations could affect the metabolism and disposition of isoniazid (INH). This study was performed to genotype NAT2 gene polymorphisms in tuberculosis (TB) patients from Chennai, India, and compare plasma INH concentrations among the different genotypes. Adult patients with TB treated in the Revised National TB Control Programme (RNTCP) in Chennai, Tamil Nadu, were genotyped for NAT2 gene polymorphism, and two-hour post-dosing INH concentrations were compared between the different genotypes. Plasma INH was determined by high-performance liquid chromatography. Genotyping of the NAT2 gene polymorphism was performed by real-time polymerase chain reaction method. Among the 326 patients genotyped, there were 189 (58%), 114 (35%) and 23 (7%) slow, intermediate and fast acetylators, respectively. The median two-hour INH concentrations in slow, intermediate and fast acetylators were 10.2, 8.1 and 4.1 μg/ml, respectively. The differences in INH concentrations among the three genotypes were significant (P<0.001). Genotyping of TB patients from south India for NAT2 gene polymorphism revealed that 58 per cent of the study population comprised slow acetylators. Two-hour INH concentrations differed significantly among the three genotypes.

Doctor pharmaceutical utilization behaviour changed by the global budget programme strategies on hypertensive outpatient prescription.

PubMed

Wei, Ching-Kuo; Wang, Shun-Mu; Yeh, Ming-Kung

2012-04-01

This study was to examine changes in doctor pharmaceutical utilization behaviour in response to Taiwan's newly implemented National Health Insurance individual hospital global budget (GB) programme and the changes in health care costs and prescription trends for hypertensive (HT) patients. We analysed hospital outpatient prescription utilization with a pre-post individual hospital GB group and comparison group (the hospitals who did not join the programme) to evaluate the impact of GB strategies on hypertensive expenditure. Descriptive analyses were performed based on the average daily medication expenditure for each prescription, and average number of items per prescription. This study reviewed 16,770,057 outpatient records and prescription records of 213,568 hypertensive patients. The average total medication expense (+17.6%), HT medication expense (+8.8%), daily medication expense (+16.3%), and daily HT medication expense (+6.3%) significantly increased after the action. After the individual hospital GB action, hospital doctors participating in action switched their patients' prescription drugs to other less expensive drugs such as rennin-angiotensin-aldosterone system inhibitors (-1.1%). The increase in volume of medications prescribed for control group were significantly larger for both alfa- and beta-adrenergic blocking agents (1.5%), and calcium channel blocking agents (3.9%). The individual hospital GB programme slowed down the trend of prescription drug cost increasing rate and reduced the prescription drug volume in hospitals. © 2010 Blackwell Publishing Ltd.

Novel Real-Time Diagnosis of the Freezing Process Using an Ultrasonic Transducer

PubMed Central

Tseng, Yen-Hsiang; Cheng, Chin-Chi; Cheng, Hong-Ping; Lee, Dasheng

2015-01-01

The freezing stage governs several critical parameters of the freeze drying process and the quality of the resulting lyophilized products. This paper presents an integrated ultrasonic transducer (UT) in a stainless steel bottle and its application to real-time diagnostics of the water freezing process. The sensor was directly deposited onto the stainless steel bottle using a sol-gel spray technique. It could operate at temperature range from −100 to 400 °C and uses an ultrasonic pulse-echo technique. The progression of the freezing process, including water-in, freezing point and final phase change of water, were all clearly observed using ultrasound. The ultrasonic signals could indicate the three stages of the freezing process and evaluate the cooling and freezing periods under various processing conditions. The temperature was also adopted for evaluating the cooling and freezing periods. These periods increased with water volume and decreased with shelf temperature (i.e., speed of freezing). This study demonstrates the effectiveness of the ultrasonic sensor and technology for diagnosing and optimizing the process of water freezing to save energy. PMID:25946629

A comparison of freezing-damage during isochoric and isobaric freezing of the potato.

PubMed

Lyu, Chenang; Nastase, Gabriel; Ukpai, Gideon; Serban, Alexandru; Rubinsky, Boris

2017-01-01

Freezing is commonly used for food preservation. It is usually done under constant atmospheric pressure (isobaric). While extending the life of the produce, isobaric freezing has detrimental effects. It causes loss of food weight and changes in food quality. Using thermodynamic analysis, we have developed a theoretical model of the process of freezing in a constant volume system (isochoric). The mathematical model suggests that the detrimental effects associated with isobaric freezing may be reduced in an isochoric freezing system. To explore this hypothesis, we performed a preliminary study on the isochoric freezing of a produce with which our group has experience, the potato. Experiments were performed in an isochoric freezing device we designed. The device is robust and has no moving parts. For comparison, we used a geometrically identical isobaric freezing device. Following freezing and thawing, the samples were weighed, examined with colorimetry, and examined with microscopy. It was found that potatoes frozen to -5 °C in an isochoric system experienced no weight loss and limited enzymatic browning. In contrast the -5 °C isobaric frozen potato experienced substantial weight loss and substantial enzymatic browning. Microscopic analysis shows that the structural integrity of the potato is maintained after freezing in the isochoric system and impaired after freezing in the isobaric system. Tissue damage during isobaric freezing is caused by the increase in extracellular osmolality and the mechanical damage by ice crystals. Our thermodynamic analysis predicts that during isochoric freezing the intracellular osmolality remains comparable to the extracellular osmolality and that isochoric systems can be designed to eliminate the mechanical damage by ice. The results of this preliminary study seem to confirm the theoretical predictions. This is a preliminary exploratory study on isochoric freezing of food. We have shown that the quality of a food product preserved by isochoric freezing is better than the quality of food preserved to the same temperature in isobaric conditions. Obviously, more extensive research remains to be done to extend this study to lower freezing temperatures and other food items.

A comparison of freezing-damage during isochoric and isobaric freezing of the potato

PubMed Central

Serban, Alexandru; Rubinsky, Boris

2017-01-01

Background Freezing is commonly used for food preservation. It is usually done under constant atmospheric pressure (isobaric). While extending the life of the produce, isobaric freezing has detrimental effects. It causes loss of food weight and changes in food quality. Using thermodynamic analysis, we have developed a theoretical model of the process of freezing in a constant volume system (isochoric). The mathematical model suggests that the detrimental effects associated with isobaric freezing may be reduced in an isochoric freezing system. To explore this hypothesis, we performed a preliminary study on the isochoric freezing of a produce with which our group has experience, the potato. Method Experiments were performed in an isochoric freezing device we designed. The device is robust and has no moving parts. For comparison, we used a geometrically identical isobaric freezing device. Following freezing and thawing, the samples were weighed, examined with colorimetry, and examined with microscopy. Results It was found that potatoes frozen to −5 °C in an isochoric system experienced no weight loss and limited enzymatic browning. In contrast the −5 °C isobaric frozen potato experienced substantial weight loss and substantial enzymatic browning. Microscopic analysis shows that the structural integrity of the potato is maintained after freezing in the isochoric system and impaired after freezing in the isobaric system. Discussion Tissue damage during isobaric freezing is caused by the increase in extracellular osmolality and the mechanical damage by ice crystals. Our thermodynamic analysis predicts that during isochoric freezing the intracellular osmolality remains comparable to the extracellular osmolality and that isochoric systems can be designed to eliminate the mechanical damage by ice. The results of this preliminary study seem to confirm the theoretical predictions. Conclusion This is a preliminary exploratory study on isochoric freezing of food. We have shown that the quality of a food product preserved by isochoric freezing is better than the quality of food preserved to the same temperature in isobaric conditions. Obviously, more extensive research remains to be done to extend this study to lower freezing temperatures and other food items. PMID:28533970

Production systems of Creole goat and their implications for a breeding programme.

PubMed

Gunia, M; Mandonnet, N; Arquet, R; de la Chevrotière, C; Naves, M; Mahieu, M; Alexandre, G

2010-12-01

The Creole goat is a local meat breed well adapted to the tropical environment of Guadeloupe, a French island in the Caribbean. A survey of 47 goat farmers was conducted in May 2008 to describe the Guadeloupean goat farming systems. It was the preliminary step for the implementation of a breeding programme for Creole goats. Farmers had 31 does on average. A small number (4%) kept only Creole goats. Most of them (62%) had a mixed herd of Creole and crossbreds. One-third of them (34%) reared only crossbred goats. Farmers appreciate the rusticity and resistance of the Creole goat but consider its growth as too slow. The most desired traits for goat selection were conformation and growth for males (77% of the answers). These traits were also important for females (30% of the answers). Maternal qualities were also frequently cited (maternal behaviour 23%, reproduction 20% and milk production 17%). Disease resistance was not seen as an important trait (10% and 7% of the answers for bucks and does, respectively). A typology constituted of five groups of farmers was also created. Farmers of three groups were retained to participate at a selection programme. They kept Creole goats and have expressed a strong willingness to join a selection programme. The results of the survey suggest that a breeding programme should mostly focus on the Creole goat as a maternal breed. Real consideration should be given to disease resistance. The Creole goat has indeed a key role to play in the sustainability of local farming systems.

Inactivation of Kudoa septempunctata in olive flounder meat by liquid freezing.

PubMed

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

2014-01-01

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

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

NASA Technical Reports Server (NTRS)

Karel, M.; Flink, J. M.

1978-01-01

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

Accurate prediction of collapse temperature using optical coherence tomography-based freeze-drying microscopy.

PubMed

Greco, Kristyn; Mujat, Mircea; Galbally-Kinney, Kristin L; Hammer, Daniel X; Ferguson, R Daniel; Iftimia, Nicusor; Mulhall, Phillip; Sharma, Puneet; Kessler, William J; Pikal, Michael J

2013-06-01

The objective of this study was to assess the feasibility of developing and applying a laboratory tool that can provide three-dimensional product structural information during freeze-drying and which can accurately characterize the collapse temperature (Tc ) of pharmaceutical formulations designed for freeze-drying. A single-vial freeze dryer coupled with optical coherence tomography freeze-drying microscopy (OCT-FDM) was developed to investigate the structure and Tc of formulations in pharmaceutically relevant products containers (i.e., freeze-drying in vials). OCT-FDM was used to measure the Tc and eutectic melt of three formulations in freeze-drying vials. The Tc as measured by OCT-FDM was found to be predictive of freeze-drying with a batch of vials in a conventional laboratory freeze dryer. The freeze-drying cycles developed using OCT-FDM data, as compared with traditional light transmission freeze-drying microscopy (LT-FDM), resulted in a significant reduction in primary drying time, which could result in a substantial reduction of manufacturing costs while maintaining product quality. OCT-FDM provides quantitative data to justify freeze-drying at temperatures higher than the Tc measured by LT-FDM and provides a reliable upper limit to setting a product temperature in primary drying. Copyright © 2013 Wiley Periodicals, Inc.

Freeze-drying of nanosuspensions, 1: freezing rate versus formulation design as critical factors to preserve the original particle size distribution.

PubMed

Beirowski, Jakob; Inghelbrecht, Sabine; Arien, Albertina; Gieseler, Henning

2011-05-01

It has been recently reported in the literature that using a fast freezing rate during freeze-drying of drug nanosuspensions is beneficial to preserve the original particle size distribution. All freezing rates studied were obtained by utilizing a custom-made apparatus and were then indirectly related to conventional vial freeze-drying. However, a standard freeze-dryer is only capable of achieving moderate freezing rates in the shelf fluid circulation system. Therefore, it was the purpose of the present study to evaluate the possibility to establish a typical freezing protocol applicable to a standard freeze-drying unit in combination with an adequate choice of cryoprotective excipients and steric stabilizers to preserve the original particle size distribution. Six different drug nanosuspensions containing itraconazole as a drug model were studied using freeze-thaw experiments and a full factorial design to reveal major factors for the stabilization of drug nanosuspensions and the corresponding interactions. In contrast to previous reports, the freezing regime showed no significant influence on preserving the original particle size distribution, suggesting that the concentrations of both the steric stabilizer and the cryoprotective agent are optimized. Moreover, it could be pinpointed that the combined effect of steric stabilizer and cryoprotectant clearly contribute to nanoparticle stability. Copyright © 2010 Wiley-Liss, Inc.

Freeze Technology for Nuclear Applications - 13590

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

Rostmark, Susanne C.; Knutsson, Sven; Lindberg, Maria

2013-07-01

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

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

PubMed Central

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

2016-01-01

Assessing freezing community response and whether freezing resistance is related to other functional traits is essential for understanding alpine community assemblages, particularly in Mediterranean environments where plants are exposed to freezing temperatures and summer droughts. Thus, we characterized the leaf freezing resistance of 42 plant species in 38 plots at Sierra de Guadarrama (Spain) by measuring their ice nucleation temperature, freezing point (FP), and low-temperature damage (LT50), as well as determining their freezing resistance mechanisms (i.e., tolerance or avoidance). The community response to freezing was estimated for each plot as community weighted means (CWMs) and functional diversity (FD), and we assessed their relative importance with altitude. We established the relationships between freezing resistance, growth forms, and four key plant functional traits (i.e., plant height, specific leaf area, leaf dry matter content (LDMC), and seed mass). There was a wide range of freezing resistance responses and more than in other alpine habitats. At the community level, the CWMs of FP and LT50 responded negatively to altitude, whereas the FD of both traits increased with altitude. The proportion of freezing-tolerant species also increased with altitude. The ranges of FP and LT50 varied among growth forms, and only leaf dry matter content was negatively correlated with freezing-resistance traits. Summer freezing events represent important abiotic filters for assemblies of Mediterranean high mountain communities, as suggested by the CWMs. However, a concomitant summer drought constraint may also explain the high freezing resistance of species that thrive in these areas and the lower FD of freezing resistance traits at lower altitudes. Leaves with high dry matter contents may maintain turgor at lower water potential and enhance drought tolerance in parallel to freezing resistance. This adaptation to drought seems to be a general prerequisite for plants found in xeric mountains. PMID:26941761

Equality and difference: persisting historical themes in health and alcohol policies affecting Indigenous Australians.

PubMed

Brady, Maggie

2007-09-01

Disseminating national health and alcohol policies to Aboriginal and Torres Strait Islander people in Australia has been a challenging task for governments and public servants. This has been for a number of reasons, including the enduring (negative) legacy of past "Aboriginal affairs" policies, the fact that Indigenous health programmes and alcohol programmes have been treated separately since the 1970s, and a more recent context in which the recognition of cultural difference was privileged. Confronted with the politics of difference, health departments were slow to examine avenues through which best practice advice emanating from WHO, and alcohol policies such as harm minimisation and early identification and treatment in primary health care, could be communicated in culturally recognisable ways to independent Indigenous services. In addition, there was hostility towards harm minimisation policies from Indigenous service providers, and Indigenous treatment programmes remained largely committed to abstinence-oriented modalities and the disease model of alcoholism, despite moves away from these approaches in the mainstream. However, genuinely innovative acute interventions and environmental controls over alcohol have been developed by Indigenous community-based organisations, approaches that are reinforced by international policy research evidence.

Survival mechanisms of vertebrate ectotherms at subfreezing temperatures: applications in cryomedicine.

PubMed

Costanzo, J P; Lee, R E; DeVries, A L; Wang, T; Layne, J R

1995-03-01

Various marine fishes, amphibians, and reptiles survive at temperatures several degrees below the freezing point of their body fluids by virtue of adaptive mechanisms that promote freeze avoidance or freeze tolerance. Freezing is avoided by a colligative depression of the blood freezing point, supercooling of the body fluids, or the biosynthesis of unique antifreeze proteins that inhibit the propagation of ice within body fluids. Conversely, freeze tolerance is an adaptation for the survival of tissue freezing under ecologically relevant thermal and temporal conditions that is conferred by the biosynthesis of permeating carbohydrate cryoprotectants and an extensive dehydration of tissues and organs. Such cryoprotective responses, invoked by the onset of freezing, mitigate the osmotic stress associated with freeze-concentration of cytoplasm, attendant metabolic perturbations, and physical damage. Cryomedical research has historically relied on mammalian models for experimentation even though endotherms do not naturally experience subfreezing temperatures. Some vertebrate ectotherms have "solved" not only the problem of freezing individual tissues and organs, but also that of simultaneously freezing all organ systems. An emerging paradigm in cryomedicine is the application of principles governing natural cold hardiness to the development of protocols for the cryopreservation of mammalian tissues and organs.

Hepatitis B vaccine freezing in the Indonesian cold chain: evidence and solutions.

PubMed

Nelson, Carib M; Wibisono, Hariadi; Purwanto, Hary; Mansyur, Isa; Moniaga, Vanda; Widjaya, Anton

2004-02-01

To document and characterize freezing temperatures in the Indonesian vaccine cold chain and to evaluate the feasibility of changes designed to reduce the occurrence of freezing. Data loggers were used to measure temperatures of shipments of hepatitis B vaccine from manufacturer to point of use. Baseline conditions and three intervention phases were monitored. During each of the intervention phases, vaccines were removed progressively from the standard 2-8 degrees C cold chain. Freezing temperatures were recorded in 75% of baseline shipments. The highest rates of freezing occurred during transport from province to district, storage in district-level ice-lined refrigerators, and storage in refrigerators in health centres. Interventions reduced freezing, without excessive heat exposure. Inadvertent freezing of freeze-sensitive vaccines is widespread in Indonesia. Simple strategies exist to reduce freezing - for example, selective transport and storage of vaccines at ambient temperatures. The use of vaccine vial monitors reduces the risk associated with heat-damaged vaccines in these scenarios. Policy changes that allow limited storage of freeze-sensitive vaccines at temperatures >2-8 degrees C would enable flexible vaccine distribution strategies that could reduce vaccine freezing, reduce costs, and increase capacity.

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.

Freezing of gait in Parkinson's disease: the paradoxical interplay between gait and cognition.

PubMed

Ricciardi, Lucia; Bloem, Bastiaan R; Snijders, Anke H; Daniele, Antonio; Quaranta, Davide; Bentivoglio, Anna Rita; Fasano, Alfonso

2014-08-01

Freezing of gait is a disabling episodic gait disturbance common in patients with Parkinson's disease. Recent evidences suggest a complex interplay between gait impairment and executive functions. Aim of our study was to evaluate whether specific motor conditions (sitting or walking) influence cognitive performance in patients with or without different types of freezing. Eight healthy controls, eight patients without freezing, nine patients with levodopa-responsive and nine patients with levodopa-resistant freezing received a clinical and neuropsychological assessment during two randomly performed conditions: at rest and during walking. At rest, patients with levodopa-resistant freezing performed worse than patients without freezing on tests of phonological fluency (p = 0.01). No differences among the four groups were detected during walking. When cognitive performances during walking were compared to the performance at rest, there was a significant decline of verbal episodic memory task (Rey Auditory Verbal Learning Test) in patients without freezing and with levodopa-responsive freezing. Interestingly, walking improved performance on the phonological fluency task in patients with levodopa-resistant freezing (p = 0.04). Compared to patients without freezing, patients with levodopa-resistant freezing perform worse when tested while seated in tasks of phonological verbal fluency. Surprisingly, gait was associated with a paradoxical improvement of phonological verbal fluency in the patients with levodopa-resistant freezing whilst walking determined a worsening of episodic memory in the other patient groups. Copyright © 2014 Elsevier Ltd. All rights reserved.

Freezing during tapping tasks in patients with advanced Parkinson's disease and freezing of gait.

PubMed

Delval, Arnaud; Defebvre, Luc; Tard, Céline

2017-01-01

Parkinson's disease patients with freezing of gait also experience sudden motor blocks (freezing) during other repetitive motor tasks. We assessed the proportion of patients with advanced PD and freezing of gait who also displayed segmental "freezing" in tapping tasks. Fifteen Parkinson's disease patients with freezing of gait were assessed. Freezing of gait was evaluated using a standardized gait trajectory with the usual triggers. Patients performed repetitive tapping movements (as described in the MDS-UPDRS task) with the hands or the feet in the presence or absence of a metronome set to 4 Hz. Movements were recorded with a video motion system. The primary endpoint was the occurrence of segmental freezing in these tapping tasks. The secondary endpoints were (i) the relationship between segmental episodic phenomena and FoG severity, and (ii) the reliability of the measurements. For the upper limbs, freezing was observed more frequently with a metronome (21% of trials) than without a metronome (5%). For the lower limbs, the incidence of freezing was higher than for the upper limbs, and was again observed more frequently in the presence of an auditory cue (47%) than in its absence (14%). Although freezing of the lower limbs was easily assessed during an MDS-UPDRS task with a metronome, it was not correlated with the severity of freezing of gait (as evaluated during a standardized gait trajectory). Only this latter was a reliable measurement in patients with advanced Parkinson's disease.

Effect of addition of wild garlic (Allium ursinum) on the quality of kefirs from sheep's milk.

PubMed

Znamirowska, Agata; Szajnar, Katarzyna; Rożek, Przemysław; Kalicka, Dorota; Kuźniar, Piotr; Hanus, Paweł; Kotula, Krzysztof; Obirek, Marcin; Kluz, Maciej

2017-01-01

Sheep’s milk has a high content of total solids, which qualifies it as a very good raw material for the production of fermented milk drinks. Currently, there are commercially produced kefirs and yogurts from sheep’s milk in the countries of the Mediterranean region. The growing interest in the consumption of these products is justified not only by their taste merits, but also because of their health-promoting proper- ties. The aim of the present study was to determine the effect of the addition of 1% of lyophilized wild garlic powder on the properties of kefirs from sheep’s milk. Sheep’s milk was pasteurized (85°C, 30 min), cooled down, enriched with 1% of freeze-dried wild garlic powder, inoculated with a Commercial VITAL kefir culture and fermented for 16 hours (26°C). The influence of wild garlic on acidity (pH, °SH), syneresis (%), texture (TPA test), colour (L*a*b*) and the sensory profile of kefirs was conducted. Wild garlic could be used as a taste and flavour modifier in the production of kefir from sheep’s milk. The addition of 1% of freeze-dried wild garlic slowed down the fermentation of kefir, changed colour and reduced syneresis. Wild garlic could be used as a valuable supplement and a modifier of taste and flavour in kefir from sheep’s milk.

Cryopreservation: Evolution of Molecular Based Strategies.

PubMed

Baust, John M; Corwin, William; Snyder, Kristi K; Van Buskirk, Robert; Baust, John G

2016-01-01

Cryopreservation (CP) is an enabling process providing for on-demand access to biological material (cells and tissues) which serve as a starting, intermediate or even final product. While a critical tool, CP protocols, approaches and technologies have evolved little over the last several decades. A lack of conversion of discoveries from the CP sciences into mainstream utilization has resulted in a bottleneck in technological progression in areas such as stem cell research and cell therapy. While the adoption has been slow, discoveries including molecular control and buffering of cell stress response to CP as well as the development of new devices for improved sample freezing and thawing are providing for improved CP from both the processing and sample quality perspectives. Numerous studies have described the impact, mechanisms and points of control of cryopreservation-induced delayed-onset cell death (CIDOCD). In an effort to limit CIDOCD, efforts have focused on CP agent and freeze media formulation to provide a solution path and have yielded improvements in survival over traditional approaches. Importantly, each of these areas, new technologies and cell stress modulation, both individually and in combination, are now providing a new foundation to accelerate new research, technology and product development for which CP serves as an integral component. This chapter provides an overview of the molecular stress responses of cells to cryopreservation, the impact of the hypothermic and cell death continuums and the targeted modulation of common and/or cell specific responses to CP in providing a path to improving cell quality.

Experimental observation of the influence of furnace temperature profile on convection and segregation in the vertical Bridgman crystal growth technique

NASA Technical Reports Server (NTRS)

Neugebauer, G. T.; Wilcox, William R.

1992-01-01

Azulene-doped naphthalene was directionally solidified during the vertical Bridgman-Stockbarger technique. Doping homogeneity and convection were determined as a function of the temperature profile in the furnace and the freezing rate. Convection velocities were two orders of magnitude lower when the temperature increased with height. Rarely was the convection pattern axisymmetric, even though the temperature varied less than 0.1 K around the circumference of the growth ampoule. Correspondingly the cross sectional variation in azulene concentration tended to be asymmetric, especially when the temperature increased with height. This cross sectional variation changed dramatically along the ingot, reflecting changes in convection presumably due to the decreasing height of the melt. Although there was large scatter and irreproducibility in the cross sectional variation in doping, this variation tended to be least when the growth rate was low and the convection was vigorous. It is expected that compositional variations would also be small at high growth rates with weak convection and flat interfaces, although this was not investigated in the present experiments. Neither rotation of the ampoule nor deliberate introduction of thermal asymmetries during solidification had a significant influence on cross sectional variations in doping. It is predicted that slow directional solidification under microgravity conditions could produce greater inhomogeneities than on Earth. Combined use of microgravity and magnetic fields would be required to achieve homogeneity when it is necessary to freeze slowly in order to avoid constitutional supercooling.

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

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

PubMed Central

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

2014-01-01

This study investigates the effects of the gelatin concentration (10-40%, w/v), freezing temperatures (from -20℃ to -50℃) and freezing methods on the structural and physical properties of gelatin matrices. To freeze gelatin, the pressure-shift freezing (PSF) is being applied at 0.1 (under atmospheric control), 50 and 100 MPa, respectively. The freezing point of gelatin solutions decrease with increasing gelatin concentrations, from -0.2℃ (10% gelatin) to -6.7℃ (40% gelatin), while the extent of supercooling did not show any specific trends. The rheological properties of the gelatin indicate that both the storage (G') and loss (G") moduli were steady in the strain amplitude range of 0.1-10%. To characterize gelatin matrices formed by the various freezing methods, the ice crystal sizes which were being determined by the scanning electron microscopy (SEM) are affected by the gelatin concentrations. The ice crystal sizes are affected by gelatin concentrations and freezing temperature, while the size distributions of ice crystals depend on the freezing methods. Smaller ice crystals are being formed with PSF rather than under the atmospheric control where the freezing temperature is above -40℃. Thus, the results of this study indicate that the PSF processing at a very low freezing temperature (-50℃) offers a potential advantage over commercial atmospheric freezing points for the formation of small ice crystals. PMID:26760743

Effects of different freezing methods on calcium enriched papaya (Carica papaya L.).

PubMed

Lovera, Nancy N; Ramallo, Laura; Salvadori, Viviana O

2018-06-01

The effect of calcium impregnation on drip loss, colour, mechanical properties, sensory perception and freezing time on frozen-thawed papaya was studied, evaluating different freezing methods: cryogenic, tunnel and household freezer freezing. Osmotic dehydration as pre-treatment was also evaluated. Freezing in liquid nitrogen was considered an inappropriate method for papaya preservation due to cracking. Calcium impregnation and osmotic dehydration increased tissue firmness and decreased freezing time (freezing time for fresh, calcium impregnated and osmo-dehydrated fruit was 23, 17 and 5 min in a tunnel and 118, 83 and 60 min in a household freezer, respectively). Calcium lactate was the most effective way to protect tissue's firmness before and after a freeze-thaw cycle (maximum stress values approx. 300-400% of the raw tissue for tunnel freezing and 260% for household freezer). Microstructure analysis showed better tissue integrity retention in papaya samples impregnated with calcium lactate than in those with calcium gluconate, after a freezing-thawing cycle, in agreement with the drip loss results. In spite of these results, consumers preferred frozen papaya without pre-treatment or impregnated with calcium gluconate.

Response of shoal grass, Halodule wrightii, to extreme winter conditions in the Lower Laguna Madre, Texas

USGS Publications Warehouse

Hicks, D.W.; Onuf, C.P.; Tunnell, J.W.

1998-01-01

Effects of a severe freeze on the shoal grass, Halodule wrightii, were documented through analysis of temporal and spatial trends in below-ground biomass. The coincidence of the second lowest temperature (-10.6??C) in 107 years of record, 56 consecutive hours below freezing, high winds and extremely low water levels exposed the Laguna Madre, TX, to the most severe cold stress in over a century. H. wrightii tolerated this extreme freeze event. Annual pre- and post-freeze surveys indicated that below-ground biomass estimated from volume was Unaffected by the freeze event. Nor was there any post-freeze change in biomass among intertidal sites directly exposed to freezing air temperatures relative to subtidal sites which remained submerged during the freezing period.

The effect of undissolved air on isochoric freezing.

PubMed

Perez, Pedro A; Preciado, Jessica; Carlson, Gary; DeLonzor, Russ; Rubinsky, Boris

2016-06-01

This study evaluates the effect of undissolved air on isochoric freezing of aqueous solutions. Isochoric freezing is concerned with freezing in a constant volume thermodynamic system. A possible advantage of the process is that it substantially reduces the percentage of ice in the system at every subzero temperature, relative to atmospheric freezing. At the pressures generated by isochoric freezing, or high pressure isobaric freezing, air cannot be considered an incompressible substance and the presence of undissolved air substantially increases the amount of ice that forms at any subfreezing temperature. This effect is measurable at air volumes as low as 1%. Therefore eliminating the undissolved air, or any separate gaseous phase, from the system is essential for retaining the properties of isochoric freezing. Copyright © 2016. Published by Elsevier Inc.

Visual Indicators on Vaccine Boxes as Early Warning Tools to Identify Potential Freeze Damage.

PubMed

Angoff, Ronald; Wood, Jillian; Chernock, Maria C; Tipping, Diane

2015-07-01

The aim of this study was to determine whether the use of visual freeze indicators on vaccines would assist health care providers in identifying vaccines that may have been exposed to potentially damaging temperatures. Twenty-seven sites in Connecticut involved in the Vaccine for Children Program participated. In addition to standard procedures, visual freeze indicators (FREEZEmarker ® L; Temptime Corporation, Morris Plains, NJ) were affixed to each box of vaccine that required refrigeration but must not be frozen. Temperatures were monitored twice daily. During the 24 weeks, all 27 sites experienced triggered visual freeze indicator events in 40 of the 45 refrigerators. A total of 66 triggered freeze indicator events occurred in all 4 types of refrigerators used. Only 1 of the freeze events was identified by a temperature-monitoring device. Temperatures recorded on vaccine data logs before freeze indicator events were within the 35°F to 46°F (2°C to 8°C) range in all but 1 instance. A total of 46,954 doses of freeze-sensitive vaccine were stored at the time of a visual freeze indicator event. Triggered visual freeze indicators were found on boxes containing 6566 doses (14.0% of total doses). Of all doses stored, 14,323 doses (30.5%) were of highly freeze-sensitive vaccine; 1789 of these doses (12.5%) had triggered indicators on the boxes. Visual freeze indicators are useful in the early identification of freeze events involving vaccines. Consideration should be given to including these devices as a component of the temperature-monitoring system for vaccines.

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.

Evaluating Long-Term Impacts of Soil-Mixing Source-Zone Treatment using Cryogenic Core Collection

DTIC Science & Technology

2017-06-01

to (a) coring equipment freezing downhole, (b) freezing or binding of the core sample in barrel, and ( c ) running out of LN in the vicinity of sampling...encountered due to (a) coring equipment freezing downhole, (b) freezing or binding of the core sample in barrel, and ( c ) running out of LN in the...equipment freezing downhole, (b) freezing or binding of the core sample in barrel, and ( c ) running out of LN in the vicinity of sampling. Downhole

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.

Application of the Quality by Design Approach to the Freezing Step of Freeze-Drying: Building the Design Space.

PubMed

Arsiccio, Andrea; Pisano, Roberto

2018-06-01

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

Freezing curve-based monitoring to quickly evaluate the viability of biological materials subject to freezing or thermal injury.

PubMed

Liu, Jing; Zhou, Yi-Xin

2003-09-01

This paper is aimed at investigating the roles of freezing dynamics of a liquid droplet to characterize the properties of the material. In particular, freezing curve-based monitoring was proposed to quickly evaluate the viability of biological materials subject to freezing, re-warming, or other kinds of injury, which is an extremely important issue in practices such as cryobiology, hyperthermia, or freshness evaluation of bio-samples. An integrated micro analysis device was fabricated which is simple in structure and cheap to make. Preliminary freezing results demonstrated that minor changes in a biological material due to freezing or warming injury might result in a significant deviation of its freezing curve from that of the intact biomaterials. Several potential thermal indexes to quantify the material features were pointed out. Further, experiments were performed on some freezing and thawing processes of small amount of water on a cooling surface to test the effects of droplet sizes, measurement sites, cooling strength, and cooling geometry, etc., on the freezing responses of a water droplet. Their implementation in developing a new micro analysis system were suggested. This freezing curve-based monitoring method may open a new strategy for the evaluation of biomaterials subject to destruction in diverse fields.

Recent developments in smart freezing technology applied to fresh foods.

PubMed

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

2017-09-02

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

Freezing tolerance of winter wheat as influenced by extended growth at low temperature and exposure to freeze-thaw cycles

USDA-ARS?s Scientific Manuscript database

As the seasons progress, autumn-planted winter wheat plants (Triticum aestivum L.) first gain, then progressively lose freezing tolerance. Exposing the plants to freeze-thaw cycles of -3/3°C results in increased ability to tolerate subsequent freezing to potentially damaging temperatures. This stu...

Host-mediated shift in the cold tolerance of an invasive insect

Treesearch

Amy C. Morey; Robert C. Venette; Erica C. Nystrom Santacruz; Laurel A. Mosca; W. D. Hutchison

2016-01-01

While many insects cannot survive the formation of ice within their bodies, a few species can. On the evolutionary continuum from freeze-intolerant (i.e., freeze-avoidant) to freeze-tolerant insects, intermediates likely exist that can withstand some ice formation, but not enough to be considered fully freeze tolerant. Theory suggests that freeze tolerance should be...

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

PubMed

Pramsohler, Manuel; Hacker, Jürgen; Neuner, Gilbert

2012-07-01

The freezing pattern and frost killing temperatures of apple (Malus domestica Borkh.) xylem were determined by differential thermal analysis and infrared differential thermal analysis (IDTA). Results from detached or attached twigs in controlled freezing experiments and during natural field freezing of trees were compared. Non-lethal freezing of apoplastic water in apple xylem as monitored during natural winter frosts in the field occurred at -1.9 ± 0.4 °C and did not change seasonally. The pattern of whole tree freezing was variable and specific to the environmental conditions. On detached twigs high-temperature freezing exotherms (HTEs) occurred 2.8 K below the temperature observed under natural frosts in the field with a seasonal mean of -4.7 ± 0.5 °C. Microporous apple xylem showed freezing without a specific pattern within a few seconds in IDTA images during HTEs, which is in contrast to macroporous xylem where a 2D freezing pattern mirrors anatomical structures. The pith tissue always remained unfrozen. Increasing twig length increased ice nucleation temperature; for increased twig diameter the effect was not significant. In attached twigs frozen in field portable freezing chambers, HTEs were recorded at a similar mean temperature (-4.6 ± 1.0 °C) to those for detached twigs. Upon lethal intracellular freezing of apple xylem parenchyma cells (XPCs) low-temperature freezing exotherms (LTEs) can be recorded. Low-temperature freezing exotherms determined on detached twigs varied significantly between a winter minimum of -36.9 °C and a summer maximum -12.7 °C. Within the temperature range wherein LTEs were recorded by IDTA in summer (-12.7 ± 0.5 to -20.3 ± 1.1 °C) various tiny clearly separated discontinuous freezing events could be detected similar to that in other species with contrasting XPC anatomy. These freezing events appeared to be initially located in the primary and only later in the secondary xylem. During the LTE no freezing events in the bark and central pith tissue were recorded. Attached twigs were exposed to various freezing temperatures at which LTEs occur. Even if 60% of XPCs were frost-damaged twigs were able to recuperate and showed full re-growth indicating a high regeneration capacity even after severe frost damage to XPCs.

Defensive freezing links Hypothalamic-Pituitary-Adrenal-axis activity and internalizing symptoms in humans.

PubMed

Niermann, Hannah C M; Figner, Bernd; Tyborowska, Anna; van Peer, Jacobien M; Cillessen, Antonius H N; Roelofs, Karin

2017-08-01

The Hypothalamic-Pituitary-Adrenal (HPA)-axis plays an important role in the expression of defensive freezing. Adaptive freezing reactivity, characterized by an immediate increase in acute stress and timely termination upon threat offset or need to act, is essential for adequate stress coping. Blunted HPA-axis activity in animals is associated with blunted freezing reactivity and internalizing symptoms. Despite their potential relevance, it remains unknown whether these mechanisms apply to humans and human psychopathology. Using a well-established method combining electrocardiography and posturography, we assessed freezing before, immediately after, and one hour after a stress induction in 92 human adolescents. In line with animal models, human adolescents showed stress-induced freezing, as quantified by relative reductions in heart rate and body sway after, as compared to before, stress. Moreover, relatively lower basal cortisol was associated with reduced stress-induced freezing reactivity (i.e., less immediate freezing and less recovery). Path analyses showed that decreased freezing recovery in individuals with reduced cortisol levels was associated with increased levels of internalizing symptoms. These findings suggest that reduced freezing recovery may be a promising marker for the etiology of internalizing symptoms. Copyright © 2017 Elsevier Ltd. All rights reserved.

Synthetic polymers enable non-vitreous cellular cryopreservation by reducing ice crystal growth during thawing.

PubMed

Deller, Robert C; Vatish, Manu; Mitchell, Daniel A; Gibson, Matthew I

2014-01-01

The cryopreservation of cells, tissue and organs is fundamental to modern biotechnology, transplantation medicine and chemical biology. The current state-of-the-art method of cryopreservation is the addition of large amounts of organic solvents such as glycerol or dimethyl sulfoxide, to promote vitrification and prevent ice formation. Here we employ a synthetic, biomimetic, polymer, which is capable of slowing the growth of ice crystals in a manner similar to antifreeze (glyco)proteins to enhance the cryopreservation of sheep and human red blood cells. We find that only 0.1 wt% of the polymer is required to attain significant cell recovery post freezing, compared with over 20 wt% required for solvent-based strategies. These results demonstrate that synthetic antifreeze (glyco)protein mimics could have a crucial role in modern regenerative medicine to improve the storage and distribution of biological material for transplantation.

Encapsulated boron as an osteoinductive agent for bone scaffolds.

PubMed

Gümüşderelioğlu, Menemşe; Tunçay, Ekin Ö; Kaynak, Gökçe; Demirtaş, Tolga T; Aydın, Seda Tığlı; Hakkı, Sema S

2015-01-01

The aim of this study was to develop boron (B)-releasing polymeric scaffold to promote regeneration of bone tissue. Boric acid-doped chitosan nanoparticles with a diameter of approx. 175 nm were produced by tripolyphosphate (TPP)-initiated ionic gelation process. The nanoparticles strongly attached via electrostatic interactions into chitosan scaffolds produced by freeze-drying with approx. 100 μm pore diameter. According to the ICP-OES results, following first 5h initial burst release, fast release of B from scaffolds was observed for 24h incubation period in conditioned medium. Then, slow release of B was performed over 120 h. The results of the cell culture studies proved that the encapsulated boron within the scaffolds can be used as an osteoinductive agent by showing its positive effects on the proliferation and differentiation of MC3T3-E1 preosteoblastic cells. Copyright © 2015 Elsevier GmbH. All rights reserved.

Magnetism at grain boundary interfacesin the colossal permittivity dielectric material; In+Nb Co-Doped Rutile

NASA Astrophysics Data System (ADS)

Berlie, Adam; Terry, Ian; Cottrell, Stephen; Hu, Wanbiao; Liu, Yun

With the emphasis in recent years on understanding novel materials with potential technological applications this work seeks to understand magnetic ordering within the colossal-permittivity material, In+Nb co-doped rutile (TiO2). Evidence for a spin-freezing transition was reported from a step like feature in the dielectic data below 50 K but this was largly glossed over. Within this work we show that below 300 K there is a slowing down of magnetic fluctuations associated with the electronic magnetism due to the defect-dipoles created by the co-doping, but the muon spectroscopy results are strongly suggestive of the behaviour being localised to the edges/interfaces of particles/grains. The TC is strongly dependent on the doping level of the samples that presents novel way to control the magnetism and ultimately magneto-electric coupling within a dielectric material.

Persistence at low temperature of the P beta' ripple in dipalmitoylphosphatidylcholine multilamellar vesicles containing either glycosphingolipids or cholesterol.

PubMed

Rock, P; Thompson, T E; Tillack, T W

1989-03-13

The disappearance and reappearance of the P beta' ripple in multilamellar liposomes of dipalmitoylphosphatidylcholine (DPPC) has been examined by freeze-etch electron microscopy. The presence of less than 10 mol% of various glycosphingolipids or cholesterol in the liposomes markedly increases the time required for ripple disappearance when the vesicles are cooled from 38 degrees C to 30 degrees C, as compared to the pure phospholipid. Once the ripples have begun to disappear in the two-component vesicles, they do not uniformly reappear until the system is heated above the main transition of DPPC and allowed to cool into the pretransition region. These results suggest that the long time for ripple disappearance in the two-component systems reflects a slow molecular reorganization process which occurs when the systems are forced to change from the P beta' gel to the L beta' gel by a temperature downshift.

Exchange bias training relaxation in spin glass/ferromagnet bilayers

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

Chi, Xiaodan; Du, An; Rui, Wenbin

2016-04-25

A canonical spin glass (SG) FeAu layer is fabricated to couple to a soft ferromagnet (FM) FeNi layer. Below the SG freezing temperature, exchange bias (EB) and training are observed. Training in SG/FM bilayers is insensitive to cooling field and may suppress the EB or change the sign of the EB field from negative to positive at specific temperatures, violating from the simple power-law or the single exponential function derived from the antiferromagnet based systems. In view of the SG nature, we employ a double decay model to distinguish the contributions from the SG bulk and the SG/FM interface tomore » training. Dynamical properties during training under different cooling fields and at different temperatures are discussed, and the nonzero shifting coefficient in the time index as a signature of slowing-down decay for SG based systems is interpreted by means of a modified Monte Carlo Metropolis algorithm.« less

Ice Melting to Release Reactants in Solution Syntheses.

PubMed

Wei, Hehe; Huang, Kai; Zhang, Le; Ge, Binghui; Wang, Dong; Lang, Jialiang; Ma, Jingyuan; Wang, Da; Zhang, Shuai; Li, Qunyang; Zhang, Ruoyu; Hussain, Naveed; Lei, Ming; Liu, Li-Min; Wu, Hui

2018-03-19

Aqueous solution syntheses are mostly based on mixing two solutions with different reactants. It is shown that freezing one solution and melting it in another solution provides a new interesting strategy to mix chemicals and to significantly change the reaction kinetics and thermodynamics. For example, a precursor solution containing a certain concentration of AgNO 3 was frozen and dropped into a reductive NaBH 4 solution at about 0 °C. The ultra-slow release of reactants was successfully achieved. An ice-melting process can be used to synthesize atomically dispersed metals, including cobalt, nickel, copper, rhodium, ruthenium, palladium, silver, osmium, iridium, platinum, and gold, which can be easily extended to other solution syntheses (such as precipitation, hydrolysis, and displacement reactions) and provide a generalized method to redesign the interphase reaction kinetics and ion diffusion in wet chemistry. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Emerging technologies in medical applications of minimum volume vitrification

PubMed Central

Zhang, Xiaohui; Catalano, Paolo N; Gurkan, Umut Atakan; Khimji, Imran; Demirci, Utkan

2011-01-01

Cell/tissue biopreservation has broad public health and socio-economic impact affecting millions of lives. Cryopreservation technologies provide an efficient way to preserve cells and tissues targeting the clinic for applications including reproductive medicine and organ transplantation. Among these technologies, vitrification has displayed significant improvement in post-thaw cell viability and function by eliminating harmful effects of ice crystal formation compared to the traditional slow freezing methods. However, high cryoprotectant agent concentrations are required, which induces toxicity and osmotic stress to cells and tissues. It has been shown that vitrification using small sample volumes (i.e., <1 μl) significantly increases cooling rates and hence reduces the required cryoprotectant agent levels. Recently, emerging nano- and micro-scale technologies have shown potential to manipulate picoliter to nanoliter sample sizes. Therefore, the synergistic integration of nanoscale technologies with cryogenics has the potential to improve biopreservation methods. PMID:21955080

Observation of giant exchange bias in bulk Mn50Ni42Sn8 Heusler alloy

NASA Astrophysics Data System (ADS)

Sharma, Jyoti; Suresh, K. G.

2015-02-01

We report a giant exchange bias (EB) field of 3520 Oe in bulk Mn50Ni42Sn8 Heusler alloy. The low temperature magnetic state of the martensite phase has been studied by DC magnetization and AC susceptibility measurements. Frequency dependence of spin freezing temperature (Tf) on critical slowing down relation and observation of memory effect in zero field cooling mode confirms the super spin glass (SSG) phase at low temperatures. Large EB is attributed to the strong exchange coupling between the SSG clusters formed by small regions of ferromagnetic order embedded in an antiferromagnetic (AFM) matrix. The temperature and cooling field dependence of EB have been studied and related to the change in unidirectional anisotropy at SSG/AFM interface. The training effect also corroborates with the presence of frozen (SSG) moments at the interface and their role in EB.

A generalized non-Gaussian consistency relation for single field inflation

NASA Astrophysics Data System (ADS)

Bravo, Rafael; Mooij, Sander; Palma, Gonzalo A.; Pradenas, Bastián

2018-05-01

We show that a perturbed inflationary spacetime, driven by a canonical single scalar field, is invariant under a special class of coordinate transformations together with a field reparametrization of the curvature perturbation in co-moving gauge. This transformation may be used to derive the squeezed limit of the 3-point correlation function of the co-moving curvature perturbations valid in the case that these do not freeze after horizon crossing. This leads to a generalized version of Maldacena's non-Gaussian consistency relation in the sense that the bispectrum squeezed limit is completely determined by spacetime diffeomorphisms. Just as in the case of the standard consistency relation, this result may be understood as the consequence of how long-wavelength modes modulate those of shorter wavelengths. This relation allows one to derive the well known violation to the consistency relation encountered in ultra slow-roll, where curvature perturbations grow exponentially after horizon crossing.

Stroboscopic Image Modulation to Reduce the Visual Blur of an Object Being Viewed by an Observer Experiencing Vibration

NASA Technical Reports Server (NTRS)

Kaiser, Mary K. (Inventor); Adelstein, Bernard D. (Inventor); Anderson, Mark R. (Inventor); Beutter, Brent R. (Inventor); Ahumada, Albert J., Jr. (Inventor); McCann, Robert S. (Inventor)

2014-01-01

A method and apparatus for reducing the visual blur of an object being viewed by an observer experiencing vibration. In various embodiments of the present invention, the visual blur is reduced through stroboscopic image modulation (SIM). A SIM device is operated in an alternating "on/off" temporal pattern according to a SIM drive signal (SDS) derived from the vibration being experienced by the observer. A SIM device (controlled by a SIM control system) operates according to the SDS serves to reduce visual blur by "freezing" (or reducing an image's motion to a slow drift) the visual image of the viewed object. In various embodiments, the SIM device is selected from the group consisting of illuminator(s), shutter(s), display control system(s), and combinations of the foregoing (including the use of multiple illuminators, shutters, and display control systems).

Improving survival and storage stability of bacteria recalcitrant to freeze-drying: a coordinated study by European culture collections.

PubMed

Peiren, Jindrich; Buyse, Joke; De Vos, Paul; Lang, Elke; Clermont, Dominique; Hamon, Sylviane; Bégaud, Evelyne; Bizet, Chantal; Pascual, Javier; Ruvira, María A; Macián, M Carmen; Arahal, David R

2015-04-01

The objective of this study is to improve the viability after freeze-drying and during storage of delicate or recalcitrant strains safeguarded at biological resource centers. To achieve this objective, a joint experimental strategy was established among the different involved partner collections of the EMbaRC project ( www.embarc.eu ). Five bacterial strains considered as recalcitrant to freeze-drying were subjected to a standardized freeze-drying protocol and to seven agreed protocol variants. Viability of these strains was determined before and after freeze-drying (within 1 week, after 6 and 12 months, and after accelerated storage) for each of the protocols. Furthermore, strains were exchanged between partners to perform experiments with different freeze-dryer-dependent parameters. Of all tested variables, choice of the lyoprotectant had the biggest impact on viability after freeze-drying and during storage. For nearly all tested strains, skim milk as lyoprotectant resulted in lowest viability after freeze-drying and storage. On the other hand, best freeze-drying and storage conditions were strain and device dependent. For Aeromonas salmonicida CECT 894(T), best survival was obtained when horse serum supplemented with trehalose was used as lyoprotectant, while Aliivibrio fischeri LMG 4414(T) should be freeze-dried in skim milk supplemented with marine broth in a 1:1 ratio. Freeze-drying Campylobacter fetus CIP 53.96(T) using skim milk supplemented with trehalose as lyoprotectant resulted in best recovery. Xanthomonas fragariae DSM 3587(T) expressed high viability after freeze-drying and storage for all tested lyoprotectants and could not be considered as recalcitrant. In contrary, Flavobacterium columnare LMG 10406(T) did not survive the freeze-drying process under all tested conditions.

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

PubMed

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

2008-06-01

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

Experimental analysis and modeling of ultrasound assisted freezing of potato spheres.

PubMed

Kiani, Hossein; Zhang, Zhihang; Sun, Da-Wen

2015-09-01

In recent years, innovative methods such as ultrasound assisted freezing have been developed in order to improve the freezing process. During freezing of foods, accurate prediction of the temperature distribution, phase ratios, and process time is very important. In the present study, ultrasound assisted immersion freezing process (in 1:1 ethylene glycol-water solution at 253.15K) of potato spheres (0.02 m diameter) was evaluated using experimental, numerical and analytical approaches. Ultrasound (25 kHz, 890 W m(-2)) was irradiated for different duty cycles (DCs=0-100%). A finite volume based enthalpy method was used in the numerical model, based on which temperature and liquid fraction profiles were simulated by a program developed using OpenFOAM® CFD software. An analytical technique was also employed to calculate freezing times. The results showed that ultrasound irradiation could decrease the characteristic freezing time of potatoes. Since ultrasound irradiation increased the heat transfer coefficient but simultaneously generated heat at the surface of the samples, an optimum DC was needed for the shortest freezing time which occurred in the range of 30-70% DC. DCs higher than 70% increased the freezing time. DCs lower than 30% did not provide significant effects on the freezing time compared to the control sample. The numerical model predicted the characteristic freezing time in accordance with the experimental results. In addition, analytical calculation of characteristic freezing time exhibited qualitative agreement with the experimental results. As the numerical simulations provided profiles of temperature and water fraction within potatoes frozen with or without ultrasound, the models can be used to study and control different operation situations, and to improve the understanding of the freezing process. Copyright © 2015 Elsevier B.V. All rights reserved.

Numerical simulation of the effect of groundwater salinity on artificial freezing wall in coastal area

NASA Astrophysics Data System (ADS)

Hu, Rui; Liu, Quan

2017-04-01

During the engineering projects with artificial ground freezing (AFG) techniques in coastal area, the freezing effect is affected by groundwater salinity. Based on the theories of artificially frozen soil and heat transfer in porous material, and with the assumption that only the variations of total dissolved solids (TDS) impact on freezing point and thermal conductivity, a numerical model of an AFG project in a saline aquifer was established and validated by comparing the simulated temperature field with the calculated temperature based on the analytic solution of rupak (reference) for single-pipe freezing temperature field T. The formation and development of freezing wall were simulated with various TDS. The results showed that the variety of TDS caused the larger temperature difference near the frozen front. With increasing TDS in the saline aquifer (1 35g/L), the average thickness of freezing wall decreased linearly and the total formation time of the freezing wall increased linearly. Compared with of the scenario of fresh-water (<1g/L), the average thickness of frozen wall decreased by 6% and the total formation time of the freezing wall increased by 8% with each increasing TDS of 7g/L. Key words: total dissolved solids, freezing point, thermal conductivity, freezing wall, numerical simulation Reference D.J.Pringel, H.Eicken, H.J.Trodahl, etc. Thermal conductivity of landfast Antarctic and Arctic sea ice[J]. Journal of Geophysical Research, 2007, 112: 1-13. Lukas U.Arenson, Dave C.Sego. The effect of salinity on the freezing of coarse- grained sand[J]. Canadian Geotechnical Journal, 2006, 43: 325-337. Hui Bing, Wei Ma. Laboratory investigation of the freezing point of saline soil[J]. Cold Regions Science and Technology, 2011, 67: 79-88.

Mechanisms of deterioration of nutrients phase 1

NASA Technical Reports Server (NTRS)

Karel, M.; Flink, J. M.

1972-01-01

Experimental methods are studied by which freeze-dried foods of improved quality are produced. Considered are: (1) Factors effecting the loss of butanol from frozen aqueous food solutions during storage; (2) a freeze-drying microscope system for observing solidification processes in organic mixtures and aqueous inorganic salt solutions; (3) browning of high quality freeze-dried foods with minimal organoleptic and nutritional detoriation; (4) retention of PVP-n-propanol in freeze-dried food models; and (5) effects of freezing rate and sucrose immersion on taste and texture of freeze-dried apple slices.

Effect of moisture content on the invertase activity of freeze-dried S. cerevisiae.

PubMed

Pitombo, R N; Spring, C; Passos, R F; Tonato, M; Vitolo, M

1994-08-01

The invertase activity of intact Saccharomyces cerevisiae submitted to freezing-thawing was affected by pH, the chemical nature of the buffer, and the freezing cooling rate (CR), leading in some cases to a complete invertase inactivation (acetate buffer, pH 4.0, CR = 0.5 degree C/min). Once established under adequate freezing conditions the invertase activity remained unchanged after freeze-drying. Nevertheless, in some cases the cell-growing capability after freeze-drying diminished around 70%, mainly if the frozen cell suspension was attained through freezing carried out at CR = 0.5 degree C/min. Water sorption isotherms of freeze-dried samples (freeze-dryer Edwards L-4KR; 30 degrees C and 0.1 mB) were determined at 10 and 25 degrees C. The monolayer moisture content (MMC) at each temperature (12.7 and 3.71 for 10 and 25 degrees C, respectively) was calculated from isotherms by applying BET and GAB models. Freeze-dried yeast with water activity (Aw) between 0 and 0.33 (about the MMC value) maintained at 25 degrees C for 235 days and at 89 degrees C for 15 min retained at least 85% of its original invertase activity (IA), whereas samples with Aw > MMC lost at least 60% of its IA. X ray diffraction showed that the freeze-dried cake before and after storage presented an amorphous structure.

Ice-Binding Proteins in Plants

PubMed Central

Bredow, Melissa; Walker, Virginia K.

2017-01-01

Sub-zero temperatures put plants at risk of damage associated with the formation of ice crystals in the apoplast. Some freeze-tolerant plants mitigate this risk by expressing ice-binding proteins (IBPs), that adsorb to ice crystals and modify their growth. IBPs are found across several biological kingdoms, with their ice-binding activity and function uniquely suited to the lifestyle they have evolved to protect, be it in fishes, insects or plants. While IBPs from freeze-avoidant species significantly depress the freezing point, plant IBPs typically have a reduced ability to lower the freezing temperature. Nevertheless, they have a superior ability to inhibit the recrystallization of formed ice. This latter activity prevents ice crystals from growing larger at temperatures close to melting. Attempts to engineer frost-hardy plants by the controlled transfer of IBPs from freeze-avoiding fish and insects have been largely unsuccessful. In contrast, the expression of recombinant IBP sequences from freeze-tolerant plants significantly reduced electrolyte leakage and enhanced freezing survival in freeze-sensitive plants. These promising results have spurred additional investigations into plant IBP localization and post-translational modifications, as well as a re-evaluation of IBPs as part of the anti-stress and anti-pathogen axis of freeze-tolerant plants. Here we present an overview of plant freezing stress and adaptation mechanisms and discuss the potential utility of IBPs for the generation of freeze-tolerant crops. PMID:29312400

Ice-Binding Proteins in Plants.

PubMed

Bredow, Melissa; Walker, Virginia K

2017-01-01

Sub-zero temperatures put plants at risk of damage associated with the formation of ice crystals in the apoplast. Some freeze-tolerant plants mitigate this risk by expressing ice-binding proteins (IBPs), that adsorb to ice crystals and modify their growth. IBPs are found across several biological kingdoms, with their ice-binding activity and function uniquely suited to the lifestyle they have evolved to protect, be it in fishes, insects or plants. While IBPs from freeze-avoidant species significantly depress the freezing point, plant IBPs typically have a reduced ability to lower the freezing temperature. Nevertheless, they have a superior ability to inhibit the recrystallization of formed ice. This latter activity prevents ice crystals from growing larger at temperatures close to melting. Attempts to engineer frost-hardy plants by the controlled transfer of IBPs from freeze-avoiding fish and insects have been largely unsuccessful. In contrast, the expression of recombinant IBP sequences from freeze-tolerant plants significantly reduced electrolyte leakage and enhanced freezing survival in freeze-sensitive plants. These promising results have spurred additional investigations into plant IBP localization and post-translational modifications, as well as a re-evaluation of IBPs as part of the anti-stress and anti-pathogen axis of freeze-tolerant plants. Here we present an overview of plant freezing stress and adaptation mechanisms and discuss the potential utility of IBPs for the generation of freeze-tolerant crops.

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

PubMed Central

2015-01-01

The ultrastructure in the beef muscle of the electro-magnetic resonance and air blast freezing during the frozen storage, and the changes in the quality characteristics after thawing were evaluated. The size of ice crystal was small and evenly formed in the initial freezing period, and it showed that the size was increased as the storage period was elapsed (p<0.05). The beef stored by the electro-magnetic resonance freezing showed the size of ice crystal with a lower rate of increase than the air blast freezing during the frozen storage. The thawing loss of beef stored by the electro-magnetic resonance freezing was significantly lower than the air blast freezing during frozen storage (p<0.05), and it showed that the thawing loss of the round was higher than the loin. Water holding capacity decreased as the storage period became longer while the electro-magnetic resonance freezing was higher than the air blast on 8 month (p<0.05). As a result of sensory evaluation, the beef stored by the electro-magnetic resonance freezing did not show the difference until 4 months, and it showed higher acceptability in comparison with the beef stored by the air blast freezing. Thus, it is considered that the freezing method has an effect on the change in the ultrastructure and quality characteristics of the beef. PMID:26761797

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

Fish antifreeze protein and the freezing and recrystallization of ice.

PubMed

Knight, C A; DeVries, A L; Oolman, L D

Antifreeze glycopeptide and peptides from the blood of polar fishes prevent the growth of ice crystals in water at temperatures down to approximately 1 degree C below freezing point, but do not appreciably influence the equilibrium freezing point. This freezing point hysteresis must be a disequilibrium effect, or it would violate Gibbs' phase rule, but the separate freezing and melting points are experimentally very definite: ice neither melts nor freezes perceptibly within the 'hysteresis gap', for periods of hours or days. We report here unusual crystal faces on ice crystals grown from solutions of very low concentrations of the anti-freeze glycopeptides and peptides. This is a clue to the mechanism of freezing inhibition, and it may be the basis of a simple, very sensitive test for antifreeze material. Very low concentrations of the antifreeze protein are also remarkably effective in preventing the recrystallization of ice.

Optimizing performance by improving core stability and core strength.

PubMed

Hibbs, Angela E; Thompson, Kevin G; French, Duncan; Wrigley, Allan; Spears, Iain

2008-01-01

Core stability and core strength have been subject to research since the early 1980s. Research has highlighted benefits of training these processes for people with back pain and for carrying out everyday activities. However, less research has been performed on the benefits of core training for elite athletes and how this training should be carried out to optimize sporting performance. Many elite athletes undertake core stability and core strength training as part of their training programme, despite contradictory findings and conclusions as to their efficacy. This is mainly due to the lack of a gold standard method for measuring core stability and strength when performing everyday tasks and sporting movements. A further confounding factor is that because of the differing demands on the core musculature during everyday activities (low load, slow movements) and sporting activities (high load, resisted, dynamic movements), research performed in the rehabilitation sector cannot be applied to the sporting environment and, subsequently, data regarding core training programmes and their effectiveness on sporting performance are lacking. There are many articles in the literature that promote core training programmes and exercises for performance enhancement without providing a strong scientific rationale of their effectiveness, especially in the sporting sector. In the rehabilitation sector, improvements in lower back injuries have been reported by improving core stability. Few studies have observed any performance enhancement in sporting activities despite observing improvements in core stability and core strength following a core training programme. A clearer understanding of the roles that specific muscles have during core stability and core strength exercises would enable more functional training programmes to be implemented, which may result in a more effective transfer of these skills to actual sporting activities.

'Scaling-up is a craft not a science': Catalysing scale-up of health innovations in Ethiopia, India and Nigeria.

PubMed

Spicer, Neil; Bhattacharya, Dipankar; Dimka, Ritgak; Fanta, Feleke; Mangham-Jefferies, Lindsay; Schellenberg, Joanna; Tamire-Woldemariam, Addis; Walt, Gill; Wickremasinghe, Deepthi

2014-11-01

Donors and other development partners commonly introduce innovative practices and technologies to improve health in low and middle income countries. Yet many innovations that are effective in improving health and survival are slow to be translated into policy and implemented at scale. Understanding the factors influencing scale-up is important. We conducted a qualitative study involving 150 semi-structured interviews with government, development partners, civil society organisations and externally funded implementers, professional associations and academic institutions in 2012/13 to explore scale-up of innovative interventions targeting mothers and newborns in Ethiopia, the Indian state of Uttar Pradesh and the six states of northeast Nigeria, which are settings with high burdens of maternal and neonatal mortality. Interviews were analysed using a common analytic framework developed for cross-country comparison and themes were coded using Nvivo. We found that programme implementers across the three settings require multiple steps to catalyse scale-up. Advocating for government to adopt and finance health innovations requires: designing scalable innovations; embedding scale-up in programme design and allocating time and resources; building implementer capacity to catalyse scale-up; adopting effective approaches to advocacy; presenting strong evidence to support government decision making; involving government in programme design; invoking policy champions and networks; strengthening harmonisation among external programmes; aligning innovations with health systems and priorities. Other steps include: supporting government to develop policies and programmes and strengthening health systems and staff; promoting community uptake by involving media, community leaders, mobilisation teams and role models. We conclude that scale-up has no magic bullet solution - implementers must embrace multiple activities, and require substantial support from donors and governments in doing so. Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.

Lessons learned from stakeholder-driven sustainability analysis of six national HIV programmes.

PubMed

Katz, Itamar; Glandon, Douglas; Wong, Wendy; Kargbo, Brima; Ombam, Regina; Singh, Shanti; Ramsammy, Leslie; Tal-Dia, Anta; Seck, Ibrahima; Osika, John S

2014-05-01

In light of the decline in donor HIV funding, HIV programmes increasingly need to assess their available and potential resources and maximize their utilization. This article presents lessons learned related to how countries have addressed the sustainability of HIV programmes in a stakeholder-driven sustainability analysis. During HIV/AIDS Programme Sustainability Analysis Tool (HAPSAT) applications in six countries (Benin, Guyana, Kenya, Lesotho, Sierra Leone and South Sudan), stakeholders identified key sustainability challenges for their HIV responses. Possible policy approaches were prepared, and those related to prioritization and resource mobilization are analysed in this article. The need to prioritize evidence-based interventions and apply efficiency measures is being accepted by countries. Five of the six countries in this study requested that the HAPSAT team prepare 'prioritization' strategies. Countries recognize the need to prepare for an alternative to 'universal access by 2015', acknowledging that their capacity might be insufficient to reach such high-coverage levels by then. There is further acceptance of the importance of reaching the most-at-risk, marginalized populations, as seen, for example, in South Sudan and Sierra Leone. However, the pace at which resources are shifting towards these populations is slow. Finally, only two of the six countries, Kenya and Benin, chose to examine options for generating additional financial resources beyond donor funding. In Kenya, three non-donor sources were recommended, yet even if all were to be implemented, it would cover only 25% of the funding needed. Countries are increasingly willing to address the challenges of HIV programme sustainability, yet in different ways and with varying urgency. To secure achievements made to date and maximize future impact, countries would benefit from strengthening their strategic plans, operational plans and funding proposals with concrete timelines and responsibilities for addressing sustainability issues.

Increased spring freezing vulnerability for alpine shrubs under early snowmelt.

PubMed

Wheeler, J A; Hoch, G; Cortés, A J; Sedlacek, J; Wipf, S; Rixen, C

2014-05-01

Alpine dwarf shrub communities are phenologically linked with snowmelt timing, so early spring exposure may increase risk of freezing damage during early development, and consequently reduce seasonal growth. We examined whether environmental factors (duration of snow cover, elevation) influenced size and the vulnerability of shrubs to spring freezing along elevational gradients and snow microhabitats by modelling the past frequency of spring freezing events. We sampled biomass and measured the size of Salix herbacea, Vaccinium myrtillus, Vaccinium uliginosum and Loiseleuria procumbens in late spring. Leaves were exposed to freezing temperatures to determine the temperature at which 50% of specimens are killed for each species and sampling site. By linking site snowmelt and temperatures to long-term climate measurements, we extrapolated the frequency of spring freezing events at each elevation, snow microhabitat and per species over 37 years. Snowmelt timing was significantly driven by microhabitat effects, but was independent of elevation. Shrub growth was neither enhanced nor reduced by earlier snowmelt, but decreased with elevation. Freezing resistance was strongly species dependent, and did not differ along the elevation or snowmelt gradient. Microclimate extrapolation suggested that potentially lethal freezing events (in May and June) occurred for three of the four species examined. Freezing events never occurred on late snow beds, and increased in frequency with earlier snowmelt and higher elevation. Extrapolated freezing events showed a slight, non-significant increase over the 37-year record. We suggest that earlier snowmelt does not enhance growth in four dominant alpine shrubs, but increases the risk of lethal spring freezing exposure for less freezing-resistant species.

Controlled crystallization of the lipophilic drug fenofibrate during freeze-drying: elucidation of the mechanism by in-line Raman spectroscopy.

PubMed

de Waard, Hans; De Beer, Thomas; Hinrichs, Wouter L J; Vervaet, Chris; Remon, Jean-Paul; Frijlink, Henderik W

2010-12-01

We developed a novel process, "controlled crystallization during freeze-drying" to produce drug nanocrystals of poorly water-soluble drugs. This process involves freeze-drying at a relatively high temperature of a drug and a matrix material from a mixture of tertiary butyl alcohol and water, resulting in drug nanocrystals incorporated in a matrix. The aim of this study was to elucidate the mechanisms that determine the size of the drug crystals. Fenofibrate was used as a model lipophilic drug. To monitor the crystallization during freeze-drying, a Raman probe was placed just above the sample in the freeze-dryer. These in-line Raman spectroscopy measurements clearly revealed when the different components crystallized during freeze-drying. The solvents crystallized only during the freezing step, while the solutes only crystallized after the temperature was increased, but before drying started. Although the solutes crystallized only after the freezing step, both the freezing rate and the shelf temperature were critical parameters that determined the final crystal size. At a higher freezing rate, smaller interstitial spaces containing the freeze-concentrated fraction were formed, resulting in smaller drug crystals (based on dissolution data). On the other hand, when the solutes crystallized at a lower shelf temperature, the degree of supersaturation is higher, resulting in a higher nucleation rate and consequently more and therefore smaller crystals. In conclusion, for the model drug fenofibrate, a high freezing rate and a relatively low crystallization temperature resulted in the smallest crystals and therefore the highest dissolution rate.

Effect of three cueing devices for people with Parkinson's disease with gait initiation difficulties.

PubMed

McCandless, Paula J; Evans, Brenda J; Janssen, Jessie; Selfe, James; Churchill, Andrew; Richards, Jim

2016-02-01

Freezing of gait (FOG) remains one of the most common debilitating aspects of Parkinson's disease and has been linked to injuries, falls and reduced quality of life. Although commercially available portable cueing devices exist claiming to assist with overcoming freezing; their immediate effectiveness in overcoming gait initiation failure is currently unknown. This study investigated the effects of three different types of cueing device in people with Parkinson's disease who experience freezing. Twenty participants with idiopathic Parkinson's disease who experienced freezing during gait but who were able to walk short distances indoors independently were recruited. At least three attempts at gait initiation were recorded using a 10 camera Qualisys motion analysis system and four force platforms. Test conditions were; Laser Cane, sound metronome, vibrating metronome, walking stick and no intervention. During testing 12 of the 20 participants had freezing episodes, from these participants 100 freezing and 91 non-freezing trials were recorded. Clear differences in the movement patterns were seen between freezing and non-freezing episodes. The Laser Cane was most effective cueing device at improving the forwards/backwards and side to side movement and had the least number of freezing episodes. The walking stick also showed significant improvements compared to the other conditions. The vibration metronome appeared to disrupt movement compared to the sound metronome at the same beat frequency. This study identified differences in the movement patterns between freezing episodes and non-freezing episodes, and identified immediate improvements during gait initiation when using the Laser Cane over the other interventions. Copyright © 2015. Published by Elsevier B.V.

Role of glucose-6-phosphate dehydrogenase in freezing-induced freezing resistance of Populus suaveolens.

PubMed

Lin, Shan-Zhi; Zhang, Zhi-Yi; Liu, Wen-Feng; Lin, Yuan-Zhen; Zhang, Qian; Zhu, Bao-Qing

2005-02-01

To explore the role of glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in the enhancement of freezing resistance induced by freezing acclimation, G6PDH was purified from the leaves of 8-week-old Populus suaveolens cuttings. The G6PDH activity in the absence or the presence of reduced dithiothreitol (DTT(red)) were determined, and the changes in superoxide dismutase (SOD), peroxides (POD) and cytosolic G6PDH activities, malondial-dehyde (MDA) content as well as freezing resistance (expressed as LT(50)) of P. suaveolens cuttings during freezing acclimation at -20 degrees C were investigated. The results showed that the purified G6PDH was probably located in the cytosol of P. suaveolens. Freezing acclimation increased the activities of SOD, POD and cytosolic G6PDH, and decreased the MDA content and LT(50) of cuttings, while 2 d of de-acclimation at 25 degrees C resulted in a decrease in SOD, POD and cytosolic G6PDH activities, and caused an increase in MDA content and LT(50). The change in cytosolic G6PDH activity was found to be closely correlated to the levels of SOD, POD and MDA, and to the degree of freezing resistance of cuttings during freezing acclimation. It is suggested that the enhancement of freezing resistance of cuttings induced by freezing acclimation is related to the distinct increase in cytosolic G6PDH activity, which may be involved in the activation of SOD and POD, and the induction of freezing resistance of cuttings.

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

PubMed Central

Liu, Mei-Zhen; Osborne, Colin P.

2013-01-01

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

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

Contact freezing of supercooled cloud droplets on collision with mineral dust particles: effect of particle size

NASA Astrophysics Data System (ADS)

Hoffmann, Nadine; Duft, Denis; Kiselev, Alexei; Leisner, Thomas

2013-04-01

The contact freezing of supercooled cloud droplets is one of the potentially important and the least investigated heterogeneous mechanism of ice formation in the tropospheric clouds [1]. On the time scales of cloud lifetime the freezing of supercooled water droplets via contact mechanism may occur at higher temperature compared to the same IN immersed in the droplet. However, the laboratory experiments of contact freezing are very challenging due to the number of factors affecting the probability of ice formation. In our experiment we study single water droplets freely levitated in the laminar flow of mineral dust particles acting as the contact freezing nuclei. By repeating the freezing experiment sufficient number of times we are able to reproduce statistical freezing behavior of large ensembles of supercooled droplets and measure the average rate of freezing events. We show that the rate of freezing at given temperature is governed only by the rate of droplet -particle collision and by the properties of the contact ice nuclei. In this contribution we investigate the relationship between the freezing probability and the size of mineral dust particle (represented by illite) and show that their IN efficiency scales with the particle size. Based on this observation, we discuss the similarity between the freezing of supercooled water droplets in immersion and contact modes and possible mechanisms of apparent enhancement of the contact freezing efficiency. [1] - K.C. Young, The role of contact nucleation in ice phase initiation in clouds, Journal of the Atmospheric Sciences 31, 1974

Ice nucleation temperature influences recovery of activity of a model protein after freeze drying.

PubMed

Cochran, Teresa; Nail, Steven L

2009-09-01

The objective of this study was to determine whether a relationship exists between ice nucleation temperature and recovery of activity of a model protein, lactate dehydrogenase, after freeze drying. Aqueous buffer systems containing 50 microg/mL of protein were frozen in vials with externally mounted thermocouples on the shelf of a freeze dryer, then freeze dried. Various methods were used to establish a wide range of ice nucleation temperatures. An inverse relationship was found between the extent of supercooling during freezing and recovery of activity in the reconstituted solution. The data are consistent with a mechanism of inactivation resulting from adsorption of protein at the ice/freeze-concentrate interface during the freezing process.

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

DOT National Transportation Integrated Search

2015-09-01

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

Ovarian tissue cryopreservation by stepped vitrification and monitored by X-ray computed tomography.

PubMed

Corral, Ariadna; Clavero, Macarena; Gallardo, Miguel; Balcerzyk, Marcin; Amorim, Christiani A; Parrado-Gallego, Ángel; Dolmans, Marie-Madeleine; Paulini, Fernanda; Morris, John; Risco, Ramón

2018-04-01

Ovarian tissue cryopreservation is, in most cases, the only fertility preservation option available for female patients soon to undergo gonadotoxic treatment. To date, cryopreservation of ovarian tissue has been carried out by both traditional slow freezing method and vitrification, but even with the best techniques, there is still a considerable loss of follicle viability. In this report, we investigated a stepped cryopreservation procedure which combines features of slow cooling and vitrification (hereafter called stepped vitrification). Bovine ovarian tissue was used as a tissue model. Stepwise increments of the Me 2 SO concentration coupled with stepwise drops-in temperature in a device specifically designed for this purpose and X-ray computed tomography were combined to investigate loading times at each step, by monitoring the attenuation of the radiation proportional to Me 2 SO permeation. Viability analysis was performed in warmed tissues by immunohistochemistry. Although further viability tests should be conducted after transplantation, preliminary results are very promising. Four protocols were explored. Two of them showed a poor permeation of the vitrification solution (P1 and P2). The other two (P3 and P4), with higher permeation, were studied in deeper detail. Out of these two protocols, P4, with a longer permeation time at -40 °C, showed the same histological integrity after warming as fresh controls. Copyright © 2018 Elsevier Inc. All rights reserved.

Cognitive Contributions to Freezing of Gait in Parkinson Disease: Implications for Physical Rehabilitation

PubMed Central

King, Laurie A.; Cohen, Rajal G.; Horak, Fay B.

2016-01-01

People with Parkinson disease (PD) who show freezing of gait also have dysfunction in cognitive domains that interact with mobility. Specifically, freezing of gait is associated with executive dysfunction involving response inhibition, divided attention or switching attention, and visuospatial function. The neural control impairments leading to freezing of gait have recently been attributed to higher-level, executive and attentional cortical processes involved in coordinating posture and gait rather than to lower-level, sensorimotor impairments. To date, rehabilitation for freezing of gait primarily has focused on compensatory mobility training to overcome freezing events, such as sensory cueing and voluntary step planning. Recently, a few interventions have focused on restitutive, rather than compensatory, therapy. Given the documented impairments in executive function specific to patients with PD who freeze and increasing evidence of overlap between cognitive and motor function, incorporating cognitive challenges with mobility training may have important benefits for patients with freezing of gait. Thus, a novel theoretical framework is proposed for exercise interventions that jointly address both the specific cognitive and mobility challenges of people with PD who freeze. PMID:26381808

Cognitive Contributions to Freezing of Gait in Parkinson Disease: Implications for Physical Rehabilitation.

PubMed

Peterson, Daniel S; King, Laurie A; Cohen, Rajal G; Horak, Fay B

2016-05-01

People with Parkinson disease (PD) who show freezing of gait also have dysfunction in cognitive domains that interact with mobility. Specifically, freezing of gait is associated with executive dysfunction involving response inhibition, divided attention or switching attention, and visuospatial function. The neural control impairments leading to freezing of gait have recently been attributed to higher-level, executive and attentional cortical processes involved in coordinating posture and gait rather than to lower-level, sensorimotor impairments. To date, rehabilitation for freezing of gait primarily has focused on compensatory mobility training to overcome freezing events, such as sensory cueing and voluntary step planning. Recently, a few interventions have focused on restitutive, rather than compensatory, therapy. Given the documented impairments in executive function specific to patients with PD who freeze and increasing evidence of overlap between cognitive and motor function, incorporating cognitive challenges with mobility training may have important benefits for patients with freezing of gait. Thus, a novel theoretical framework is proposed for exercise interventions that jointly address both the specific cognitive and mobility challenges of people with PD who freeze. © 2016 American Physical Therapy Association.

Experiments with a homologous, inactivated canine parvovirus vaccine in vaccination programmers for dogs.

PubMed

Wilson, J H; Hermann-Dekkers, W M

1982-01-01

The significance of canine parvovirus (CPV) infections as a permanent threat susceptible dogs, in particular pups, made the authors develop three liquid homologous inactivated adjuvant CPV vaccines that were compatible with existing canine vaccines and could be incorporated in current vaccination programmes. On vaccine (Kavak Parvo) contained only the CPV component, the second product (Kavak i-LP) also contained two inactivated leptospiral antigens, and the third vaccine (Kavak i-HLP) contained in addition an inactivated canine hepatitis virus. This paper reports on the studies conducted to test the safety and efficacy of the three products. They were used as such and as diluents for freeze dried vaccines containing live attenuated measles, distemper, and hepatitis viruses. The study was performed in a breeding kennel where all dogs were free from CPV antibodies and the nonvaccinated sentinels remained so for the course of the study. All vaccines proved to be safe in dogs of all ages, including pregnant bitches. The efficacy of the CPV component was studied both by monitoring antibody titres for more than a year and by challenge exposure of some dogs to virulent CPV. The results obtained from these studies prove that the CPV component used in the three vaccines can be incorporated as indicated in the recommended canine vaccination programmes. The observations that the inactivated CPV and hepatitis components do induce an active immunity in pups that are still protected by low levels of maternally derived antibodies against these viruses, make those vaccines very suitable in breeding kennels. Additional studies on a comparative basis are being continued in edemically CPV infected breeding kennels to quantify the significance of these observations in these special conditions.

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.

N-acetyltransferase gene polymorphisms & plasma isoniazid concentrations in patients with tuberculosis

PubMed Central

Hemanth Kumar, A. K.; Ramesh, K.; Kannan, T.; Sudha, V.; Haribabu, Hemalatha; Lavanya, J.; Swaminathan, Soumya; Ramachandran, Geetha

2017-01-01

Background & objectives: Variations in the N-acetyltransferase (NAT2) gene among different populations could affect the metabolism and disposition of isoniazid (INH). This study was performed to genotype NAT2 gene polymorphisms in tuberculosis (TB) patients from Chennai, India, and compare plasma INH concentrations among the different genotypes. Methods: Adult patients with TB treated in the Revised National TB Control Programme (RNTCP) in Chennai, Tamil Nadu, were genotyped for NAT2 gene polymorphism, and two-hour post-dosing INH concentrations were compared between the different genotypes. Plasma INH was determined by high-performance liquid chromatography. Genotyping of the NAT2 gene polymorphism was performed by real-time polymerase chain reaction method. Results: Among the 326 patients genotyped, there were 189 (58%), 114 (35%) and 23 (7%) slow, intermediate and fast acetylators, respectively. The median two-hour INH concentrations in slow, intermediate and fast acetylators were 10.2, 8.1 and 4.1 μg/ml, respectively. The differences in INH concentrations among the three genotypes were significant (P<0.001). Interpretation & conclusions: Genotyping of TB patients from south India for NAT2 gene polymorphism revealed that 58 per cent of the study population comprised slow acetylators. Two-hour INH concentrations differed significantly among the three genotypes. PMID:28574024

The single nucleotide polymorphism Gly482Ser in the PGC-1α gene impairs exercise-induced slow-twitch muscle fibre transformation in humans.

PubMed

Steinbacher, Peter; Feichtinger, René G; Kedenko, Lyudmyla; Kedenko, Igor; Reinhardt, Sandra; Schönauer, Anna-Lena; Leitner, Isabella; Sänger, Alexandra M; Stoiber, Walter; Kofler, Barbara; Förster, Holger; Paulweber, Bernhard; Ring-Dimitriou, Susanne

2015-01-01

PGC-1α (peroxisome proliferator-activated receptor γ co-activator 1α) is an important regulator of mitochondrial biogenesis and a master regulator of enzymes involved in oxidative phosphorylation. Recent evidence demonstrated that the Gly482Ser single nucleotide polymorphism (SNP) in the PGC-1α gene affects insulin sensitivity, blood lipid metabolism and binding to myocyte enhancer factor 2 (MEF2). Individuals carrying this SNP were shown to have a reduced cardiorespiratory fitness and a higher risk to develop type 2 diabetes. Here, we investigated the responses of untrained men with the Gly482Ser SNP to a 10 week programme of endurance training (cycling, 3 x 60 min/week, heart rate at 70-90% VO2peak). Quantitative data from analysis of biopsies from vastus lateralis muscle revealed that the SNP group, in contrast to the control group, lacked a training-induced increase in content of slow contracting oxidative fibres. Capillary supply, mitochondrial density, mitochondrial enzyme activities and intramyocellular lipid content increased similarly in both groups. These results indicate that the impaired binding of MEF2 to PGC-1α in humans with this SNP impedes exercise-induced fast-to-slow muscle fibre transformation.

Muscle atrophy associated with microgravity in rat: Basic data for countermeasures

NASA Astrophysics Data System (ADS)

Falempin, M.; Mounier, Y.

Morphological, contractile properties and myosin heavy chain (MHC) composition of rat soleus muscles were studied after 2 weeks of unloading (HS) and after 2 weeks of HS associated with selective deafferentation (HS + DEAF) at the level L4 and L5. The same significant reductions in muscle mass and tetanic tension were found after HS and HS + DEAF. However, the transformation of the slow-twitch soleus muscle towards a faster type characterized by a decrease in twitch time parameters and an increase in fast-twitch type MHC isoforms in HS did not appear in HS + DEAF conditions. Our results also showed that a pattern similar to firing rate of motoneurones innervating slow-twitch muscles inhibited the slow to fast fiber changes observed during HS. Nevertheless, neither the loss of mass or force output in the HS muscles were prevented by electrostimulation. Immobilization in a stretched position during HS maintained the muscle wet weight, mechanical and electrophoretical characteristics close to control values. We concluded that the decrease in mechanical strains imposed on the muscle during unloading was the main factor for the development of atrophy, while the kinetic changes might be predominantly modulated by the nervous command. These basic data suggested that some experimental conditions such as electrostimulation or stretching, could participate in countermeasure programmes.

The Single Nucleotide Polymorphism Gly482Ser in the PGC-1α Gene Impairs Exercise-Induced Slow-Twitch Muscle Fibre Transformation in Humans

PubMed Central

Steinbacher, Peter; Feichtinger, René G.; Kedenko, Lyudmyla; Kedenko, Igor; Reinhardt, Sandra; Schönauer, Anna-Lena; Leitner, Isabella; Sänger, Alexandra M.; Stoiber, Walter; Kofler, Barbara; Förster, Holger; Paulweber, Bernhard; Ring-Dimitriou, Susanne

2015-01-01

PGC-1α (peroxisome proliferator-activated receptor γ co-activator 1α) is an important regulator of mitochondrial biogenesis and a master regulator of enzymes involved in oxidative phosphorylation. Recent evidence demonstrated that the Gly482Ser single nucleotide polymorphism (SNP) in the PGC-1α gene affects insulin sensitivity, blood lipid metabolism and binding to myocyte enhancer factor 2 (MEF2). Individuals carrying this SNP were shown to have a reduced cardiorespiratory fitness and a higher risk to develop type 2 diabetes. Here, we investigated the responses of untrained men with the Gly482Ser SNP to a 10 week programme of endurance training (cycling, 3 x 60 min/week, heart rate at 70-90% VO2peak). Quantitative data from analysis of biopsies from vastus lateralis muscle revealed that the SNP group, in contrast to the control group, lacked a training-induced increase in content of slow contracting oxidative fibres. Capillary supply, mitochondrial density, mitochondrial enzyme activities and intramyocellular lipid content increased similarly in both groups. These results indicate that the impaired binding of MEF2 to PGC-1α in humans with this SNP impedes exercise-induced fast-to-slow muscle fibre transformation. PMID:25886402

Representative Values of Icing-Related Variables Aloft in Freezing Rain and Freezing Drizzle

DOT National Transportation Integrated Search

1996-03-01

Radiosonde and surface observations in freezing rain (ZR) and freezing drizzle (ZL), and a limited number of aircraft measurements in ZR, have been examined for information on the magnitude and altitude dependence of meteorological variables associat...

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

EPA Science Inventory

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

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

Demonstrating Functional Equivalence of Pilot and Production Scale Freeze-Drying of BCG

PubMed Central

ten Have, R.; Reubsaet, K.; van Herpen, P.; Kersten, G.; Amorij, J.-P.

2016-01-01

Process analytical technology (PAT)-tools were used to monitor freeze-drying of Bacille Calmette-Guérin (BCG) at pilot and production scale. Among the evaluated PAT-tools, there is the novel use of the vacuum valve open/close frequency for determining the endpoint of primary drying at production scale. The duration of primary drying, the BCG survival rate, and the residual moisture content (RMC) were evaluated using two different freeze-drying protocols and were found to be independent of the freeze-dryer scale evidencing functional equivalence. The absence of an effect of the freeze-dryer scale on the process underlines the feasibility of the pilot scale freeze-dryer for further BCG freeze-drying process optimization which may be carried out using a medium without BCG. PMID:26981867

Demonstrating Functional Equivalence of Pilot and Production Scale Freeze-Drying of BCG.

PubMed

Ten Have, R; Reubsaet, K; van Herpen, P; Kersten, G; Amorij, J-P

2016-01-01

Process analytical technology (PAT)-tools were used to monitor freeze-drying of Bacille Calmette-Guérin (BCG) at pilot and production scale. Among the evaluated PAT-tools, there is the novel use of the vacuum valve open/close frequency for determining the endpoint of primary drying at production scale. The duration of primary drying, the BCG survival rate, and the residual moisture content (RMC) were evaluated using two different freeze-drying protocols and were found to be independent of the freeze-dryer scale evidencing functional equivalence. The absence of an effect of the freeze-dryer scale on the process underlines the feasibility of the pilot scale freeze-dryer for further BCG freeze-drying process optimization which may be carried out using a medium without BCG.

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.

MAL62 overexpression and NTH1 deletion enhance the freezing tolerance and fermentation capacity of the baker's yeast in lean dough.

PubMed

Sun, Xi; Zhang, Cui-Ying; Wu, Ming-Yue; Fan, Zhi-Hua; Liu, Shan-Na; Zhu, Wen-Bi; Xiao, Dong-Guang

2016-04-04

Trehalose is related to several types of stress responses, especially freezing response in baker's yeast (Saccharomyces cerevisiae). It is desirable to manipulate trehalose-related genes to create yeast strains that better tolerate freezing-thaw stress with improved fermentation capacity, which are in high demand in the baking industry. The strain overexpressing MAL62 gene showed increased trehalose content and cell viability after prefermention-freezing and long-term frozen. Deletion of NTH1 in combination of MAL62 overexpression further strengthens freezing tolerance and improves the leavening ability after freezing-thaw stress. The mutants of the industrial baker's yeast with enhanced freezing tolerance and leavening ability in lean dough were developed by genetic engineering. These strains had excellent potential industrial applications.

Freeze/Thaw-Induced Embolism: Probability of Critical Bubble Formation Depends on Speed of Ice Formation

DOE PAGES

Sevanto, Sanna; Holbrook, N. Michele; Ball, Marilyn C.

2012-06-06

Bubble formation in the conduits of woody plants sets a challenge for uninterrupted water transportation from the soil up to the canopy. Freezing and thawing of stems has been shown to increase the number of air-filled (embolized) conduits, especially in trees with large conduit diameters. Despite numerous experimental studies, the mechanisms leading to bubble formation during freezing have not been addressed theoretically. We used classical nucleation theory and fluid mechanics to show which mechanisms are most likely to be responsible for bubble formation during freezing and what parameters determine the likelihood of the process. Our results confirm the common assumptionmore » that bubble formation during freezing is most likely due to gas segregation by ice. If xylem conduit walls are not permeable to the salts expelled by ice during the freezing process, osmotic pressures high enough for air seeding could be created. The build-up rate of segregated solutes in front of the ice-water interface depends equally on conduit diameter and freezing velocity. Therefore, bubble formation probability depends on these variables. The dependence of bubble formation probability on freezing velocity means that the experimental results obtained for cavitation threshold conduit diameters during freeze/thaw cycles depend on the experimental setup; namely sample size and cooling rate. The velocity dependence also suggests that to avoid bubble formation during freezing trees should have narrow conduits where freezing is likely to be fast (e.g., branches or outermost layer of the xylem). Avoidance of bubble formation during freezing could thus be one piece of the explanation why xylem conduit size of temperate and boreal zone trees varies quite systematically.« less

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

Rapid freezing without cooling equilibration in canine sperm.

PubMed

Kim, Suhee; Lee, Yongcheol; Yang, Honghyun; Kim, Yong-Jun

2012-01-01

The aim of this study was to develop a rapid method of canine semen freezing without cooling equilibration using treatment with different cryoprotectant agents (CPAs) and freezing in liquid nitrogen (LN(2)) vapor in a 0.5-mL straw via modifying vitrification. Ejaculates from eight beagle dogs were frozen with different CPAs (CPA-free, 5% glycerol, 5% ethylene glycol, and 10% ethylene glycol) and freezing times (direct plunging into LN(2) or freezing for 1, 2, 3, or 10 min in LN(2) vapor before plunging into LN(2)). Frozen-thawed sperm were evaluated for motility, viability, normal morphology, and plasma- and acrosome-membrane integrities. The 5% glycerol treatment resulted in improved sperm motility, plasma-membrane integrity and acrosome-membrane integrity (P<0.05). Freezing in LN(2) vapor showed improved sperm motility, viability, and plasma membrane integrity (P<0.05), and freezing for more than 2 min in LN(2) vapor increased acrosome-membrane integrity compared with direct plunging into LN(2) (P<0.05). The direct plunging into LN(2) showed no motile sperm. However, freezing for more than 2 min in LN(2) vapor increased the total abnormalities compared to direct plunging into LN(2) (P<0.05). In conclusion, use of 5% glycerol and freezing in LN(2) vapor were essential for the rapid freezing of canine sperm without cooling equilibration. In particular, holding for 2 min in LN(2) vapor was sufficient to yield successful rapid freezing. This rapid freezing method is simple and effective in canine sperm and would be helpful to offer information for trial of vitrification in large volumes of canine sperm. Copyright © 2012 Elsevier B.V. All rights reserved.

Freeze/Thaw-induced embolism: probability of critical bubble formation depends on speed of ice formation.

PubMed

Sevanto, Sanna; Holbrook, N Michele; Ball, Marilyn C

2012-01-01

Bubble formation in the conduits of woody plants sets a challenge for uninterrupted water transportation from the soil up to the canopy. Freezing and thawing of stems has been shown to increase the number of air-filled (embolized) conduits, especially in trees with large conduit diameters. Despite numerous experimental studies, the mechanisms leading to bubble formation during freezing have not been addressed theoretically. We used classical nucleation theory and fluid mechanics to show which mechanisms are most likely to be responsible for bubble formation during freezing and what parameters determine the likelihood of the process. Our results confirm the common assumption that bubble formation during freezing is most likely due to gas segregation by ice. If xylem conduit walls are not permeable to the salts expelled by ice during the freezing process, osmotic pressures high enough for air seeding could be created. The build-up rate of segregated solutes in front of the ice-water interface depends equally on conduit diameter and freezing velocity. Therefore, bubble formation probability depends on these variables. The dependence of bubble formation probability on freezing velocity means that the experimental results obtained for cavitation threshold conduit diameters during freeze/thaw cycles depend on the experimental setup; namely sample size and cooling rate. The velocity dependence also suggests that to avoid bubble formation during freezing trees should have narrow conduits where freezing is likely to be fast (e.g., branches or outermost layer of the xylem). Avoidance of bubble formation during freezing could thus be one piece of the explanation why xylem conduit size of temperate and boreal zone trees varies quite systematically.

Mutagenic effect of freezing on mitochondrial DNA of Saccharomyces cerevisiae.

PubMed

Stoycheva, T; Venkov, P; Tsvetkov, Ts

2007-06-01

Although suggested in some studies, the mutagenic effect of freezing has not been proved by induction and isolation of mutants. Using a well-defined genetic model, we supply in this communication evidence for the mutagenic effect of freezing on mitochondrial DNA (mtDNA) of the yeast Saccharomyces cerevisiae. The cooling for 2 h at +4 degrees C, followed by freezing for 1 h at -10 degrees C and 16 h at -20 degrees C resulted in induction of respiratory mutations. The immediate freezing in liquid nitrogen was without mutagenic effect. The study of the stepwise procedure showed that the induction of respiratory mutants takes place during the freezing at -10 and -20 degrees C of cells pre-cooled at +4 degrees C. The genetic crosses of freeze-induced mutants evidenced their mitochondrial rho- origin. The freeze-induced rho- mutants are most likely free of simultaneous nuclear mutations. The extracellular presence of cryoprotectants did not prevent the mutagenic effect of freezing while accumulation of cryoprotectors inside cells completely escaped mtDNA from cryodamage. Although the results obtained favor the notion that the mutagenic effect of freezing on yeast mtDNA is due to formation and growth of intracellular ice crystals, other reasons, such as impairment of mtDNA replication or elevated levels of ROS production are discussed as possible explanations of the mutagenic effect of freezing. It is concluded that: (i) freezing can be used as a method for isolation of mitochondrial mutants in S. cerevisiae and (ii) given the substantial development in cryopreservation of cells and tissues, special precautions should be made to avoid mtDNA damage during the cryopreservation procedures.

SENSITIVE TO FREEZING2 Aids in Resilience to Salt and Drought in Freezing-Sensitive Tomato

DOE PAGES

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

2016-09-06

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

SENSITIVE TO FREEZING2 Aids in Resilience to Salt and Drought in Freezing-Sensitive Tomato

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

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

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

SENSITIVE TO FREEZING2 Aids in Resilience to Salt and Drought in Freezing-Sensitive Tomato1[OPEN

PubMed Central

Hersh, Hope Lynn

2016-01-01

SENSITIVE TO FREEZING2 (SFR2) is crucial for protecting chloroplast membranes following freezing in Arabidopsis (Arabidopsis thaliana). It has been shown that SFR2 homologs are present in all land plants, including freezing-sensitive species, raising the question of SFR2 function beyond freezing tolerance. Similar to freezing, salt and drought can cause dehydration. Thus, it is hypothesized that in freezing-sensitive plants SFR2 may play roles in their resilience to salt or drought. To test this hypothesis, SlSFR2 RNAi lines were generated in the cold/freezing-sensitive species tomato (Solanum lycopersicum [M82 cv]). Hypersensitivity to salt and drought of SlSFR2-RNAi lines was observed. Higher tolerance of wild-type tomatoes was correlated with the production of trigalactosyldiacylglycerol, a product of SFR2 activity. Tomato SFR2 in vitro activity is Mg2+-dependent and its optimal pH is 7.5, similar to that of Arabidopsis SFR2, but the specific activity of tomato SFR2 in vitro is almost double that of Arabidopsis SFR2. When salt and drought stress were applied to Arabidopsis, no conditions could be identified at which SFR2 was induced prior to irreversibly impacting plant growth, suggesting that SFR2 protects Arabidopsis primarily against freezing. Discovery of tomato SFR2 function in drought and salt resilience provides further insights into general membrane lipid remodeling-based stress tolerance mechanisms and together with protection against freezing in freezing-resistant plants such as Arabidopsis, it adds lipid remodeling as a possible target for the engineering of abiotic stress-resilient crops. PMID:27600812

Arginine and proline applied as food additives stimulate high freeze tolerance in larvae of Drosophila melanogaster.

PubMed

Koštál, Vladimír; Korbelová, Jaroslava; Poupardin, Rodolphe; Moos, Martin; Šimek, Petr

2016-08-01

The fruit fly Drosophila melanogaster is an insect of tropical origin. Its larval stage is evolutionarily adapted for rapid growth and development under warm conditions and shows high sensitivity to cold. In this study, we further developed an optimal acclimation and freezing protocol that significantly improves larval freeze tolerance (an ability to survive at -5°C when most of the freezable fraction of water is converted to ice). Using the optimal protocol, freeze survival to adult stage increased from 0.7% to 12.6% in the larvae fed standard diet (agar, sugar, yeast, cornmeal). Next, we fed the larvae diets augmented with 31 different amino compounds, administered in different concentrations, and observed their effects on larval metabolomic composition, viability, rate of development and freeze tolerance. While some diet additives were toxic, others showed positive effects on freeze tolerance. Statistical correlation revealed tight association between high freeze tolerance and high levels of amino compounds involved in arginine and proline metabolism. Proline- and arginine-augmented diets showed the highest potential, improving freeze survival to 42.1% and 50.6%, respectively. Two plausible mechanisms by which high concentrations of proline and arginine might stimulate high freeze tolerance are discussed: (i) proline, probably in combination with trehalose, could reduce partial unfolding of proteins and prevent membrane fusions in the larvae exposed to thermal stress (prior to freezing) or during freeze dehydration; (ii) both arginine and proline are exceptional among amino compounds in their ability to form supramolecular aggregates which probably bind partially unfolded proteins and inhibit their aggregation under increasing freeze dehydration. © 2016. Published by The Company of Biologists Ltd.

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

None, None

This 3-year project has studied how aerosol pollution influences glaciated clouds. The tool applied has been an 'aerosol-cloud model'. It is a type of Cloud-System Resolving Model (CSRM) modified to include 2-moment bulk microphysics and 7 aerosol species, as described by Phillips et al. (2009, 2013). The study has been done by, first, improving the model and then performing sensitivity studies with validated simulations of a couple of observed cases from ARM. These are namely the Tropical Warm Pool International Cloud Experiment (TWP-ICE) over the tropical west Pacific and the Cloud and Land Surface Interaction Campaign (CLASIC) over Oklahoma. Duringmore » the project, sensitivity tests with the model showed that in continental clouds, extra liquid aerosols (soluble aerosol material) from pollution inhibited warm rain processes for precipitation production. This promoted homogeneous freezing of cloud droplets and aerosols. Mass and number concentrations of cloud-ice particles were boosted. The mean sizes of cloud-ice particles were reduced by the pollution. Hence, the lifetime of glaciated clouds, especially ice-only clouds, was augmented due to inhibition of sedimentation and ice-ice aggregation. Latent heat released from extra homogeneous freezing invigorated convective updrafts, and raised their maximum cloud-tops, when aerosol pollution was included. In the particular cases simulated in the project, the aerosol indirect effect of glaciated clouds was twice than of (warm) water clouds. This was because glaciated clouds are higher in the troposphere than water clouds and have the first interaction with incoming solar radiation. Ice-only clouds caused solar cooling by becoming more extensive as a result of aerosol pollution. This 'lifetime indirect effect' of ice-only clouds was due to higher numbers of homogeneously nucleated ice crystals causing a reduction in their mean size, slowing the ice-crystal process of snow production and slowing sedimentation. In addition to the known indirect effects (glaciation, riming and thermodynamic), new indirect effects were discovered and quantified due to responses of sedimentation, aggregation and coalescence in glaciated clouds to changing aerosol conditions. In summary, the change in horizontal extent of the glaciated clouds ('lifetime indirect effects'), especially of ice-only clouds, was seen to be of higher importance in regulating aerosol indirect effects than changes in cloud properties ('cloud albedo indirect effects').« less

Surface Temperature and Precipitation Affecting GPS Signals Before the 2009 L'Aquila Earthquake (Central Italy).

NASA Astrophysics Data System (ADS)

Crescentini, L.; Amoruso, A.; Chiaraluce, L.

2017-12-01

This work focuses on GPS time series recorded before the Mw 6.1 earthquake which struck Central Italy in April 2009. It shows how environmental noise effects may be subtle and relevant when investigating relatively small strain signals and how the availability of data from weather stations and water level sensors co-located with GPS stations may provide critical information which must be taken into consideration while dealing with deformation signals.The preparatory phase of a large earthquake may include both seismic (foreshocks) and aseismic (slow slip event, SSE) deforming episodes but, unlike afterslip, no slow event has yet been recorded before moderate earthquakes, even when they occurred close to high-sensitivity strain meters. An exception to this seems to be represented by the 2009 earthquake. The main shock was preceded by a foreshock sequence lasting 6 months; it has been claimed that an analysis of continuous GPS data shows that during the foreshock sequence a 5.9 Mw SSE occurred along a decollement located beneath the reactivated normal fault system. This hypothesized SSE, that started in the middle of February 2009 and lasted for almost two weeks, would have eventually loaded the largest foreshock and the main shock.We show that the strain signal that the SSE would have generated at two laser strainmeters operating at about 20 km NE from the SSE source was essentially undetected. On the contrary, a transient signal is present in temperature and precipitation time series recorded close to the GPS station, MTTO, that has largest signal referred to the SSE, implying that these contaminated the GPS record. This interpretation is corroborated by the strong similarity, during the coldest winter months, between the displacement data of MTTO and a linear combination of filtered temperature and precipitation data, mimicking simple heat conduction and snow accumulation/removal processes. Such a correlation between displacement and environmental data is missing during the other seasons, when it neither snows nor freezes. This lack of correlation seems to exclude simple thermal expansion of the bedrock or the pillar monument itself as a local source of deformation. We hypothesize that thermal effects might be caused by ground freezing and consequent water expansion (about 9%) when liquid water is converted into ice.

Evidence of cyclical light/dark-regulated expression of freezing tolerance in young winter wheat plants.

PubMed

Skinner, Daniel Z; Bellinger, Brian; Hiscox, William; Helms, Gregory L

2018-01-01

The ability of winter wheat (Triticum aestivum L.) plants to develop freezing tolerance through cold acclimation is a complex rait that responds to many environmental cues including day length and temperature. A large part of the freezing tolerance is conditioned by the C-repeat binding factor (CBF) gene regulon. We investigated whether the level of freezing tolerance of 12 winter wheat lines varied throughout the day and night in plants grown under a constant low temperature and a 12-hour photoperiod. Freezing tolerance was significantly greater (P<0.0001) when exposure to subfreezing temperatures began at the midpoint of the light period, or the midpoint of the dark period, compared to the end of either period, with an average of 21.3% improvement in survival. Thus, freezing survival was related to the photoperiod, but cycled from low, to high, to low within each 12-hour light period and within each 12-hour dark period, indicating ultradian cyclic variation of freezing tolerance. Quantitative real-time PCR analysis of expression levels of CBF genes 14 and 15 indicated that expression of these two genes also varied cyclically, but essentially 180° out of phase with each other. Proton nuclear magnetic resonance analysis (1H-NMR) showed that the chemical composition of the wheat plants' cellular fluid varied diurnally, with consistent separation of the light and dark phases of growth. A compound identified as glutamine was consistently found in greater concentration in a strongly freezing-tolerant wheat line, compared to moderately and poorly freezing-tolerant lines. The glutamine also varied in ultradian fashion in the freezing-tolerant wheat line, consistent with the ultradian variation in freezing tolerance, but did not vary in the less-tolerant lines. These results suggest at least two distinct signaling pathways, one conditioning freezing tolerance in the light, and one conditioning freezing tolerance in the dark; both are at least partially under the control of the CBF regulon.

A study of the impact of freezing on the lyophilization of a concentrated formulation with a high fill depth.

PubMed

Liu, Jinsong; Viverette, Todd; Virgin, Marlin; Anderson, Mitch; Paresh, Dalal

2005-01-01

The objective of this study was to evaluate the impact of freezing on the lyophilization of a concentrated formulation with a high fill depth. A model system consisting of a 15-mL fill of 15% (w/w) sulfobutylether 7-beta-cyclodextrin (SBECD) solution in a 30-mL vial was selected for this study. Various freezing methods including single-step freezing, two-step freezing with a super-cooling holding, annealing, vacuum-induced freezing, changing ice habit using tert-butyl-alcohol (TBA), ice nucleation with silver iodide (AgI), as well as combinations of some of the methods, were used in the lyophilization of this model system. This work demonstrated that the freezing process had a significant impact on primary drying rate and product quality of a concentrated formulation with a high fill depth. Annealing, vacuum-induced freezing, and addition of either TBA or an ice nucleating agent (AgI) to the formulation accelerated the subsequent ice sublimation process. Two-step freezing or addition of TBA improved the product quality by eliminating vertical heterogeneity within the cake. The combination of two-step freezing in conjunction with an annealing step was shown to be a method of choice for freezing in the lyophilization of a product with a high fill depth. In addition to being an effective method of freezing, it is most applicable for scaling up. An alternative approach is to add a certain amount of TBA to the formulation, if the TBA-formulation interaction or regulatory concerns can be demonstrated as not being an issue. An evaluation of vial size performed in this study showed that although utilizing large-diameter vials to reduce the fill depth can greatly shorten the cycle time of a single batch, it will substantially decrease the product throughput in a large-scale freeze-dryer.

Paper-based Synthetic Gene Networks

PubMed Central

Pardee, Keith; Green, Alexander A.; Ferrante, Tom; Cameron, D. Ewen; DaleyKeyser, Ajay; Yin, Peng; Collins, James J.

2014-01-01

Synthetic gene networks have wide-ranging uses in reprogramming and rewiring organisms. To date, there has not been a way to harness the vast potential of these networks beyond the constraints of a laboratory or in vivo environment. Here, we present an in vitro paper-based platform that provides a new venue for synthetic biologists to operate, and a much-needed medium for the safe deployment of engineered gene circuits beyond the lab. Commercially available cell-free systems are freeze-dried onto paper, enabling the inexpensive, sterile and abiotic distribution of synthetic biology-based technologies for the clinic, global health, industry, research and education. For field use, we create circuits with colorimetric outputs for detection by eye, and fabricate a low-cost, electronic optical interface. We demonstrate this technology with small molecule and RNA actuation of genetic switches, rapid prototyping of complex gene circuits, and programmable in vitro diagnostics, including glucose sensors and strain-specific Ebola virus sensors. PMID:25417167

Using a fast dual-wavelength imaging ellipsometric system to measure the flow thickness profile of an oil thin film

NASA Astrophysics Data System (ADS)

Kuo, Chih-Wei; Han, Chien-Yuan; Jhou, Jhe-Yi; Peng, Zeng-Yi

2017-11-01

Dual-wavelength light sources with stroboscopic illumination technique were applied in a process of photoelastic modulated ellipsometry to retrieve two-dimensional ellipsometric parameters of thin films on a silicon substrate. Two laser diodes were alternately switched on and modulated by a programmable pulse generator to generate four short pulses at specific temporal phase angles in a modulation cycle, and short pulses were used to freeze the intensity variation of the PEM modulated signal that allows ellipsometric images to be captured by a charge-coupled device. Although the phase retardation of a photoelastic modulator is related to the light wavelength, we employed an equivalent phase retardation technique to avoid any setting from the photoelastic modulator. As a result, the ellipsometric parameters of different wavelengths may be rapidly obtained using this dual-wavelength ellipsometric system every 4 s. Both static and dynamic experiments are demonstrated in this work.

Paper-based synthetic gene networks.

PubMed

Pardee, Keith; Green, Alexander A; Ferrante, Tom; Cameron, D Ewen; DaleyKeyser, Ajay; Yin, Peng; Collins, James J

2014-11-06

Synthetic gene networks have wide-ranging uses in reprogramming and rewiring organisms. To date, there has not been a way to harness the vast potential of these networks beyond the constraints of a laboratory or in vivo environment. Here, we present an in vitro paper-based platform that provides an alternate, versatile venue for synthetic biologists to operate and a much-needed medium for the safe deployment of engineered gene circuits beyond the lab. Commercially available cell-free systems are freeze dried onto paper, enabling the inexpensive, sterile, and abiotic distribution of synthetic-biology-based technologies for the clinic, global health, industry, research, and education. For field use, we create circuits with colorimetric outputs for detection by eye and fabricate a low-cost, electronic optical interface. We demonstrate this technology with small-molecule and RNA actuation of genetic switches, rapid prototyping of complex gene circuits, and programmable in vitro diagnostics, including glucose sensors and strain-specific Ebola virus sensors.

Polymer-dispersed liquid crystal elastomers

NASA Astrophysics Data System (ADS)

Rešetič, Andraž; Milavec, Jerneja; Zupančič, Blaž; Domenici, Valentina; Zalar, Boštjan

2016-10-01

The need for mechanical manipulation during the curing of conventional liquid crystal elastomers diminishes their applicability in the field of shape-programmable soft materials and future applications in additive manufacturing. Here we report on polymer-dispersed liquid crystal elastomers, novel composite materials that eliminate this difficulty. Their thermal shape memory anisotropy is imprinted by curing in external magnetic field, providing for conventional moulding of macroscopically sized soft, thermomechanically active elastic objects of general shapes. The binary soft-soft composition of isotropic elastomer matrix, filled with freeze-fracture-fabricated, oriented liquid crystal elastomer microparticles as colloidal inclusions, allows for fine-tuning of thermal morphing behaviour. This is accomplished by adjusting the concentration, spatial distribution and orientation of microparticles or using blends of microparticles with different thermomechanical characteristics. We demonstrate that any Gaussian thermomechanical deformation mode (bend, cup, saddle, left and right twist) of a planar sample, as well as beat-like actuation, is attainable with bilayer microparticle configurations.

A controlled rate freeze/thaw system for cryopreservation of biological materials

NASA Technical Reports Server (NTRS)

Anselmo, V. J.; Harrison, R. G.

1979-01-01

A system which allows programmable temperature-time control for a 5 cc sample volume of an arbitrary biological material was constructed. Steady state and dynamic temperature control was obtained by supplying heat to the sample volume through resistive elements constructed as an integral part of the sample container. For cooling purposes, this container was totally immersed into a cold heat sink. Sample volume thermodynamic property data were obtained by measurements of heater power and heat flux through the container walls. Using a mixture of dry ice and alcohol at -79 C, sample volume was controlled from +40 C to -60 C at rates from steady state to + or - 65 C/min. Steady state temperature precision was better than 0.2 C while the dynamic capability depends on the temperature rate of change as well as the thermal mass of the sample and the container.

A freeze-stable formulation for DTwP and DTaP vaccines.

PubMed

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

2014-01-01

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

Indirect contact freeze water desalination for an ice maker machine - CFD simulation

NASA Astrophysics Data System (ADS)

Jayakody, Harith; Al-Dadah, Raya; Mahmoud, Saad

2017-11-01

To offer for potable water shortages, sea water desalination is a potential solution for the global rising demand for fresh water. The latent heat of fusion is about one-seventh the latent heat of vaporisation, thus indicating the benefit of lower energy consumption for the freeze desalination process. Limited literature is reported on computational fluid dynamics (CFD) on freeze desalination. Therefore, analysing and investigating thermodynamic processes are easily conducted by the powerful tool of CFD. A single unit of ice formation in an ice maker machine was modelled using ANSYS Fluent software three-dimensionally. Energy, species transport and solidification/melting modules were used in building the CFD model. Parametric analysis was conducted using the established CFD model to predict the effects of freezing temperature and the geometry of the ice maker machine; on ice production and the freezing time. Lower freezing temperatures allowed more ice production and faster freezing. Increasing the diameter and the length of the freezing tube enabled more ice to be produced.

Characterization of freezing precipitation events through other meteorological variables and their recent changes over Northern Extratropics

NASA Astrophysics Data System (ADS)

Groisman, P. Y.; Yin, X.; Bulygina, O.

2017-12-01

Freezing precipitation events intertwine with agriculture, recreation, energy consumption, and seasonal transportation cycles of human activities. Using supplementary synoptic reports at 1,500 long-term stations of North America and Northern Eurasia, we created climatology of freezing precipitation near the surface and found significant changes (increases) in these occurrences in the past decade at high latitudes/elevations (Groisman et al. 2016; updated). Firstly, we document narrow boundaries of near surface temperature and humidity fields when freezing precipitation events occur; these are necessary but insufficient conditions of their occurrence. Secondly, using the upper air data at the sites collocated with in situ observations of freezing events, we quantify the typical pattern of lower troposphere temperature anomalies during freezing events: At the same locations and Julian days, the presence of freezing event at the surface is associated with significantly warmer temperatures in the lower troposphere; comparison of temperatures at nearest days before and after the freezing events with days during these events also shows statistically significant positive temperature anomalies in the lower troposphere to 500 hPa (on average, +3 to 4 °C) In the days with freezing events, vertical air temperature gradients between surface and 850 hPa become less than usual with frequent inversions, when the tropospheric air is warmer than at the surface. The above features of the lower tropospheric temperature, near-surface temperature and humidity represent a combination of weather conditions conducive for precipitation, if it happens, falling in the freezing rain form. The in situ reports of freezing events at synoptic stations allow us to estimate temporary and spatial distributions of such "special weather conditions". Thus, a posteriori high probability of freezing events under these weather conditions invokes similar probabilities of freezing rain over the ungauged terrain, where we do not have special synoptic reports but can reproduce these "special weather conditions" from less sophisticated observational networks and/or reanalyses. Reference: Groisman et al. 2016: Recent changes in the frequency of freezing precipitation in North America and Northern Eurasia. Environ Res Lett 11 045007.

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.

Faculty development to support interprofessional education in healthcare professions: A realist synthesis.

PubMed

Watkins, Katherine Dolan

2016-11-01

Interprofessional collaboration is expected of healthcare providers to effect positive patient care experiences, reduce healthcare costs, and improve population health. While interprofessional education (IPE) is essential to graduate collaboration-ready healthcare professionals, faculty have limited experience and expertise in facilitating IPE, slowing adoption of this strategy. Faculty who are expected to develop, implement, and facilitate IPE activities in health professions need support and training to be successful. Faculty development programmes specific to IPE are examined through a comprehensive realist synthesis. The review began by identification of the mechanisms underpinning the intervention and then continued through a search for evidence relevant to the identified mechanisms. From 1,749 citations reviewed, 15 articles and book chapters were synthesised. The findings demonstrate that through the mechanisms-roles and role modelling, valuing diversity, reflection, group process, and knowledge, skills, and attitudes for IPE-positive outcomes can be achieved. Outcomes of increasing capacity and sustainability of IPE programmes, forming networks of individuals concerned with IPE, and evaluating and assessing of outcomes of IPE, may all be achieved through these mechanisms. The contextual factors include attitudes and expectations, programme logistics, leadership, and commitment, which interact with the mechanisms to impact the outcomes. Multiple context-mechanism-outcome configurations were revealed and analysed which help to explain how faculty development for IPE works in varying settings.

Improvement of the Error-detection Mechanism in Adults with Dyslexia Following Reading Acceleration Training.

PubMed

Horowitz-Kraus, Tzipi

2016-05-01

The error-detection mechanism aids in preventing error repetition during a given task. Electroencephalography demonstrates that error detection involves two event-related potential components: error-related and correct-response negativities (ERN and CRN, respectively). Dyslexia is characterized by slow, inaccurate reading. In particular, individuals with dyslexia have a less active error-detection mechanism during reading than typical readers. In the current study, we examined whether a reading training programme could improve the ability to recognize words automatically (lexical representations) in adults with dyslexia, thereby resulting in more efficient error detection during reading. Behavioural and electrophysiological measures were obtained using a lexical decision task before and after participants trained with the reading acceleration programme. ERN amplitudes were smaller in individuals with dyslexia than in typical readers before training but increased following training, as did behavioural reading scores. Differences between the pre-training and post-training ERN and CRN components were larger in individuals with dyslexia than in typical readers. Also, the error-detection mechanism as represented by the ERN/CRN complex might serve as a biomarker for dyslexia and be used to evaluate the effectiveness of reading intervention programmes. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Current Status of the Milky Stork Re-introduction Programme in Malaysia and Its Challenges

PubMed Central

Ismail, Ahmad; Rahman, Faid

2016-01-01

This review discussed the current status of the Milky Stork Re-introduction Programme in Malaysia and the challenges it faced. Although it has continued for almost seven years, more challenges appeared as time elapsed mainly due to the arising conflicts between the implementation of conservation policy versus the development projects in Kuala Gula. Hence, the released population is struggling to adapt mainly due to the reduction of suitable habitat for nesting and disturbed foraging areas by the continuous anthropogenic activities. Furthermore, the lack of appropriate training among captive storks prior to being released also slows the adaptation of the birds in their new habitat. The increasing pattern of pollution in the area is also highlighted. Several suggestions were given to help improve the current re-introduction programme. These include improvements to the captive training method, improvement of the existing enclosure’s condition and environment, protection of remaining mangrove forest, creation of a buffer zone to mitigate the increasing pollution level in the area, close monitoring of the released population, and maintaining continuous support and awareness among the public. Considering the ongoing anthropogenic activities that may impair the status of Kuala Gula as an important bird sanctuary, emphasis should be given to achieve sustainable development throughout the area. PMID:27688848

Forgetting motor programmes: retrieval dynamics in procedural memory.

PubMed

Tempel, Tobias; Frings, Christian

2014-01-01

When motor sequences are stored in memory in a categorised manner, selective retrieval of some sequences can induce forgetting of the non-retrieved sequences. We show that such retrieval-induced forgetting (RIF) occurs not only in cued recall but also in a test assessing memory indirectly by providing novel test cues without involving recall of items. Participants learned several sequential finger movements (SFMs), each consisting of the movement of two fingers of either the left or the right hand. Subsequently, they performed retrieval practice on half of the sequences of one hand. A final task then required participants to enter letter dyads. A subset of these dyads corresponded to the previously learned sequences. RIF was present in the response times during the entering of the dyads. The finding of RIF in the slowed-down execution of motor programmes overlapping with initially trained motor sequences suggests that inhibition resolved interference between procedural representations of the acquired motor sequences of one hand during retrieval practice.

Care during freeze-drying of bovine pericardium tissue to be used as a biomaterial: a comparative study.

PubMed

Polak, Roberta; Pitombo, Ronaldo N M

2011-10-01

Bovine pericardium (BP) tissue is widely used in the manufacture of bioprosthetics. The effects of freeze-drying on the BP tissue have been studied by some researchers in order to decrease their cytotoxicity due to preservation in formaldehyde solution, and to increase the lifetime of the product in storage. This study was undertaken in order to study the effect of freeze-drying in the structure of BP. To perform this study BP samples were freeze-dried in two different types of freeze-dryers available in our laboratory: a laboratory freeze-dryer, in which it was not possible to control parameters and a pilot freeze-dryer, wherein all parameters during freezing and drying were controlled. After freeze-drying processes, samples were analyzed by SEM, Raman spectroscopy, tensile strength, water uptake tests and TEM. In summary, it has been demonstrated that damages occur in collagen fibers by the loss of bulk water of collagen structure implicating in a drastic decreasing of BP mechanical properties due to its structural alterations. Moreover, it was proven that the collagen fibrils suffered breakage at some points, which can be attributed to the uncontrolled parameters during drying. Copyright © 2011 Elsevier Inc. All rights reserved.

Perspective: Surface freezing in water: A nexus of experiments and simulations

NASA Astrophysics Data System (ADS)

Haji-Akbari, Amir; Debenedetti, Pablo G.

2017-08-01

Surface freezing is a phenomenon in which crystallization is enhanced at a vapor-liquid interface. In some systems, such as n-alkanes, this enhancement is dramatic and results in the formation of a crystalline layer at the free interface even at temperatures slightly above the equilibrium bulk freezing temperature. There are, however, systems in which the enhancement is purely kinetic and only involves faster nucleation at or near the interface. The first, thermodynamic, type of surface freezing is easier to confirm in experiments, requiring only the verification of the existence of crystalline order at the interface. The second, kinetic, type of surface freezing is far more difficult to prove experimentally. One material that is suspected of undergoing the second type of surface freezing is liquid water. Despite strong indications that the freezing of liquid water is kinetically enhanced at vapor-liquid interfaces, the findings are far from conclusive, and the topic remains controversial. In this perspective, we present a simple thermodynamic framework to understand conceptually and distinguish these two types of surface freezing. We then briefly survey fifteen years of experimental and computational work aimed at elucidating the surface freezing conundrum in water.

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

PubMed

Slotsbo, Stine; Hansen, Lars Monrad; Jordaens, Kurt; Backeljau, Thierry; Malmendal, Anders; Nielsen, Niels Chr; Holmstrup, Martin

2012-04-01

Cold tolerance and metabolic responses to freezing of three slug species common in Scandinavia (Arion ater, Arion rufus and Arion lusitanicus) are reported. Autumn collected slugs were cold acclimated in the laboratory and subjected to freezing conditions simulating likely winter temperatures in their habitat. Slugs spontaneously froze at about -4 °C when cooled under dry conditions, but freezing of body fluids was readily induced at -1 °C when in contact with external ice crystals. All three species survived freezing for 2 days at -1 °C, and some A. rufus and A. lusitanicus also survived freezing at -2 °C. (1)H NMR spectroscopy revealed that freezing of body fluids resulted in accumulation of lactate, succinate and glucose. Accumulation of lactate and succinate indicates that ATP production occurred via fermentative pathways, which is likely a result of oxygen depletion in frozen tissues. Glucose increased from about 6 to 22 μg/mg dry tissue upon freezing in A. rufus, but less so in A. ater and A. lusitanicus. Glucose may thus act as a cryoprotectant in these slugs, although the concentrations are not as high as reported for other freeze tolerant invertebrates. Copyright © 2012 Elsevier Inc. All rights reserved.

To freeze or not to freeze embryos: clarity, confusion and conflict.

PubMed

Goswami, Mohar; Murdoch, Alison P; Haimes, Erica

2015-06-01

Although embryo freezing is a routine clinical practice, there is little contemporary evidence on how couples make the decision to freeze their surplus embryos, or of their perceptions during that time. This article describes a qualitative study of 16 couples who have had in vitro fertilisation (IVF) treatment. The study question was 'What are the personal and social factors that patients consider when deciding whether to freeze embryos?' We show that while the desire for a baby is the dominant drive, couples' views revealed more nuanced and complex considerations in the decision-making process. It was clear that the desire to have a baby influenced couples' decision-making and that they saw freezing as 'part of the process'. However, there were confusions associated with the term 'freezing' related to concerns about the safety of the procedure. Despite being given written information, couples were confused about the practical aspects of embryo freezing, which suggests they were preoccupied with the immediate demands of IVF. Couples expressed ethical conflicts about freezing 'babies'. We hope the findings from this study will inform clinicians and assist them in providing support to couples confronted with this difficult decision-making.

Silicon-based Porous Ceramics via Freeze Casting of Preceramic Polymers

NASA Astrophysics Data System (ADS)

Naviroj, Maninpat

Freeze casting is a technique for processing porous materials that has drawn significant attention for its effectiveness in producing a variety of tailorable pore structures for ceramics, metals, and polymers. With freeze casting, pores are generated based on a solidification process where ice crystals act as a sacrificial template which can eventually be sublimated to create pores. While the majority of freeze-casting studies have been performed using conventional ceramic suspensions, this work explores an alternative processing route by freeze casting with preceramic polymer solutions. Significant differences exist between freeze casting of a particulate suspension and a polymeric solution. These changes affect the processing method, solidification behavior, and pore structure, thereby introducing new challenges and possibilities for the freeze-casting technique. The first part of this study explored the processing requirements involved with freeze casting of preceramic polymers, along with methods to control the resulting pore structure. Solvent choice, freezing front velocity, and polymer concentration were used as processing variables to manipulate the pore structures. A total of seven organic solvents were freeze cast with a polymethylsiloxane preceramic polymer to produce ceramics with isotropic, dendritic, prismatic, and lamellar pore morphologies. Changes in freezing front velocity and polymer concentration were shown to influence pore size, shape, and connectivity. Differences between suspension- and solution-based samples freeze cast under equivalent conditions were also investigated. Certain solidification microstructures were strongly affected by the presence of suspended particles, creating differences between pore structures generated from the same solvents. Additionally, processing of solution-based samples were found to be the more facile technique. Compressive strength and water permeability of dendritic and lamellar structures were analyzed to determine functional differences between the pore structures. Results show that dendritic structures were up to 30 times stronger, while lamellar structures provided higher permeability constants. A change in freezing front velocity was shown to significantly affect permeability but not compressive strength. Finally, improved pore alignment along the freezing direction was achieved by controlling the nucleation and growth of solvent crystals through the use of a grain-selection template. Dendritic samples freeze cast with a template showed substantial increase in pore alignment, as determined by image analysis and permeability tests, with the permeability constant increasing by up to 6-fold when compared to a control sample.

Modification of physical properties of freeze-dried rice

NASA Technical Reports Server (NTRS)

Huber, C. S.

1971-01-01

Freeze cycling process consists of alternately freezing and thawing precooked rice for two cycles, rice is then frozen and freeze-dehydrated in vacuum sufficient to remove water from rice by sublimitation. Process modifies rice grain structure and porosity, enabling complete rehydration in one minute in hot water.

9 CFR 590.534 - Freezing facilities.

Code of Federal Regulations, 2010 CFR

2010-01-01

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

9 CFR 590.536 - Freezing operations.

Code of Federal Regulations, 2010 CFR

2010-01-01

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

The role of calcium and calmodulin in freezing-induced freezing resistance of Populus tomentosa cuttings.

PubMed

Lin, Shan-Zhi; Zhang, Zhi-Yi; Lin, Yuan-Zhen; Zhang, Qian; Guo, Huan

2004-02-01

To explore the role of calcium-calmodulin messenger system in the transduction of low temperature signal in woody plants, Populus tomentosa cuttings after being treated with CaCl(2) (10 mmol/L), Ca(2+) chelator EGTA (3 mmol/L), Ca(2+) channel inhibitor LaCl(3) (100 mmol/L) or CaM antagonist CPZ (50 mmol/L) were used for freezing acclimation at -3 degrees C. The changes in the calmodulin (CaM) and malonaldehyde (MDA) contents, the activities of superoxide dismutase (SOD), peroxidase (POD) and Ca(2+)-dependent adenosinetriphosphatase (Ca(2+)-ATPase) of mitochondrial membrane as well as freezing resistance (expressed as LT(50)) of cuttings were investigated to elucidate the physiological mechanisms by which trees adapt to freezing. The results showed that freezing acclimation increased the CaM content, the activities of SOD, POD and Ca(2+)-ATPase of mitochondrial membrane as well as freezing resistance of cuttings, and decreased the MDA content as compared with control cuttings. Treatment with CaCl(2) at the time of freezing acclimation enhanced the effect of freezing acclimation on the above-mentioned indexes, but this enhancement was abolished by Ca(2+)chelator EGTA, Ca(2+) channel inhibitor LaCl(3) or CaM antagonist CPZ, indicating that the calcium-calmodulin messenger system was involved in the course of freezing resistance development. The presence of CaCl(2) at the same time of freezing acclimation also reduced the degree of decline in CaM content, and in SOD, POD and Ca(2+)-ATPase activities caused by freezing stress at -14 degrees C, and enhanced the level of increase in CaM content, and in SOD, POD and Ca(2+)-ATPase activity in the recovery periods at 25 degrees C . The change in CaM content was found to be closely correlated to the levels of SOD, POD and Ca(2+)-ATPase, and to the degree of freezing resistance of cuttings during freezing acclimation either with or without CaCl(2) treatment. It was suggested that the increase of CaM content induced by CaCl(2) treatment promote the formation of Ca(2+)-CaM complexes, which effectively activates the activities of SOD, POD and mitochondrial Ca(2+)-ATPase and then further result in the adaptive changes associated with the development and enhancement of freezing resistance. Thus, It could be concluded that Ca(2+)-calmodulin may be involved in the regulation of the increase in SOD, POD and Ca(2+)-ATPase activities, and the induction of freezing resistance of cuttings.

Effects of warming rate, temperature, and antifreeze proteins on the survival of mouse spermatozoa frozen at an optimal rate.

PubMed

Koshimoto, Chihiro; Mazur, Peter

2002-08-01

We have recently reported that the survival of mouse spermatozoa is decreased when they are warmed at a suboptimal rate after being frozen at an optimal rate. We proposed that this drop in survival is caused by physical damage derived from the recrystallization of extracellular ice during slow warming. The first purpose of the present study was to determine the temperatures over which the decline in survival occurs during slow warming and the kinetics of the decline at fixed subzero temperatures. The second purpose was to examine the effects of antifreeze proteins (AFP) on the survival of slowly warmed mouse spermatozoa, the rationale being that AFP have the property of inhibiting ice recrystallization. With respect to the first point, a substantial loss in motility occurred when slow warming was continued to higher than -50 degrees C and the survival of the sperm decreased with an increase in the temperature at which slow warming was terminated. In contrast, the motility of sperm that were warmed rapidly to these temperatures remained high initially but dropped with increased holding time. At -30 degrees C, most of the drop occurred in 5 min. These results are consistent with the hypothesis that damage develops as a consequence of the recrystallization of the external ice. AFP ought to inhibit such recrystallization, but we found that the addition of AFP-I, AFP-III, and an antifreeze glycoprotein at concentrations of 1-100 microg/ml did not protect the frozen-thawed cells; rather it led to a decrease in survival that was proportional to the concentration. There was no decrease in survival from exposure to the AFP in the absence of freezing. AFP are known to produce changes in the structure and habit of ice crystals, and some have reported deleterious consequences associated with those structural changes. We suggest that such changes may be the basis of the adverse effects of AFP on the survival of the sperm, especially since mouse sperm are exquisitely sensitive to a variety of mechanical stresses.

Intact preservation of environmental samples by freezing under an alternating magnetic field.

PubMed

Morono, Yuki; Terada, Takeshi; Yamamoto, Yuhji; Xiao, Nan; Hirose, Takehiro; Sugeno, Masaya; Ohwada, Norio; Inagaki, Fumio

2015-04-01

The study of environmental samples requires a preservation system that stabilizes the sample structure, including cells and biomolecules. To address this fundamental issue, we tested the cell alive system (CAS)-freezing technique for subseafloor sediment core samples. In the CAS-freezing technique, an alternating magnetic field is applied during the freezing process to produce vibration of water molecules and achieve a stable, super-cooled liquid phase. Upon further cooling, the temperature decreases further, achieving a uniform freezing of sample with minimal ice crystal formation. In this study, samples were preserved using the CAS and conventional freezing techniques at 4, -20, -80 and -196 (liquid nitrogen) °C. After 6 months of storage, microbial cell counts by conventional freezing significantly decreased (down to 10.7% of initial), whereas that by CAS-freezing resulted in minimal. When Escherichia coli cells were tested under the same freezing conditions and storage for 2.5 months, CAS-frozen E. coli cells showed higher viability than the other conditions. In addition, an alternating magnetic field does not impact on the direction of remanent magnetization in sediment core samples, although slight partial demagnetization in intensity due to freezing was observed. Consequently, our data indicate that the CAS technique is highly useful for the preservation of environmental samples. © 2014 Society for Applied Microbiology and John Wiley & Sons Ltd.

Freeze for action: neurobiological mechanisms in animal and human freezing

PubMed Central

2017-01-01

Upon increasing levels of threat, animals activate qualitatively different defensive modes, including freezing and active fight-or-flight reactions. Whereas freezing is a form of behavioural inhibition accompanied by parasympathetically dominated heart rate deceleration, fight-or-flight reactions are associated with sympathetically driven heart rate acceleration. Despite the potential relevance of freezing for human stress-coping, its phenomenology and neurobiological underpinnings remain largely unexplored in humans. Studies in rodents have shown that freezing depends on amygdala projections to the brainstem (periaqueductal grey). Recent neuroimaging studies in humans have indicated that similar brain regions may be involved in human freezing. In addition, flexibly shifting between freezing and active defensive modes is critical for adequate stress-coping and relies on fronto-amygdala connections. This review paper presents a model detailing these neural mechanisms involved in freezing and the shift to fight-or-flight action. Freezing is not a passive state but rather a parasympathetic brake on the motor system, relevant to perception and action preparation. Study of these defensive responses in humans may advance insights into human stress-related psychopathologies characterized by rigidity in behavioural stress reactions. The paper therefore concludes with a research agenda to stimulate translational animal–human research in this emerging field of human defensive stress responses. This article is part of the themed issue ‘Movement suppression: brain mechanisms for stopping and stillness’. PMID:28242739

Structurally Caused Freezing Point Depression of Biological Tissues

PubMed Central

Bloch, Rene; Walters, D. H.; Kuhn, Werner

1963-01-01

When investigating the freezing behaviour (by thermal analysis) of the glycerol-extracted adductor muscle of Mytilus edulis it was observed that the temperature of ice formation in the muscular tissue was up to 1.5°C lower than the freezing point of the embedding liquid, a 0.25 N KCl solution with pH = 4.9 with which the tissue had been equilibrated prior to the freezing experiment. A smaller freezing point depression was observed if the pH values of the embedding 0.25 N KCl solution were above or below pH = 4.9. Reasoning from results obtained previously in analogous experiments with artificial gels, the anomalous freezing depression is explained by the impossibility of growing at the normal freezing temperature regular macroscopic crystals inside the gel, due to the presence of the gel network. The freezing temperature is here determined by the size of the microprisms penetrating the meshes of the network at the lowered freezing temperature. This process leads finally to an ice block of more or less regular structure in which the filaments are embedded. Prerequisite for this hindrance of ideal ice growth is a sufficient tensile strength of the filamental network. The existence of structurally caused freezing point depression in biological tissue is likely to invalidate many conclusions reported in the literature, in which hypertonicity was deduced from cryoscopic data. PMID:13971682

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

PubMed

Pittermann, Jarmila; Sperry, John

2003-09-01

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

Evaluation and Validation of the Messinger Freezing Fraction

NASA Technical Reports Server (NTRS)

Anderson, David N.; Tsao, Jen-Ching

2005-01-01

One of the most important non-dimensional parameters used in ice-accretion modeling and scaling studies is the freezing fraction defined by the heat-balance analysis of Messinger. For fifty years this parameter has been used to indicate how rapidly freezing takes place when super-cooled water strikes a solid body. The value ranges from 0 (no freezing) to 1 (water freezes immediately on impact), and the magnitude has been shown to play a major role in determining the physical appearance of the accreted ice. Because of its importance to ice shape, this parameter and the physics underlying the expressions used to calculate it have been questioned from time to time. Until now, there has been no strong evidence either validating or casting doubt on the current expressions. This paper presents experimental measurements of the leading-edge thickness of a number of ice shapes for a variety of test conditions with nominal freezing fractions from 0.3 to 1.0. From these thickness measurements, experimental freezing fractions were calculated and compared with values found from the Messinger analysis as applied by Ruff. Within the experimental uncertainty of measuring the leading-edge thickness, agreement of the experimental and analytical freezing fraction was very good. It is also shown that values of analytical freezing fraction were entirely consistent with observed ice shapes at and near rime conditions: At an analytical freezing fraction of unity, experimental ice shapes displayed the classic rime shape, while for conditions producing analytical freezing fractions slightly lower than unity, glaze features started to appear.

Freeze Tolerance of Nine Zoysiagrass Cultivars Using Natural Cold Acclimation and Freeze Chambers

USDA-ARS?s Scientific Manuscript database

Winter hardiness of zoysiagrass (Zoysia spp.) cultivars is an important attribute throughout the biogeographical transition zone, thus the inability to withstand freezing temperatures may limit the use of these cultivars. The objective of this research was to determine the freeze tolerance (LT50) of...

77 FR 12240 - Application(s) for Duty-Free Entry of Scientific Instruments

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-29

... Institute, 5353 Parkside Dr MC 19-RE, Jupiter, FL 33458. Instrument: Freeze Fracture/Freeze Etch device... localization of membrane proteins using freeze fracture replica immuno- gold labeling, including all kinds of receptors and channels. Because freeze-fracture replica immuno-gold labeling has a high sensitivity for the...

Effect of Warming Rate on the Survival of Vitrified Mouse Oocytes and on the Recrystallization of Intracellular Ice1

PubMed Central

Seki, Shinsuke; Mazur, Peter

2008-01-01

Successful cryopreservation demands there be little or no intracellular ice. One procedure is classical slow equilibrium freezing, and it has been successful in many cases. However, for some important cell types, including some mammalian oocytes, it has not. For the latter, there are increasing attempts to cryopreserve them by vitrification. However, even if intracellular ice formation (IIF) is prevented during cooling, it can still occur during the warming of a vitrified sample. Here, we examine two aspects of this occurrence in mouse oocytes. One took place in oocytes that were partly dehydrated by an initial hold for 12 min at −25°C. They were then cooled rapidly to −70°C and warmed slowly, or they were warmed rapidly to intermediate temperatures and held. These oocytes underwent no IIF during cooling but blackened from IIF during warming. The blackening rate increased about 5-fold for each five-degree rise in temperature. Upon thawing, they were dead. The second aspect involved oocytes that had been vitrified by cooling to −196°C while suspended in a concentrated solution of cryoprotectants and warmed at rates ranging from 140°C/min to 3300°C/min. Survivals after warming at 140°C/min and 250°C/min were low (<30%). Survivals after warming at ≥2200°C/min were high (80%). When warmed slowly, they were killed, apparently by the recrystallization of previously formed small internal ice crystals. The similarities and differences in the consequences of the two types of freezing are discussed. PMID:18562703

Investigation of alternative organic solvents and methods for the preparation of long-circulating and pH-sensitive liposomes containing cisplatin.

PubMed

Giuberti, Cristiane dos Santos; Boratto, Fernanda Alves; Degobert, Ghania; Silveira, Josianne Nicácio; Oliveira, Mônica Cristina

2013-09-01

Recent studies using long-circulating and pH-sensitive liposomes containing cisplatin (SpHL-CDDP) have resulted in a formulation with improved pharmacokinetic, toxicity and tumor localization properties. In this study, SpHL-CDDP were prepared in both laboratory and pilot scales. This study evaluated the possibility of using the dehydration-rehydration method, as well as using alternative organic solvents (ethyl acetate/ethanol mixtures at 2:1 and 1:1 volume ratios), for the preparation of liposomes by the reverse-phase evaporation (REV) method. The influence of different concentrations of cisplatin (CDDP) (2.0, 1.0, 0.5 and 0.25 mg/mL) on the entrapment percentage and size of SpHL-CDDP was also investigated. In addition, carbohydrates were tested as cryoprotectants in a freeze-thaw study as a pretest to screen the type to be used in the freeze-drying process. A decrease in the encapsulation percentage of CDDP and an increase in the vesicle diameter could be observed for both liposome formulations prepared with ethyl acetate:ethanol mixtures, as compared with REV liposomes prepared with ethyl ether. It is important to note that after applying either quick or slow cooling, the mean diameter of SpHL (empty liposomes) proved to be similar when in the presence of cryoprotectants. In sum, the optimal processing conditions were achieved when using a 0.5 mg/mL CDDP solution, ethyl ether and the REV method, resulting in liposomal dispersions of mean diameters and homogeneities that were deemed suitable for intravenous administration.

Kinetics and Pathways for the Debromination of Polybrominated Diphenyl Ethers by Bimetallic and Nanoscale Zerovalent Iron: Effects of Particle Properties and Catalyst

PubMed Central

Zhuang, Yuan; Jin, Luting; Luthy, Richard G.

2012-01-01

Polybrominated diphenyl ethers (PBDEs) are recognized as a new class of widely-distributed and persistent contaminants for which effective treatment and remediation technologies are needed. In this study, two kinds of commercially available nanoscale Fe° slurries (Nanofer N25 and N25S), a freeze-dried laboratory-synthesized Fe° nanoparticle (nZVI), and their palladized forms were used to investigate the effect of particle properties and catalyst on PBDE debromination kinetics and pathways. Nanofers and their palladized forms were found to debrominate PBDEs effectively. The laboratory-synthesized Fe° nanoparticles also debrominated PBDEs, but were slower due to deactivation by the freeze-drying and stabilization processes in the laboratory synthesis. An organic modifier, polyacrylic acid (PAA), bound on N25S slowed PBDE debromination by a factor of three to four compared to N25. The activity of palladized nZVI (nZVI/Pd) was optimized at 0.3 Pd/Fe wt% in our system. N25 could debrominate selected environmentally-abundant PBDEs, including BDE 209, 183, 153, 99, and 47, to end products di-BDEs, mono-BDEs and diphenyl ether (DE) in one week, while nZVI/Pd (0.3 Pd/Fe wt%) mainly resulted in DE as a final product. Step-wise major PBDE debromination pathways by unamended and palladized Fe° are described and compared. Surface precursor complex formation is an important limiting factor for palladized Fe° reduction as demonstrated by PBDE pathways where steric hindrance and rapid sequential debromination of adjacent bromines play an important role. PMID:22732301

The western painted turtle genome, a model for the evolution of extreme physiological adaptations in a slowly evolving lineage

PubMed Central

2013-01-01

Background We describe the genome of the western painted turtle, Chrysemys picta bellii, one of the most widespread, abundant, and well-studied turtles. We place the genome into a comparative evolutionary context, and focus on genomic features associated with tooth loss, immune function, longevity, sex differentiation and determination, and the species' physiological capacities to withstand extreme anoxia and tissue freezing. Results Our phylogenetic analyses confirm that turtles are the sister group to living archosaurs, and demonstrate an extraordinarily slow rate of sequence evolution in the painted turtle. The ability of the painted turtle to withstand complete anoxia and partial freezing appears to be associated with common vertebrate gene networks, and we identify candidate genes for future functional analyses. Tooth loss shares a common pattern of pseudogenization and degradation of tooth-specific genes with birds, although the rate of accumulation of mutations is much slower in the painted turtle. Genes associated with sex differentiation generally reflect phylogeny rather than convergence in sex determination functionality. Among gene families that demonstrate exceptional expansions or show signatures of strong natural selection, immune function and musculoskeletal patterning genes are consistently over-represented. Conclusions Our comparative genomic analyses indicate that common vertebrate regulatory networks, some of which have analogs in human diseases, are often involved in the western painted turtle's extraordinary physiological capacities. As these regulatory pathways are analyzed at the functional level, the painted turtle may offer important insights into the management of a number of human health disorders. PMID:23537068

An in vitro analysis of disintegration times of different formulations of olanzapine orodispersible tablet: a preliminary report.

PubMed

Hobbs, David; Karagianis, Jamie; Treuer, Tamas; Raskin, Joel

2013-12-01

Orodispersible tablets (ODTs) are tablet or wafer forms of medication that disintegrate in the mouth, aided only by saliva. ODTs rely on different fast dissolve/disintegration manufacturing technologies. Disintegration time differences for several olanzapine ODT forms were investigated. Risperdal M-Tab(®) was included as a non-olanzapine ODT comparator. Eleven olanzapine ODT examples and orodispersible risperidone strengths were evaluated in vitro for formulation composition, manufacturing method, disintegration and dissolution characteristics, and formulation differences in comparison with freeze dried Zydis(®) ODT. Automated dissolution test equipment captured ODT dissolution rates by measuring real-time release of active ingredient. A high-speed video camera was used to capture tablet disintegration times in warm simulated saliva. The main outcome measure was the disintegration and dissolution characteristics of the ODT formulations. The ODT manufacturing method was associated with time to disintegrate; the fastest were freeze dried tablets, followed by soft compressed tablets and then hard/dense tablets. Olanzapine Zydis(®) was the only ODT that completely disintegrated in less than 4 s for all strengths (5, 10, 15, and 20 mg), followed by 5-mg Prolanz FAST(®) (12 s) and then risperidone ODT 4 mg (40 s). Reasons for slow dissolution of the olanzapine generics may include low product potency, excipient binding, excipient solubility, active ingredient particle size and incomplete disintegration. Differences in the formulation and manufacturing process of olanzapine ODTs appear to have a strong influence on the disintegration time of the active compound; differences that may potentially impact their use in clinical practice.

Disruption of self-organized actions in monkeys with progressive MPTP-induced parkinsonism. I. Effects of task complexity.

PubMed

Pessiglione, Mathias; Guehl, Dominique; Hirsch, Etienne C; Féger, Jean; Tremblay, Léon

2004-01-01

Parkinson's disease (PD) is characterized by motor symptoms, usually accompanied by cognitive deficits. The question addressed in this study is whether complexity of routine actions can exacerbate parkinsonian disorders that are often considered to be motor symptoms. To examine this question, we trained four vervet monkeys (Cercopithecus aethiops) to perform three multiple-choice retrieval tasks. In order of ascending complexity, rewards were freely available (task 1), covered with transparent sliding plaques (task 2), and covered with opaque sliding plaques cued by symbols (task 3). Thus, from task 1 to task 2 we added a motor difficulty--the recall of context-adapted movement; and from task 2 to task 3 we added a cognitive difficulty: the recall of symbol-reward associations. The more complex the task, the longer it took to learn, but after extensive training the performance was stable in all tasks, with similar retrieval durations. The monkeys then received systemic 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) injections (0.3-0.4 mg/kg) every 4-7 days, until the first motor symptoms appeared. In the course of MPTP intoxication, the behavioural performance declined while the motor symptoms were absent or mild--the retrieval duration increased, and non-initiated choices and hesitations between choices became frequent. Interestingly, this decline was in proportion to task complexity, and was particularly pronounced with the cognitive difficulty. Furthermore, freezing appeared only with the cognitive difficulty. We therefore suggest that everyday cognitive difficulties may exacerbate hypokinesia (lack of initiation, abnormal slowness) and executive disorders (hesitations, freezing) in the early stages of human PD.

Quantitative evaluation of colloidal stability of antibody solutions using PEG-induced liquid-liquid phase separation.

PubMed

Wang, Ying; Latypov, Ramil F; Lomakin, Aleksey; Meyer, Julie A; Kerwin, Bruce A; Vunnum, Suresh; Benedek, George B

2014-05-05

Colloidal stability of antibody solutions, i.e., the propensity of the folded protein to precipitate, is an important consideration in formulation development of therapeutic monoclonal antibodies. In a protein solution, different pathways including crystallization, colloidal aggregation, and liquid-liquid phase separation (LLPS) can lead to the formation of precipitates. The kinetics of crystallization and aggregation are often slow and vary from protein to protein. Due to the diverse mechanisms of these protein condensation processes, it is a challenge to develop a standardized test for an early evaluation of the colloidal stability of antibody solutions. LLPS would normally occur in antibody solutions at sufficiently low temperature, provided that it is not preempted by freezing of the solution. Poly(ethylene glycol) (PEG) can be used to induce LLPS at temperatures above the freezing point. Here, we propose a colloidal stability test based on inducing LLPS in antibody solutions and measuring the antibody concentration of the dilute phase. We demonstrate experimentally that such a PEG-induced LLPS test can be used to compare colloidal stability of different antibodies in different solution conditions and can be readily applied to high-throughput screening. We have derived an equation for the effects of PEG concentration and molecular weight on the results of the LLPS test. Finally, this equation defines a binding energy in the condensed phase, which can be determined in the PEG-induced LLPS test. This binding energy is a measure of attractive interactions between antibody molecules and can be used for quantitative characterization of the colloidal stability of antibody solutions.

Consequences of metaphase II oocyte cryopreservation on mRNA content.

PubMed

Chamayou, S; Bonaventura, G; Alecci, C; Tibullo, D; Di Raimondo, F; Guglielmino, A; Barcellona, M L

2011-04-01

We studied the consequences of freezing/thawing processes on mRNA contents in MII oocytes after slow-freezing/rapid thawing (SF/RT) and vitrification/warming (V/W) protocols, and compared the results to fresh MII oocytes. We quantified the nuclear transcript mRNA responsible for the translation of proteins belonging either to trans-regulatory protein family or to functional structural proteins such as proteins involved in DNA structural organization (NAP1L1, TOP1, H1F0H1), chromosomal structure maintenance (SMC, SCC3, RAD21, SMC1A, SMC1B, STAG3, REC8), mitochondrial energetic pathways (ATP5GJ, SDHC), cell cycle regulation and processes (CLTA, MAPK6, CKS2) and staminal cell potency-development competence stage (DPPA3, OCT4, FOXJ2). Surplus MII oocytes were donated from patients in IVF cycles and divided in three groups of 15 oocytes. Group 1 was comprised of non-cryopreserved oocytes and Groups 2 and 3 underwent SF/RT and V/W procedures, respectively. There was an overall decrease of mRNA extracted from cryopreserved oocytes compared to control group. Only 39.4% of mRNA content were preserved after SF/RT while 63.3% of mRNA content were maintained after V/W. Oocyte cryopreservation is associated with molecular injury associated with the decrease of stored mRNA. However the V/W protocol is more conservative than SF/RT resulting in a level of mRNA sufficient to maintain biologic functions in the subsequent fertilized oocyte. Copyright © 2011 Elsevier Inc. All rights reserved.

Direct Measurement of Water States in Cryopreserved Cells Reveals Tolerance toward Ice Crystallization

PubMed Central

Huebinger, Jan; Han, Hong-Mei; Hofnagel, Oliver; Vetter, Ingrid R.; Bastiaens, Philippe I.H.; Grabenbauer, Markus

2016-01-01

Complex living systems such as mammalian cells can be arrested in a solid phase by ultrarapid cooling. This allows for precise observation of cellular structures as well as cryopreservation of cells. The state of water, the main constituent of biological samples, is crucial for the success of cryogenic applications. Water exhibits many different solid states. If it is cooled extremely rapidly, liquid water turns into amorphous ice, also called vitreous water, a glassy and amorphous solid. For cryo-preservation, the vitrification of cells is believed to be mandatory for cell survival after freezing. Intracellular ice crystallization is assumed to be lethal, but experimental data on the state of water during cryopreservation are lacking. To better understand the water conditions in cells subjected to freezing protocols, we chose to directly analyze their subcellular water states by cryo-electron microscopy and tomography, cryoelectron diffraction, and x-ray diffraction both in the cryofixed state and after warming to different temperatures. By correlating the survival rates of cells with their respective water states during cryopreservation, we found that survival is less dependent on ice-crystal formation than expected. Using high-resolution cryo-imaging, we were able to directly show that cells tolerate crystallization of extra- and intracellular water. However, if warming is too slow, many small ice crystals will recrystallize into fewer but bigger crystals, which is lethal. The applied cryoprotective agents determine which crystal size is tolerable. This suggests that cryoprotectants can act by inhibiting crystallization or recrystallization, but they also increase the tolerance toward ice-crystal growth. PMID:26541066

Freezer or non-freezer: clinical assessment of freezing of gait.

PubMed

Snijders, Anke H; Haaxma, Charlotte A; Hagen, Yolien J; Munneke, Marten; Bloem, Bastiaan R

2012-02-01

Freezing of gait (FOG) is both common and debilitating in patients with Parkinson's disease (PD). Future pathophysiology studies will depend critically upon adequate classification of patients as being either 'freezers' or 'non-freezers'. This classification should be based ideally upon objective confirmation by an experienced observer during clinical assessment. Given the known difficulties to elicit FOG when examining patients, we aimed to investigate which simple clinical test would be the most sensitive to provoke FOG objectively. We examined 50 patients with PD, including 32 off-state freezers (defined as experiencing subjective 'gluing of the feet to the floor'). Assessment including a FOG trajectory (three trials: normal speed, fast speed, and with dual tasking) and several turning variants (180° vs. 360° turns; leftward vs. rightward turns; wide vs. narrow turning; and slow vs. fast turns). Sensitivity of the entire assessment to provoke FOG in subjective freezers was 0.74, specificity was 0.94. The most effective test to provoke FOG was rapid 360° turns in both directions and, if negative, combined with a gait trajectory with dual tasking. Repeated testing improved the diagnostic yield. The least informative tests included wide turns, 180° turns or normal speed full turns. Sensitivity to provoke objective FOG in subjective freezers was 0.65 for the rapid full turns in both directions and 0.63 for the FOG trajectory. The most efficient way to objectively ascertain FOG is asking patients to repeatedly make rapid 360° narrow turns from standstill, on the spot and in both directions. Copyright © 2011 Elsevier Ltd. All rights reserved.

Long Pulse Operation on Tore-Supra: Towards Steady State

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

Moreau, P.; Bucalossi, J.; Brosset, C.

The experimental programme of Tore Supra is devoted to the study of technology and physics issues associated to long-duration high performance discharges. This new domain of operation requires simultaneously and in steady state: heat removal capability, particle exhaust, fully non-inductive current drive, advanced technology integration and real time plasma control. The long discharge allows for addressing new time scale physic such as the wall particle retention and erosion. Moreover, the physics of fully non-inductive discharges is full of novelty, namely: the MHD stability, the slow spontaneous oscillation of the central electron temperature or the outstanding inward particle pinch.

Surface freezing of water.

PubMed

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

2016-01-01

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

An Investigation of the Cryogenic Freezing of Water in Non-Metallic Pipelines

NASA Astrophysics Data System (ADS)

Martin, C. I.; Richardson, R. N.; Bowen, R. J.

2004-06-01

Pipe freezing is increasingly used in a range of industries to solve otherwise intractable pipe line maintenance and servicing problems. This paper presents the interim results from an experimental study on deliberate freezing of polymeric pipelines. Previous and contemporary works are reviewed. The object of the current research is to confirm the feasibility of ice plug formation within a polymeric pipe as a method of isolation. Tests have been conducted on a range of polymeric pipes of various sizes. The results reported here all relate to freezing of horizontal pipelines. In each case the process of plug formation was photographed, the frozen plug pressure tested and the pipe inspected for signs of damage resulting from the freeze procedure. The time to freeze was recorded and various temperatures logged. These tests have demonstrated that despite the poor thermal and mechanical properties of the polymers, freezing offers a viable alternative method of isolation in polymeric pipelines.

Frost heave susceptibility of saturated soil under constant rate of freezing

NASA Astrophysics Data System (ADS)

Ryokai, K.; Iguro, M.; Yoneyama, K.

Introduced are the results of experiments carried out to quantitatively obtain the frost heave pressure and displacement of soil subjected to artificial freezing or freezing around in-ground liquefied natural gas storage tanks. This experiment is conducted to evaluate the frost heave susceptibility of saturated soil under overconsolidation. In other words, this experiment was carried out to obtain the relation of the over-burden pressure and freezing rate to the frost heave ratio by observing the frost heave displacement and freezing time of specimens by freezing the specimens at a constant freezing rate under a constant overburden pressure, while letting water freely flow in and out of the system. Introduced are the procedures for frost heave test required to quantitatively obtain the frost heave displacement and pressure of soil. Furthermore, the relation between the frost heave susceptibility and physical properties of soil obtained by this test is reported.

Contact Freezing of Water by Salts.

PubMed

Niehaus, Joseph; Cantrell, Will

2015-09-03

Water is unlikely to crystallize homogeneously at temperatures greater than -34 °C. Freezing at higher temperatures is heterogeneous-catalyzed by the presence of a second substance. If that substance is at an air-water interface, then the mode is called contact freezing, and it typically will trigger nucleation at a higher temperature than if the substance were wholly immersed within the liquid. We find that the impact of salt particles initiates freezing in experiments using water droplets at supercoolings of 9 to 16 °C. These results show that contact freezing nuclei need not be effective as immersion mode nuclei. We discuss our results in the context of proposed mechanisms of contact freezing. Finally, we use the time scales for diffusion of heat and of ions and the propagation of a sound wave through the droplet to estimate that contact freezing occurs within 10 ns of impact.

Cold hardiness in molluscs

NASA Astrophysics Data System (ADS)

Ansart, Armelle; Vernon, Philippe

2003-05-01

Molluscs inhabit all types of environments: seawater, intertidal zone, freshwater and land, and of course may have to deal with subzero temperatures. Ectotherm animals survive cold conditions by avoiding it by extensive supercooling (freezing avoidant species) or by bearing the freezing of their extracellular body fluids (freezing tolerant species). Although some studies on cold hardiness are available for intertidal molluscs, they are scarce for freshwater and terrestrial ones. Molluscs often exhibit intermediary levels of cold hardiness, with a moderate or low ability to supercool and a limited survival to the freezing of their tissues. Several factors could be involved: their dependence on water, their ability to enter dormancy, the probability of inoculative freezing in their environment, etc. Size is an important parameter in the development of cold hardiness abilities: it influences supercooling ability in land snails, which are rather freezing avoidant and survival to ice formation in intertidal organisms, which generally tolerate freezing.

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

PubMed

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

2016-11-01

The effects of industrial pre-freezing processing and freezing handling on the contents of glucosinolates and antioxidants (vitamin C, polyphenols, carotenoid and chlorophyll), as well as the antioxidant capacity in broccoli (Brassica oleracea L. var. italica) florets were investigated in the present study. Our results showed that the glucosinolate accumulations were significantly decreased after pre-freezing processing, whereas elevated levels of phenols, carotenoids, chlorophyll, and also antioxidant capacity were observed in frozen broccoli florets. The contents of vitamin C remained constant during above mentioned processing. In conclusion, the current industrial freezing processing method is a good practice for the preservation of main antioxidant nutrients in broccoli florets, although some improvements in pre-freezing processing, such as steam blanching and ice-water cooling, are needed to attenuate the decrease in glucosinolate content. Copyright © 2016 Elsevier Ltd. All rights reserved.

Two-dimensional freezing criteria for crystallizing colloidal monolayers

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

Wang Ziren; Han Yilong; Alsayed, Ahmed M.

Video microscopy was employed to explore crystallization of colloidal monolayers composed of diameter-tunable microgel spheres. Two-dimensional (2D) colloidal liquids were frozen homogenously into polycrystalline solids, and four 2D criteria for freezing were experimentally tested in thermal systems for the first time: the Hansen-Verlet freezing rule, the Loewen-Palberg-Simon dynamical freezing criterion, and two other rules based, respectively, on the split shoulder of the radial distribution function and on the distribution of the shape factor of Voronoi polygons. Importantly, these freezing criteria, usually applied in the context of single crystals, were demonstrated to apply to the formation of polycrystalline solids. At themore » freezing point, we also observed a peak in the fluctuations of the orientational order parameter and a percolation transition associated with caged particles. Speculation about these percolated clusters of caged particles casts light on solidification mechanisms and dynamic heterogeneity in freezing.« less

Effect of pasteurisation and freezing method on bioactive compounds and antioxidant activity of strawberry pulp.

PubMed

Gonçalves, Gilma Auxiliadora Santos; Resende, Nathane Silva; Carvalho, Elisângela Elena Nunes; Resende, Jaime Vilela de; Vilas Boas, Eduardo Valério de Barros

2017-09-01

This study evaluated the stability of strawberry pulp subjected to three factors, pasteurisation (pasteurised and unpasteurised), freezing method (static air and forced air) and storage time (0, 2, 4 and 6 months). Pasteurisation favoured vitamin C retention during storage but enhanced the total loss of phenolics without affecting anthocyanin levels. Freezing by forced air was more effective in retaining phenolics during the first 4 months of storage, although the freezing method did not affect the anthocyanin levels. Processing and storage reduced the levels of individual phenolics. Freezing by forced air was more effective than static air in retaining antioxidant activity of the pulp. Polyphenol oxidase and peroxidase enzyme levels were relatively stable and independent of pasteurisation, freezing and storage time. Even after 6 months of frozen storage, strawberry pulp is a significant source of nutrients and bioactive compounds and retains high antioxidant capacity independent of pasteurisation and freezing method.

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) ...

7 CFR 929.11 - To can, freeze, or dehydrate.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 8 2010-01-01 2010-01-01 false To can, freeze, or dehydrate. 929.11 Section 929.11... LONG ISLAND IN THE STATE OF NEW YORK Order Regulating Handling Definitions § 929.11 To can, freeze, or dehydrate. To can, freeze, or dehydrate means to convert cranberries into canned, frozen, or dehydrated...

Applications of Simulator Freeze to Carrier Glideslope Tracking Instruction.

DTIC Science & Technology

1982-07-01

Showing Datum Bars and Meatball . .. .. .. ... .. ... .... 19 4 Freezes Per Trial Averaged Across Freeze Conditions and Across 4-Trial Blocks of Training...algorithm linearly increased the criterion in meatball units from 1.0 at 6000 feet from the ramp to 1.5 at the ramp. "Freezes" did not occur beyond 6000

7 CFR 305.18 - Quick freeze treatment schedule.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 5 2010-01-01 2010-01-01 false Quick freeze treatment schedule. 305.18 Section 305.18... SERVICE, DEPARTMENT OF AGRICULTURE PHYTOSANITARY TREATMENTS Quick Freeze Treatments § 305.18 Quick freeze treatment schedule. (a) T110. (1) Initially, lower the commodity's temperature to 0 °F or below. (2) Hold...

The equilibrated state of freezing as a basis for distinguishing lethal stresses of freezing in plants

USDA-ARS?s Scientific Manuscript database

A model for coordination of stresses that limit winterhardiness in plants based on the thermodynamic equilibrated state of freezing and melting provides a rational basis for distinction of freeze-induced energies which can stress and injure living organisms in various ways. The departure from equili...

7 CFR 58.620 - Freezing and packaging rooms.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 3 2013-01-01 2013-01-01 false Freezing and packaging rooms. 58.620 Section 58.620... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Rooms and Compartments § 58.620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall be...

7 CFR 58.620 - Freezing and packaging rooms.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 3 2014-01-01 2014-01-01 false Freezing and packaging rooms. 58.620 Section 58.620... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Rooms and Compartments § 58.620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall be...

7 CFR 58.620 - Freezing and packaging rooms.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 3 2012-01-01 2012-01-01 false Freezing and packaging rooms. 58.620 Section 58.620... Specifications for Dairy Plants Approved for USDA Inspection and Grading Service 1 Rooms and Compartments § 58.620 Freezing and packaging rooms. The rooms used for freezing and packaging frozen desserts shall be...

Freezing temperature of finger skin.

PubMed

Wilson, O; Goldman, R F; Molnar, G W

1976-10-01

In 45 subjects, 154 frostnips of the finger were induced by cooling in air at -15 degrees C with various wind speeds. The mean supercooled skin temperature at which frostnip appeared was -9.4 degrees C. The mean skin temperature rise due to heat of fusion at ice crystallization was 5.3 degrees C. The skin temperature rose to what was termed the apparent freezing point. The relation of this point to the supercooled skin temperature was analyzed for the three wind speeds used. An apparent freezing point for a condition of no supercooling was calculated, estimating the highest temperature at which skin freezes at a given wind speed. The validity of the obtained differences in apparent freezing point was tested by an analysis of covariance. Although not statistically significant, the data suggest that the apparent freezing point with no supercooling decreases with increasing wind velocity. The highest calculated apparent freezing point at -15 degrees C and 6.8 m/s was 1.2 degrees C lower than the true freezing point for skin previously determined in brine, which is a statistically significant difference.

A Vivens Ex Vivo Study on the Synergistic Effect of Electrolysis and Freezing on the Cell Nucleus.

PubMed

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

2015-01-01

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

Inner ear tissue preservation by rapid freezing: Improving fixation by high-pressure freezing and hybrid methods

PubMed Central

Bullen, A.; Taylor, R.R.; Kachar, B.; Moores, C.; Fleck, R.A.; Forge, A.

2014-01-01

In the preservation of tissues in as ‘close to life’ state as possible, rapid freeze fixation has many benefits over conventional chemical fixation. One technique by which rapid freeze-fixation can be achieved, high pressure freezing (HPF), has been shown to enable ice crystal artefact-free freezing and tissue preservation to greater depths (more than 40 μm) than other quick-freezing methods. Despite increasingly becoming routine in electron microscopy, the use of HPF for the fixation of inner ear tissue has been limited. Assessment of the quality of preservation showed routine HPF techniques were suitable for preparation of inner ear tissues in a variety of species. Good preservation throughout the depth of sensory epithelia was achievable. Comparison to chemically fixed tissue indicated that fresh frozen preparations exhibited overall superior structural preservation of cells. However, HPF fixation caused characteristic artefacts in stereocilia that suggested poor quality freezing of the actin bundles. The hybrid technique of pre-fixation and high pressure freezing was shown to produce cellular preservation throughout the tissue, similar to that seen in HPF alone. Pre-fixation HPF produced consistent high quality preservation of stereociliary actin bundles. Optimising the preparation of samples with minimal artefact formation allows analysis of the links between ultrastructure and function in inner ear tissues. PMID:25016142

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.

Application of freeze-drying technology in manufacturing orally disintegrating films.

PubMed

Liew, Kai Bin; Odeniyi, Michael Ayodele; Peh, Kok-Khiang

2016-01-01

Freeze drying technology has not been maximized and reported in manufacturing orally disintegrating films. The aim of this study was to explore the freeze drying technology in the formulation of sildenafil orally disintegrating films and compare the physical properties with heat-dried orally disintegrating film. Central composite design was used to investigate the effects of three factors, namely concentration of carbopol, wheat starch and polyethylene glycol 400 on the tensile strength and disintegration time of the film. Heat-dried films had higher tensile strength than films prepared using freeze-dried method. For folding endurance, freeze-dried films showed improved endurance than heat-dried films. Moreover, films prepared using freeze-dried methods were thicker and had faster disintegration time. Formulations with higher amount of carbopol and starch showed higher tensile strength and thickness whereas formulations with higher PEG 400 content showed better flexibility. Scanning electron microscopy showed that the freeze-dried films had more porous structure compared to the heat-dried film as a result of the release of water molecule from the frozen structure when it was subjected to freeze drying process. The sildenafil film was palatable. The dissolution profiles of freeze-dried and heat-dried films were similar to Viagra® with f2 of 51.04 and 65.98, respectively.

Freezing: an underutilized food safety technology?

PubMed

Archer, Douglas L

2004-01-15

Freezing is an ancient technology for preserving foods. Freezing halts the activities of spoilage microorganisms in and on foods and can preserve some microorganisms for long periods of time. Frozen foods have an excellent overall safety record. The few outbreaks of food-borne illness associated with frozen foods indicate that some, but not all human pathogens are killed by commercial freezing processes. Freezing kills microorganisms by physical and chemical effects and possibly through induced genetic changes. Research is needed to better understand the physical and chemical interactions of various food matrices with the microbial cell during freezing and holding at frozen temperatures. The literature suggests that many pathogenic microorganisms may be sublethally injured by freezing, so research should be done to determine how to prevent injured cells from resuscitating and becoming infectious. Studies on the genetics of microbial stress suggest that the induction of resistance to specific stresses may be counteracted by, for example, simple chemicals. Research is needed to better understand how resistance to the lethal effect of freezing is induced in human pathogens and means by which it can be counteracted in specific foods. Through research, it seems possible that freezing may in the future be used to reliably reduce populations of food-borne pathogens as well as to preserve foods.

The Use of High-resolution Infrared Thermography (HRIT) for the Study of Ice Nucleation and Ice Propagation in Plants

PubMed Central

Wisniewski, Michael; Neuner, Gilbert; Gusta, Lawrence V.

2015-01-01

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events often have devastating effects on agricultural production and can also play an important role in shaping community structure in natural populations of plants, especially in alpine, sub-arctic, and arctic ecosystems. Therefore, a better understanding of the freezing process in plants can play an important role in the development of methods of frost protection and understanding mechanisms of freeze avoidance. Here, we describe a protocol to visualize the freezing process in plants using high-resolution infrared thermography (HRIT). The use of this technology allows one to determine the primary sites of ice formation in plants, how ice propagates, and the presence of ice barriers. Furthermore, it allows one to examine the role of extrinsic and intrinsic nucleators in determining the temperature at which plants freeze and evaluate the ability of various compounds to either affect the freezing process or increase freezing tolerance. The use of HRIT allows one to visualize the many adaptations that have evolved in plants, which directly or indirectly impact the freezing process and ultimately enables plants to survive frost events. PMID:25992743

The use of high-resolution infrared thermography (HRIT) for the study of ice nucleation and ice propagation in plants.

PubMed

Wisniewski, Michael; Neuner, Gilbert; Gusta, Lawrence V

2015-05-08

Freezing events that occur when plants are actively growing can be a lethal event, particularly if the plant has no freezing tolerance. Such frost events often have devastating effects on agricultural production and can also play an important role in shaping community structure in natural populations of plants, especially in alpine, sub-arctic, and arctic ecosystems. Therefore, a better understanding of the freezing process in plants can play an important role in the development of methods of frost protection and understanding mechanisms of freeze avoidance. Here, we describe a protocol to visualize the freezing process in plants using high-resolution infrared thermography (HRIT). The use of this technology allows one to determine the primary sites of ice formation in plants, how ice propagates, and the presence of ice barriers. Furthermore, it allows one to examine the role of extrinsic and intrinsic nucleators in determining the temperature at which plants freeze and evaluate the ability of various compounds to either affect the freezing process or increase freezing tolerance. The use of HRIT allows one to visualize the many adaptations that have evolved in plants, which directly or indirectly impact the freezing process and ultimately enables plants to survive frost events.

Water-cooled probe technique for the study of freeze lining formation

NASA Astrophysics Data System (ADS)

Verscheure, Karel; Campforts, Mieke; Verhaeghe, Frederik; Boydens, Eddy; Blanpain, Bart; Wollants, Patrick; van Camp, Maurits

2006-12-01

Furnace protection by water-cooled freeze linings becomes increasingly important as the metal producing industry attempts to achieve higher process intensities. Systematic investigations of the growth and the resulting microstructure and compositional profile of freeze linings are necessary to understand the behavior of freeze linings, their relation with the industrial process, and their interaction with the wall cooling system. We have developed a technique based on the submergence of a water-cooled probe into a liquid slag bath. Freeze linings of two industrial nonferrous slags have been produced using this technique and their growth, microstructural, and compositional profiles as a function of submergence time were determined. Thermodynamic equilibrium for the investigated slag systems was calculated and compared with the observed microstructures. The freeze linings form in approximately 15 minutes. Close to the water cooling, the freeze linings are predominantly amorphous in structure. With increasing distance from the water cooling, the proportion of crystalline phases increases and bath material is entrapped in the microstructure. Cellular crystals are observed close to the bath. The freeze linings exhibit an approximate homogeneous composition. The results demonstrate that the technique is a successful tool in obtaining information on the growth, microstructure, and composition of freeze linings in industrial water-cooled furnaces.

Freeze-dried spermatozoa: A future tool?

PubMed

Olaciregui, M; Gil, L

2017-04-01

Cryopreservation has been routinely used to preserve sperm of human and different animal species. However, frozen sperm storage for a long time brings many inconveniences because of liquid nitrogen. Many attempts have been made to overcome the disadvantages of the current cryopreservation method. Freeze-drying has been proposed as alternative method for sperm preservation to achieve the ability to store sperm doses indefinitely at ambient temperature or in ordinary refrigerators. At present, it has been reported successfully sperm freeze-drying on many animal species including canine and feline. It is well known that during freeze-drying process, sperm DNA could be damaged, but if suitable protection is provided, the sperm nucleus could preserve the ability to activate the oocyte and embryos could be generated by intracytoplasmic sperm injection (ICSI). Many factors influence the freeze-drying efficacy, so current researches have been conducted to find strategies to control these factors to maintain the sperm DNA integrity. This review describes the latest method of sperm freeze-drying for practical application in preserving and transporting genetic resources. In addition, the approaches to improve the efficiency of the technique were studied. We demonstrated that the DNA integrity of freeze-dried dog sperm is affected by the composition of the freeze-drying solution as well as the temperature and period of storage. Further studies are necessary to refine freeze-drying protocol in order to protect the DNA and maintain the sperm functionality and obtain offspring from freeze-dried sperm. © 2016 Blackwell Verlag GmbH.

Role of cells in freezing-induced cell-fluid-matrix interactions within engineered tissues.

PubMed

Seawright, Angela; Ozcelikkale, Altug; Dutton, Craig; Han, Bumsoo

2013-09-01

During cryopreservation, ice forms in the extracellular space resulting in freezing-induced deformation of the tissue, which can be detrimental to the extracellular matrix (ECM) microstructure. Meanwhile, cells dehydrate through an osmotically driven process as the intracellular water is transported to the extracellular space, increasing the volume of fluid for freezing. Therefore, this study examines the effects of cellular presence on tissue deformation and investigates the significance of intracellular water transport and cell-ECM interactions in freezing-induced cell-fluid-matrix interactions. Freezing-induced deformation characteristics were examined through cell image deformetry (CID) measurements of collagenous engineered tissues embedded with different concentrations of MCF7 breast cancer cells versus microspheres as their osmotically inactive counterparts. Additionally, the development of a biophysical model relates the freezing-induced expansion of the tissue due to the cellular water transport and the extracellular freezing thermodynamics for further verification. The magnitude of the freezing-induced dilatation was found to be not affected by the cellular water transport for the cell concentrations considered; however, the deformation patterns for different cell concentrations were different suggesting that cell-matrix interactions may have an effect. It was, therefore, determined that intracellular water transport during freezing was insignificant at the current experimental cell concentrations; however, it may be significant at concentrations similar to native tissue. Finally, the cell-matrix interactions provided mechanical support on the ECM to minimize the expansion regions in the tissues during freezing.

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

NASA Astrophysics Data System (ADS)

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

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

Effects of freezing conditions on quality changes in blueberries.

PubMed

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

2018-03-12

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

Involvement of Multiple Types of Dehydrins in the Freezing Response in Loquat (Eriobotrya japonica)

PubMed Central

Xu, Hongxia; Yang, Yong; Xie, Li; Li, Xiaoying; Feng, Chao; Chen, Junwei; Xu, Changjie

2014-01-01

Dehydrins (DHNs) are a family of plant proteins typically induced in response to stress conditions that cause cellular dehydration, such as low temperatures, high salinity, and drought. Loquat (Eriobotrya japonica) is a perennial fruit crop that blossoms during winter. Loquat fruitlets are frequently injured by freezing. To evaluate the role of the EjDHNs in freezing resistance in loquat fruitlets, two cultivars of loquat, the freezing-sensitive ‘Ninghaibai’ (FS-NHB) and the freezing-tolerant ‘Jiajiao’ (FT-JJ), were analyzed under induced freezing stress. Freezing stress led to obvious accumulation of reactive oxygen species and considerable lipid peroxidation in membranes during the treatment period. Both these phenomena were more pronounced in ‘FS-NHB’ than in ‘FS-JJ.’ Immunogold labeling of dehydrin protein was performed. DHN proteins were found to be concentrated mainly in the vicinity of the plasma membrane, and the density of the immunogold labeling was significantly higher after freezing treatment, especially in the more freezing-tolerant cultivar ‘FT-JJ.’ Seven DHNs, showing four different structure types, were obtained from loquat fruitlets and used to study the characteristics of different EjDHN proteins. These DHN proteins are all highly hydrophilic, but they differ significantly in size, ranging from 188 to 475 amino acids, and in biochemical properties, such as theoretical pI, aliphatic index, and instability index. Freezing treatment resulted in up-regulation of the expression levels of all seven EjDHNs, regardless of structure type. The accumulation of the transcripts of these EjDHN genes was much more pronounced in ‘FT-JJ’ than in ‘FS-NHB.’ Altogether, this study provides evidence that EjDHNs are involved in the cryoprotection of the plasma membrane during freeze-induced dehydration in loquat fruitlets. PMID:24498141

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.

"On" freezing in Parkinson's disease: resistance to visual cue walking devices.

PubMed

Kompoliti, K; Goetz, C G; Leurgans, S; Morrissey, M; Siegel, I M

2000-03-01

To measure "on" freezing during unassisted walking (UW) and test if two devices, a modified inverted stick (MIS) and a visual laser beam stick (LBS) improved walking speed and number of "on" freezing episodes in patients with Parkinson's disease (PD). Multiple visual cues can overcome "off' freezing episodes and can be useful in improving gait function in parkinsonian patients. These devices have not been specifically tested in "on" freezing, which is unresponsive to pharmacologic manipulations. Patients with PD, motor fluctuations and freezing while "on," attempted walking on a 60-ft track with each of three walking conditions in a randomized order: UW, MIS, and LBS. Total time to complete a trial, number of freezes, and the ratio of walking time to the number of freezes were compared using Friedman's test. Twenty-eight patients with PD, mean age 67.81 years (standard deviation [SD] 7.54), mean disease duration 13.04 years (SD 7.49), and mean motor Unified Parkinson's Disease Rating Scale score "on" 32.59 (SD 10.93), participated in the study. There was a statistically significant correlation of time needed to complete a trial and number of freezes for all three conditions (Spearman correlations: UW 0.973, LBS 0.0.930, and MIS 0.842). The median number of freezes, median time to walk in each condition, and median walking time per freeze were not significantly different in pairwise comparisons of the three conditions (Friedman's test). Of the 28 subjects, six showed improvement with the MIS and six with the LBS in at least one outcome measure. Assisting devices, specifically based on visual cues, are not consistently beneficial in overcoming "on" freezing in most patients with PD. Because this is an otherwise untreatable clinical problem and because occasional subjects do respond, cautious trials of such devices under the supervision of a health professional should be conducted to identify those patients who might benefit from their long-term use.

Freezing of gait in PD: prospective assessment in the DATATOP cohort.

PubMed

Giladi, N; McDermott, M P; Fahn, S; Przedborski, S; Jankovic, J; Stern, M; Tanner, C

2001-06-26

To study the development of freezing of gait in PD. Freezing of gait is a common, disabling, and poorly understood symptom in PD. The authors analyzed data from 800 patients with early PD from the Deprenyl and Tocopherol Antioxidative Therapy of Parkinsonism (DATATOP) clinical trial who were assigned either placebo, deprenyl, tocopherol, or the combination of deprenyl and tocopherol. The primary outcome measure was the time from randomization until the freezing of gait score on the Unified Parkinson's Disease Rating Scale (UPDRS) became positive. Fifty-seven patients (7.1%) had freezing of gait at study entry and 193 (26%) of the remaining patients experienced the symptom by the end of the follow-up period. Those with freezing of gait at baseline had significantly more advanced disease than those without the symptom, as measured by total UPDRS and Hoehn and Yahr stage. High baseline risk factors for developing freezing of gait during the follow-up period were the onset of PD with a gait disorder; higher scores of rigidity, postural instability, bradykinesia and speech; and longer disease duration. In contrast, tremor was strongly associated with a decreased risk for freezing of gait. At the end of follow-up, the signs most strongly associated with the freezing phenomenon were gait, balance, and speech disorders, not rigidity or bradykinesia. Deprenyl treatment was strongly associated with a decreased risk for developing freezing of gait; tocopherol had no effect. Freezing of gait is directly related to duration of PD. Risk factors at onset of disease are the absence of tremor and PD beginning as a gait disorder. The development of freezing of gait in the course of the illness is strongly associated with the development of balance and speech problems, less so with the worsening of bradykinesia, and is not associated with the progression of rigidity. These results support the concept that the freezing phenomenon is distinct from bradykinesia. Deprenyl, in the absence of L-dopa, was found to be an effective prophylactic treatment and should be considered for patients with PD who have an onset of gait difficulty.

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

ERIC Educational Resources Information Center

Lahne, Jacob B.; Schmidt, Shelly J.

2010-01-01

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

The 2007 California Citrus Freeze: Vulnerability, Poverty, and Unemployment Issues of Farmworkers

ERIC Educational Resources Information Center

Orozco, Graciela Leon

2010-01-01

In January of 2007, freezing temperatures destroyed citrus and vegetable crops in California. By May 2007, more than 9,000 freeze-related unemployment applications had been fled. Through face-to-face interviews, this study documents the experience of 63 farmworkers to find out how they survived the freeze and accessed services. Findings revealed…

Method of freezing living cells and tissues with improved subsequent survival

DOEpatents

Senkan, Selim M.; Hirsch, Gerald P.

1980-01-01

This invention relates to an improved method for freezing red blood cells, ther living cells, or tissues with improved subsequent survival, wherein constant-volume freezing is utilized that results in significantly improved survival compared with constant-pressure freezing; optimization is attainable through the use of different vessel geometries, cooling baths and warming baths, and sample concentrations.

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

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Cell-freezing apparatus and reagents for in vitro... Establishments That Manufacture Blood and Blood Products § 864.9225 Cell-freezing apparatus and reagents for in vitro diagnostic use. (a) Identification. Cell-freezing apparatus and reagents for in vitro diagnostic...

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

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Cell-freezing apparatus and reagents for in vitro... Establishments That Manufacture Blood and Blood Products § 864.9225 Cell-freezing apparatus and reagents for in vitro diagnostic use. (a) Identification. Cell-freezing apparatus and reagents for in vitro diagnostic...