9 CFR 327.21 - Inspection procedures for chilled fresh and frozen boneless manufacturing meat.

Code of Federal Regulations, 2011 CFR

2011-01-01

... fresh and frozen boneless manufacturing meat. 327.21 Section 327.21 Animals and Animal Products FOOD... MEAT AND POULTRY PRODUCTS INSPECTION AND VOLUNTARY INSPECTION AND CERTIFICATION IMPORTED PRODUCTS § 327.21 Inspection procedures for chilled fresh and frozen boneless manufacturing meat. (a) Definitions...

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

PubMed Central

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

2016-01-01

Background Extracellular vesicles or microparticles exhibiting procoagulant and thrombogenic activity may contribute to the haemostatic potential of fresh frozen plasma. Materials and methods Fresh frozen plasma was prepared from platelet-rich plasma at 20 °C (Group-1 donors) or directly from whole blood at 4 °C (Group-2 donors). Each unit was aseptically divided into three parts, stored frozen for specific periods of time, and analysed by flow cytometry for procoagulant activity immediately after thaw or following post-thaw storage for 24 h at 4 °C. Donors’ haematologic, biochemical and life-style profiles as well as circulating microparticles were analysed in parallel. Results Circulating microparticles exhibited a considerable interdonor but not intergroup variation. Fresh frozen plasma units were enriched in microparticles compared to plasma in vivo. Duration of storage significantly affected platelet- and red cell-derived microparticles. Fresh frozen plasma prepared directly from whole blood contained more residual platelets and more platelet-derived microparticles compared to fresh frozen plasma prepared from platelet-rich plasma. Consequently, there was a statistically significant difference in total, platelet- and red cell-derived microparticles between the two preparation protocols over storage time in the freezer. Preservation of the thawed units for 24 h at 4 °C did not significantly alter microparticle accumulation. Microparticle accumulation and anti-oxidant capacity of fresh frozen plasma was positively or negatively correlated, respectively, with the level of circulating microparticles in individual donors. Discussion The preparation protocol and the duration of storage in the freezer, independently and in combination, influenced the accumulation of microparticles in fresh frozen plasma units. In contrast, storage of thawed units for 24 h at 4 °C had no significant effect on the concentration of microparticles. PMID:27136430

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

PubMed

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

2016-05-01

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

Comparison of human papillomavirus detection between freshly frozen tissue and paraffin embedded tissue of invasive cervical cancer

PubMed Central

2010-01-01

Background Human Papillomavirus (HPV) detection results comparing paraffin embedded cervical tissue and other cervical specimens have been done with varying degrees of agreement. However, studies comparing freshly frozen specimens and paraffin embedded specimens of invasive cervical carcinomas are lacking. The aim of the study was to compare HPV detection using SPF10 broad-spectrum primers PCR followed by DEIA and genotyping by LiPA25 (version 1) between freshly frozen cervical tissue samples and paraffin embedded blocks of cervical tissue from the same patient. There were 171 pairs of paraffin embedded and freshly frozen samples analyzed from cervical carcinoma cases from Kampala, Uganda. Results 88.9% (95% CI: 83.2%-93.2%) of paraffin embedded samples were HPV positive compared with 90.1% (95% CI: 84.6%-94.1%) of freshly frozen samples, giving an overall agreement in HPV detection between fresh tissue and paraffin embedded tissue at 86.0% (95% CI: 79.8%-90.8%). Although the proportion of HPV positive cases in freshly frozen tissue was higher than those in paraffin blocks, the difference was not statistically significant (p > 0.05). In both types of tissues, single HPV infections were predominant, with HPV16 accounting for 47% of positive cases. Comparison in the overall agreement, taking into accounts not only positivity in general, but also HPV types, showed a 65% agreement (complete agreement of 59.7%, partial agreement of 5.3%) and complete disagreement of 35.0%. HPV detection in squamous cell carcinomas (SCC) and adenocarcinomas (ADC) was similar in fresh tissue or paraffin blocks (p ≥ 0.05). p16 immunostaining in samples that had at least one HPV negative results showed that 24 out of 25 cases had an over-expressed pattern. Conclusions HPV DNA detection was lower among ADC as compared to SCC. However, such differences were minimized when additional p16 testing was added, suggesting that the technical issues may largely explain the HPV negative cases. PMID:20846370

Storage of Unfed and Leftover Mothers' Own Milk.

PubMed

Fogleman, April D; Meng, Ting; Osborne, Jason; Perrin, Maryanne T; Jones, Frances; Allen, Jonathan C

The objective was to examine the bacteriological and immunological properties of freshly expressed, previously frozen, and leftover mothers' own milk during storage. In the first of two pilot studies, 12 mother-infant dyads participated. The milk studied included freshly expressed unfed and freshly expressed leftover milk. Milk samples were stored at 24°C, 4°C, or -20°C. In the second pilot study, 11 mother-infant dyads participated. The milk studied included milk that had been previously frozen, including previously frozen leftover milk. Milk samples were stored at 24°C and 4°C. After storage in both studies, the milk was analyzed for bacteriological and immunological properties. Bacteriological and immunological characteristics of freshly expressed unfed and freshly expressed leftover milk and previously frozen unfed and previously frozen leftover milk remained stable during storage at 4°C for at least 6 days. The quality of all groups of mothers' milk declined when stored at 24°C for longer than 3 hours. While this study provides evidence that human milk might be safe at longer storage times, storage guidelines should not be revised until more research is performed. This study serves as a call to action for more research on the topic of human milk storage, specifically leftover human milk. The study provides information to inform future study designs on the topic of unpasteurized human milk storage. More research is needed regarding leftover human milk storage with a greater number of participants, determination of the quality of human milk, and the storage of human milk in a real-life setting.

The Development of Alternative Food Cost Indexes

DTIC Science & Technology

1974-11-01

Frozen frufts, vegetables , fruit juices , and cake mixes are added and the quantity of purchased bread increased in this FCI. Comment was made that some... Grain , Frozen Vegetables , Mixed, Frozen Beans, Green, Frozen Peaches, Canned Apples, Fresh Oranges, Fresh Strawberries, Frozen Juice , Pineapple...also falls to recognize such accepted food service practices as the use of frozen fruits , vegetables , and juices , or prepared cake mixes, Tj,e

Post-operative infection with fresh frozen allograft: reported outcomes of a hospital-based bone bank over 14 years.

PubMed

Man, Wing Yum; Monni, Toni; Jenkins, Ruth; Roberts, Paul

2016-06-01

Femoral head bone allografts have traditionally been used to provide mechanical stability to areas of bony deficiency, or for its osteoinductive and osteoconductive properties. Concerns have been raised over increased infection rates following the use of fresh-frozen graft tissue. This retrospective study aims to investigate the outcomes of fresh frozen femoral heads kept in a regulated, non-commercial bone bank at a university teaching hospital.The local bone bank database was used to identify released femoral heads during a 14 year study period (September 1999-December 2013) whereby a retrospective review of patient records was undertaken to determine clinical outcome. During the observed study period, 427 femoral heads were released from cold storage. Of these, 270 femoral heads had a mean follow-up of 347 days. 157 femoral heads were excluded due to insufficient follow-up data (n = 132) or discarded due to breaks in the cold chain prior to use (n = 25). Of the 270 included femoral heads, 231 (85.6 %) had no reported complications with good graft incorporation. In the remaining 39 with reported complications, only 5 (2.6 %) developed a postoperative infection. Our findings suggest that the use of fresh frozen allograft does not materially increase the risk of post-operative bacterial infection. Our reported post-operative infection rates are comparable with infection rates of other similar studies on fresh frozen allograft use.

Descriptive texture analyses of cooked broiler breast fillets with the wooden condition after fresh and frozen storage

USDA-ARS?s Scientific Manuscript database

The objective of this study was to evaluate the effects of the wooden breast condition on the texture of cooked broiler breast fillets (Pectoralis major) after fresh and frozen storage. Texture characteristics of normal (NORM) and severe wooden breast (WB) fillets were studied by both sensory descr...

[Influence of S-nitrosoglutathione on agglutination and nitric oxide concentration in frozen platelets].

PubMed

Wu, Tao; Liu, Jing-Han; Li, Hui; Zhou, Wu; Wang, Shu-Ying

2012-04-01

The aim of this study was to investigate the influence of S-nitrosoglutathione (GSNO) on agglutination and nitric oxide (NO) concentration in frozen platelets. The agglutination of platelets was detected by using platelet agglutination apparatus, the level of NO in platelets was detected by the nitrate enzyme reduction method. The results showed that the rates of agglutination in freeze platelets and frozen platelets treated with GSNO were (35.47 ± 2.93) and (24.43 ± 3.07), which were significantly lower than that in fresh liquid platelets (63.44 ± 2.96). The level of NO concentration in frozen platelets was (22.16 ± 6.38), which was significantly lower than that in fresh liquid platelets (31.59 ± 16.88). The level of NO concentration in frozen platelets treated with GSNO was (45.64 ± 6.31), which was significantly higher than that in fresh liquid platelets (P < 0.01). It is concluded that GSNO increases the concentration of NO in frozen platelets, inhibits platelet activation and maintains platelet function, thus GSNO can be used as a frozen protective agent.

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

PubMed

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

2010-04-01

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

Biobanking of fresh frozen tissue from clinical surgical specimens: transport logistics, sample selection, and histologic characterization.

PubMed

Botling, Johan; Micke, Patrick

2011-01-01

Access to high-quality fresh frozen tissue is critical for translational cancer research and molecular -diagnostics. Here we describe a workflow for the collection of frozen solid tissue samples derived from fresh human patient specimens after surgery. The routines have been in operation at Uppsala University Hospital since 2001. We have integrated cryosection and histopathologic examination of each biobank sample into the biobank manual. In this way, even small, macroscopically ill-defined lesions can be -procured without a diagnostic hazard due to the removal of uncharacterized tissue from a clinical -specimen. Also, knowledge of the histomorphology of the frozen tissue sample - tumor cell content, stromal components, and presence of necrosis - is pivotal before entering a biobank case into costly molecular profiling studies.

Using rewards-based incentives to increase purchase of fruit and vegetables in lower-income households: design and start-up of a randomized trial.

PubMed

Phipps, Etienne J; Wallace, Samantha L; Stites, Shana D; Uplinger, Nadine; Brook Singletary, S; Hunt, Lacy; Axelrod, Saul; Glanz, Karen; Braitman, Leonard E

2013-05-01

To report the design and baseline results of a rewards-based incentive to promote purchase of fruit and vegetables by lower-income households. A four-phase randomized trial with wait-listed controls. In a pilot study, despite inadequate study coupon use, purchases of fresh fruit (but not vegetables) increased, but with little maintenance. In the present study, credits on the study store gift card replace paper coupons and a tapering phase is added. The primary outcome is the number of servings of fresh and frozen fruit and vegetables purchased per week. A large full-service supermarket located in a predominantly minority community in Philadelphia, Pennsylvania, USA. Fifty-eight households, with at least one child living in the home. During the baseline period, households purchased an average of 3·7 servings of fresh vegetables and an average of less than 1 serving of frozen vegetables per week. Households purchased an average of 1·9 servings of fresh fruit per week, with little to no frozen fruit purchases. Overall, the range of fresh and frozen produce purchased during this pre-intervention period was limited. At baseline, produce purchases were small and of limited variety. The study will contribute to understanding the impact of financial incentives on increasing the purchases of healthier foods by lower-income populations.

Urine phenobarbital drug screening: potential use for compliance assessment in neonates.

PubMed

Guillet, Ronnie; Kwon, Jennifer M; Chen, Sixaio; McDermott, Michael P

2012-02-01

This study was done to determine if urine phenobarbital measurements provide a reliable indicator of presence of the drug in neonates. Urine was collected from neonates treated with phenobarbital for clinical indications within 4 to 6 hours of clinically indicated collection of serum phenobarbital levels. Urine samples were also collected from control neonates not treated with phenobarbital. One aliquot was assayed fresh, another frozen at -30°C and assayed 1 to 3 months later. Phenobarbital was assayed using the ONLINE TDM Roche/Hitachi automated clinical chemistry analyzer. Serum and urine concentrations were compared as were fresh and frozen urine measurements. Serum phenobarbital ranged from 5.6 to 52.7 μg/mL. Matched urine samples were 56.6 ± 12.5% of the serum level. Frozen samples were 98.3 ± 8.0% of the fresh samples. Urine phenobarbital concentrations, either fresh or frozen, can be used in neonates as a noninvasive estimate of drug levels.

Cryopreservation of epididymal sperm.

PubMed

Patrizio, P

2000-11-27

The advent of ICSI and the perfecting of freezing protocols for sperm samples that in the pre-ICSI era would not have been frozen, allows now routine cryopreservation of epididymal sperm regardless of their quality and quantity. There are two methods to retrieve epididymal sperm: microsurgical epididymal sperm aspiration (MESA) and percutaneous epididymal sperm aspiration (PESA). The majority of the literature has focused on the technique of MESA to obtain sperm on the claim that the amount of sperm retrieved with PESA might not be sufficient to allow cryopreservation. However, there are no data on cryopreservation and ICSI with epididymal sperm collected with PESA technique. In this study, a total of 68 consecutive cycles of PESA, of which 46 were performed with fresh epididymal sperm and 22 with frozen/thawed specimens were retrospectively analyzed. In the fresh epididymal group (n = 46), 446 eggs were injected and 207 cleaving embryos were obtained (fertilization rate of 46%). In the cryopreserved epididymal sperm group (n = 22), 216 eggs were injected and 115 cleaving embryos were obtained (fertilization rate of 53%, P = NS). There were 18 pregnancies (39%) with 17 (37%) delivered/ongoing in the fresh group, while there were 11 (50%) with 9 (41%) delivered/ongoing in the frozen group (P = NS). Epididymal sperm for cryopreservation was available in 44 of the 46 PESA cycles. Additionally, in the fresh group, 19 couples had excess embryos for cryopreservation while in the frozen group, ten couples had excess embryos for cryopreservation. A total of 17 frozen embryo transfer with epididymal sperm from PESA were analyzed. Of these, 12 FET were from embryos from the fresh epididymal group and three pregnancies with livebirths (25%) were recorded. Five FET were performed with extra embryos from frozen epididymal sperm and two (40%) pregnancies with livebirths were obtained. In summary, these data show that epididymal sperm obtained by PESA can be successfully cryopreserved in order to avoid future retrievals procedures and fertilization and pregnancy rates are similar between fresh and cryopreserved epididymal sperm. It is also reported for the first time that the transfer of frozen embryos obtained with either fresh or frozen thawed epididymal sperm leads to the same pregnancy'and delivery rate.

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

PubMed

Kulíková, Barbora; Oravcová, Marta; Baláži, Andrej; Supuka, Peter; Chrenek, Peter

2017-10-01

In this study, fresh and frozen-thawed semen of Nitra and Zobor rabbit breeds were evaluated for potential inter-breed or inter-male differences in sperm quality traits. Individual male semen from four rabbits of each breed were diluted (v:v; 1:1) in a freezing medium composed of a commercial diluent, 16% of dimethyl sulphoxide (DMSO), 4% of Ficoll 70 and 2% of sucrose and frozen in liquid nitrogen vapours before being plunged into liquid nitrogen. Different motility traits, viability and plasma membrane integrity of fresh and frozen-thawed semen were evaluated in vitro using computer-assisted sperm analysis and flow cytometry. To evaluate the sperm fertilization ability, artificial insemination of fresh and frozen-thawed sperm was performed. Our results showed the effect of breed (P ≤ 0.05) on frozen-thawed sperm viability and plasma membrane integrity. Moreover, individual variability in semen quality among the rabbits was revealed (0.31 to 0.71 among quality traits). Our results thereby confirmed that the cryopreservation procedure could not ensure comparable sperm post-thaw survival for different breeds or males. Nevertheless, correlations between numbers of fresh total motile and progressively moving sperm and several quality parameters measured post thawing were revealed. Therefore, we suggest that the objective assessment of fresh rabbit sperm motility may be an effective indicator of frozen-thawed semen quality. Consequently, regular semen assessment is required in order to preserve good-quality insemination doses from native breeds.

Effect of Frozen Human Epidermis Storage Duration and Cryoprotectant on Barrier Function using Two Model Compounds

PubMed Central

Barbero, Ana M.; Frasch, H. Frederick

2015-01-01

Skin is commonly stored frozen and then thawed prior to use for in-vitro permeation experiments. Does frozen storage of skin alter its barrier property? Numerous studies have found contradictory answers to this question. In this study, the steady state flux and lag time of diethyl phthalate (DEP) were measured for fresh human skin and skin frozen at −85 °C for 1, 2, 3, 6, 9, 12, and 18 months, with 10% glycerol as cryoprotective agent. No significant differences in steady state flux were found between fresh and previously frozen samples (P = 0.6). For lag time, a significant (P = 0.002) difference was found among all groups but comparisons with fresh skin were not significant. Does glycerol have a cryoprotective effect? The steady state flux and lag time of DEP and caffeine were measured through human skin stored at −85 °C for up to 12 months with and without 10 % glycerol. No significant differences in steady state flux or lag time were found between samples stored with or without glycerol for either DEP or caffeine (P ≥ 0.17). These findings support the use of frozen skin to measure the passive permeation of chemicals in studies unconcerned with viability and metabolism. PMID:26606593

Effect of Canning and Freezing on the Nutritional Content of Apricots.

PubMed

Adkison, Erin Claire; Biasi, William B; Bikoba, Veronique; Holstege, Dirk M; Mitcham, Elizabeth J

2018-05-22

The effect of commercial canning and freezing on the nutritional content of fresh apricots was investigated. Processed samples were analyzed post-processing and after 3 months of storage and compared directly to fresh apricots from the same source. Vitamin C, beta-carotene, total phenols, and antioxidants were quantified. Compared to fresh, canned apricots initially exhibited similar levels of antioxidants, a 17% increase in beta-carotene, and a 48% increase in phenols, while vitamin C was reduced by 37%. After 3 months of storage, antioxidant levels were 47% higher than fresh. Vitamin C did not change significantly following storage and beta-carotene decreased by 15%. The canned apricot fruit packed in light syrup did not have higher total soluble solids (TSS) levels indicating no increase in fruit sugar content. Frozen apricots exhibited large increases in antioxidants (529%), beta-carotene (35%), vitamin C (3,370%), and phenols (406%) compared to fresh. After 3 months of storage, frozen apricots decreased in vitamin C (29%) and phenols (17%), but remained 2,375% and 318% higher than fresh, respectively. Beta-carotene increased during storage, reaching levels 56% higher than fresh while antioxidant activity was unchanged. This study demonstrates that key nutrients in canned and frozen apricots are retained or amplified upon processing, with the exception of vitamin C in canned apricots. The routine addition of citric and ascorbic acid to fruit prior to freezing resulted in significantly higher antioxidants, vitamin C, and phenols. Consumers eating canned or frozen apricots can feel confident of similar or superior nutritional content as compared to fresh apricots. The apricot industry is limited by the short shelf life of the fruit and consumer belief that processed produce is not as nutritious as fresh. Assessing the nutritional content of canned and frozen apricots and determining that processed apricots can deliver nearly comparable nutrient levels to fresh apricots provides the evidence needed to dispel these misconceptions and potentially increase demand for processed apricots among consumers. © 2018 Institute of Food Technologists®.

In Vivo Efficacy of Fresh vs. Frozen Osteochondral Allografts in the Goat at 6 Months is Associated with PRG4 Secretion

PubMed Central

Pallante-Kichura, Andrea L.; Chen, Albert C.; Temple-Wong, Michele M.; Bugbee, William D.; Sah, Robert L.

2014-01-01

The long-term efficacy of osteochondral allografts is due to the presence of viable chondrocytes within graft cartilage. Chondrocytes in osteochondral allografts, especially those at the articular surface that normally produce the lubricant proteoglycan-4 (PRG4), are susceptible to storage-associated death. The hypothesis of this study was that the loss of chondrocytes within osteochondral grafts leads to decreased PRG4 secretion, after graft storage and subsequent implant. The objectives were to determine the effect of osteochondral allograft treatment (FROZEN vs. FRESH) on secretion of functional PRG4 after (i) storage, and (ii) 6months in vivo in adult goats. FROZEN allograft storage reduced PRG4 secretion from cartilage by ~85% compared to FRESH allograft storage. After 6months in vivo, the PRG4-secreting function of osteochondral allografts was diminished with prior FROZEN storage by ~81% versus FRESH allografts and by ~84% versus non-operated control cartilage. Concomitantly, cellularity at the articular surface in FROZEN allografts was ~96% lower than FRESH allografts and non-operated cartilage. Thus, the PRG4-secreting function of allografts appears to be maintained in vivo based on its state after storage. PRG4 secretion may be not only a useful marker of allograft performance, but also a biological process protecting the articular surface of grafts following cartilage repair. PMID:23362152

Effect of freezing method and frozen storage duration on lamb sensory quality.

PubMed

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

2012-01-01

This study assessed the effect of three freezing methods with three frozen storage durations (1, 3, and 6 months) on the sensory quality of lamb. Methods were: air blast freezer, freezing tunnel+air blast freezer, and nitrogen chamber+air blast freezer. Meat was frozen after 48 h of ageing (0-4°C). Fresh meat (72 h ageing at 2-4°C) was used as control. Sensory analyses (trained panel and consumer tests) were performed on loin chops (Longissimus lumborum) after 24 h of thawing. Results from the trained panel test showed that freezing (method and/or storage duration) had no significant effect. Consumers found that freezing affected sensory quality. Cluster analysis for overall acceptability divided the population into four classes with different preference patterns, and none of them showed a significant preference for fresh meat. The small differences between fresh and thawed meat shown in this study should not give consumers concerns about buying frozen meat or consuming thawed meat. Copyright © 2011. Published by Elsevier Ltd.

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

PubMed

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

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

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

PubMed

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

2009-02-01

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

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

PubMed

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

2015-04-01

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

Comparative performance of isolation methods using Preston broth, Bolton broth and their modifications for the detection of Campylobacter spp. from naturally contaminated fresh and frozen raw poultry meat.

PubMed

Seliwiorstow, T; De Zutter, L; Houf, K; Botteldoorn, N; Baré, J; Van Damme, I

2016-10-03

The performance of different isolation methods was evaluated for the detection of Campylobacter from naturally contaminated raw poultry meat. Therefore, fresh and frozen poultry meat samples were analysed using the standard procedure (ISO 10272-1:2006), enrichment in Preston broth, and enrichment in modified Bolton broth (supplemented with (i) potassium clavulanate (C-BB), (ii) triclosan (T-BB), (iii) polymyxin B (P-BB)). The enrichment cultures were streaked onto both modified charcoal cefoperazone deoxycholate agar (mCCDA) and RAPID'Campylobacter agar (RCA). Moreover, direct plating on mCCDA and RCA was performed to quantify Campylobacter. In total, 33 out of 59 fresh retail meat samples (55.9%) were Campylobacter positive. For both fresh and frozen poultry meat samples, enrichment in Bolton broth (ISO 10272-1:2006) resulted in a higher number of positive samples than enrichment in Preston broth. Supplementation of Bolton broth with potassium clavulanate (C-BB) and triclosan (T-BB) enhanced the Campylobacter recovery from fresh poultry meat compared to non-supplemented Bolton broth, although the use of C-BB was less applicable than T-BB for Campylobacter recovery from frozen samples. Additionally, the use of RCA resulted in a higher isolation rate compared to mCCDA. The present study demonstrates the impact of culture medium on the recovery of Campylobacter from fresh and frozen naturally contaminated poultry meat samples and can support laboratories in choosing the most appropriate culturing method to detect Campylobacter. Copyright © 2016 Elsevier B.V. All rights reserved.

RNA-Seq-based toxicogenomic assessment of fresh frozen and formalin-fixed tissues yields similar mechanistic insights.

PubMed

Auerbach, Scott S; Phadke, Dhiral P; Mav, Deepak; Holmgren, Stephanie; Gao, Yuan; Xie, Bin; Shin, Joo Heon; Shah, Ruchir R; Merrick, B Alex; Tice, Raymond R

2015-07-01

Formalin-fixed, paraffin-embedded (FFPE) pathology specimens represent a potentially vast resource for transcriptomic-based biomarker discovery. We present here a comparison of results from a whole transcriptome RNA-Seq analysis of RNA extracted from fresh frozen and FFPE livers. The samples were derived from rats exposed to aflatoxin B1 (AFB1 ) and a corresponding set of control animals. Principal components analysis indicated that samples were separated in the two groups representing presence or absence of chemical exposure, both in fresh frozen and FFPE sample types. Sixty-five percent of the differentially expressed transcripts (AFB1 vs. controls) in fresh frozen samples were also differentially expressed in FFPE samples (overlap significance: P < 0.0001). Genomic signature and gene set analysis of AFB1 differentially expressed transcript lists indicated highly similar results between fresh frozen and FFPE at the level of chemogenomic signatures (i.e., single chemical/dose/duration elicited transcriptomic signatures), mechanistic and pathology signatures, biological processes, canonical pathways and transcription factor networks. Overall, our results suggest that similar hypotheses about the biological mechanism of toxicity would be formulated from fresh frozen and FFPE samples. These results indicate that phenotypically anchored archival specimens represent a potentially informative resource for signature-based biomarker discovery and mechanistic characterization of toxicity. Copyright © 2014 John Wiley & Sons, Ltd.

Fresh frozen cadaver workshops for advanced vascular surgical training.

PubMed

Jansen, Shirley; Cowie, Margaret; Linehan, John; Hamdorf, Jeffery M

2014-11-01

Reduction in working hours, streamlined training schemes and increasing use of endovascular techniques has meant a reduction in operative experience for newer vascular surgical trainees, especially those exposures which are not routinely performed such as thoracoabdominal, thoracotomy and retroperitoneal aortic, for example. This paper describes an Advanced Anatomy of Exposure course which was designed and convened at the Clinical Training & Evaluation Centre in Western Australia and uses fresh frozen cadavers. Feedback was obtained from the participants who attended over three courses by questionnaire. Feedback was strongly positive for the course meeting both its learning outcomes and personal learning objectives, and in addition, making a significant contribution to specialty skills. Most participants thought the fresh frozen cadaveric model significantly improved the learning objectives for training. The fresh frozen cadaver is an excellent teaching model highly representative of the living open surgical scenario where advanced trainees and newly qualified consultants can improve their operative confidence and consequently patient safety in vascular surgery. An efficient fresh frozen cadaver teaching programme can benefit many health professionals simultaneously maximizing the use of donated human tissue. © 2013 Royal Australasian College of Surgeons.

Influence of cryopreservation on perinatal outcome after blastocyst- vs cleavage-stage embryo transfer: systematic review and meta-analysis.

PubMed

Alviggi, C; Conforti, A; Carbone, I F; Borrelli, R; de Placido, G; Guerriero, S

2018-01-01

To compare the perinatal outcomes of singleton pregnancies resulting from blastocyst- vs cleavage-stage embryo transfer and to assess whether they differ between fresh and frozen embryo transfer cycles. A systematic review of the literature was carried out using the Scopus, MEDLINE and ISI Web of Science databases with no time restriction. We included only peer-reviewed articles involving humans, in which perinatal outcomes of singleton pregnancies after blastocyst-stage embryo transfer were compared with those after cleavage-stage embryo transfer. Primary outcomes were preterm birth before 37 weeks and low birth weight (< 2500 g). Secondary outcomes were very preterm birth before 32 weeks, very low birth weight (< 1500 g), small-for-gestational-age (SGA), large-for-gestational-age (LGA), perinatal mortality and congenital anomaly. A meta-analysis was performed using a random-effects model. Three subgroups were evaluated: fresh only, frozen only and fresh plus frozen embryo transfer cycles. From a total of 3928 articles identified, 14 were selected for qualitative/quantitative analysis. Significantly higher incidences of preterm birth < 37 weeks (11 studies, n = 106 629 participants; risk ratio (RR), 1.15 (95% CI, 1.05 - 1.25); P = 0.002) and very preterm birth < 32 weeks (seven studies, n = 103 742; RR, 1.16 (95% CI, 1.02-1.31); P = 0.03) were observed after blastocyst- than after cleavage-stage embryo transfer in fresh cycles. However, the risk of preterm and very preterm birth was similar after blastocyst- and cleavage-stage transfers in frozen and fresh plus frozen cycles. Overall effect size analysis revealed fewer SGA deliveries after blastocyst- compared with cleavage-stage transfer in fresh cycles but a similar number in frozen cycles. Conversely, more LGA deliveries were observed after blastocyst- compared with cleavage-stage transfer in frozen cycles (two studies, n = 39 044; RR, 1.18 (95% CI, 1.09-1.27); P < 0.0001) and no differences between the two groups in fresh cycles (four studies, n = 42 982; RR, 1.14 (95% CI, 0.97-1.35); P = 0.11). There were no differences with respect to low birth weight, very low birth weight or congenital anomalies between blastocyst- and cleavage-stage transfers irrespective of the cryopreservation method employed. Only one study reported a higher incidence of perinatal mortality after blastocyst- vs cleavage-stage embryo transfer in frozen cycles, while no differences were found in fresh cycles. Our results suggest that cryopreservation of embryos can influence outcome of pregnancy conceived following blastocyst- vs cleavage-stage embryo transfer in terms of preterm birth, very preterm birth, LGA, SGA and perinatal mortality. Caution should be exercised in interpreting these findings given the low level of evidence and wide heterogeneity of the studies. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd. Copyright © 2017 ISUOG. Published by John Wiley & Sons Ltd.

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

PubMed

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

2018-05-01

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

48 CFR 52.232-25 - Prompt payment.

Code of Federal Regulations, 2012 CFR

2012-10-01

... edible fresh or frozen poultry meat, any perishable poultry meat food product, fresh eggs, and any...) For fresh or frozen fish, as defined in section 204(3) of the Fish and Seafood Promotion Act of 1986..., cheese, certain processed cheese products, butter, yogurt, ice cream, mayonnaise, salad dressings, and...

48 CFR 52.232-25 - Prompt payment.

Code of Federal Regulations, 2014 CFR

2014-10-01

... edible fresh or frozen poultry meat, any perishable poultry meat food product, fresh eggs, and any...) For fresh or frozen fish, as defined in section 204(3) of the Fish and Seafood Promotion Act of 1986..., cheese, certain processed cheese products, butter, yogurt, ice cream, mayonnaise, salad dressings, and...

48 CFR 52.232-25 - Prompt payment.

Code of Federal Regulations, 2013 CFR

2013-10-01

... edible fresh or frozen poultry meat, any perishable poultry meat food product, fresh eggs, and any...) For fresh or frozen fish, as defined in section 204(3) of the Fish and Seafood Promotion Act of 1986..., cheese, certain processed cheese products, butter, yogurt, ice cream, mayonnaise, salad dressings, and...

In vitro fertilization outcomes after fresh and frozen blastocyst transfer in South Asian compared with Caucasian women.

PubMed

Shah, Meera Sridhar; Caballes, Marissa; Lathi, Ruth Bunker; Baker, Valerie Lynn; Westphal, Lynn Marie; Milki, Amin A

2016-06-01

To study pregnancy outcomes between South Asian and Caucasian women undergoing frozen blastocyst transfer cycles. Retrospective cohort study. Not applicable. Caucasian and South Asian patients undergoing frozen blastocyst transfer between January 2011 and December 2014. Not applicable. Live birth rate. A total of 196 Caucasian and 117 South Asian women were included in our study. Indians were on average 2.2 years younger than Caucasian women (34.9 vs. 37.1 years), and were more likely to be nulliparous (59% vs. 43%). All other baseline characteristics were similar. In women undergoing their first frozen ET cycle, implantation rate (49% vs. 47%), clinical pregnancy rate (PR; 54% vs. 49%), and live birth rate (43% vs. 43%) were similar between South Asians and Caucasians, respectively. In patients who underwent a prior fresh blastocyst transfer, the live birth rate was significantly lower in South Asian versus Caucasian women (21% vs. 37%). Our data demonstrate that IVF outcomes are better in frozen versus fresh cycles among South Asian women. The IVF clinics may wish to consider these findings when counseling South Asian patients about the timing of ET. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

The effects of sulfur mustard exposure and freezing on transdermal penetration of tritiated water through ex vivo pig skin.

PubMed

Payne, O J; Graham, S J; Dalton, C H; Spencer, P M; Mansson, R; Jenner, J; Azeke, J; Braue, E

2013-02-01

The percutaneous absorption of tritiated water ((3)H(2)O) through sulfur mustard (SM) exposed abdominal pig skin was measured using in vitro Franz-type static diffusion cells. The barrier function to water permeation following exposure to liquid SM for 8 min and excision 3h later did not change significantly. A small, but statistically significant difference (P<0.05) in steady state penetration (Jss), permeability coefficient (Kp) and lag time (t(L)) of (3)H(2)O was observed between fresh skin and skin stored frozen (-20 °C) for up to two weeks. Steady-state penetration and Kp values were significantly higher (P < 0.05) in skin stored frozen compared with fresh skin. Fresh naïve skin had an average Kp of 1.65 × 10(-3) cm h(-1), whereas frozen naïve skin was 2.04 × 10(-3) cm h(-1). Fresh SM exposed skin had a mean Kp of 1.72 × 10(-3) cm h(-1), whereas frozen SM exposed skin was 2.31 × 10(-3) cm h(-1). Lag times were also shorter (P<0.05) in skin that had been stored frozen. Frozen, SM-exposed porcine abdominal skin may be used for in vitro penetration studies, but effects of treatment and storage on the barrier layer should be taken into account. Copyright © 2012 Elsevier Ltd. All rights reserved.

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.

Association of birth defects with the mode of assisted reproductive technology in a Chinese data-linkage cohort.

PubMed

Yu, Hui-Ting; Yang, Qing; Sun, Xiao-Xi; Chen, Guo-Wu; Qian, Nai-Si; Cai, Ren-Zhi; Guo, Han-Bing; Wang, Chun-Fang

2018-05-01

To evaluate the impact of assisted reproductive technology (ART) on the offspring of Chinese population. Retrospective, data-linkage cohort. Not applicable. Live births resulting from ART or natural conception. None. Birth defects coded according to ICD-10. Births after ART were more likely to be female and multiple births, especially after intracytoplasmic sperm injection (ICSI). ART was associated with a significantly increased risk of birth defects, especially, among singleton births, a significantly increased risk in fresh-embryo cycles after in vitro fertilization (IVF) and frozen-embryo cycles after ICSI. Associations between ART and multiple defects, between ART and gastrointestinal malformation, genital organs malformation, and musculoskeletal malformation among singleton births, and between ART and cardiac septa malformation among multiple births were observed. This study suggests that ART increases the risk of birth defects. Subgroup analyses indicate higher risk for both fresh and frozen embryos, although nonsignificantly for frozen embryos after IVF and for fresh embryos were presented with low power. Larger sample size research is needed to clarify effects from fresh- or frozen-embryo cycles after IVF and ICSI. Copyright © 2018 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

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

PubMed

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

2018-04-23

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

Fresh-frozen Complete Extensor Mechanism Allograft versus Autograft Reconstruction in Rabbits

PubMed Central

Chen, Guanyin; Zhang, Hongtao; Ma, Qiong; Zhao, Jian; Zhang, Yinglong; Fan, Qingyu; Ma, Baoan

2016-01-01

Different clinical results have been reported in the repair of extensor mechanism disruption using fresh-frozen complete extensor mechanism (CEM) allograft, creating a need for a better understanding of fresh-frozen CME allograft reconstruction. Here, we perform histological and biomechanical analyses of fresh-frozen CEM allograft or autograft reconstruction in an in vivo rabbit model. Our histological results show complete incorporation of the quadriceps tendon into the host tissues, patellar survival and total integration of the allograft tibia, with relatively fewer osteocytes, into the host tibia. Vascularity and cellularity are reduced and delayed in the allograft but exhibit similar distributions to those in the autograft. The infrapatellar fat pad provides the main blood supply, and the lowest cellularity is observed in the patellar tendon close to the tibia in both the allograft and autograft. The biomechanical properties of the junction of quadriceps tendon and host tissues and those of the allograft patellar tendon are completely and considerably restored, respectively. Therefore, fresh-frozen CEM allograft reconstruction is viable, but the distal patellar tendon and the tibial block may be the weak links of the reconstruction. These findings provide new insight into the use of allograft in repairing disruption of the extensor mechanism. PMID:26911538

Fresh-frozen Complete Extensor Mechanism Allograft versus Autograft Reconstruction in Rabbits.

PubMed

Chen, Guanyin; Zhang, Hongtao; Ma, Qiong; Zhao, Jian; Zhang, Yinglong; Fan, Qingyu; Ma, Baoan

2016-02-25

Different clinical results have been reported in the repair of extensor mechanism disruption using fresh-frozen complete extensor mechanism (CEM) allograft, creating a need for a better understanding of fresh-frozen CME allograft reconstruction. Here, we perform histological and biomechanical analyses of fresh-frozen CEM allograft or autograft reconstruction in an in vivo rabbit model. Our histological results show complete incorporation of the quadriceps tendon into the host tissues, patellar survival and total integration of the allograft tibia, with relatively fewer osteocytes, into the host tibia. Vascularity and cellularity are reduced and delayed in the allograft but exhibit similar distributions to those in the autograft. The infrapatellar fat pad provides the main blood supply, and the lowest cellularity is observed in the patellar tendon close to the tibia in both the allograft and autograft. The biomechanical properties of the junction of quadriceps tendon and host tissues and those of the allograft patellar tendon are completely and considerably restored, respectively. Therefore, fresh-frozen CEM allograft reconstruction is viable, but the distal patellar tendon and the tibial block may be the weak links of the reconstruction. These findings provide new insight into the use of allograft in repairing disruption of the extensor mechanism.

Effect of Age and Bulls on Fresh Semen Quality and Frozen Semen Production of Holstein Bulls in Indonesia

NASA Astrophysics Data System (ADS)

Argiris, A.; Ondho, Y. S.; Santoso, S. I.; Kurnianto, E.

2018-02-01

Artificial Insemination is a compatible method of reproduction in an effort to increase dairy productivity. Artificial Insemination Center as a producer of frozen semen was required to maximize bulls in producing high quality frozen semen optimally. The purpose of this research was to determine effect of age and bulls on fresh semen quality and frozen semen production of Holstein bulls in Indonesia. The research was conducted at Lembang and Singosari AI Centers. The material used were 24.634 data of qualified fresh semen and frozen semen production from 81 Holstein Bulls aged 1-9 years that used as frozen semen producer in period of 2008 to 2016. The variables observed in this research were data of age of bulls, fresh semen volume (mL); sperm motility (%); mass movement; concentrations (million/mL) and frozen semen doses at each production at age of bulls. Nested design was applied to obtain and analyze data. Results showed that Age and bulls have significant effect (P<0,01) to volume, mass, motility, concentration and frozen semen production. Increasing the age of bulls resulted in increase semen volume until 7-year-old, while semen concentration decreased from 3 years old with increasing age. Frozen semen production, mass movement and motility shown the same relative value on 3-9 years old except on 1 to 2 years old had increase. Bulls would produce frozen semen optimally on 3-9 years old. Indeed, with knowledge of this factor, AI Centre might adapt management of AI bulls to improve semen production.

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

PubMed

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

2015-12-01

First, to assess if there are any differences in birth weight or gestational length in newborns from egg-donation pregnancies delivering singletons, originating from either fresh or frozen-thawed embryos when they were developed and delivered within the same mothers. Second, to determine if there are any clinical, phenotypic, or laboratory factors influencing this relationship, including the origin of the oocyte (same or different donor), the order of the children (first fresh or first frozen-thawed embryo transfer), the embryo freezing technique (vitrification or slow freezing), the in vitro embryo culture length, and the duration that embryos remained frozen. Retrospective cohorts study. University-affiliated infertility centers. A total of 360 women undergoing oocyte donation (OD), delivering (>28 weeks) at least two babies, each one from a single pregnancy, originating from at least one fresh and one frozen-thawed embryo transfer, controlling maternal and laboratory characteristics, to test the effect of embryo freezing on children size (n = 731). None. Birth weight, gestational age, weight percentile, being large for gestational age (LGA), small for gestational age (SGA), size out of normal range (ONR = LGA + SGA), and macrosomy. From fresh versus thawed embryos, respectively, mean birth weight of children was 3,183.7 g versus 3,226.4 g, gestational age was 272.1 days versus 268.8 days, and mean weight percentiles were 47.6 versus 50.1. The proportions and corresponding odds ratios (ORs) from fresh versus thawed embryos, respectively, were for LGA 13.6% versus 11.3% (OR 0.81), for SGA 9.4% versus 12.5% (OR 1.37), for ONR 23.1% versus 23.8% (OR 1.04), and for macrosomy 0.3% versus 0.8% (OR 3.1). After adjusting for clinically relevant variables, the ORs were for LGA 0.96, for SGA 1.40, for ONR 1.20, and for macrosomy not computable. None of the stated measures were significantly different. Also, independent analyses run on the origin of the oocytes, cryopreservation technique, cleavage stage of the embryos, and time that embryos remained frozen did not reveal any significant trends. This study comparing siblings from OD cycles, and eliminating the independent variables that affect early events in pregnancy, revealed no difference in duration of gestation and live birth weights between fetuses obtained after the replacement of fresh or frozen embryos. Moreover, no clinical, phenotypic, or laboratory factors appeared to be relevant, once statistically controlled. Copyright © 2015 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

The effect of different levels of sunflower head pith addition on the properties of model system emulsions prepared from fresh and frozen beef.

PubMed

Sariçoban, Cemalettin; Yilmaz, Mustafa Tahsin; Karakaya, Mustafa; Tiske, Sümeyra Sultan

2010-01-01

The effect of sunflower head pith on the functional properties of emulsions was studied by using a model system. Oil/water (O/W) model emulsion systems were prepared from fresh and frozen beef by the addition of the pith at five concentrations. Emulsion capacity (EC), stability (ES), viscosity (EV), colour and flow properties of the prepared model system emulsions were analyzed. The pith addition increased the EC and ES and the highest EC and ES values were reached when 5% of pith added; however, further increase in the pith concentration caused an inverse trend in these values. Fresh beef emulsions had higher EC and ES values than did frozen beef emulsions. One percent pith concentration was the critic level for the EV values of fresh beef emulsions. EV values of the emulsions reached a maximum level at 5% pith level, followed by a decrease at 7% pit level.

Isothiocyanate concentrations and interconversion of sulforaphane to erucin in human subjects after consumption of commercial frozen broccoli compared to fresh broccoli.

PubMed

Saha, Shikha; Hollands, Wendy; Teucher, Birgit; Needs, Paul W; Narbad, Arjan; Ortori, Catharine A; Barrett, David A; Rossiter, John T; Mithen, Richard F; Kroon, Paul A

2012-12-01

Sulforaphane (a potent anticarcinogenic isothiocyanate derived from glucoraphanin) is widely considered responsible for the protective effects of broccoli consumption. Broccoli is typically purchased fresh or frozen and cooked before consumption. We compared the bioavailability and metabolism of sulforaphane from portions of lightly cooked fresh or frozen broccoli, and investigated the bioconversion of sulforaphane to erucin. Eighteen healthy volunteers consumed broccoli soups produced from fresh or frozen broccoli florets that had been lightly cooked and sulforaphane thio-conjugates quantified in plasma and urine. Sulforaphane bioavailability was about tenfold higher for the soups made from fresh compared to frozen broccoli, and the reduction was shown to be due to destruction of myrosinase activity by the commercial blanching-freezing process. Sulforaphane appeared in plasma and urine in its free form and as several thio-conjugates forms. Erucin N-acetyl-cysteine conjugate was a significant urinary metabolite, and it was shown that human gut microflora can produce sulforaphane, erucin, and their nitriles from glucoraphanin. The short period of blanching used to produce commercial frozen broccoli destroys myrosinase and substantially reduces sulforaphane bioavailability. Sulforaphane was converted to erucin and excreted in urine, and it was shown that human colonic flora were capable of this conversion. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Freezing does not alter multiscale tendon mechanics and damage mechanisms in tension.

PubMed

Lee, Andrea H; Elliott, Dawn M

2017-12-01

It is common in biomechanics to use previously frozen tissues, where it is assumed that the freeze-thaw process does not cause consequential mechanical or structural changes. We have recently quantified multiscale tendon mechanics and damage mechanisms using previously frozen tissue, where damage was defined as an irreversible change in the microstructure that alters the macroscopic mechanical parameters. Because freezing has been shown to alter tendon microstructures, the objective of this study was to determine if freezing alters tendon multiscale mechanics and damage mechanisms. Multiscale testing using a protocol that was designed to evaluate tendon damage (tensile stress-relaxation followed by unloaded recovery) was performed on fresh and previously frozen rat tail tendon fascicles. At both the fascicle and fibril levels, there was no difference between the fresh and frozen groups for any of the parameters, suggesting that there is no effect of freezing on tendon mechanics. After unloading, the microscale fibril strain fully recovered, and interfibrillar sliding only partially recovered, suggesting that the tendon damage is localized to the interfibrillar structures and that mechanisms of damage are the same in both fresh and previously frozen tendons. © 2017 New York Academy of Sciences.

The effects of soy on freezable bread dough: a magnetic resonance study.

PubMed

Simmons, Amber L; Vodovotz, Yael

2012-11-15

Hygroscopic soy ingredients were hypothesised to slow the rate of water migration in unleavened bread dough during frozen storage. Thawed soy (18% dry weight) and wheat dough samples were assessed using non-destructive nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) for up to 8 wks frozen storage time. MRI suggested a spatially homogeneous, net increase in proton mobility with frozen storage and, with solution state proton NMR, distinct "free" and "bound" states were discerned. T(2) relaxation times of the majority proton population suggested increased mobility with frozen storage time, and statistical difference from the fresh sample was seen later for the soy samples than the wheat samples. As seen by (13)C-solid state NMR, the crystallinity of the starch was not affected by either soy addition or frozen storage. In conclusion, addition of soy to bakery products led to slightly enhanced preservation of "fresh" characteristics of the dough during frozen storage. Copyright © 2012 Elsevier Ltd. All rights reserved.

Soybeans vs. textured soy proteins as meat extenders. Cooking losses, palatability, and thiamin content of freshly cooked and frozen meat loaves.

PubMed

Ali, F S; Perry, A K; Van Duyne, F O

1982-10-01

The effects of replacing 30 percent of the ground beef in meat loaves with boiled ground soybeans or textured soy protein (TSP) were studied. The substitutions were equally effective in increasing yields of freshly cooked, raw frozen and cooked, and cooked, frozen, and reheated meat loaves. Scores for several palatability characteristics, including general desirability, were higher for meat loaves containing soybeans than for loaves containing TSP. The mixtures and loaves contained similar amounts of protein and thiamin. Cooking losses and mean palatability scores indicated advantages for freezing raw loaf mixtures rather than cooked loaves.

High quality copy number and genotype data from FFPE samples using Molecular Inversion Probe (MIP) microarrays

PubMed Central

Wang, Yuker; Carlton, Victoria EH; Karlin-Neumann, George; Sapolsky, Ronald; Zhang, Li; Moorhead, Martin; Wang, Zhigang C; Richardson, Andrea L; Warren, Robert; Walther, Axel; Bondy, Melissa; Sahin, Aysegul; Krahe, Ralf; Tuna, Musaffe; Thompson, Patricia A; Spellman, Paul T; Gray, Joe W; Mills, Gordon B; Faham, Malek

2009-01-01

Background A major challenge facing DNA copy number (CN) studies of tumors is that most banked samples with extensive clinical follow-up information are Formalin-Fixed Paraffin Embedded (FFPE). DNA from FFPE samples generally underperforms or suffers high failure rates compared to fresh frozen samples because of DNA degradation and cross-linking during FFPE fixation and processing. As FFPE protocols may vary widely between labs and samples may be stored for decades at room temperature, an ideal FFPE CN technology should work on diverse sample sets. Molecular Inversion Probe (MIP) technology has been applied successfully to obtain high quality CN and genotype data from cell line and frozen tumor DNA. Since the MIP probes require only a small (~40 bp) target binding site, we reasoned they may be well suited to assess degraded FFPE DNA. We assessed CN with a MIP panel of 50,000 markers in 93 FFPE tumor samples from 7 diverse collections. For 38 FFPE samples from three collections we were also able to asses CN in matched fresh frozen tumor tissue. Results Using an input of 37 ng genomic DNA, we generated high quality CN data with MIP technology in 88% of FFPE samples from seven diverse collections. When matched fresh frozen tissue was available, the performance of FFPE DNA was comparable to that of DNA obtained from matched frozen tumor (genotype concordance averaged 99.9%), with only a modest loss in performance in FFPE. Conclusion MIP technology can be used to generate high quality CN and genotype data in FFPE as well as fresh frozen samples. PMID:19228381

Treatment of Articular Cartilage Defects in the Goat with Frozen Versus Fresh Osteochondral Allografts: Effects on Cartilage Stiffness, Zonal Composition, and Structure at Six Months

PubMed Central

Pallante, Andrea L.; Görtz, Simon; Chen, Albert C.; Healey, Robert M.; Chase, Derek C.; Ball, Scott T.; Amiel, David; Sah, Robert L.; Bugbee, William D.

2012-01-01

Background: Understanding the effectiveness of frozen as compared with fresh osteochondral allografts at six months after surgery and the resultant consequences of traditional freezing may facilitate in vivo maintenance of cartilage integrity. Our hypothesis was that the state of the allograft at implantation affects its performance after six months in vivo. Methods: The effect of frozen as compared with fresh storage on in vivo allograft performance was determined for osteochondral allografts that were transplanted into seven recipient goats and analyzed at six months. Allograft performance was assessed by examining osteochondral structure (cartilage thickness, fill, surface location, surface degeneration, and bone-cartilage interface location), zonal cartilage composition (cellularity, matrix content), and cartilage biomechanical function (stiffness). Relationships between cartilage stiffness or cartilage composition and surface degeneration were assessed with use of linear regression. Results: Fresh allografts maintained cartilage load-bearing function, while also maintaining zonal organization of cartilage cellularity and matrix content, compared with frozen allografts. Overall, allograft performance was similar between fresh allografts and nonoperative controls. However, cartilage stiffness was approximately 80% lower (95% confidence interval [CI], 73% to 87%) in the frozen allografts than in the nonoperative controls or fresh allografts. Concomitantly, in frozen allografts, matrix content and cellularity were approximately 55% (95% CI, 22% to 92%) and approximately 96% (95% CI, 94% to 99%) lower, respectively, than those in the nonoperative controls and fresh allografts. Cartilage stiffness correlated positively with cartilage cellularity and matrix content, and negatively with surface degeneration. Conclusions: Maintenance of cartilage load-bearing function in allografts is associated with zonal maintenance of cartilage cellularity and matrix content. In this animal model, frozen allografts displayed signs of failure at six months, with cartilage softening, loss of cells and matrix, and/or graft subsidence, supporting the importance of maintaining cell viability during allograft storage and suggesting that outcomes at six months may be indicative of long-term (dys)function. Clinical Relevance: Fresh versus frozen allografts represent the “best versus worst” conditions with respect to chondrocyte viability, but “difficult versus simple” with respect to acquisition and distribution. The outcomes described from these two conditions expand the current understanding of in vivo cartilage remodeling and describe structural properties (initial graft subsidence), which may have implications for impending graft failure. PMID:23138239

Colour and oxidative stability of mince produced from fresh and frozen/thawed fallow deer (Dama dama) meat.

PubMed

Chakanya, Chido; Arnaud, Elodie; Muchenje, Voster; Hoffman, Louwrens C

2017-04-01

Colour and oxidative stability of minced meat from fresh and frozen/thawed fallow deer was investigated. For the seven fallow deer harvested, half of the meat was minced fresh and half was frozen (-20°C) for 2months under vacuum prior to grinding. Surface colour attributes, myoglobin content and surface redox forms, pH and lipid oxidation of the mince were measured during eight days of display storage. Proximate composition was determined in mince on day 0, fatty acid composition on day 0 and 8. Freezing had no effect on the proximate composition or fatty acid composition of the mince. Frozen meat mince had lower (P≤0.05) total myoglobin content but higher (P≤0.05) decrease in redness (a*) during display storage, higher (P≤0.05) accumulation of metmyoglobin at the surface from day 2 and higher (P≤0.05) TBARS values. Results showed shorter colour and oxidative stability for frozen meat mince as compared to mince from fresh meat. Display storage however did not affect fatty acid composition. Copyright © 2016 Elsevier Ltd. All rights reserved.

Detection of Cryptosporidium oocysts in fresh and frozen cattle faeces: comparison of three methods.

PubMed

Brook, E J; Christley, R M; French, N P; Hart, C A

2008-01-01

The aim of this study was to compare the performance of three commonly used screening tests for Cryptosporidium oocysts in fresh and frozen cattle faeces. Twenty-nine freshly voided faecal samples were collected from calves from three farms in the northwest of England. Three diagnostic tests for Cryptosporidium were carried out on each sample both before and after freezing - the modified Ziehl-Neelsen (MZN) and auramine phenol (APh) stains and a commercial enzyme immunoassay (EIA) kit, the ProSpecT Cryptosporidium Microplate assay (Remel, Lenexa, KS). Twelve samples were deemed positive by the reference standard (polymerase chain reaction, PCR). There were some discrepancies between the results of the screening tests and the levels of agreement were quantified. The sensitivity and specificity of each method was determined, with PCR as the gold standard. Sensitivity and specificity of the MZN stain was optimized when samples with fewer than two oocyst-like bodies were classified as negative. All three screening methods used were effective in detecting Cryptosporidium infection in both fresh and frozen calf faeces. This study has highlighted the value of determining characteristics of tests used for diagnosis and epidemiological studies.

Effect of species, breed, and age on bacterial load in bovine and bubaline semen

PubMed Central

Sannat, Chandrahas; Nair, Ajit; Sahu, S. B.; Sahasrabudhe, S. A.; Kumar, Ashish; Gupta, Amit Kumar; Shende, R. K.

2015-01-01

Aim: The present study was conducted to investigate the effect of species, breed and age on bacterial load in fresh and frozen semen of Cattle and Buffalo bull. Materials and Methods: Present study covered 56 cow and 10 buffalo bulls stationed at Central Semen Station Anjora, Durg (Chhattisgarh). Impact of breeds on bacterial load in semen was assessed using six breeds of cattle viz. Sahiwal, Gir, Red Sindhi, Tharparkar, Jersey and Holstein Friesian (HF) cross. Cow bulls were categorized into four different groups based on their age (<4 years, 4-5 years, 5-6 years and > 6 years) to study variation among age groups. Bacterial load was measured in fresh and frozen semen samples from these bulls using the standard plate count (SPC) method and count was expressed as colony forming unit (CFU) per ml of semen. Results: Higher bacterial load was reported in fresh (2.36 × 104 ± 1943 CFU/ml) and frozen (1.00 × 10 ± 90 CFU/ml) semen of cow bulls as compared to buffalo bulls (1.95 × 104 ± 2882 and 7.75 × 102 ± 160 CFU/ml in fresh and frozen semen, respectively). Jersey bull showed significantly higher bacterial count (p < 0.05) both in fresh (4.07 × 104 ± 13927 CFU/ml) and frozen (1.92 × 103 ± 178 CFU/ml) semen followed by HF cross, Sahiwal, Gir, Red Sindhi and Tharparkar bull. Bulls aged < 4 years and more than 6 years yielded increased bacterial load in their semen. Although a minor variation was reported between species and among age groups, no significant differences were measured. Conclusion: Bacterial load in semen did not differ significantly between species and age groups; however significant variation was reported among different breeds. Bulls of Jersey breed showed significantly higher bacterial load in semen as compared to the crossbred and indigenous bull. PMID:27047115

Histology as a Valid Tool To Differentiate Fresh from Frozen-Thawed Marinated Fish.

PubMed

Meistro, Serena; Pezzolato, Marzia; Muscolino, Daniele; Giarratana, Filippo; Baioni, Elisa; Panebianco, Antonio; Bozzetta, Elena

2016-08-01

European Commission Regulation (EU) 1276/2011 requires that fishery products intended for raw consumption be frozen at -20°C for not less than 24 h or at -35°C for at least 15 h in order to kill viable parasites other than trematodes. But because marinating processes are not always effective in destroying nematode larvae, raw marinated fish preparations should be frozen before consumption. This study evaluated the performance of a standardized histological method to distinguish between fresh and frozen-thawed raw marinated fish. Sixty anchovy (Engraulis encrasicolus) fillets were sampled: 30 were marinated at +4°C for 24 h, and 30 were frozen at -20°C for 24 h before being marinated for 24 h. All 60 samples were fixed in formalin, processed for paraffin embedding, cut, and stained with hematoxylin and eosin. The slide preparations were examined microscopically by three independent histopathologists and classified as frozen-thawed or negative according to standard operating procedure criteria in use at our laboratory. Performance evaluation of the method showed 100% sensitivity (95% confidence interval [CI], 88.4 to 100%) and 100% specificity (95% CI, 88.4 to 100%), and the interrater agreement (Cohen's kappa) was 1 (95% CI, 0.85 to 1). Histology proved a valid and reliable tool to distinguish fresh from frozen-thawed marinated fish. It can be applied to deliver safe raw fishery products to consumers in order to minimize the risk of anisakidosis.

Risk of ectopic pregnancy lowest with transfer of single frozen blastocyst.

PubMed

Li, Z; Sullivan, E A; Chapman, M; Farquhar, C; Wang, Y A

2015-09-01

What type of transferred embryo is associated with a lower rate of ectopic pregnancy? The lowest risk of ectopic pregnancy was associated with the transfer of blastocyst, frozen and single embryo compared with cleavage stage, fresh and multiple embryos. Ectopic pregnancy is a recognized complication following assisted reproductive technology (ART) treatment. It has been estimated that the rate of ectopic pregnancy is doubled in pregnancies following ART treatment compared with spontaneous pregnancies. However, it was not clear whether the excess rate of ectopic pregnancy following ART treatment is related to the underlying demographic factors of women undergoing ART treatment, the number of embryos transferred or the developmental stage of the embryo. A population-based cohort study of pregnancies following autologous treatment cycles between January 2009 and December 2011 were obtained from the Australian and New Zealand Assisted Reproduction Technology Database (ANZARD). The ANZARD collects ART treatment information and clinical outcomes annually from all fertility centres in Australia and New Zealand. Between 2009 and 2011, a total of 44 102 pregnancies were included in the analysis. The rate of ectopic pregnancy was compared by demographic and ART treatment factors. Generalized linear regression of Poisson distribution was used to estimate the likelihood of ectopic pregnancy. Odds ratios, adjusted odds ratios (AOR) and 95% confidence intervals (CI) were calculated. The overall rate of ectopic pregnancy was 1.4% for women following ART treatment in Australia and New Zealand. Pregnancies following single embryo transfers had 1.2% ectopic pregnancies, significantly lower than double embryo transfers (1.8%) (P < 0.01). The highest ectopic pregnancy rate was 1.9% for pregnancies from transfers of fresh cleavage embryo, followed by transfers of frozen cleavage embryo (1.7%), transfers of fresh blastocyst (1.3%), and transfers of frozen blastocyst (0.8%). Compared with fresh blastocyst transfer, the likelihood of ectopic pregnancy was 30% higher for fresh cleavage stage embryo transfers (AOR 1.30, 95% CI 1.07-1.59) and was consistent across subfertility groups. Transfer of frozen blastocyst was associated with a significantly decreased risk of ectopic pregnancy (AOR 0.70, 95% CI 0.54-0.91) compared with transfer of fresh blastocyst. A limitation of this population-based study is the lack of information available on clinical- specific protocols and processes for embryo transfer (i.e. embryo quality, cryopreservation protocol, transfer techniques, etc.) and the potential impact on outcomes. The lowest risk of ectopic pregnancy was associated with the transfer of a single frozen blastocyst. This finding adds to the increasing evidence of better perinatal outcomes following frozen embryo transfers. The approach of freezing all embryos in the initiated fresh cycle and transfer of a single frozen blastocyst in the subsequent thaw cycle may improve the overall pregnancy and birth outcomes following ART treatment, in part by reducing the ectopic pregnancy rate. There is no funding for this study. Authors declared no competing interest related to this study. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Comparison of the Skin Penetration of 3 Metabolically Stable Chemicals Using Fresh and Frozen Human Skin.

PubMed

Jacques-Jamin, Carine; Duplan, Hélène; Rothe, Helga; Vaillant, Ophelie; Eilstein, Joan; Grégoire, Sebastien; Cubberley, Richard; Lange, Daniela; Ellison, Corie; Klaric, Martina; Hewitt, Nicola; Schepky, Andreas

2017-01-01

The Cosmetics Europe ADME Task Force is developing in vitro and in silico tools for predicting skin and systemic concentrations after topical application of cosmetic ingredients. There are conflicting reports as to whether the freezing process affects the penetration of chemicals; therefore, we evaluated whether the storage of human skin used in our studies (8-12 weeks at -20°C) affected the penetration of model chemicals. Finite doses of trans-cinnamic acid (TCA), benzoic acid (BA), and 6-methylcoumarin (6MC) (non-volatile, non-protein reactive and metabolically stable in skin) were applied to fresh and thawed frozen skin from the same donors. The amounts of chemicals in different skin compartments were analysed after 24 h. Although there were some statistical differences in some parameters for 1 or 2 donors, the penetration of TCA, BA, and 6MC was essentially the same in fresh and frozen skin, i.e., there were no biologically relevant differences in penetration values. Statistical differences that were evident indicated that penetration was marginally lower in frozen than in fresh skin, indicating that the barrier function of the skin was not lost. The penetration of the 3 chemicals was essentially unaffected by freezing the skin at -20°C for up to 12 weeks. © 2017 S. Karger AG, Basel.

Contraction-free, fume-fixed longitudinal sections of fresh frozen muscle

NASA Technical Reports Server (NTRS)

Riley, Danny A.; Slocum, Glenn R.

1988-01-01

Contraction damage occurring when longitudinal frozen sections of fresh unfixed muscles are thawed on microscope slides has limited histological examination of this tissue mainly to cross sections. Longitudinally oriented sections are advantageous for investigating properties that vary along the length of the muscle fibers. A fume fixation technique has been developed for preventing contraction of thick longitudinal frozen sections. The technique is compatible with histochemical staining of enzymes.

Implications of the pH and temperature of diluted, cooled boar semen on fresh and frozen-thawed sperm motility characteristics.

PubMed

Purdy, P H; Tharp, N; Stewart, T; Spiller, S F; Blackburn, H D

2010-10-15

Boar semen is typically collected, diluted and cooled for AI use over numerous days, or frozen immediately after shipping to capable laboratories. The storage temperature and pH of the diluted, cooled boar semen could influence the fertility of boar sperm. Therefore, the purpose of this study was to determine the effects of pH and storage temperature on fresh and frozen-thawed boar sperm motility end points. Semen samples (n = 199) were collected, diluted, cooled and shipped overnight to the National Animal Germplasm Program laboratory for freezing and analysis from four boar stud facilities. The temperature, pH and motility characteristics, determined using computer automated semen analysis, were measured at arrival. Samples were then cryopreserved and post-thaw motility determined. The commercial stud was a significant source of variation for mean semen temperature and pH, as well as total and progressive motility, and numerous other sperm motility characteristics. Based on multiple regression analysis, pH was not a significant source of variation for fresh or frozen-thawed boar sperm motility end points. However, significant models were derived which demonstrated that storage temperature, boar, and the commercial stud influenced sperm motility end points and the potential success for surviving cryopreservation. We inferred that maintaining cooled boar semen at approximately 16 °C during storage will result in higher fresh and frozen-thawed boar sperm quality, which should result in greater fertility. Copyright © 2010 Elsevier Inc. All rights reserved.

Conventional freezing plus high pressure-low temperature treatment: Physical properties, microbial quality and storage stability of beef meat.

PubMed

Fernández, Pedro P; Sanz, Pedro D; Molina-García, Antonio D; Otero, Laura; Guignon, Bérengère; Vaudagna, Sergio R

2007-12-01

Meat high-hydrostatic pressure treatment causes severe decolouration, preventing its commercialisation due to consumer rejection. Novel procedures involving product freezing plus low-temperature pressure processing are here investigated. Room temperature (20°C) pressurisation (650MPa/10min) and air blast freezing (-30°C) are compared to air blast freezing plus high pressure at subzero temperature (-35°C) in terms of drip loss, expressible moisture, shear force, colour, microbial quality and storage stability of fresh and salt-added beef samples (Longissimus dorsi muscle). The latter treatment induced solid water transitions among ice phases. Fresh beef high pressure treatment (650MPa/20°C/10min) increased significantly expressible moisture while it decreased in pressurised (650MPa/-35°C/10min) frozen beef. Salt addition reduced high pressure-induced water loss. Treatments studied did not change fresh or salt-added samples shear force. Frozen beef pressurised at low temperature showed L, a and b values after thawing close to fresh samples. However, these samples in frozen state, presented chromatic parameters similar to unfrozen beef pressurised at room temperature. Apparently, freezing protects meat against pressure colour deterioration, fresh colour being recovered after thawing. High pressure processing (20°C or -35°C) was very effective reducing aerobic total (2-log(10) cycles) and lactic acid bacteria counts (2.4-log(10) cycles), in fresh and salt-added samples. Frozen+pressurised beef stored at -18°C during 45 days recovered its original colour after thawing, similarly to just-treated samples while their counts remain below detection limits during storage.

Effect of cryopreservation on IL-4, IFNgamma and IL-6 production of porcine peripheral blood lymphocytes.

PubMed

Li, Xiujin; Zhong, Zhenyu; Liang, Shuang; Wang, Xingxing; Zhong, Fei

2009-12-01

Cryopreservation of animal or human peripheral blood mononuclear cells (PBMC) is a commonly used technique. Effects of cryopreservation on functional capacity, especially the cytokine production of human PBMCs, have been extensively defined. However, certain animals, such as livestock, are a shortage of these information. Here we investigated the effects of cryopreservation on cytokine (IL-4, IFNgamma and IL-6) production of porcine PBMC. The porcine PBMCs were cryopreserved at -196 degrees C for a variety time periods for 2, 5, 25 and 50 days. Viability and cytokine production of the porcine PBMCs were measured before and after cryopreservation. The results showed that about 90% cell recovery rate was obtained at each storage time, indicating that about 10% loss of PBMCs in this short-term cryopreservation was due to the freezing process rather than the duration of cryopreservation. The fresh or frozen resting porcine PBMCs produced little cytokines in the absence of stimulation. However, three cytokines were apparently increased after PMA stimulation in both fresh and frozen porcine PBMCs. The sensitivity of frozen cells to PMA simulation for IFNgamma and IL-6 production was different from that of the fresh ones. IFNgamma production from the frozen PBMCs was significantly higher than that from the fresh ones (P<0.01). In contrast, IL-6 level from the frozen sample was significantly lower than that from the fresh one (P<0.05). Those results indicate that cryopreservation can increase the sensitivity of porcine PBMCs stimulated by PMA for IFNgamma production but not for IL-6 production. There was no significant difference of IL-4 production between fresh and frozen cells either stimulated (P>0.05) or un-stimulated (P>0.05).

Effect of incorporation of natural chemicals in water ice-glazing on freshness and shelf-life of Pacific saury (Cololabis saira) during -18 °C frozen storage.

PubMed

Luo, Haibo; Wang, Weihua; Chen, Wei; Tang, Haiqing; Jiang, Li; Yu, Zhifang

2018-07-01

Microbial spoilage and lipid oxidation are two major factors causing freshness deterioration of Pacific saury (Cololabis saira) during frozen storage. To provide a remedy, the effects of several natural chemicals incorporated alone or in combination in traditional water ice-glazing on the freshness and shelf-life of Pacific saury during frozen storage at -18 °C were investigated. Pacific sauries were subjected to individual quick freezing followed immediately by dipping into cold tap water (control) or solutions containing nisin, chitosan, phytic acid (single-factor experiment) or their combinations ((L 9 (3 4 ) orthogonal experiment) for 10 s at 1 °C and then packaged in polypropylene bags before frozen storage at -18 °C. The storage duration tested was up to 12 months. All ice-glazing treatments with individual chemicals could significantly (P < 0.05) inhibit the accumulation of thiobarbituric acid-reactive substances (TBARS), total volatile basic nitrogen (TVB-N) and histamine as well as the increase in bacterial total viable count (TVC) compared with controls, while the combination treatments gave even better effects. The L 9 (3 4 ) orthogonal experiment showed that the optimal combination was A 2 B 1 C 2 (i.e. 0.5 g L -1 nisin, 5 g L -1 chitosan and 0.2 g L -1 phytic acid). The TBARS, TVB-N, histamine and TVC values in A 2 B 1 C 2 -treated samples remained far below the maximum acceptable limit for good-freshness fish after 12 months of frozen storage at -18 °C. The incorporation of natural chemicals tested herein in ice-glazing could inhibit microbial spoilage and lipid oxidation and therefore maintain the freshness of Pacific saury during frozen storage. Under the optimal conditions, the shelf-life of Pacific saury could be extended up to 12 months at -18 °C. The study indicated that the combination treatment with natural chemicals could be commercially utilized to maintain the freshness and prolong the shelf-life of Pacific saury. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

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.

An improved cryopreservation method for porcine buccal mucosa in ex vivo drug permeation studies using Franz diffusion cells.

PubMed

Amores, Sonia; Domenech, José; Colom, Helena; Calpena, Ana C; Clares, Beatriz; Gimeno, Álvaro; Lauroba, Jacinto

2014-08-18

The use of isolated animal models to assess percutaneous absorption of molecules is frequently reported. The porcine buccal mucosa has been proposed as a substitute for the buccal mucosa barrier on ex vivo permeability studies avoiding unnecessary sacrifice of animals. But it is not always easy to obtain fresh buccal mucosa. Consequently, human and porcine buccal mucosa is sometimes frozen and stored in liquid nitrogen, but this procedure is not always feasible. One cheaper and simpler alternative is to freeze the buccal mucosa of freshly slaughtered pigs in a mechanical freezer, using DMSO and albumin as cryoprotective agents. This study compared the ex vivo permeability parameters of propranolol hydrochloride through porcine buccal mucosa using a Franz diffusion cell system and HPLC as detection method. The freezing effects on drug permeability parameters were evaluated. Equally histological studies were performed. Furthermore, the use of the parameter transmucosal water loss (TMWL) as an indicator of the buccal mucosa integrity was evaluated just as transepidermal water loss (TEWL) is utilized for skin integrity. The results showed no difference between fresh and frozen mucosal flux, permeability coefficient or lag time of propranolol. However, statistical significant difference in TMWL between fresh and frozen mucosa was observed. Copyright © 2014 Elsevier B.V. All rights reserved.

DNA methylation profiling of genomic DNA isolated from urine in diabetic chronic kidney disease: A pilot study

PubMed Central

Sexton-Oates, Alexandra; Carmody, Jake; Ekinci, Elif I.; Dwyer, Karen M.; Saffery, Richard

2018-01-01

Aim To characterise the genomic DNA (gDNA) yield from urine and quality of derived methylation data generated from the widely used Illuminia Infinium MethylationEPIC (HM850K) platform and compare this with buffy coat samples. Background DNA methylation is the most widely studied epigenetic mark and variations in DNA methylation profile have been implicated in diabetes which affects approximately 415 million people worldwide. Methods QIAamp Viral RNA Mini Kit and QIAamp DNA micro kit were used to extract DNA from frozen and fresh urine samples as well as increasing volumes of fresh urine. Matched buffy coats to the frozen urine were also obtained and DNA was extracted from the buffy coats using the QIAamp DNA Mini Kit. Genomic DNA of greater concentration than 20μg/ml were used for methylation analysis using the HM850K array. Results Irrespective of extraction technique or the use of fresh versus frozen urine samples, limited genomic DNA was obtained using a starting sample volume of 5ml (0–0.86μg/mL). In order to optimize the yield, we increased starting volumes to 50ml fresh urine, which yielded only 0–9.66μg/mL A different kit, QIAamp DNA Micro Kit, was trialled in six fresh urine samples and ten frozen urine samples with inadequate DNA yields from 0–17.7μg/mL and 0–1.6μg/mL respectively. Sufficient genomic DNA was obtained from only 4 of the initial 41 frozen urine samples (10%) for DNA methylation profiling. In comparison, all four buffy coat samples (100%) provided sufficient genomic DNA. Conclusion High quality data can be obtained provided a sufficient yield of genomic DNA is isolated. Despite optimizing various extraction methodologies, the modest amount of genomic DNA derived from urine, may limit the generalisability of this approach for the identification of DNA methylation biomarkers of chronic diabetic kidney disease. PMID:29462136

Biomechanical measurements of stiffness and strength for five types of whole human and artificial humeri.

PubMed

Aziz, Mina S R; Nicayenzi, Bruce; Crookshank, Meghan C; Bougherara, Habiba; Schemitsch, Emil H; Zdero, Radovan

2014-05-01

The human humerus is the third largest longbone and experiences 2-3% of all fractures. Yet, almost no data exist on its intact biomechanical properties, thus preventing researchers from obtaining a full understanding of humerus behavior during injury and after being repaired with fracture plates and nails. The aim of this experimental study was to compare the biomechanical stiffness and strength of "gold standard" fresh-frozen humeri to a variety of humerus models. A series of five types of intact whole humeri were obtained: human fresh-frozen (n = 19); human embalmed (n = 18); human dried (n = 15); artificial "normal" (n = 12); and artificial "osteoporotic" (n = 12). Humeri were tested under "real world" clinical loading modes for shear stiffness, torsional stiffness, cantilever bending stiffness, and cantilever bending strength. After removing geometric effects, fresh-frozen results were 585.8 ± 181.5 N/mm2 (normalized shear stiffness); 3.1 ± 1.1 N/(mm2 deg) (normalized torsional stiffness); 850.8 ± 347.9 N/mm2 (normalized cantilever stiffness); and 8.3 ± 2.7 N/mm2 (normalized cantilever strength). Compared to fresh-frozen values, statistical equivalence (p ≥ 0.05) was obtained for all four test modes (embalmed humeri), 1 of 4 test modes (dried humeri), 1 of 4 test modes (artificial "normal" humeri), and 1 of 4 test modes (artificial "osteoporotic" humeri). Age and bone mineral density versus experimental results had Pearson linear correlations ranging from R = -0.57 to 0.80. About 77% of human humeri failed via a transverse or oblique distal shaft fracture, whilst 88% of artificial humeri failed with a mixed transverse + oblique fracture. To date, this is the most comprehensive study on the biomechanics of intact human and artificial humeri and can assist researchers to choose an alternate humerus model that can substitute for fresh-frozen humeri.

DNA methylation profiling of genomic DNA isolated from urine in diabetic chronic kidney disease: A pilot study.

PubMed

Lecamwasam, Ashani; Sexton-Oates, Alexandra; Carmody, Jake; Ekinci, Elif I; Dwyer, Karen M; Saffery, Richard

2018-01-01

To characterise the genomic DNA (gDNA) yield from urine and quality of derived methylation data generated from the widely used Illuminia Infinium MethylationEPIC (HM850K) platform and compare this with buffy coat samples. DNA methylation is the most widely studied epigenetic mark and variations in DNA methylation profile have been implicated in diabetes which affects approximately 415 million people worldwide. QIAamp Viral RNA Mini Kit and QIAamp DNA micro kit were used to extract DNA from frozen and fresh urine samples as well as increasing volumes of fresh urine. Matched buffy coats to the frozen urine were also obtained and DNA was extracted from the buffy coats using the QIAamp DNA Mini Kit. Genomic DNA of greater concentration than 20μg/ml were used for methylation analysis using the HM850K array. Irrespective of extraction technique or the use of fresh versus frozen urine samples, limited genomic DNA was obtained using a starting sample volume of 5ml (0-0.86μg/mL). In order to optimize the yield, we increased starting volumes to 50ml fresh urine, which yielded only 0-9.66μg/mL A different kit, QIAamp DNA Micro Kit, was trialled in six fresh urine samples and ten frozen urine samples with inadequate DNA yields from 0-17.7μg/mL and 0-1.6μg/mL respectively. Sufficient genomic DNA was obtained from only 4 of the initial 41 frozen urine samples (10%) for DNA methylation profiling. In comparison, all four buffy coat samples (100%) provided sufficient genomic DNA. High quality data can be obtained provided a sufficient yield of genomic DNA is isolated. Despite optimizing various extraction methodologies, the modest amount of genomic DNA derived from urine, may limit the generalisability of this approach for the identification of DNA methylation biomarkers of chronic diabetic kidney disease.

21 CFR 640.34 - Processing.

Code of Federal Regulations, 2010 CFR

2010-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.34 Processing. (a) Plasma. Plasma shall be... collecting, processing, and storage system unless the product is to be stored as Liquid Plasma. (b) Fresh Frozen Plasma. Fresh frozen plasma shall be prepared from blood collected by a single uninterrupted...

21 CFR 640.34 - Processing.

Code of Federal Regulations, 2012 CFR

2012-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.34 Processing. (a) Plasma. Plasma shall be... collecting, processing, and storage system unless the product is to be stored as Liquid Plasma. (b) Fresh Frozen Plasma. Fresh frozen plasma shall be prepared from blood collected by a single uninterrupted...

21 CFR 640.34 - Processing.

Code of Federal Regulations, 2011 CFR

2011-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.34 Processing. (a) Plasma. Plasma shall be... collecting, processing, and storage system unless the product is to be stored as Liquid Plasma. (b) Fresh Frozen Plasma. Fresh frozen plasma shall be prepared from blood collected by a single uninterrupted...

21 CFR 640.34 - Processing.

Code of Federal Regulations, 2013 CFR

2013-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.34 Processing. (a) Plasma. Plasma shall be... collecting, processing, and storage system unless the product is to be stored as Liquid Plasma. (b) Fresh Frozen Plasma. Fresh frozen plasma shall be prepared from blood collected by a single uninterrupted...

21 CFR 640.34 - Processing.

Code of Federal Regulations, 2014 CFR

2014-04-01

... STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.34 Processing. (a) Plasma. Plasma shall be... collecting, processing, and storage system unless the product is to be stored as Liquid Plasma. (b) Fresh Frozen Plasma. Fresh frozen plasma shall be prepared from blood collected by a single uninterrupted...

High pressure treatments combined with sodium lactate to inactivate Escherichia coli O157:H7 and spoilage microbiota in cured beef carpaccio.

PubMed

Masana, Marcelo Oscar; Barrio, Yanina Ximena; Palladino, Pablo Martín; Sancho, Ana Maria; Vaudagna, Sergio Ramón

2015-04-01

High-pressure treatments (400 and 600 MPa) combined with the addition of sodium lactate (1 and 3%) were tested to reduce Escherichia coli O157:H7 (STEC O157) and spoilage microbiota contamination in a manufactured cured beef carpaccio in fresh or frozen conditions. Counts of spoilage microorganisms and STEC O157 were also examined during the curing step to prepare the carpaccio. STEC O157 counts remained almost unchanged through the curing process performed at 1 ± 1 °C for 12 days, with a small decrease in samples with 3% of sodium lactate. High-pressure treatments at 600 MPa for 5 min achieved an immediate reduction of up to 2 logarithmic units of STEC O157 in frozen carpaccio, and up to 1.19 log in fresh condition. Counts of spoilage bacteria diminished below detection limits in fresh or frozen carpaccio added with sodium lactate by the application of 400 and 600 MPa. Maximum injury on STEC O157 cells was observed at 600 MPa in carpaccio in fresh condition without added sodium lactate. Lethality of high-pressure treatments on STEC O157 was enhanced in frozen carpaccio, while the addition of sodium lactate at 3% reduced the lethality on STEC O157 in frozen samples, and the degree of injury in fresh carpaccio. Copyright © 2014 Elsevier Ltd. All rights reserved.

48 CFR 552.232-25 - Prompt Payment.

Code of Federal Regulations, 2012 CFR

2012-10-01

... further defined in Pub. L. 98-181, including any edible fresh or frozen poultry meat, any perishable... later than, the 7th day after product delivery. (B) For fresh or frozen fish, as defined in section 204... invoice has been received. Liquid milk, cheese, certain processed cheese products, butter, yogurt, ice...

48 CFR 552.232-25 - Prompt Payment.

Code of Federal Regulations, 2013 CFR

2013-10-01

... further defined in Pub. L. 98-181, including any edible fresh or frozen poultry meat, any perishable... later than, the 7th day after product delivery. (B) For fresh or frozen fish, as defined in section 204... invoice has been received. Liquid milk, cheese, certain processed cheese products, butter, yogurt, ice...

48 CFR 552.232-25 - Prompt Payment.

Code of Federal Regulations, 2014 CFR

2014-10-01

... further defined in Pub. L. 98-181, including any edible fresh or frozen poultry meat, any perishable... later than, the 7th day after product delivery. (B) For fresh or frozen fish, as defined in section 204... invoice has been received. Liquid milk, cheese, certain processed cheese products, butter, yogurt, ice...

Isochoric and isobaric freezing of fish muscle.

PubMed

Năstase, Gabriel; Lyu, Chenang; Ukpai, Gideon; Şerban, Alexandru; Rubinsky, Boris

2017-04-01

We have recently shown that, a living organism, which succumbs to freezing to -4 °C in an isobaric thermodynamic system (constant atmospheric pressure), can survive freezing to -4 °C in an isochoric thermodynamic system (constant volume). It is known that the mechanism of cell damage in an isobaric system is the freezing caused increase in extracellular osmolality, and, the consequent cell dehydration. An explanation for the observed survival during isochoric freezing is the thermodynamic modeling supported hypothesis that, in the isochoric frozen solution the extracellular osmolality is comparable to the cell intracellular osmolality. Therefore, cells in the isochoric frozen organism do not dehydrate, and the tissue maintains its morphological integrity. Comparing the histology of: a) fresh fish white muscle, b) fresh muscle frozen to -5 °C in an isobaric system and c) fresh muscle frozen to -5 °C I in an isochoric system, we find convincing evidence of the mechanism of cell dehydration during isobaric freezing. In contrast, the muscle tissue frozen to -5 °C in an isochoric system appears morphologically identical to fresh tissue, with no evidence of dehydration. This is the first experimental evidence in support of the hypothesis that in isochoric freezing there is no cellular dehydration and therefore the morphology of the frozen tissue remains intact. Copyright © 2017 Elsevier Inc. All rights reserved.

The effect of cellular isolation and cryopreservation on the expression of markers identifying subsets of regulatory T cells.

PubMed

Zhang, Weiying; Nilles, Tricia L; Johnson, Jacquett R; Margolick, Joseph B

2016-04-01

The role of CD4(+) regulatory T cells (Tregs) and their subsets during HIV infection is controversial. Cryopreserved peripheral blood mononuclear cells (PBMC) are an important source for assessing number and function of Tregs. However, it is unknown if PBMC isolation and cryopreservation affect the expression of CD120b and CD39, markers that identify specific subsets of Tregs. HIV-uninfected (HIV-) and -infected (HIV+) men were randomly selected from the Multicenter AIDS Cohort Study (MACS). Percentages of CD120b(+) and CD39(+) Tregs measured by flow cytometry in whole blood and in corresponding fresh and cryopreserved PBMC were compared. Percentages of CD120b(+) Tregs were significantly lower in a) fresh PBMC relative to whole blood, and b) freshly thawed frozen PBMC relative to fresh PBMC when the recovery of viable cryopreserved cells was low. When present, low expression of CD120b in frozen PBMC was reversible by 4h of in vitro culture. In contrast, expression of CD39 on Tregs was not affected by isolation and/or cryopreservation of PBMC, or by relative recovery of cryopreserved PBMC. These findings were unaffected by the HIV status of the donor. The data suggest that percentages of CD120b(+) Tregs and CD39(+) Tregs can be validly measured in either whole blood or PBMC (fresh and frozen) in HIV- and HIV+ men. However, for measurement of CD120b(+) Tregs one type of sample should be used consistently within a given study, and thawed frozen cells may require in vitro culture if recovery of viable cells is low. Copyright © 2016 Elsevier B.V. All rights reserved.

Radioprotection provides functional mechanics but delays healing of irradiated tendon allografts after ACL reconstruction in sheep.

PubMed

Seto, Aaron U; Culp, Brian M; Gatt, Charles J; Dunn, Michael

2013-12-01

Successful protection of tissue properties against ionizing radiation effects could allow its use for terminal sterilization of musculoskeletal allografts. In this study we functionally evaluate Achilles tendon allografts processed with a previously developed radioprotective treatment based on (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) crosslinking and free radical scavenging using ascorbate and riboflavin, for ovine anterior cruciate ligament reconstruction. Arthroscopic anterior cruciate ligament (ACL) reconstruction was performed using double looped allografts, while comparing radioprotected irradiated and fresh frozen allografts after 12 and 24 weeks post-implantation, and to control irradiated grafts after 12 weeks. Radioprotection was successful at preserving early subfailure mechanical properties comparable to fresh frozen allografts. Twelve week graft stiffness and anterior-tibial (A-T) translation for radioprotected and fresh frozen allografts were comparable at 30 % of native stiffness, and 4.6 and 5 times native A-T translation, respectively. Fresh frozen allograft possessed the greatest 24 week peak load at 840 N and stiffness at 177 N/mm. Histological evidence suggested a delay in tendon to bone healing for radioprotected allografts, which was reflected in mechanical properties. There was no evidence that radioprotective treatment inhibited intra-articular graft healing. This specific radioprotective method cannot be recommended for ACL reconstruction allografts, and data suggest that future efforts to improve allograft sterilization procedures should focus on modifying or eliminating the pre-crosslinking procedure.

Construct Validity of Fresh Frozen Human Cadaver as a Training Model in Minimal Access Surgery

PubMed Central

Macafee, David; Pranesh, Nagarajan; Horgan, Alan F.

2012-01-01

Background: The construct validity of fresh human cadaver as a training tool has not been established previously. The aims of this study were to investigate the construct validity of fresh frozen human cadaver as a method of training in minimal access surgery and determine if novices can be rapidly trained using this model to a safe level of performance. Methods: Junior surgical trainees, novices (<3 laparoscopic procedure performed) in laparoscopic surgery, performed 10 repetitions of a set of structured laparoscopic tasks on fresh frozen cadavers. Expert laparoscopists (>100 laparoscopic procedures) performed 3 repetitions of identical tasks. Performances were scored using a validated, objective Global Operative Assessment of Laparoscopic Skills scale. Scores for 3 consecutive repetitions were compared between experts and novices to determine construct validity. Furthermore, to determine if the novices reached a safe level, a trimmed mean of the experts score was used to define a benchmark. Mann-Whitney U test was used for construct validity analysis and 1-sample t test to compare performances of the novice group with the benchmark safe score. Results: Ten novices and 2 experts were recruited. Four out of 5 tasks (nondominant to dominant hand transfer; simulated appendicectomy; intracorporeal and extracorporeal knot tying) showed construct validity. Novices’ scores became comparable to benchmark scores between the eighth and tenth repetition. Conclusion: Minimal access surgical training using fresh frozen human cadavers appears to have construct validity. The laparoscopic skills of novices can be accelerated through to a safe level within 8 to 10 repetitions. PMID:23318058

Reconstruction of the extensor mechanism with fresh-frozen tendon allograft in total knee arthroplasty.

PubMed

Llombart Blanco, Rafael; Valentí, Andrés; Díaz de Rada, Pablo; Mora, Gonzalo; Valentí, Juan R

2014-11-01

Patellar tendon rupture after total knee replacement is a rare and highly limiting injury with multifactorial aetiology. Many reconstruction techniques have been described with not very predictable results. The use of allografts has been accepted as a suitable solution. A series of seven patients with patellar tendon rupture treated with fresh-frozen tendon allograft reconstruction after knee arthroplasty is presented. Median follow-up is 25 months (20-31). Functional assessment improved, and the knee society score and knee functional score improved from 26 and 16 to 82 and 55, respectively. Median extension lag was 5° (0°-20°), with a median range of motion of 95° (70-100). Radiological study showed a rise of the patella of 22.26 mm. The use of fresh-frozen allografts as a solution to patellar tendon ruptures after knee arthroplasty seems to provide acceptable results. Increased patellar height does not seem to affect functionality. Case series, Level IV.

9 CFR 319.145 - Italian sausage products.

Code of Federal Regulations, 2011 CFR

2011-01-01

... containing at least 85 percent meat, or combination of meat and fat, with the total fat content constituting... fresh or frozen pork, or pork and pork fat, and may contain Mechanically Separated (Species) in... Sausage” shall be prepared with fresh or frozen beef or beef and beef fat. “Italian Veal Sausage” or...

Bacteria-killing ability of fresh blood plasma compared to frozen blood plasma

DOE PAGES

Jacobs, Anne C.; Fair, Jeanne Marie

2015-10-09

In recent years, the bacteria-killing assay (BKA) has become a popular technique among ecoimmunologists. New variations of that assay allow researchers to use smaller volumes of blood, an important consideration for those working on small-bodied animals. However, this version of the assay requires access to a lab with a nanodrop spectrophotometer, something that may not be available in the field. One possible solution is to freeze plasma for transport; however, this assumes that frozen plasma samples will give comparable results to fresh ones. Here, we tested this assumption using plasma samples from three species of birds: chickens (Gallus gallus), ash-throatedmore » flycatchers (Myiarchus cinerascens), and western bluebirds (Sialia mexicana). Chicken plasma samples lost most or all of their bacterial killing ability after freezing. This did not happen in flycatchers and bluebirds; however, frozen plasma did not produce results comparable to those obtained using fresh plasma. Finally, we caution researchers using the BKA to use fresh samples whenever possible, and to validate the use of frozen samples on a species-by-species basis.« less

Bacteria-killing ability of fresh blood plasma compared to frozen blood plasma.

PubMed

Jacobs, Anne C; Fair, Jeanne M

2016-01-01

In recent years, the bacteria-killing assay (BKA) has become a popular technique among ecoimmunologists. New variations of that assay allow researchers to use smaller volumes of blood, an important consideration for those working on small-bodied animals. However, this version of the assay requires access to a lab with a nanodrop spectrophotometer, something that may not be available in the field. One possible solution is to freeze plasma for transport; however, this assumes that frozen plasma samples will give comparable results to fresh ones. We tested this assumption using plasma samples from three species of birds: chickens (Gallus gallus), ash-throated flycatchers (Myiarchus cinerascens), and western bluebirds (Sialia mexicana). Chicken plasma samples lost most or all of their bacterial killing ability after freezing. This did not happen in flycatchers and bluebirds; however, frozen plasma did not produce results comparable to those obtained using fresh plasma. We caution researchers using the BKA to use fresh samples whenever possible, and to validate the use of frozen samples on a species-by-species basis. Copyright © 2015 Elsevier Inc. All rights reserved.

Bacteria-killing ability of fresh blood plasma compared to frozen blood plasma

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

Jacobs, Anne C.; Fair, Jeanne Marie

In recent years, the bacteria-killing assay (BKA) has become a popular technique among ecoimmunologists. New variations of that assay allow researchers to use smaller volumes of blood, an important consideration for those working on small-bodied animals. However, this version of the assay requires access to a lab with a nanodrop spectrophotometer, something that may not be available in the field. One possible solution is to freeze plasma for transport; however, this assumes that frozen plasma samples will give comparable results to fresh ones. Here, we tested this assumption using plasma samples from three species of birds: chickens (Gallus gallus), ash-throatedmore » flycatchers (Myiarchus cinerascens), and western bluebirds (Sialia mexicana). Chicken plasma samples lost most or all of their bacterial killing ability after freezing. This did not happen in flycatchers and bluebirds; however, frozen plasma did not produce results comparable to those obtained using fresh plasma. Finally, we caution researchers using the BKA to use fresh samples whenever possible, and to validate the use of frozen samples on a species-by-species basis.« less

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

PubMed

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

1988-01-01

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

Detection and Characterization of Circulating Tumour Cells from Frozen Peripheral Blood Mononuclear Cells

PubMed Central

Lu, David; Graf, Ryon P.; Harvey, Melissa; Madan, Ravi A.; Heery, Christopher; Marte, Jennifer; Beasley, Sharon; Tsang, Kwong Y.; Krupa, Rachel; Louw, Jessica; Wahl, Justin; Bales, Natalee; Landers, Mark; Marrinucci, Dena; Schlom, Jeffrey; Gulley, James L.; Dittamore, Ryan

2015-01-01

Retrospective analysis of patient tumour samples is a cornerstone of clinical research. CTC biomarker characterization offers a non-invasive method to analyse patient samples. However, current CTC technologies require prospective blood collection, thereby reducing the ability to utilize archived clinical cohorts with long-term outcome data. We sought to investigate CTC recovery from frozen, archived patient PBMC pellets. Matched samples from both mCRPC patients and mock samples, which were prepared by spiking healthy donor blood with cultured prostate cancer cell line cells, were processed “fresh” via Epic CTC Platform or from “frozen” PBMC pellets. Samples were analysed for CTC enumeration and biomarker characterization via immunofluorescent (IF) biomarkers, fluorescence in-situ hybridization (FISH) and CTC morphology. In the frozen patient PMBC samples, the median CTC recovery was 18%, compared to the freshly processed blood. However, abundance and localization of cytokeratin (CK) and androgen receptor (AR) protein, as measured by IF, were largely concordant between the fresh and frozen CTCs. Furthermore, a FISH analysis of PTEN loss showed high concordance in fresh vs. frozen. The observed data indicate that CTC biomarker characterization from frozen archival samples is feasible and representative of prospectively collected samples. PMID:28936240

Nitrite-embedded packaging film effects on fresh and frozen beef color development and stability as influenced by meat age and muscle type.

PubMed

Claus, James R; Du, Chen

2013-11-01

Muscles (Longissimus lumborum, LL; Psoas major, PM, semitendinosus, ST) were aged (2, 9d postmortem), cut into steaks, anaerobically packaged (nitrite-embedded film, NEF), and displayed (fresh, 19d; frozen, 39d). Fresh NEF increased (P<0.05) in redness (first 48h). Upon opening fresh NEF (d 6) and overwrapping in PVC film, redness declined (P<0.05). NEF cooked LL had more red surface compared to non-NEF. Meat age influenced NEF color. Intact NEF maintained acceptable red color throughout display. Residual nitrite and nitrate associated with fresh NEF and nitrate in NEF cooked LL were found (P<0.05) in the outer layer. Consideration should be given to providing sufficient time for nitric oxide myoglobin development when using NEF which may be influenced by meat age and muscle differences. NEF packaging has potential to extend fresh beef color display life. NEF appears to offer the opportunity to display bright red beef in frozen display by limiting typical effects of photooxidation. Copyright © 2013 Elsevier Ltd. All rights reserved.

The Change of Total Anthocyanins in Blueberries and Their Antioxidant Effect After Drying and Freezing

PubMed Central

Srzednicki, George

2004-01-01

This study examined the effects of freezing, storage, and cabinet drying on the anthocyanin content and antioxidant activity of blueberries (Vaccinium corymbosum L). Fresh samples were stored for two weeks at 5°C while frozen samples were kept for up to three months at −20°C. There were two drying treatments, one including osmotic pretreatment followed by cabinet drying and the other involving only cabinet drying. Total anthocyanins found in fresh blueberries were 7.2 ± 0.5 mg/g dry matter, expressed as cyanidin 3-rutinoside equivalents. In comparison with fresh samples, total anthocyanins in untreated and pretreated dried blueberries were significantly reduced to 4.3 ± 0.1 mg/g solid content, 41% loss, and 3.7 ± 0.2 mg/g solid content, 49% loss, respectively. Osmotic treatment followed by a thermal treatment had a greater effect on anthocyanin loss than the thermal treatment alone. In contrast, the frozen samples did not show any significant decrease in anthocyanin level during three months of storage. Measurement of the antioxidant activity of anthocyanin extracts from blueberries showed there was no significant difference between fresh, dried, and frozen blueberries. PMID:15577185

Effect of short- and long-term heat stress on the conception risk of dairy cows under natural service and artificial insemination breeding programs.

PubMed

Schüller, L-K; Burfeind, O; Heuwieser, W

2016-04-01

The objectives of this retrospective study were to examine the effect of heat stress on natural service and artificial insemination (AI) breeding methods. We investigated the influence of short- and long-term heat stress on the conception risk (CR) of dairy cows bred by natural service or by AI with frozen-thawed or fresh semen. In addition, the relationship between breeding method and parity was determined. Cows bred by AI with frozen-thawed semen exposed to long-term heat stress (mean temperature-humidity index ≥73 in the period 21d before breeding) were 63% less likely to get pregnant compared with cows not exposed to heat stress. Cows bred by AI with fresh semen were 80% less likely to get pregnant during periods of short-term heat stress than during periods without heat stress. Furthermore, multiparous cows bred by AI with frozen-thawed or fresh semen were 22 and 67% less likely to get pregnant, respectively, than primiparous cows. No influence of heat stress or parity was noted on the CR of cows bred by natural service. The present study indicates that the likelihood of dairy cows becoming pregnant is reduced by short- and long-term heat stress depending on the type of semen employed. In particular, CR of cows inseminated with fresh semen is negatively affected by short-term heat stress and CR of cows inseminated with frozen-thawed semen is negatively affected by long-term heat stress. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

9 CFR 94.7 - Disposal of animals, meats, and other articles ineligible for importation.

Code of Federal Regulations, 2014 CFR

2014-01-01

..., AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR DISEASE, AND BOVINE SPONGIFORM ENCEPHALOPATHY... importation. (a) Ruminants and swine, and fresh (chilled or frozen) meats, prohibited importation under §§ 94... swine, and fresh (chilled or frozen) meats, prohibited importation under §§ 94.1, 94.8, 94.9, 94.10, 94...

Patient and cycle characteristics predicting high pregnancy rates with single-embryo transfer: an analysis of the Society for Assisted Reproductive Technology outcomes between 2004 and 2013.

PubMed

Mersereau, Jennifer; Stanhiser, Jamie; Coddington, Charles; Jones, Tiffany; Luke, Barbara; Brown, Morton B

2017-11-01

To analyze factors associated with high live birth rate and low multiple birth rate in fresh and frozen-thawed assisted reproductive technology (ART) cycles. Retrospective cohort analysis. Not applicable. The study population included 181,523 women undergoing in vitro fertilization with autologous fresh first cycles, 27,033 with fresh first oocyte donor cycles, 37,658 with fresh second cycles, and 35,446 with frozen-thawed second cycles. None. Live birth rate and multiple birth rate after single-embryo transfer (SET) and double embryo transfer (DET) were measured, in addition to cycle characteristics. In patients with favorable prognostic factors, including younger maternal age, transfer of a blastocyst, and additional embryos cryopreserved, the gain in the live birth rate from SET to DET was approximately 10%-15%; however, the multiple birth rate increased from approximately 2% to greater than 49% in both autologous and donor fresh and frozen-thawed transfer cycles. This study reports a 10%-15% reduction in live birth rate and a 47% decrement in multiple birth rate with SET compared with DET in the setting of favorable patient prognostic factors. Our findings present an opportunity to increase the rate of SET across the United States and thereby reduce the multiple birth rate and its associated poor perinatal outcomes with assisted reproductive technology pregnancies. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Biomechanical measurements of stopping and stripping torques during screw insertion in five types of human and artificial humeri.

PubMed

Aziz, Mina Sr; Tsuji, Matthew Rs; Nicayenzi, Bruce; Crookshank, Meghan C; Bougherara, Habiba; Schemitsch, Emil H; Zdero, Radovan

2014-05-01

During orthopedic surgery, screws are inserted by "subjective feel" in humeri for fracture fixation, that is, stopping torque, while trying to prevent accidental over-tightening that causes screw-bone interface failure, that is, stripping torque. However, no studies exist on stopping torque, stripping torque, or stopping/stripping torque ratio in human or artificial humeri. This study evaluated five types of humeri, namely, human fresh-frozen (n = 19), human embalmed (n = 18), human dried (n = 15), artificial "normal" (n = 13), and artificial "osteoporotic" (n = 13). An orthopedic surgeon used a torque screwdriver to insert 3.5-mm-diameter cortical screws into humeral shafts and 6.5-mm-diameter cancellous screws into humeral heads by "subjective feel" to obtain stopping and stripping torques. The five outcome measures were raw and normalized stopping torque, raw and normalized stripping torque, and stopping/stripping torque ratio. Normalization was done as raw torque/screw-bone interface area. For "gold standard" fresh-frozen humeri, cortical screw tests yielded averages of 1312 N mm (raw stopping torque), 30.4 N/mm (normalized stopping torque), 1721 N mm (raw stripping torque), 39.0 N/mm (normalized stripping torque), and 82% (stopping/stripping torque ratio). Similarly, fresh-frozen humeri gave cancellous screw average results of 307 N mm (raw stopping torque), 0.9 N/mm (normalized stopping torque), 392 N mm (raw stripping torque), 1.2 N/mm (normalized stripping torque), and 79% (stopping/stripping torque ratio). Of the five cortical screw parameters for fresh-frozen humeri versus other groups, statistical equivalence (p ≥ 0.05) occurred in four cases (embalmed), three cases (dried), four cases (artificial "normal"), and four cases (artificial "osteoporotic"). Of the five cancellous screw parameters for fresh-frozen humeri versus other groups, statistical equivalence (p ≥ 0.05) occurred in five cases (embalmed), one case (dried), one case (artificial "normal"), and zero cases (artificial "osteoporotic"). Stopping/stripping torque ratios were relatively constant for all groups at 77%-88% (cortical screws) and 79%-92% (cancellous screws). © IMechE 2014.

Isochoric and isobaric freezing of fish muscle

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

Năstase, Gabriel; Department of Building Services, University of Transilvania, Braşov, Braşov, 500152; Lyu, Chenang

We have recently shown that, a living organism, which succumbs to freezing to −4 °C in an isobaric thermodynamic system (constant atmospheric pressure), can survive freezing to −4 °C in an isochoric thermodynamic system (constant volume). It is known that the mechanism of cell damage in an isobaric system is the freezing caused increase in extracellular osmolality, and, the consequent cell dehydration. An explanation for the observed survival during isochoric freezing is the thermodynamic modeling supported hypothesis that, in the isochoric frozen solution the extracellular osmolality is comparable to the cell intracellular osmolality. Therefore, cells in the isochoric frozen organism do not dehydrate, andmore » the tissue maintains its morphological integrity. Comparing the histology of: a) fresh fish white muscle, b) fresh muscle frozen to −5 °C in an isobaric system and c) fresh muscle frozen to −5 °C I in an isochoric system, we find convincing evidence of the mechanism of cell dehydration during isobaric freezing. In contrast, the muscle tissue frozen to −5 °C in an isochoric system appears morphologically identical to fresh tissue, with no evidence of dehydration. This is the first experimental evidence in support of the hypothesis that in isochoric freezing there is no cellular dehydration and therefore the morphology of the frozen tissue remains intact. - Highlights: • Preservation of fish muscle at, subfreezing temperatures, in an isochoric system, is demonstrated. • Experiments were performed to an average pressure of 41.3 MPa and temperatures of −5 °C. • Isochoric subfreezing temperature is a new preservation method that does not require the.use of cryoprotectants. • No cellular dehydration and therefore the morphology of the frozen tissue remains intact.« less

A comparison of biochemical pregnancy rates between women who underwent IVF and fertile controls who conceived spontaneously†.

PubMed

Zeadna, Atif; Son, Weon Young; Moon, Jeong Hee; Dahan, Michael H

2015-04-01

Does IVF affect the biochemical pregnancy rate? The likelihood of an early pregnancy loss may be lower and is certainly not higher in IVF cycles when compared with published rates of biochemical pregnancy in fertile women ≤42 years old. The use of gonadotrophins to stimulate multi-folliculogenesis alters endometrial expression of genes and proteins, compared with unstimulated cycles. Exogenous estrogen and progesterone taken for endometrial preparation in frozen embryo transfer cycles, also cause changes in endometrial gene and protein expression .These endometrial alterations may compromise the ability of embryos to develop once implanted, possibly increasing the biochemical pregnancy rate. This is a retrospective study, involving 1636 fresh and 188 frozen, single embryo transfer (SET) IVF cycles performed between August 2008 and December 2012. The biochemical pregnancy rate of the 1824 combined IVF and frozen cycles were compared with fertile controls, derived from the three prospective studies in the medical literature that evaluate this rate. Subjects ≤42-years old, who underwent a SET, as part of a fresh or thawed IVF cycle were considered for inclusion. Each subject is represented only once. The biochemical pregnancy rates were compared with those of historical standard, fertile populations with spontaneous conceptions. The pregnancy rates per transfer for fresh and frozen IVF cycles were similar at 39 and 40%, respectively. There was also no significant difference in the likelihood of pregnancy outcomes (clinical, biochemical and ectopic pregnancy) between fresh IVF and frozen cycles (85.4 versus 85.6%, 13.8 versus 14.8%, 0.5 versus 0%, P = 0.82). However, pregnancy rates decreased in older patients when compared with younger ones P < 0.0001. The biochemical pregnancy rate for fresh and frozen IVF cycles combined was 13.8% of all pregnancies. IVF and frozen cycles were combined as the IVF group treated with hormones for further comparison with the fertile control group. The biochemical pregnancy rate (14%) in the IVF group was lower than the rate based on the total fertile group (18%), P = 0.01 and differed significantly from the rate in two out of the three studies used to establish the normative rate. The age ranges of the IVF and fertile controls were 21-42 years. The mean age in the IVF population was 34.8 years, as compared with 29 years, 29, 4 years and 30.6 years (Zinaman) in the three published studies (mean: 29.4 years). This is a retrospective study and it was impossible to recruit an in-house biochemical pregnancy control population. Lower early pregnancy wastage after IVF may be due to the opportunity to select the embryo for transfer. This finding should be confirmed in further studies but supports the idea that embryo selection is an important step. None. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

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

PubMed

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

1998-09-01

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

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

PubMed

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

2016-09-19

Embryo culture media used for IVF treatment might affect fetal growth and thus birthweight of the newborns. A retrospective study was conducted in South China using data from 2370 singleton neonates born after IVF/ICSI between 2009 and 2012. Two culture media, i.e., either Vitrolife or SAGE were used as embryo culture media during the study period. Neonates' birthweights were compared between the two embryo culture media groups. Among the 2370 singletons, 1755 cases came from fresh cleavage embryo transfer while 615 were from frozen-thawed cleavage embryo transfer. Within the fresh embryo transfer newborns, no statistical difference was observed in either birthweight (mean ± SD: 3196.0 ± 468.9 versus 3168.4 ± 462.0g, p > 0.05) or adjusted birthweight controlled for gestational age and gender (z-score mean ± SD: 0.11 ± 1.02 versus 0.11 ± 0.99 g, P > 0.05) between the Vitrolife (n = 419) and the SAGE group (n = 1336). Likewise within frozen embryo transfer neotates, no statistical difference of the birthweight (3300.6 ± 441.3 vs.3256.0 ± 466.7 g, P > 0.05) and adjusted birthweight (0.30 ± 0.99 g versus 0.29 ± 0.97 g, P > 0.05) was found between the Vitrolife (n = 202) and the SAGE group (n = 413). The sex ratio [OR1.17, 95 % CI (0.94-1.46)/OR1.1, 95 % CI (0.78-1.54)], rate of small for gestational age [OR1.14, 95 % CI (0.82-1.59)/OR1.06, 95 % CI (0.56-2.02)] and large for gestational age [OR1.07, 95 % CI (0.64-1.76)/OR0.98, 95 % CI (0.47-2.02)] in fresh and frozen-thawed subgourps are all comparable respectively between the two culture media. No group differences were found in the rate of low birthweight and macosomia. Multiple linear regression analysis demonstrated that maternal weight, gestational age, frozen-thawed embryo transfer and infant gender were significantly related to neonatal birthweight (P < 0.001). It appears that embryos cultured in SAGE or Vitrolife media after fresh or frozen-thawed cleavage embryo transfer did not affect neonate's birthweight.

Postoperative acute kidney injury following intraoperative blood product transfusions during cardiac surgery.

PubMed

Kindzelski, Bogdan A; Corcoran, Philip; Siegenthaler, Michael P; Horvath, Keith A

2018-01-01

This study explored the nature of the association between intraoperative usage of red blood cell, fresh frozen plasma, cryoprecipitate or platelet transfusions and acute kidney injury. A total of 1175 patients who underwent cardiac surgery between 2008 and 2013 were retrospectively analyzed. We assessed the association between: (1) preoperative patient characteristics and acute kidney injury, (2) intraoperative blood product usage and acute kidney injury, (3) acute kidney injury and 30-day mortality or re-hospitalization. In our cohort of 1175 patients, 288 patients (24.5%) developed acute kidney injury. This included 162 (13.8%), 69 (5.9%) and 57 (4.9%) developing stage 1, stage 2 or stage 3 acute kidney injury, respectively. Increased red blood cell, fresh frozen plasma or platelet transfusions increased the odds of developing acute kidney injury. Specifically, every unit of red blood cells, fresh frozen plasma or platelets transfused was associated with an increase in the covariate-adjusted odds ratio of developing ⩾ stage 2 kidney injury of 1.18, 1.19 and 1.04, respectively. Intraoperative blood product transfusions were independently associated with an increased odds of developing acute kidney injury following cardiac surgery. Further randomized studies are needed to better define intraoperative transfusion criteria.

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

PubMed

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

2017-10-01

Cryopreservation is an advanced method used to protect germplasm in liquid nitrogen. Honey bees are of special interest to protect because of their pollination activity and critical role in agriculture. There has been important progress in the cryopreservation of honey bee germplasm in recent years, leading to practical recovery of genetic material for breeding purposes following freezing. However, there remains room for improvement and the goal of the present study was to evaluate the effect of different "extenders" added post-thaw on the fertilization rate of cryopreserved honey bee semen. The purpose of adding extender post-thaw was to dilute the cryoprotectant to remove chemicals after centrifugation because of potential adverse effects. The control consisted of frozen-thawed semen without the addition of an extender; treatment groups included the addition of one of the following extenders: glucose solution, fresh ram semen plasma, fresh honey bee semen plasma, extender solution. All of the above treatments and frozen-thawed control were compared to fresh semen. For each group, 15 virgin queens were instrumentally inseminated with the semen-diluent solution and introduced into nucleus colonies to determine the brood patterns of the queens. Percentages of worker brood produced in the fresh semen, frozen-thawed semen control, glucose, fresh ram semen plasma, fresh honey bee semen plasma, and extender solution supplemented groups were 98.±1.1%, 47.0 ± 0.9%, 3.0 ± 0.8%, 0.3 ± 0.1%, 48.1 ± 4.1% and 40.3 ± 2.4%, respectively. Similiarly, spermatozoa numbers in the spermathecae of the same treatment groups were 3.6 × 10 6 , 1.6 × 10 6 , 7.3 × 10 5 , 4.7 × 10 5 , 8.1 × 10 5 , and 4.6 × 10 5 spermatozoa for the same treatment, respectively. The differences in both worker brood percentage and sperm count in the spermatheca were statistically significant (P < 0.01) among all treatment groups, except the frozen-thawed control group and fresh drone semen plasma group. We found a positive correlation between sperm count in the spermatheca and the percentage of worker brood (r = 0.91). With the exception of fresh honey bee semen plasma, the fertility rate was reduced following the addition of various plasmas and diluents post-freezing. Copyright © 2017 Elsevier Inc. All rights reserved.

Survey of Salmonella and Campylobacter contamination of whole, raw poultry on retail sale in Wales in 2003.

PubMed

Meldrum, R J; Tucker, I D; Smith, R M M; Edwards, C

2005-07-01

A survey of the Salmonella and Campylobacter contamination of raw, whole chickens available to consumers in Wales was performed between March and December 2003. In total, 736 samples were taken, and overall contamination rates of 73.1% for Campylobacter and 5.7% for Salmonella were found. This survey follows a survey performed during 2001 to 2002 by Welsh local authorities and the National Public Health Service for Wales that established updated baseline rates for both pathogens in raw, whole chicken available to consumers in Wales. This survey indicated no difference in Campylobacter rates between fresh and frozen samples or between samples taken from retailers and local butchers, but significant differences existed in Salmonella rates between fresh and frozen samples and between those sampled from retailers and butchers, with frozen chickens and samples taken from retailers having significantly higher rates. However, the difference in Salmonella isolation rate between retailers and butchers was found to be due to the differences in the proportions of fresh and frozen chickens sampled from these locations, with a significantly higher number of frozen chickens (with a higher Salmonella rate) being sampled from retailers.

9 CFR 94.2 - Fresh (chilled or frozen) products (other than meat), and milk and milk products of ruminants and...

Code of Federal Regulations, 2010 CFR

2010-01-01

... (other than meat), and milk and milk products of ruminants and swine. 94.2 Section 94.2 Animals and... (other than meat), and milk and milk products of ruminants and swine. (a) The importation of fresh (chilled or frozen) products (other than meat and milk and milk products) derived from ruminants or swine...

9 CFR 319.141 - Fresh pork sausage.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Fresh pork sausage. 319.141 Section 319.141 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... Sausage § 319.141 Fresh pork sausage. “Fresh Pork Sausage” is sausage prepared with fresh pork or frozen...

9 CFR 319.141 - Fresh pork sausage.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Fresh pork sausage. 319.141 Section 319.141 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... Sausage § 319.141 Fresh pork sausage. “Fresh Pork Sausage” is sausage prepared with fresh pork or frozen...

9 CFR 319.141 - Fresh pork sausage.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Fresh pork sausage. 319.141 Section 319.141 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... Sausage § 319.141 Fresh pork sausage. “Fresh Pork Sausage” is sausage prepared with fresh pork or frozen...

9 CFR 319.142 - Fresh beef sausage.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 9 Animals and Animal Products 2 2013-01-01 2013-01-01 false Fresh beef sausage. 319.142 Section 319.142 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... Sausage § 319.142 Fresh beef sausage. “Fresh Beef Sausage” is sausage prepared with fresh beef or frozen...

9 CFR 319.142 - Fresh beef sausage.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 9 Animals and Animal Products 2 2014-01-01 2014-01-01 false Fresh beef sausage. 319.142 Section 319.142 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... Sausage § 319.142 Fresh beef sausage. “Fresh Beef Sausage” is sausage prepared with fresh beef or frozen...

9 CFR 319.142 - Fresh beef sausage.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 9 Animals and Animal Products 2 2012-01-01 2012-01-01 false Fresh beef sausage. 319.142 Section 319.142 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... Sausage § 319.142 Fresh beef sausage. “Fresh Beef Sausage” is sausage prepared with fresh beef or frozen...

The influence of the type of embryo culture medium on neonatal birthweight after single embryo transfer in IVF.

PubMed

Vergouw, Carlijn G; Kostelijk, E Hanna; Doejaaren, Els; Hompes, Peter G A; Lambalk, Cornelis B; Schats, Roel

2012-09-01

Does the type of medium used to culture fresh and frozen-thawed embryos influence neonatal birthweight after single embryo transfer (SET) in IVF? A comparison of two commercially available culture media showed no significant influence on mean birthweight and mean birthweight adjusted for gestational age, gender and parity (z-scores) of singletons born after a fresh or frozen-thawed SET. Furthermore, we show that embryo freezing and thawing cycles may lead to a significantly higher mean birthweight. Animal studies have shown that culture media constituents are responsible for changes in birthweight of offspring. In human IVF, there is still little knowledge of the effect of medium type on birthweight. Until now, only a small number of commercially available culture media have been investigated (Vitrolife, Cook(®) Medical and IVF online medium). Our study adds new information: it has a larger population of singleton births compared with the previously published studies, it includes outcomes of other media types (HTF and Sage(®)), not previously analysed, and it includes data on frozen-thawed SETs. This study was a retrospective analysis of birthweights of singleton newborns after fresh (Day 3) or frozen-thawed (Day 5) SET cycles, using embryos cultured in either of two different types of commercially available culture media, between 2008 and 2011. Before January 2009, a single-step culture medium was used: human tubal fluid (HTF) with 4 mg/ml human serum albumin. From January 2009 onwards, a commercially available sequential medium was introduced: Sage(®), Quinn's advantage protein plus medium. Singletons born after a fresh SET (99 embryos cultured in HTF and 259 in Sage(®)) and singletons born after a frozen-thawed SET (32 embryos cultured in HTF only, 41 in HTF and Sage(®) and 86 in Sage(®) only) were analysed. Only patients using autologous gametes without the use of a gestational carrier were considered. Also excluded were (vanishing) twins, triplets, babies with congenital or chromosomal abnormalities and babies born before 22 weeks of gestation. Analysis of 358 singletons born after a fresh SET and 159 singletons born after a frozen-thawed SET showed no significant difference between the HTF and Sage(®) groups in terms of birthweight. Gestational age, parity and gender of the baby were significantly related to birthweight in multiple linear regression analyses, and other possible confounding factors included maternal age, BMI and smoking, the number of blastomeres in the transferred embryo and the type of culture medium. Maternal age, BMI and smoking, gestational age at birth, gender of the baby and the percentage of firstborns did not differ significantly between the HTF and Sage(®) groups; however, among the fresh embryos, those cultured in Sage(®) had significantly more blastomeres at the time of embryo transfer compared with the embryos cultured in HTF. Birthweights adjusted for gestational age and gender or gestational age and parity (z-scores) were not significantly different between the HTF and Sage(®) groups for fresh or frozen-thawed SETs. Mean birthweight, as well as the mean birthweight among firstborns and the mean birthweights adjusted for gestational age and gender or parity (z-scores) were significantly higher in the cryopreservation group compared with the fresh embryo transfer group. Our study is limited by its retrospective design and only two commercially available types of culture media were tested. More research is necessary to investigate the potential influence of culture media on gene expression. Although our data do not indicate the major influences of the HTF and Sage(®) culture media on birthweight, our results cannot be extrapolated to other culture media types. Furthermore, there remains a potential influence of embryo culture environment on epigenetic variation not represented by birthweight differences but by more subtle features.

Fresh embryo transfer versus frozen embryo transfer in in vitro fertilization cycles: a systematic review and meta-analysis.

PubMed

Roque, Matheus; Lattes, Karinna; Serra, Sandra; Solà, Ivan; Geber, Selmo; Carreras, Ramón; Checa, Miguel Angel

2013-01-01

To examine the available evidence to assess if cryopreservation of all embryos and subsequent frozen embryo transfer (FET) results in better outcomes compared with fresh transfer. Systematic review and meta-analysis. Centers for reproductive care. Infertility patient(s). An exhaustive electronic literature search in MEDLINE, EMBASE, and the Cochrane Library was performed through December 2011. We included randomized clinical trials comparing outcomes of IVF cycles between fresh and frozen embryo transfers. The outcomes of interest were ongoing pregnancy rate, clinical pregnancy rate, and miscarriage. We included three trials accounting for 633 cycles in women aged 27-33 years. Data analysis showed that FET resulted in significantly higher ongoing pregnancy rates and clinical pregnancy rates. Our results suggest that there is evidence that IVF outcomes may be improved by performing FET compared with fresh embryo transfer. This could be explained by a better embryo-endometrium synchrony achieved with endometrium preparation cycles. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Embryo quality is the main factor affecting cumulative live birth rate after elective single embryo transfer in fresh stimulation cycles.

PubMed

Niinimäki, Maarit; Veleva, Zdravka; Martikainen, Hannu

2015-11-01

The study was aimed to evaluate which factors affect the cumulative live birth rate after elective single embryo transfer in women younger than 36 years. Additionally, number of children in women with more than one delivery per ovum pick-up after fresh elective single embryo transfer and subsequent frozen embryo transfers was assessed. Retrospective cohort study analysing data of a university hospital's infertility clinic in 2001-2010. A total of 739 IVF/ICSI cycles with elective single embryo transfer were included. Analyses were made per ovum pick-up including fresh and subsequent frozen embryo transfers. Factors affecting cumulative live birth rates were examined in uni- and multivariate analyses. A secondary endpoint was the number of children born after all treatments. In the fresh cycles, the live birth rate was 29.2% and the cumulative live birth rate was 51.3%, with a twin rate of 3.4%. In the multivariate analysis, having two (odds ratio (OR) 1.73; 95% confidence interval (CI) 1.12-2.67) or ≥3 top embryos (OR 2.66; 95% CI 1.79-3.95) was associated with higher odds for live birth after fresh and frozen embryo cycles. Age, body mass index, duration of infertility, diagnosis or total gonadotropin dose were not associated with the cumulative live birth rate. In cycles with one top embryo, the cumulative live birth rate was 40.2%, whereas it was 64.1% in those with at least three top embryos. Of women who had a live birth in the fresh cycle, 20.4% had more than one child after all frozen embryo transfers. Among women with three or more top embryos after ovum pick-up, 16.1% gave birth to more than one child. The cumulative live birth rate in this age group varies from 40% to 64% and is dependent on the quality of embryos. Women with three or more top embryos have good chance of having more than one child per ovum pick-up without elevated risk of multiple pregnancies. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Can the Society for Assisted Reproductive Technology Clinic Outcome Reporting System (SART CORS) be used to accurately report clinic total reproductive potential (TRP)?

PubMed

Stern, Judy E; Hickman, Timothy N; Kinzer, Donna; Penzias, Alan S; Ball, G David; Gibbons, William E

2012-04-01

To assess whether total reproductive potential (TRP), the chance of a live birth from each fresh cycle (fresh cycle plus frozen transfers), could be calculated from the national Society for Assisted Reproductive Technology Clinic Outcome Reporting System (SART CORS) database and whether information not available in SART CORS resulted in significant changes to the TRP calculation. Retrospective study using SART CORS and clinic data. Three assisted reproductive technology clinics. Women undergoing ART. None. Two- and three-year TRPs for 2005 and 2006 were calculated according to patient age at cycle start by linking fresh to frozen cycles up to first live birth. Clinic records were used to adjust for (remove) frozen cycles that used more than one fresh cycle as a source of embryos and for any embryos donated to other patients or research or shipped to another facility before a live birth. TRP was higher than fresh per-cycle rates for most ages at all clinics, although accuracy was compromised when there were fewer than 20 cycles per category. Two- and 3-year TRPs differed in only 2 of 24 calculations. Adjusted TRPs differed less than three percentage points from unadjusted TRPs when volume was sufficient. Clinic TRP can be calculated from SART CORS. Data suggest that calculations of clinic TRP from the national dataset would be meaningful. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

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

PubMed

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

2018-01-01

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

Classification of fresh and frozen-thawed pork muscles using visible and near infrared hyperspectral imaging and textural analysis.

PubMed

Pu, Hongbin; Sun, Da-Wen; Ma, Ji; Cheng, Jun-Hu

2015-01-01

The potential of visible and near infrared hyperspectral imaging was investigated as a rapid and nondestructive technique for classifying fresh and frozen-thawed meats by integrating critical spectral and image features extracted from hyperspectral images in the region of 400-1000 nm. Six feature wavelengths (400, 446, 477, 516, 592 and 686 nm) were identified using uninformative variable elimination and successive projections algorithm. Image textural features of the principal component images from hyperspectral images were obtained using histogram statistics (HS), gray level co-occurrence matrix (GLCM) and gray level-gradient co-occurrence matrix (GLGCM). By these spectral and textural features, probabilistic neural network (PNN) models for classification of fresh and frozen-thawed pork meats were established. Compared with the models using the optimum wavelengths only, optimum wavelengths with HS image features, and optimum wavelengths with GLCM image features, the model integrating optimum wavelengths with GLGCM gave the highest classification rate of 93.14% and 90.91% for calibration and validation sets, respectively. Results indicated that the classification accuracy can be improved by combining spectral features with textural features and the fusion of critical spectral and textural features had better potential than single spectral extraction in classifying fresh and frozen-thawed pork meat. Copyright © 2014 Elsevier Ltd. All rights reserved.

The effect of flash-freezing temperature on stallion sperm DNA structure.

PubMed

Serafini, R; Varner, D D; Bissett, W; Blanchard, T L; Teague, S R; Love, C C

2017-06-01

The effect of flash-freezing storage temperature on stallion sperm DNA has not been evaluated. Commonly, sperm are flash-frozen at various temperatures to preserve sperm DNA prior to analysis. It is unclear whether the temperature at which sperm are frozen and stored may affect the results of DNA assays. In this study, the neutral comet assay was used to evaluate the effect of flash-freezing storage temperature (freezer [-60 °C], dry ice [-78.5 °C], liquid nitrogen [-196 °C]) compared to fresh sperm DNA structure. In addition, intra- and inter-assay and intra- and inter-stallion variabilities were determined. All comet tail measures were higher following any flash-freezing method, as compared to fresh sperm DNA (P < 0.05), with no difference among flash-frozen treatments (P > 0.05). For most comet variables, intra- and inter-assay variabilities were <10%. Intra- and inter-stallion variabilities revealed that comet head length (HL) and width (CW) were less variable as compared to comet tail values, i.e., % comet tail DNA (T-DNA), tail length (TL), tail moment (OTM), and tail migration (TM). Certain comet tail values in fresh (% T-DNA, and OTM) and flash-frozen sperm (OTM, % T-DNA, TL, and TM) were correlated to the Sperm Chromatin Structure Assay (SCSA) variable, COMP-α t . The comet tail measures were negatively correlated to % morphologically normal sperm (P < 0.05) and positively correlated to % abnormal heads and premature germ cells (P < 0.05). Variables COMP-α t and % total sperm motility were not correlated to any morphologic sperm feature in this group of stallions (P > 0.05). While significant differences in the structure of the sperm DNA were identified in the flash-frozen as compared to the fresh sperm DNA with the neutral comet assay, it cannot be assumed that these changes are fertility limiting. Copyright © 2017. Published by Elsevier Inc.

Validation of freezing tissues and cells for analysis of DNA strand break levels by comet assay

PubMed Central

Jackson, Petra

2013-01-01

The comet analysis of DNA strand break levels in tissues and cells has become a common method of screening for genotoxicity. The large majority of published studies have used fresh tissues and cells processed immediately after collection. However, we have used frozen tissues and cells for more than 10 years, and we believe that freezing samples improve efficiency of the method. We compared DNA strand break levels measured in fresh and frozen bronchoalveolar cells, and lung and liver tissues from mice exposed to the known mutagen methyl methanesulphonate (0, 25, 75, 112.5mg/kg). We used a high-throughput comet protocol with fully automated scoring of DNA strand break levels. The overall results from fresh and frozen samples were in agreement [R 2 = 0.93 for %DNA in tail (%TDNA) and R 2 = 0.78 for tail length (TL)]. A slightly increased %TDNA was observed in lung and liver tissue from vehicle controls; and TL was slightly reduced in bronchoalveolar lavage cells from the high-dose group. In our comet protocol, a small block of tissue designated for comet analysis is frozen immediately at tissue collection and kept deep frozen until rapidly homogenised and embedded in agarose. To demonstrate the feasibility of long-term freezing of samples, we analysed the day-to-day variation of our internal historical negative and positive comet assay controls collected over a 10-year period (1128 observations, 11 batches of frozen untreated and H2O2-treated A549 lung epithelial cells). The H2O2 treatment explained most of the variation 57–77% and the day-to-day variation was only 2–12%. The presented protocol allows analysis of samples collected over longer time span, at different locations, with reduced variation by reducing number of electrophoreses and is suitable for both toxicological and epidemiological studies. The use of frozen tissues; however, requires great care during preparation before analysis, with handling as a major risk factor. PMID:24136994

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

PubMed

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

2010-08-01

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

Method for Differentiation between Fresh and Frozen-thawed Fish

NASA Astrophysics Data System (ADS)

Kitamikado, Manabu; Yoshioka, Keiko

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

The use of frozen plasma samples in thromboelastometry.

PubMed

Schoergenhofer, Christian; Buchtele, Nina; Schwameis, Michael; Bartko, Johann; Jilma, Bernd; Jilma-Stohlawetz, Petra

2017-11-01

Thromboelastometry is increasingly used in the clinical and scientific setting. The use of frozen plasma samples may be useful in overcoming certain limitations such as local and timely availability. Whole blood (WB) samples of 20 healthy volunteers were obtained, and plasma was generated. NATEM (n = 20), EXTEM (n = 20) and INTEM (n = 8) analyses were performed in WB, fresh plasma and frozen and thawed plasma. Dabigatran (500, 1000 ng/ml), rivaroxaban (100, 200 ng/ml) or alteplase (333 ng/ml) were added ex vivo to WB, and thromboelastometry was performed in WB and in frozen and thawed plasma samples. Clot formation time, mean clot firmness and the area under the curve were significantly altered in plasma compared to WB. In INTEM and EXTEM analysis, clotting time (CT) was comparable between WB (100%) and fresh (INTEM 114% and EXTEM 93%, ratio of the means) and frozen plasma samples (85 and 99%), whereas in NATEM analysis, the CT increased in fresh (193%) and frozen plasma samples (130%). Dabigatran dose-dependently increased the CT approximately 5- and 9-fold in WB and even more pronounced 10- and 26-fold in plasma. Accordingly, rivaroxaban dose-dependently increased the CT 2- and 2.7-fold in WB, and 3.5- and 4-fold in plasma samples. Hyperfibrinolysis was achieved by addition of alteplase in all WB samples and was reproducible in plasma samples. In conclusion, thromboelastometry, especially INTEM and EXTEM analyses, is possible using frozen and stored plasma samples with comparable results to the corresponding whole blood samples.

Effect of blanching treatments on antioxidant activity of frozen green capsicum (Capsicum annuum L. var bell pepper) using radical scavenging activity (DPPH) assay

NASA Astrophysics Data System (ADS)

Azizzuddin, Norafida; Abdullah, Aminah

2016-11-01

Blanching treatments are needed to deactivate enzymes in frozen vegetables. Antioxidant activity using DPPH radical scavenging activity assay were evaluated in steaming, boiling water, and microwave blanching at different temperature, time and microwave power level on frozen green capsicum. Green capsicum was chosen for frozen treatment compared to other capsicum with different maturity index because of the firm texture. The objective of this study was to compare the antioxidant activity of frozen green capsicum between conventional and Oxi Count Kit® assay for DPPH radical scavenging activity. Results showed frozen green capsicum blanched using microwave at high level/90 seconds (sample J) contained higher level of DPPH in both conventional method and Oxi Count Kit® compared to other treatments. However, there were no significant differences between sample J and fresh sample (sample A). Overall, the sequences from highest to lowest in blanching treatments for both DPPH conventional method, and DPPH Oxi Count Kit® were J (microwave high level/90 seconds) > A (Fresh) > H (Microwave Medium Level/120 seconds) > D (Boiling Water 80°C/150 seconds) > K (Microwave High Level/120 seconds) > I (Microwave Medium Level/150 seconds) > F (Microwave Low Level/150 seconds)> B (Steam 100°C/150 seconds) > E (Boiling Water 100°C /120 seconds) > G (Microwave Low Level /180 seconds)> C (Steam 100°C/180 seconds). Almost all frozen green capsicum samples showed no significant differences for comparison between test using DPPH conventional method and Oxi Count Kit®. Frozen storage for 0, and 3rd months showed no significant differences which indicate no changes on antioxidant activity during frozen storage at -18°C.

An evaluation of Brix refractometry instruments for measurement of colostrum quality in dairy cattle.

PubMed

Bielmann, V; Gillan, J; Perkins, N R; Skidmore, A L; Godden, S; Leslie, K E

2010-08-01

Acquisition of high quality colostrum is an important factor influencing neonatal calf health. Many methods have been used to assess the Ig concentration of colostrum; however, improved, validated evaluation tools are needed. The aims of this study were to evaluate both optical and digital Brix refractometer instruments for the measurement of Ig concentration of colostrum as compared with the gold standard radial immunodiffusion assay laboratory assessment and to determine the correlation between Ig measurements taken from fresh and frozen colostrum samples for both Brix refractometer instruments. This research was completed using 288 colostrum samples from 3 different farms. It was concluded that the optical and digital Brix refractometers were highly correlated for both fresh and frozen samples (r=0.98 and r=0.97, respectively). Correlation between both refractometer instruments for fresh and frozen samples and the gold standard radial immunodiffusion assay were determined to be very similar, with a correlation coefficient between 0.71 and 0.74. Both instruments exhibited excellent test characteristics, indicating an appropriate cut-off point of 22% Brix score for the identification of good quality colostrum. Copyright (c) 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

PubMed

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

2017-02-01

The aim of this study was to evaluate the fertility response of artificial insemination (AI) methods with fresh and frozen sperm in sheep. In experiment 1, one hundred and fifty fat tailed Zandi ewes were assigned into 3 equal groups and inseminated with three AI methods consisting of vaginal, laparoscopic and trans-cervical AI with fresh semen. In experiment 2, a factorial study (3 AI methods × 2 extenders) was used to analyze the effects of three AI methods and two freezing extenders containing soybean lecithin (SL) or Egg yolk (EY) on reproductive performance of 300 fat tailed Zandi ewes. Also, total motility, progressive motility, viability and lipid peroxidation of semen were evaluated after freeze-thawing in two extenders. In result, there was no significant difference among three AI methods when fresh semen was used. In experiment 2, the highest percentage of pregnancy rate, parturition rate and lambing rate were obtained in laparoscopic AI group (P < 0.05). Although pregnancy rate, parturition rate and lambing rate in trans-cervical group were higher (P < 0.05) than vaginal group, the results were not as high as laparoscopic group. No difference was observed between SL and EY extenders and their performance was close to each other. It can be concluded that although no difference was observed on reproductive performance for fresh semen, trans-cervical AI was more efficient than vaginal method when frozen-thawed semen was used, but its efficiency was not as high as laparoscopic method. Also, SL extender can be an efficient alternative extender to preserve ram sperm during cryopreservation procedure without adverse effects of EY. Copyright © 2016. Published by Elsevier Inc.

Live Birth from Slow-Frozen Rabbit Oocytes after In Vivo Fertilisation

PubMed Central

Jiménez-Trigos, Estrella; Vicente, José S.; Marco-Jiménez, Francisco

2013-01-01

In vivo fertilisation techniques such as intraoviductal oocyte transfer have been considered as alternatives to bypass the inadequacy of conventional in vitro fertilisation in rabbit. There is only one study in the literature, published in 1989, that reports live offspring from cryopreserved rabbit oocytes. The aim of the present study was to establish the in vivo fertilisation procedure to generate live offspring with frozen oocytes. First, the effect of two recipient models (i) ovariectomised or (ii) oviduct ligated immediately after transfer on the ability of fresh oocytes to fertilise were compared. Second, generation of live offspring from slow-frozen oocytes was carried out using the ligated oviduct recipient model. Throughout the experiment, recipients were artificially inseminated 9 hours prior to oocyte transfer. In the first experiment, two days after unilateral transfer of fresh oocytes, oviducts and uterine horns were flushed to assess embryo recovery rates. The embryo recovery rates were low compared to control in both ovariectomised and ligated oviduct groups. However, ligated oviduct recipient showed significantly (P<0.05) higher embryo recovery rates compared to ovariectomised and control-transferred. In the second experiment, using bilateral oviduct ligation model, all females that received slow-frozen oocytes became pregnant and delivered a total of 4 live young naturally. Thus, in vivo fertilisation is an effective technique to generate live offspring using slow-frozen oocytes in rabbits. PMID:24358281

Effect of cryopreservation and transplantation on the expression of kit ligand and anti-Mullerian hormone in human ovarian tissue.

PubMed

David, Anu; Van Langendonckt, Anne; Gilliaux, Sébastien; Dolmans, Marie-Madeleine; Donnez, Jacques; Amorim, Christiani A

2012-04-01

Although cryopreservation and transplantation of ovarian tissue represent a promising alternative to safeguard fertility in cancer patients, low recovery rates of oocytes aspirated from antral follicles and a significant number of empty follicles have been observed in women with transplanted frozen-thawed ovarian tissue. In order to understand how freezing and/or grafting may affect follicular development, the follicular expression of kit ligand (KL) and anti-Müllerian hormone (AMH), two key factors activating and inhibiting follicle growth, were assessed after long-term grafting in severe combined immunodeficient (SCID) mice. Ovarian biopsies from eight patients were used for fresh and frozen-thawed tissue xenografting in 13 SCID mice for a period of 28 weeks, including 2 weeks of gonadotrophin stimulation. KL, AMH and proliferating cell nuclear antigen (PCNA) immunostaining were quantified before and after grafting in the two treatment groups (fresh and frozen-thawed grafted ovarian tissue). Lower expression of KL was found in primordial and primary follicles after grafting of both fresh and frozen-thawed tissue. Consistent expression of AMH was found in most growing follicles at a similar rate in both graft types. In fresh and frozen-thawed grafts, 13-14% of primordial follicles were PCNA-positive, indicating a similar maintenance of quiescent follicles despite follicle activation. Grafting and/or gonadotrophin stimulation appear to affect the follicular expression of KL, which may alter oocyte quality. AMH expression in growing follicles after ovarian tissue transplantation may be one of the factors contributing to the preservation of resting follicles in 28-week-old grafts.

Characterizing the glycocalyx of poultry spermatozoa: III. Semen cryopreservation methods alter the carbohydrate component of rooster sperm membrane glycoconjugates.

PubMed

Peláez, J; Bongalhardo, D C; Long, J A

2011-02-01

The carbohydrate-rich zone on the sperm surface is essential for inmunoprotection in the female tract and early gamete interactions. We recently have shown the glycocalyx of chicken sperm to be extensively sialylated and to contain residues of mannose, glucose, galactose, fucose, N-acetyl-galactosamine, N-acetyl-glucosamine, and N-acetyl-lactosamine. Our objective here was to evaluate the effects of 3 different cryopreservation methods on the sperm glycocalyx. Semen from roosters was pooled, diluted, cooled to 5°C, and aliquoted for cryopreservation using 6% dimethylacetamide (DMA), 11% dimethylsulfoxide (DMSO), or 11% glycerol (GOH). For the DMA method, semen was equilibrated for 1 min with cryoprotectant and rapidly frozen by dropping 25-µL aliquots into liquid nitrogen. For the other methods, semen was equilibrated for either 1 min (DMSO) or 20 min (GOH), loaded into straws, and frozen with a programmable freezer. Thawing rates mimicked the freezing rates (e.g., rapid for DMA; moderate for DMSO and GOH). Aliquots of thawed and fresh, unfrozen semen were incubated with 1 of 12 fluorescein isothiocyanate-conjugated lectins and counterstained with propidium iodide, and mean fluorescence intensity (MFI) was assessed by flow cytometry. For each lectin, the MFI of propidium iodide-negative (viable sperm) was compared among the fresh and frozen-thawed treatments (n = 5). For sperm frozen with GOH and DMA, the MFI of most lectins was similar (P > 0.05) to that of fresh sperm, whereas only 5 of 12 lectins were similar between fresh and DMSO-frozen sperm. Sperm from all 3 methods had higher (P < 0.05) MFI for lectins specific for N-acetyl-glucosamine and β-galactose than did fresh sperm. Fewer sperm were damaged (P < 0.001) with GOH than with DMA or DMSO, and membrane integrity was correlated with MFI for 9 of 12 lectins (P < 0.05). These data indicate that surface carbohydrates are altered during cryopreservation, and that cryoprotectant type and freezing-thawing rates affect the degree of modification. Although the glycoconjugates have not yet been identified, it is likely that these cryopreservation-induced changes contribute to the reduced fertility of frozen-thawed chicken semen.

Healthy Snacks for Kids: 10 Child-Friendly Tips

MedlinePlus

... Dip graham cracker sticks or fresh fruit in yogurt. Spread peanut butter on celery, apples or bananas. ... child's sweet tooth with low-fat puddings, frozen yogurt or frozen fruit bars. Serve smoothies made with ...

Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

PubMed Central

Rungsiwiwut, Ruttachuk; Numchaisrika, Pranee; Ahnonkitpanit, Vichuda; Isarasena, Nipan; Virutamasen, Pramuan

2012-01-01

Abstract Human embryonic stem (hES) cells are considered to be a potential source for the therapy of human diseases, drug screening, and the study of developmental biology. In the present study, we successfully derived hES cell lines from blastocysts developed from frozen and fresh embryos. Seventeen- to eighteen-year-old frozen embryos were thawed, cultured to the blastocyst stage, and induced to form hES cells using human foreskin fibroblasts. The Chula2.hES cell line and the Chula4.hES and Chula5.hES cell lines were derived from blastocysts developed from frozen and fresh embryos, respectively. The cell lines expressed pluripotent markers, including alkaline phosphatase (AP), Oct3/4, stage-specific embryonic antigen (SSEA)-4, and tumor recognition antigen (TRA)-1-60 and TRA-1-81 as detected with immunocytochemistry. The real-time polymerase chain reaction (RT-PCR) results showed that the cell lines expressed pluripotent genes, including OCT3/4, SOX2, NANOG, UTF, LIN28, REX1, NODAL, and E-Cadherin. In addition, the telomerase activities of the cell lines were higher than in the fibroblast cells. Moreover, the cell lines differentiated into all three germ layers both in vitro and in vivo. The cell lines had distinct identities, as revealed with DNA fingerprinting, and maintained their normal karyotype after a long-term culture. This study is the first to report the successful derivation of hES cell lines in Thailand and that frozen embryos maintained their pluripotency similar to fresh embryos, as shown by the success of hES cell derivation, even after years of cryopreservation. Therefore, embryos from prolonged cryopreservation could be an alternative source for embryonic stem cell research. PMID:23514952

The impact of milk handling procedures on Ostertagia ostertagi antibody ELISA test results.

PubMed

Vanderstichel, Raphaël; Dohoo, Ian; Stryhn, Henrik

2010-04-19

The impact of various milk handling stressors were analyzed using a commercially available enzyme-linked immunosorbent assay (ELISA) test measuring Ostertagia ostertagi antibodies in milk from dairy cattle (Svanovir). An indirect ELISA has the ability to determine the amount of milk production losses related to intestinal parasitism. The ELISA test recommends fresh defatted milk, however, milk collected from Dairy Herd Improvement (DHI) programs in North America undergo many stressors, including, heating, freezing and are not defatted. Normalized optical density ratios (ODRs) were compared between fresh defatted milk and milk subjected to one or more stressors with a linear mixed model accounting for differences in variation between the fresh and the frozen samples. Concordance correlation coefficients were also analyzed for comparisons to other similar studies. After accounting for random cow and container effects, the treatment factors interacted with each other (p<0.001). Biologically interesting contrasts were created to explain the interaction. The estimated difference in ODR between the milk samples handled according to recommendations of the manufacturers of Svanovir and the whole milk samples that were subjected to the most extreme treatment (heated, frozen, thawed, and re-frozen for 4 weeks) was 0.062 (p<0.001). This difference represented less than 5% of the range, and was thus considered biologically negligible. Frozen whole milk processed by DHI programs, the most likely method of collecting on-farm samples in North America, will likely yield reliable results for the indirect ELISA tests, particularly, Svanovir.

Improvement of Blood-Brain Barrier Integrity in Traumatic Brain Injury and Hemorrhagic Shock Following Treatment With Valproic Acid and Fresh Frozen Plasma.

PubMed

Nikolian, Vahagn C; Dekker, Simone E; Bambakidis, Ted; Higgins, Gerald A; Dennahy, Isabel S; Georgoff, Patrick E; Williams, Aaron M; Andjelkovic, Anuska V; Alam, Hasan B

2018-01-01

Combined traumatic brain injury and hemorrhagic shock are highly lethal. Following injuries, the integrity of the blood-brain barrier can be impaired, contributing to secondary brain insults. The status of the blood-brain barrier represents a potential factor impacting long-term neurologic outcomes in combined injuries. Treatment strategies involving plasma-based resuscitation and valproic acid therapy have shown efficacy in this setting. We hypothesize that a component of this beneficial effect is related to blood-brain barrier preservation. Following controlled traumatic brain injury, hemorrhagic shock, various resuscitation and treatment strategies were evaluated for their association with blood-brain barrier integrity. Analysis of gene expression profiles was performed using Porcine Gene ST 1.1 microarray. Pathway analysis was completed using network analysis tools (Gene Ontology, Ingenuity Pathway Analysis, and Parametric Gene Set Enrichment Analysis). Female Yorkshire swine were subjected to controlled traumatic brain injury and 2 hours of hemorrhagic shock (40% blood volume, mean arterial pressure 30-35 mmHg). Subjects were resuscitated with 1) normal saline, 2) fresh frozen plasma, 3) hetastarch, 4) fresh frozen plasma + valproic acid, or 5) hetastarch + valproic acid (n = 5 per group). After 6 hours of observation, brains were harvested for evaluation. Immunofluoroscopic evaluation of the traumatic brain injury site revealed significantly increased expression of tight-junction associated proteins (zona occludin-1, claudin-5) following combination therapy (fresh frozen plasma + valproic acid and hetastarch + valproic acid). The extracellular matrix protein laminin was found to have significantly improved expression with combination therapies. Pathway analysis indicated that valproic acid significantly modulated pathways involved in endothelial barrier function and cell signaling. Resuscitation with fresh frozen plasma results in improved expression of proteins essential for blood-brain barrier integrity. The addition of valproic acid provides significant improvement to these protein expression profiles. This is likely secondary to activation of key pathways related to endothelial functions.

Survey of total mercury in some edible fish and shellfish species collected in Canada in 2002.

PubMed

Dabeka, R; McKenzie, A D; Forsyth, D S; Conacher, H B S

2004-05-01

Total mercury was measured in the edible portions of 244 selected fish and shellfish purchased in Canada at the retail level. By species, average mercury concentrations ranged from 0.011 microg g(-1) for oysters to 1.82 microg g(-1) for swordfish. The predatory fish contained the highest concentrations of mercury: swordfish (mean 1.82 microg g(-1), range 0.40-3.85 microg g(-1)), marlin (1.43, 0.34-3.19 microg g(-1)), shark (1.26, 0.087-2.73 microg g(-1)), and canned, fresh and frozen tuna (0.35, 0.020-2.12 microg g(-1)). Levels of mercury in the fresh and frozen tuna contained a mean of 0.93 microg g(-1) (range 0.077-2.12 microg g(-1)) and were substantially higher than in the canned tuna (0.15, 0.02-0.59 microg g(-1)). In the canned tuna, mercury concentrations varied with subspecies, with the highest average concentrations being found in Albacore tuna (mean 0.26 microg g(-1), range 0.19-0.38 microg g(-1)) and the lowest (0.047, 0.025-0.069 microg g(-1)) in five samples for which the subspecies of tuna were not identified. Mean concentrations of mercury in swordfish and fresh and frozen tuna were up to three times higher than reported for the USA. Dietary intake estimations found that provided fresh and frozen tuna, marlin, swordfish or shark are consumed once a month or less, the dietary intakes of total mercury by women of child-bearing age, averaged over 1 month, would fall below the Joint FAO/WHO Expert Committee on Food Additives provisional tolerable weekly intake for total mercury. The current Canadian advisory to children and women of child-bearing age is to limit their consumption of fresh and frozen tuna, swordfish and shark to no more than one meal per month.

[Relationship between mitochondrial DNA copy number, membrane potential of human embryo and embryo morphology].

PubMed

Zhao, H; Teng, X M; Li, Y F

2017-11-25

Objective: To explore the relationship between the embryo with the different morphological types in the third day and its mitochondrial copy number, the membrane potential. Methods: Totally 117 embryos with poor development after normal fertilization and were not suitable transferred in the fresh cycle and 106 frozen embryos that were discarded voluntarily by infertility patients with in vitro fertilization-embryo transfer after successful pregnancy were selected. According to evaluation of international standard in embryos, all cleavage stage embryos were divided into class Ⅰ frozen embryo group ( n= 64), class Ⅱ frozen embryo group ( n= 42) and class Ⅲ fresh embryonic group (not transplanted embryos; n= 117). Real-time PCR and confocal microscopy methods were used to detect mitochondrial DNA (mtDNA) copy number and the mitochondrial membrane potential of a single embryo. The differences between embryo quality and mtDNA copy number and membrane potential of each group were compared. Results: The copy number of mtDNA and the mitochondrial membrane potential in class Ⅲ fresh embryonic group [(1.7±1.0)×10(5) copy/μl, 1.56±0.32] were significantly lower than those in class Ⅰ frozen embryo group [(3.4±1.7)×10(5) copy/μl, 2.66±0.21] and class Ⅱ frozen embryo group [(2.6±1.2)×10(5) copy/μl, 1.80±0.32; all P< 0.05]. The copy number of mtDNA and the mitochondrial membrane potential in classⅠ frozen embryo group were significantly higher than those in classⅡ frozen embryo group (both P< 0.05). Conclusion: The mtDNA copy number and the mitochondrial membrane potential of embryos of the better quality embryo are higher.

Laser-assisted in vitro fertilization facilitates fertilization of vitrified-warmed C57BL/6 mouse oocytes with fresh and frozen-thawed spermatozoa, producing live pups.

PubMed

Woods, Stephanie E; Qi, Peimin; Rosalia, Elizabeth; Chavarria, Tony; Discua, Allan; Mkandawire, John; Fox, James G; García, Alexis

2014-01-01

The utility of cryopreserved mouse gametes for reproduction of transgenic mice depends on development of assisted reproductive technologies, including vitrification of unfertilized mouse oocytes. Due to hardening of the zona pellucida, spermatozoa are often unable to penetrate vitrified-warmed (V-W) oocytes. Laser-assisted in vitro fertilization (LAIVF) facilitates fertilization by allowing easier penetration of spermatozoa through a perforation in the zona. We investigated the efficiency of V-W C57BL/6NTac oocytes drilled by the XYClone laser, compared to fresh oocytes. By using DAP213 for cryoprotection, 83% (1,470/1,762) of vitrified oocytes were recovered after warming and 78% were viable. Four groups were evaluated for two-cell embryo and live offspring efficiency: 1) LAIVF using V-W oocytes, 2) LAIVF using fresh oocytes, 3) conventional IVF using V-W oocytes and 4) conventional IVF using fresh oocytes. First, the groups were tested using fresh C57BL/6NTac spermatozoa (74% motile, 15 million/ml). LAIVF markedly improved the two-cell embryo efficiency using both V-W (76%, 229/298) and fresh oocytes (69%, 135/197), compared to conventional IVF (7%, 12/182; 6%, 14/235, respectively). Then, frozen-thawed C57BL/6NTac spermatozoa (35% motile, 15 million/ml) were used and LAIVF was again found to enhance fertilization efficiency, with two-cell embryo rates of 87% (298/343) using V-W oocytes (P<0.05, compared to fresh spermatozoa), and 73% (195/266) using fresh oocytes. Conventional IVF with frozen-thawed spermatozoa using V-W (6%, 10/168) and fresh (5%, 15/323) oocytes produced few two-cell embryos. Although live offspring efficiency following embryo transfer was greater with conventional IVF (35%, 18/51; LAIVF: 6%, 50/784), advantage was seen with LAIVF in live offspring obtained from total oocytes (5%, 50/1,010; conventional IVF: 2%, 18/908). Our results demonstrated that zona-drilled V-W mouse oocytes can be used for IVF procedures using both fresh and frozen-thawed spermatozoa, producing live pups. The ability to cryopreserve mouse gametes for LAIVF may facilitate management of large-scale transgenic mouse production facilities.

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

PubMed

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

2018-05-21

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

Ex vivo rabbit cornea diffusion studies with a soluble insert of moxifloxacin.

PubMed

Sebastián-Morelló, María; Calatayud-Pascual, María Aracely; Rodilla, Vicent; Balaguer-Fernández, Cristina; López-Castellano, Alicia

2018-02-01

The objective of this research was to develop and evaluate an ocular insert for the controlled drug delivery of moxifloxacin which could perhaps be used in the treatment of corneal keratitis or even bacterial endophthalmitis. We have evaluated the ex vivo ocular diffusion of moxifloxacin through rabbit cornea, both fresh and preserved under different conditions. Histological studies were also carried out. Subsequently, drug matrix inserts were prepared using bioadhesive polymers. The inserts were evaluated for their physicochemical parameters. Ophthalmic ex vivo permeation of moxifloxacin was carried out with the most promising insert. The formulate insert was thin and provided higher ocular diffusion than commercial formulations. Ocular diffusion studies revealed significant differences between fresh and frozen corneas. Histological examinations also showed differences in the thickness of stroma between fresh and frozen corneas. The ophthalmic insert we have developed allows a larger quantity of moxifloxacin to permeate through the cornea than existing commercial formulations of the drug. Ocular delivery of moxifloxacin with this insert could be a new approach for the treatment of eye diseases.

Evaluation of the qualitative and quantitative effectiveness of three media of centrifugation (Maxifreeze, Cushion Fluid Equine, and PureSperm 100) in preparation of fresh or frozen-thawed brown bear spermatozoa.

PubMed

Nicolas, M; Alvarez, M; Borragán, S; Martinez-Pastor, F; Chamorro, C A; Alvarez-Rodriguez, M; de Paz, P; Anel, L

2012-04-01

Centrifugation is a crucial procedure in sperm cryopreservation protocols of brown bear (Ursus arctos), because the semen must be processed to increase sperm concentration and/or clean urine-contaminated samples. The efficacy of three media for centrifugation (Maxifreeze [IMV technologies, L'Aigle, France], Cushion Fluid Equine (Minitübe, Tiefenbach, Germany), and PureSperm [Nidacon, Gothenburg, Sweden]) on the quality of bear spermatozoa was evaluated. In experiment one, two cushioned media used for protecting against mechanical stress during centrifugation were analyzed. In experiment two, a density gradient based on PureSperm was assessed in relation to the maximum retrieval and the quality of fresh spermatozoa, and the freezability of the spermatozoa selected in this density gradient was studied in experiment three. Finally, the selection of frozen-thawed sperm using PureSperm was analyzed in experiment four. Our results indicate that the use of dense isotonic cushion solutions (Maxifreeze, Cushion Fluid Equine) in centrifugation did not improve the quality of recovered spermatozoa compared with standard centrifugation. However, a density gradient prepared with PureSperm improved the quality of spermatozoa in fresh semen and frozen-thawed semen, but the spermatozoa selected from the fresh sample with this density gradient did not show a better resistance to freezing with this density gradient in comparison with the control sample. Copyright © 2012 Elsevier Inc. All rights reserved.

Isolation, culture and characterisation of somatic cells derived from semen and milk of endangered sheep and eland antelope.

PubMed

Nel-Themaat, L; Gómez, M C; Damiani, P; Wirtu, G; Dresser, B L; Bondioli, K R; Lyons, L A; Pope, C E; Godke, R A

2007-01-01

Semen and milk are potential sources of somatic cells for genome banks. In the present study, we cultured and characterised cells from: (1) cooled sheep milk; (2) fresh, cooled and frozen-thawed semen from Gulf Coast native (GCN) sheep (Ovis aries); and (3) fresh eland (Taurotragus oryx) semen. Cells attached to the culture surface from fresh (29%), cooled (43%) and slow-frozen (1 degrees C/min; 14%) ram semen, whereas no attachment occurred in the fast-frozen (10 degrees C/min) group. Proliferation occurred in fresh (50%) and cooled (100%) groups, but no cells proliferated after passage 1 (P1). Eland semen yielded cell lines (100%) that were cryopreserved at P1. In samples from GCN and cross-bred milk, cell attachment (83% and 95%, respectively) and proliferation (60% and 37%, respectively) were observed. Immunocytochemical detection of cytokeratin indicated an epithelial origin of semen-derived cells, whereas milk yielded either fibroblasts, epithelial or a mixture of cell types. Deoxyribonucleic acid microsatellite analysis using cattle-derived markers confirmed that eland cells were from the semen donor. Eland epithelial cells were transferred into eland oocytes and 12 (71%), six (35%) and two (12%) embryos cleaved and developed to morulae or blastocyst stages, respectively. In conclusion, we have developed a technique for obtaining somatic cells from semen. We have also demonstrated that semen-derived cells can serve as karyoplast donors for nuclear transfer.

CO₂ processing and hydration of fruit and vegetable tissues by clathrate hydrate formation.

PubMed

Takeya, Satoshi; Nakano, Kohei; Thammawong, Manasikan; Umeda, Hiroki; Yoneyama, Akio; Takeda, Tohoru; Hyodo, Kazuyuki; Matsuo, Seiji

2016-08-15

CO2 hydrate can be used to preserve fresh fruits and vegetables, and its application could contribute to the processing of carbonated frozen food. We investigated water transformation in the frozen tissue of fresh grape samples upon CO2 treatment at 2-3 MPa and 3°C for up to 46 h. Frozen fresh bean, radish, eggplant and cucumber samples were also investigated for comparison. X-ray diffraction indicated that after undergoing CO2 treatment for several hours, structure I CO2 hydrate formed within the grape tissue. Phase-contrast X-ray imaging using the diffraction-enhanced imaging technique revealed the presence of CO2 hydrate within the intercellular spaces of these tissues. The carbonated produce became effervescent because of the dissociation of CO2 hydrate through the intercellular space, especially above the melting point of ice. In addition, suppressed metabolic activity resulting from CO2 hydrate formation, which inhibits water and nutrient transport through intercellular space, can be expected. Copyright © 2016 Elsevier Ltd. All rights reserved.

Fresh, frozen, or ambient food equivalents and their impact on food waste generation in Dutch households.

PubMed

Janssen, Anke M; Nijenhuis-de Vries, Mariska A; Boer, Eric P J; Kremer, Stefanie

2017-09-01

In Europe, it is estimated that more than 50% of total food waste - of which most is avoidable - is generated at household level. Little attention has been paid to the impact on food waste generation of consuming food products that differ in their method of food preservation. This exploratory study surveyed product-specific possible impacts of different methods of food preservation on food waste generation in Dutch households. To this end, a food waste index was calculated to enable relative comparisons of the amounts of food waste from the same type of foods with different preservation methods on an annual basis. The results show that, for the majority of frozen food equivalents, smaller amounts were wasted compared to their fresh or ambient equivalents. The waste index (WI) proposed in the current paper confirms the hypothesis that it may be possible to reduce the amount of food waste at household level by encouraging Dutch consumers to use (certain) foods more frequently in a frozen form (instead of fresh or ambient). However, before this approach can be scaled to population level, a more detailed understanding of the underlying behavioural causes with regard to food provisioning and handling and possible interactions is required. Copyright © 2017. Published by Elsevier Ltd.

Blood collection, components preparation and distribution in Iran, 2008-2012.

PubMed

Omidkhoda, Azadeh; Amini Kafi-Abad, Sedigheh; Pourfatollah, Ali Akbar; Maghsudlu, Mahtab

2016-02-01

The information about the dynamics of blood collection, components preparation and distribution in Iran was measured and compared during 2008-2012. The survey instruments were based on collecting data from all 220 blood collections and blood processing centers over the country, registering them in the validated data base and reporting them to headquarter of Iranian Blood Transfusion Organization. Total blood collection increased during this period, and in 2012 represented a 12.6 percent increase compared to that in 2008. On average, red blood cells, fresh frozen plasma and platelet concentrate were prepared from 95.5 ± 2.4, 81 ± 3.8 and 47 ± 8.8 percent of all whole blood collection. From 2008 to 2011, the distribution of whole blood and fresh frozen plasma revealed different patterns. For whole blood, declines were noted, while for fresh frozen plasma increases were reported. In addition the distribution of red blood cells and platelet concentrate did not change considerably. Also between 2008 and 2012, the mean percentage of outdated and discarded units was 3.6 ± 1 and 5.2 ± 4.6. This study as a first national survey provides comprehensive information about the blood supply, components preparation and distribution, and helps to define strategy for the future. Copyright © 2016 Elsevier Ltd. All rights reserved.

A Multivariate Evaluation of Factors Affecting the Quality of Freshly Frozen Tissue Specimens.

PubMed

Wang, Tong-Hong; Chen, Chin-Chuan; Liang, Kung-Hao; Chen, Chi-Yuan; Chuang, Wen-Yu; Ueng, Shir-Hwa; Chu, Pao-Hsien; Huang, Chung-Guei; Chen, Tse-Ching; Hsueh, Chuen

2017-08-01

Well-prepared and preserved freshly frozen specimens are indispensable materials for clinical studies. To manage specimen quality and to understand the factors potentially affecting specimen quality during preservation processes, we analyzed the quality of RNA and genomic DNA of various tissues collected between 2002 and 2011 in Linkou Chang Gung Memorial Hospital, Taiwan. During this period, a total of 1059 freshly frozen specimens from eight major cancer categories were examined. It was found that preservation duration, organ origin, and tissue type could all influence the quality of RNA samples. The increased preservation period correlated with decreased RNA quality; the brain, breast, and stomach RNA specimens displayed faster degradation rates than those of other organs, and RNA specimens isolated from tumor tissues were apparently more stable than those of other tissues. These factors could all be used as quality predictors of RNA quality. In contrast, almost all analyses revealed that the genomic DNA samples had good quality, which was not influenced by the aforementioned factors. The results assisted us in determining preservation factors that affect specimen quality, which could provide evidence for improving processes of sample collection and preservation. Furthermore, the results are also useful for researchers to adopt as the evaluation criteria for choosing specimen collection and preservation strategies.

Circumferential thoracolumbar corrective fusion with an anterior interbody fresh-frozen femoral head allograft for osteoporotic lower acute kyphosis: a case report

PubMed Central

2009-01-01

Introduction Lower acute kyphosis (LAK) is a postural deformity caused by severe osteoporotic vertebral collapse at the thoracolumbar junction. Corrective surgery is indicated for severe cases, but no case report using a fresh-frozen femoral head allograft was found in the English literature. Case presentation A 69-year-old Japanese woman with severe LAK with osteoporotic vertebral fractures from T11 to L2 complained of severe back pain and difficulty in walking. The rigid kyphosis measured 74° from T10 to L3. The patient underwent an anterior release and interbody fusion using a fresh-frozen femoral head allograft (T11-L3) and a posterior instrumented fusion (T10-L3). Postoperatively, kyphosis was corrected to 28°, and the patient's symptoms were alleviated. The allograft bone was fully incorporated 1 year postoperatively. A new vertebral fracture at T10 occurred after 2 years, resulting in a slight loss of correction. A kyphosis angle of 35° at 2 years was maintained at 12 years (age, 81 years). She remained free of back pain and able to walk without a cane over the 12-year follow-up. Conclusion For treatment of severe osteoporotic LAK, anterior reconstruction is essential to obtain good spinal alignment and prevent recurrence. A fresh-frozen femoral head allograft, in combination with rigid posterior instrumented fixation, fulfills this function. PMID:19946544

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

PubMed

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

2012-12-01

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

An approach to successful freezing of demi-embryos derived from day-7 bovine embryos.

PubMed

Niemann, H; Brem, G; Sacher, B; Smidt, D; Kräusslich, H

1986-04-01

The developmental capacity of frozen/thawed bisected embryos (n = 33) derived from day-7 bovine embryos was investigated and compared to ordinary embryos after freezing and thawing (n = 28) and to freshly bisected embryos (n = 19). The freezing and thawing protocol was identical for ordinary and demi-embryos. The percentage of intact embryos classified as excellent, good, or poor after thawing was 92.9 and 96.3% for ordinary and demi-embryos, respectively. Pregnancy rates of 53.8 (8 15 ), 46.2 (6 13 ), and 47.5% (9 19 ) were obtained when frozen/thawed ordinary embryos and frozen/thawed demi-embryos classified as excellent or good and sealed with an additional zona pellucida from hatched pig blastocysts or freshly bisected embryos were transferred. One pair of identical twins resulted from the transfer of frozen/thawed demi-embryos sealed with an additional zona pellucida. Transfer of four frozen/thawed demi-embryos without an additional zona pellucida led to one pregnancy. In contrast, demi-embryos derived from frozen/thawed ordinary embryos (n = 8) as well as frozen/thawed demi-embryos classified as poor (n = 6) did not result in any pregnancies although two halves were transferred per recipient. It is concluded that sealing the punctured zona pellucida improves the developmental capacity of frozen/thawed demi-embryos derived from day-7 bovine embryos, and freezing demi-embryos is more efficient compared to the splitting of frozen/thawed ordinary embryos.

Does a frozen embryo transfer ameliorate the effect of elevated progesterone seen in fresh transfer cycles?

PubMed

Healy, Mae Wu; Patounakis, George; Connell, Matt T; Devine, Kate; DeCherney, Alan H; Levy, Michael J; Hill, Micah J

2016-01-01

To compare the effect of progesterone (P) on the day of trigger in fresh assisted reproduction technology (ART) transfer cycles versus its effect on subsequent frozen embryo transfer (FET) cycles. Retrospective cohort study. Large private ART practice. Fresh autologous and FET cycles from 2011-2013. None. Live birth. A paired analysis of patients who underwent both a fresh transfer and subsequent FET cycle and an unpaired analysis of data from all fresh transfer cycles and all FET cycles were performed. We analyzed 1,216 paired and 4,124 unpaired cycles, and P was negatively associated with birth in fresh but not FET cycles in all analyses. Interaction testing of P and cycle type indicated P had a different association with birth in fresh versus FET cycles. When P was ≥ 2 ng/mL at the time of trigger, live birth was more likely in FET versus fresh cycles in the paired analysis (47% vs. 10%), in the unpaired analysis (51% vs. 14%), and in unpaired, good blastocyst only transfer subgroup (51% vs. 29%). Live birth was similar in FET cycles, with P ≥ 2 ng/mL versus P < 2 ng/mL (51% vs. 49%). Conversely, live birth was lower in fresh cycles, with P ≥ 2 ng/mL versus P <2 ng/mL (15% vs. 45%). Elevated P levels on the day of trigger during the initial fresh cycle were negatively associated with live birth in the fresh transfer cycles but not in subsequent FET cycles. Freezing embryos and performing a subsequent FET cycle ameliorates the effect of elevated P on live-birth rates. Published by Elsevier Inc.

Cooking can decrease mercury contamination of a mushroom meal: Cantharellus cibarius and Amanita fulva.

PubMed

Falandysz, Jerzy; Drewnowska, Małgorzata

2017-05-01

Mushrooms (Cantharellus cibarius and Amanita fulva) were blanched (parboiled) and pickled using different treatment conditions with the aim of carrying out the study into effect on removal of toxic mercury (Hg) accumulated in flesh. Blanching of fresh sliced C. cibarius caused leaching of Hg by approximately 15%, while loss of up to 35% was observed for sliced, deep-frozen fruit bodies. The rate of Hg leaching from the C. cibarius in practice was the same when blanched for 5 or 15 min irrespective of potable or deionized water used. Pickling of blanched C. cibarius with a diluted vinegar marinade had only a minor, if any, effect on removal of Hg and was without effect on blanched caps of A. fulva. Mercury was better extracted by boiling water from the fresh caps of A. fulva (56 ± 2% of the initial level in fresh caps) than from the fresh or frozen fruit bodies of C. cibarius. Total leaching rate of Hg from a pickled C. cibarius when fresh fruit bodies were processed was between 15 ± 5 and 37 ± 7% (median range 13-34%), and when deep-frozen fruit bodies were processed, it was between 37 ± 7 and 39 ± 8% (median range 34-39%). Pickling of the caps of A. fulva with diluted vinegar did not increase leaching of Hg. Blanching of mushrooms before future culinary use is a simple procedure recommended in reduction of contamination with Hg of cooked mushroom meal. Pickling had little if any effect on further removal of Hg from the initially blanched mushrooms.

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

PubMed

Bowker, B; Zhuang, H

2017-09-01

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

Cost-effectiveness and budget impact study of solvent/detergent (SD) treated plasma (octaplasLG®) versus fresh-frozen plasma (FFP) in any patient receiving transfusion in Canada.

PubMed

Huisman, Eline L; van Eerd, Margreet C; Ouwens, J N Mario; de Peuter, Maria A

2014-08-01

The objectives of this study were to evaluate the cost-effectiveness and budget impact of octaplasLG(®) compared with fresh-frozen plasma (FFP) in all patients receiving a transfusion in Canada. A decision analytic framework was used to model acute and long-term complications that could follow plasma transfusion. Over a life time horizon, the cost with octaplasLG(®) were CA$612.91, which is CA$303.14 less than those with FFP. OctaplasLG(®) resulted in 0.021 quality adjusted life years (QALYs) gained in comparison with FFP. Because of higher efficacy and lower costs, octaplasLG(®) is expected to be the dominant treatment option over FFP in Canada. Copyright © 2013 Elsevier Ltd. All rights reserved.

Evaluation of fresh and frozen-thawed semen of individual ganders by assessment of spermatozoa motility and morphology.

PubMed

Łukaszewicz, Ewa; Kruszyński, Wojciech

2003-04-01

Individual differences in gander Anser anser L. reaction to semen collection procedure, quality and quantity of fresh semen and its susceptibility to the freezing process are discussed. Semen was collected individually by dorso-abdominal massage, from 1-year old White Koluda ganders (n = 12) every 2-3 days. Ganders' reactions to massage were observed during the entire reproductive cycle (from 11 February to 13 June, from every male 40 semen collections were performed). For individual evaluation and freezing purpose semen was collected 13 times from every male. In the fresh semen, the following parameters were evaluated: ejaculate volume, color, density, blood or fecal contamination, motility, concentration and morphology of spermatozoa. Motility and spermatozoa morphology were evaluated in the frozen-thawed semen. Semen diluted in 2:1 ratio with EK diluent was frozen with 6% of dimethyl-formamide (DMF) to -140 degrees C at a rate 60 degrees C/min. Semen was thawed by placing the straws in a 60 degrees C water-bath for 4-5 s. Ten out of 12 ganders had from 67.5 to 100.0% positive reactions resulting in semen ejaculation. Significant (P < or = 0.01) differences in fresh semen quality of particular ganders were observed for all evaluated traits. In 1-year-old gander semen morphologically intact spermatozoa constitute only 27.8-45.2% of all cells. Therefore, the sperm quality factor (SQF), proposed by the authors, which includes ejaculate volume, sperm concentration and the percentage of live normal spermatozoa, seems to be a good predictor of gander semen fertilizing ability. The SQF of individual ganders varied from 7.7 to 11.5. The percentage of live normal spermatozoa in the frozen-thawed semen depended mainly on fresh semen quality. In relation to the fresh semen average from 57.2 to 63.2% of spermatozoa survived freezing process and from 23.9 to 38.5% remained morphologically intact.

Systematic evaluation of RNA quality, microarray data reliability and pathway analysis in fresh, fresh frozen and formalin-fixed paraffin-embedded tissue samples.

PubMed

Wimmer, Isabella; Tröscher, Anna R; Brunner, Florian; Rubino, Stephen J; Bien, Christian G; Weiner, Howard L; Lassmann, Hans; Bauer, Jan

2018-04-20

Formalin-fixed paraffin-embedded (FFPE) tissues are valuable resources commonly used in pathology. However, formalin fixation modifies nucleic acids challenging the isolation of high-quality RNA for genetic profiling. Here, we assessed feasibility and reliability of microarray studies analysing transcriptome data from fresh, fresh-frozen (FF) and FFPE tissues. We show that reproducible microarray data can be generated from only 2 ng FFPE-derived RNA. For RNA quality assessment, fragment size distribution (DV200) and qPCR proved most suitable. During RNA isolation, extending tissue lysis time to 10 hours reduced high-molecular-weight species, while additional incubation at 70 °C markedly increased RNA yields. Since FF- and FFPE-derived microarrays constitute different data entities, we used indirect measures to investigate gene signal variation and relative gene expression. Whole-genome analyses revealed high concordance rates, while reviewing on single-genes basis showed higher data variation in FFPE than FF arrays. Using an experimental model, gene set enrichment analysis (GSEA) of FFPE-derived microarrays and fresh tissue-derived RNA-Seq datasets yielded similarly affected pathways confirming the applicability of FFPE tissue in global gene expression analysis. Our study provides a workflow comprising RNA isolation, quality assessment and microarray profiling using minimal RNA input, thus enabling hypothesis-generating pathway analyses from limited amounts of precious, pathologically significant FFPE tissues.

Cumulative live birth rates after one ART cycle including all subsequent frozen-thaw cycles in 1050 women: secondary outcome of an RCT comparing GnRH-antagonist and GnRH-agonist protocols.

PubMed

Toftager, M; Bogstad, J; Løssl, K; Prætorius, L; Zedeler, A; Bryndorf, T; Nilas, L; Pinborg, A

2017-03-01

Are cumulative live birth rates (CLBRs) similar in GnRH-antagonist and GnRH-agonist protocols for the first ART cycle including all subsequent frozen-thaw cycles from the same oocyte retrieval? The chances of at least one live birth following utilization of all fresh and frozen embryos after the first ART cycle are similar in GnRH-antagonist and GnRH-agonist protocols. Reproductive outcomes of ART treatment are traditionally reported as pregnancies per cycle or per embryo transfer. However, the primary concern is the overall chance of a live birth. After the first ART cycle with fresh embryo transfer, we found live birth rates (LBRs) of 22.8% and 23.8% (P = 0.70) for the GnRH-antagonist and GnRH-agonist protocols, respectively. But with CLBRs including both fresh and frozen embryos from the first oocyte retrieval, chances of at least one live birth increases. There are no previous randomized controlled trials (RCTs) comparing CLBRs in GnRH-antagonist versus GnRH-agonist protocols. Previous studies on CLBR are either retrospective cohort studies including multiple fresh cycles or RCTs comparing single embryo transfer (SET) with double embryo transfer (DET). CLBR was a secondary outcome in a Phase IV, dual-center, open-label, RCT including 1050 women allocated to a short GnRH-antagonist or a long GnRH-agonist protocol in a 1:1 ratio over a 5-year period using a web-based concealed randomization code. The minimum follow-up time from the first IVF cycle was 2 years. The aim was to compare CLBR between the two groups following utilization of all fresh and frozen embryos from the first ART cycle. All women referred for their first ART cycle at two public fertility clinics, <40 years of age were approached. A total of 1050 subjects were allocated to treatment and 1023 women started standardized ART protocols with recombinant human follitropin-β (rFSH) stimulation. Day-2 SET was planned and additional embryos were frozen and used in subsequent frozen-thawed cycles. All pregnancies generated from oocyte retrieval during the first IVF cycle including fresh and frozen-thaw cycles were registered. Ongoing pregnancy was determined by ultrasonography at gestational week 7-9 and live birth was irrespective of the duration of gestation. CLBR was defined as at least one live birth per allocated woman after fresh and frozen cycles. Subjects were censored out after the first live birth. Cox proportional hazard model was used to evaluate the relative prognostic significance of female age, BMI, the number of retrieved oocytes and the diagnosis of infertility in relation to the CLBR. Baseline characteristics were similar and equal proportions of patients continued with frozen-thaw (frozen embryo transfer, FET) cycles after their fresh ART cycle in the GnRH-antagonist and GnRH-agonist arms. When combining all fresh and frozen-thaw embryo transfers from first oocyte retrieval with a minimum of 2-year follow-up, the CLBR was 34.1% (182/534) in the GnRH-antagonist group versus 31.2% (161/516) in the GnRH-agonist group (odds ratio (OR):1.14; 95% CI: 0.88-1.48, P = 0.32). Mean time to the first live birth was 11.0 months in the GnRH-antagonist group compared to 11.5 months in the GnRH-agonist group (P < 0.01). The total number of deliveries from all FET cycles where embryos were thawed were higher in the antagonist group 64/330 (19.4%) compared to the agonist group 43/355 (12.1%) ((OR): 1.74; 95% CI: 1.14-2.66, P = 0.01). The evaluation of prognostic factors showed that more retrieved oocytes were associated with a significantly higher CLBR in both treatment groups. For the subgroup of obese women (BMI >30 kg/m2), the CLBR was significantly higher in the GnRH-antagonist group (P = 0.02). The duration of the trial is a possible limitation with introduction of new methods as 'Freeze all' and 'GnRH-agonist triggering', but as these treatments were used in only few women, a systematic bias is not likely. Blastocyst culture of surplus embryos for freezing was introduced to both groups simultaneously, thereby minimizing the risk of bias. Furthermore, with a minimum of 2-year follow-up, a minority (<1%) still had cryopreserved embryos and no live birth at the end of the trial. The post hoc prognostic covariate analyses with multiple strata should be interpreted with caution. Finally, the physicians were not blinded to GnRH treatment group after randomization. With the improvement of embryo culture, freezing and thawing methods as well as a strategy of elective SET, CLBR until first live birth provides an all-inclusive success rate for ART. When comparing GnRH-antagonist and GnRH-agonist protocols, we find similar CLBRs, despite more oocytes being retrieved in the GnRH-agonist protocol. An unrestricted research grant is funded by Merck Sharp & Dohme Corp., a subsidiary of Merck & Co., Inc., Kenilworth, NJ, USA (MSD). The funders had no influence on the data collection, analyses or conclusions of the study. No conflict of interests to declare. EudraCT #: 2008-005452-24. ClinicalTrial.gov: NCT00756028. 18 September 2008. 14 January 2009. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Nuclear Matrix Proteins in Disparity of Prostate Cancer

DTIC Science & Technology

2013-09-01

nuclear coactivator-3 (NCOA3). 5 Methods Patients and Prostate Cancer Specimens Fresh, flash -frozen specimens were obtained from age- (50 to...for reliable data interpretation. Gene Array Analysis Total RNA isolated from LCM-procured normal epithelium and tumor cells from flash -frozen...PCR Briefly, RNA was extracted from matched LCM procured normal epithelium and tumor cells of age-, tumor grade-matched flash -frozen sections (n=24

Treatment of plasminogen deficiency patients with fresh frozen plasma.

PubMed

Kızılocak, Hande; Ozdemir, Nihal; Dikme, Gürcan; Koç, Begüm; Atabek, Ayşe Ayzıt; Çokuğraş, Haluk; İskeleli, Güzin; Dönmez-Demir, Buket; Christiansen, Nina Merete; Ziegler, Maike; Ozdağ, Hilal; Schuster, Volker; Celkan, Tiraje

2018-02-01

Congenital plasminogen (Plg) deficiency leads to the development of ligneous membranes on mucosal surfaces. Here, we report our experience with local and intravenous fresh frozen plasma (FFP). We retrospectively reviewed medical files of 17 patients and their eight first-degree relatives. Conjunctivitis was the main complaint. Thirteen patients were treated both with intravenous and conjunctival FFP. Venous thrombosis did not develop in any. Genetic evaluation revealed heterogeneous mutations as well as polymorphisms. Diagnosis and treatment of Plg deficiency is challenging; topical and intravenous FFP may be an alternative treatment. © 2017 Wiley Periodicals, Inc.

The effect of different solvents and number of extraction steps on the polyphenol content and antioxidant capacity of basil leaves (Ocimum basilicum L.) extracts.

PubMed

Złotek, Urszula; Mikulska, Sylwia; Nagajek, Małgorzata; Świeca, Michał

2016-09-01

The objectives of this study were to determine best conditions for the extraction of phenolic compounds from fresh, frozen and lyophilized basil leaves. The acetone mixtures with the highest addition of acetic acid extracted most of the phenolic compounds when fresh and freeze-dried material have been used. The three times procedure was more effective than once shaking procedure in most of the extracts obtained from fresh basil leaves - unlike the extracts derived from frozen material. Surprisingly, there were not any significant differences in the content of phenolics between the two used procedures in the case of lyophilized basil leaves used for extraction. Additionally, the positive correlation between the phenolic compounds content and antioxidant activity of the studied extracts has been noted. It is concluded that the acetone mixtures were more effective than the methanol ones for polyphenol extraction. The number of extraction steps in most of the cases was also a statistically significant factor affecting the yield of phenolic extraction as well as antioxidant potential of basil leaf extracts.

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.

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

PubMed

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

2015-05-01

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

Economic evaluation of pooled solvent/detergent treated plasma versus single donor fresh-frozen plasma in patients receiving plasma transfusions in the United States.

PubMed

Huisman, Eline L; de Silva, Shamika U; de Peuter, Maria A

2014-08-01

This study assessed the cost-effectiveness of Octaplas™ versus fresh frozen plasma (FFP) in patients receiving plasma transfusions in the United States (US). Acute and long-term complications of plasma transfusions were modelled in a decision tree followed by a Markov model, using a healthcare payer perspective. Over a lifetime time horizon, patients receiving Octaplas™ accumulate slightly more life years (0.00613 [95% uncertainty interval (95%UI): 0.00166-0.01561]) and quality-adjusted life years (QALY) (0.023 [95%UI: 0.012-0.044]) at lower cost compared with those treated with FFP. Octaplas™ demonstrated to be the dominant treatment option over FFP (95%UI: Dominant-US$ 15,764/QALY). Copyright © 2014 Elsevier Ltd. All rights reserved.

Characterization of alkaline phosphatase activity in seminal plasma and in fresh and frozen-thawed stallion spermatozoa.

PubMed

Bucci, Diego; Giaretta, Elisa; Spinaci, Marcella; Rizzato, Giovanni; Isani, Gloria; Mislei, Beatrice; Mari, Gaetano; Tamanini, Carlo; Galeati, Giovanna

2016-01-15

Alkaline phosphatase (AP) has been studied in several situations to elucidate its role in reproductive biology of the male from different mammalian species; at present, its role in horse sperm physiology is not clear. The aim of the present work was to measure AP activity in seminal plasma and sperm extracts from freshly ejaculated as well as in frozen-thawed stallion spermatozoa and to verify whether relationship exists between AP activity and sperm quality parameters. Our data on 40 freshly ejaculated samples from 10 different stallions demonstrate that the main source of AP activity is seminal plasma, whereas sperm extracts contribution is very low. In addition, we found that AP activity at physiological pH (7.0) is significantly lower than that observed at pH 8.0, including the optimal AP pH (pH 10.0). Alkaline phosphatase did not exert any effect on sperm-oocyte interaction assessed by heterologous oocyte binding assay. Additionally, we observed a thermal stability of seminal plasma AP, concluding that it is similar to that of bone isoforms. Positive correlations were found between seminal plasma AP activity and sperm concentration, whereas a negative correlation was present between both spermatozoa extracts and seminal plasma AP activity and seminal plasma protein content. A significant decrease in sperm extract AP activity was found in frozen-thawed samples compared with freshly ejaculated ones (n = 21), concomitantly with the decrease in sperm quality parameters. The positive correlation between seminal plasma AP activity measured at pH 10 and viability of frozen-thawed spermatozoa suggests that seminal plasma AP activity could be used as an additional predictive parameter for stallion sperm freezability. In conclusion, we provide some insights into AP activity in both seminal plasma and sperm extracts and describe a decrease in AP after freezing and thawing. Copyright © 2016 Elsevier Inc. All rights reserved.

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.

The influence of female age on the cumulative live-birth rate of fresh cycles and subsequent frozen cycles using vitrified blastocysts in hyper-responders.

PubMed

Wu, Cheng-Hsuan; Lee, Tsung-Hsien; Chen, Hsiu-Hui; Chen, Chung-I; Huang, Chun-Chia; Lee, Maw-Sheng

2015-10-01

The aim of this research was to study the influence of female age on the cumulative live-birth rate of fresh and subsequent frozen cycles using vitrified blastocysts of the same cohort in hyper-responders. This was a retrospective study of 1137 infertile women undergoing their first in vitro fertilization treatment between 2006 and 2013. The main outcome measure was cumulative live births among the fresh and all vitrified blastocyst transfers combined after the same stimulation cycle. The results were also analyzed according to age (i.e., <35 years, 35-39 years, and ≥ 40 years). The mean number of retrieved oocytes was 19.9 ± 8.5 oocytes. The cumulative pregnancy rate was 89.2% and the cumulative live-birth rate was 73.3%. The cumulative live-birth rate declined from 73.9% for women younger than 35 years old to 67.3% for women 35-39 years old to 57.9% for women 40 years or older. Combined fresh and vitrified blastocyst transfer cycles can result in a high cumulative live-birth rate. The cumulative live-birth rates among older women are lower than the rates among younger women when autologous oocytes are used. Copyright © 2015. Published by Elsevier B.V.

Determination and assessment of total mercury levels in local, frozen and canned fish in Lebanon.

PubMed

Obeid, Pierre J; El-Khoury, Bilal; Burger, Joanne; Aouad, Samer; Younis, Mira; Aoun, Amal; El-Nakat, John Hanna

2011-01-01

Fish is an important constituent of the Lebanese diet. However, very little attention in our area is given to bring awareness regarding the effect of the toxicity of mercury (Hg) mainly through fish consumption. This study aimed to report analytical data on total mercury levels in several fish species for the first time in thirty years and to also made individuals aware of the presence and danger from exposure to mercury through fish consumption. Fish samples were selected from local Lebanese markets and fisheries and included 94 samples of which were fresh, frozen, processed, and canned fish. All values were reported as microgram of mercury per gram of fish based on wet weight. The level of mercury ranged from 0.0190 to 0.5700 microg/g in fresh samples, 0.0059 to 0.0665 microg/g in frozen samples, and 0.0305 to 0.1190 microg/g in canned samples. The data clearly showed that higher levels of mercury were detected in local fresh fish as opposed to other types thus placing consumers at higher risk from mercury exposure. Moreover, the data revealed that Mallifa (yellowstripe barracuda/Sphyraena chrysotaenia), Sargous (white seabream/Diplodus sargus), Ghobbos (bogue/Boops boops), and shrimp (Penaeus sp.) were among the types containing the highest amounts of mercury. On the other hand, processed fish such as fish fillet, fish burger, small shrimp and crab are found to contain lower levels of mercury and are associated with lower exposure risks to mercury. Lebanese population should therefore, be aware to consume limited amounts of fresh local fish to minimize exposure to mercury.

Splitting blood and blood product packaging reduces donor exposure for patients undergoing cardiopulmonary bypass.

PubMed

Nuszkowski, M M; Jonas, R A; Zurakowski, D; Deutsch, N

2015-11-01

Cardiopulmonary bypass for congenital heart surgery requires packed red cells (PRBC) and fresh frozen plasma (FFP) to be available, both for priming of the circuit as well as to replace blood loss. This study examines the hypothesis that splitting one unit of packed red blood cells and one unit of fresh frozen plasma into two half units reduces blood product exposure and wastage in the Operating Room. Beginning August 2013, the blood bank at Children's National Medical Center began splitting one unit of packed red blood cells (PRBC) and one unit of fresh frozen plasma (FFP) for patients undergoing cardiopulmonary bypass (CPB). The 283 patients who utilized CPB during calendar year 2013 were divided into 2 study groups: before the split and after the split. The principal endpoints were blood product usage and donor exposure intra-operatively and within 72 hours post-operatively. There was a significant decrease in median total donor exposures for FFP and cryoprecipitate from 5 to 4 per case (p = 0.007, Mann-Whitney U-test). However, there was no difference in the volume of blood and blood products used; in fact, there was a significant increase in the amount of FFP that was wasted with the switch to splitting the unit of FFP. We found that modification of blood product packaging can decrease donor exposure. Future investigation is needed as to how to modify packaging to minimize wastage. © The Author(s) 2015.

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.

C1q deficiency: identification of a novel missense mutation and treatment with fresh frozen plasma.

PubMed

Topaloglu, Rezan; Taskiran, Ekim Z; Tan, Cagman; Erman, Baran; Ozaltin, Fatih; Sanal, Ozden

2012-07-01

A Turkish patient with C1q deficiency presented with a lupus-like disease, and a new missense mutation at A chain is presented. To characterize the genetic defect, all exons of the genes for the A, B, and C chains of C1q were sequenced in the patient. This revealed a missense mutation in the collagen-like domain of the A chain, p.Gly31 Arg. No other sequence variants, including the common silent mutations, were found in the three chains. Exon 1 of the C1q A chain was sequenced in 105 samples from healthy controls for this particular mutation. None of these carried the mutation. The C1q-deficient patient was treated with fresh frozen plasma infusions. Our findings showed that Turkish patients may have different mutations than the previously described common mutation, and once again, not only nonsense mutations but also missense mutations cause hereditary C1q deficiency. Regular fresh frozen plasma infusions to the patient have been clinically and therapeutically successful.

Effects of three different preservation methods on the mechanical properties of human and bovine cortical bone.

PubMed

Unger, Stefan; Stefan, Unger; Blauth, Michael; Michael, Blauth; Schmoelz, Werner; Werner, Schmoelz

2010-12-01

In the development of new strategies for fracture fixation, new methods have to be tested biomechanically under in vitro conditions before clinical trials can be performed. The gold standard for laboratory evaluations is fresh-frozen specimen. As the availability of fresh-frozen specimens is limited and since their use bears infectious risks, specimens treated with various chemical embalming fluids are also used. These preservation methods may alter the mechanical properties of the specimens used. Therefore, the aims of the present study were to determine the effects of three different preservation methods (formalin fixation (FO), Thiel-fixation (TH), and alcohol-glycerine fixation (AG)) on the elastic and postyield mechanical properties of cortical bone and to compare these properties to those of fresh-frozen (FF) specimens. Cylindrical cortical specimens (diameter 3mm, length 60 mm) were obtained from human femurs (n=48) and bovine tibiae (n=40). Before specimen immersion in different fixation fluids, bone mineral density (BMD) as well as the initial Young's modulus was determined. The Young's modulus was determined in a nondestructive bending test, and measurements were repeated after 6 months of immersion in fixative solution. Subsequent to the nondestructive test, a destructive 3-point bending test was conducted to assess the postyield and fracture properties. The BMD as well as the initial Young's modulus showed no significant differences between the four test groups. After 6 months in fixative solution, the Young's modulus was significantly lowered in human Thiel specimens and only showed minor changes in formalin- and alcohol-glycerine-treated specimens. The plastic energy absorption of human and bovine specimens was altered significantly. Formalin as well as alcohol-glycerine fixation yielded a significant decrease in plastic energy absorption, whereas Thiel fixation significantly increased the plastic energy absorption. Because of the significantly altered plastic mechanical properties of cortical bone, the use fresh-frozen bone specimens is recommended in biomechanical studies investigating failure loads of orthopaedic implants. The use of embalmed specimens should be restricted to pilot tests. Copyright © 2010 Elsevier Inc. All rights reserved.

Pronuclear formation by ICSI using chemically activated ovine oocytes and zona pellucida bound sperm.

PubMed

Hernández-Pichardo, J E; Ducolomb, Y; Romo, S; Kjelland, M E; Fierro, R; Casillas, F; Betancourt, M

2016-01-01

In order to improve ICSI, appropiate sperm selection and oocyte activation is necessary. The objective of the present study was to determine the efficiency of fertilization using ICSI with chemically activated ovine oocytes and sperm selected by swim up (SU) or swim up + zona pellucida (SU + ZP) binding. Experiment 1, 4-20 replicates with total 821 in vitro matured oocytes were chemically activated with ethanol, calcium ionophore or ionomycin, to determine oocyte activation (precense of one PN). Treatments showed similar results (54, 47, 42 %, respectively) but statistically differents ( P  < 0.05) than mechanical activated oocytes in sham, ICSI and sham injection (13, 25, 32 %, respectively) (10-17 replicates; n  = 429). Experiment 2: Twelve ejaculates and 28 straws of semen were used (11-19 replicates). Sperm were selected by SU in BSA-TCM 199-H medium. A total of 2,294 fresh sperm and 2,760 from frozen-thawed semen were analyzed after SU or SU + ZP binding. Fresh sperm selected by SU showed acrosome reaction (AR) of 59 %, the sperm selected by SU + ZP binding increased AR to 91 %. In comparison, the AR of frozen-thawed sperm using SU or SU + ZP binding was 77 and 86 %, respectively ( P  < 0.05). Experiment 3: fertilization in 200 mechanical activativated oocytes (17 replicates) was 4 %, but fertilization increased in ethanol activated oocytes after ICSI (12-28 %) (5-6 replicates). When fresh sperm only selected by SU were injected to 123 oocytes, a fertilization rate (28 %) was achieved; in sperm selected by SU + ZP was 25 % (73 oocytes). In comparison, in frozen-thawed sperm selected by SU, fertilization was 13 % (70 oocytes), whereas sperm from SU + ZP binding displayed 12 % (51 oocytes) ( P  > 0.05). Chemical activation induces higher ovine oocyte activation than mechanical activation. Ethanol slightly displays higher oocyte activation than calcium ionophore and ionomicine. Sperm selection with SU + ZP increased AR/A and AR/D rates in comparison with SU in fresh and frozen-thawed sperm. According to this, in terms of fertilization rates, chemical activation after ICSI increased oocyte PN formation compared to mechanical activation. Also, fresh sperm treated with SU and SU + ZP were significantly different than frozen-thawed sperm, but between sperm treatments no significant differences were obtained.

Preparation and Immunoaffinity Depletion of Fresh Frozen Tissue Homogenates for Mass Spectrometry-Based Proteomics in the Context of Drug Target/Biomarker Discovery.

PubMed

Prieto, DaRue A; Chan, King C; Johann, Donald J; Ye, Xiaoying; Whitely, Gordon; Blonder, Josip

2017-01-01

The discovery of novel drug targets and biomarkers via mass spectrometry (MS)-based proteomic analysis of clinical specimens has proven to be challenging. The wide dynamic range of protein concentration in clinical specimens and the high background/noise originating from highly abundant proteins in tissue homogenates and serum/plasma encompass two major analytical obstacles. Immunoaffinity depletion of highly abundant blood-derived proteins from serum/plasma is a well-established approach adopted by numerous researchers; however, the utilization of this technique for immunodepletion of tissue homogenates obtained from fresh frozen clinical specimens is lacking. We first developed immunoaffinity depletion of highly abundant blood-derived proteins from tissue homogenates, using renal cell carcinoma as a model disease, and followed this study by applying it to different tissue types. Tissue homogenate immunoaffinity depletion of highly abundant proteins may be equally important as is the recognized need for depletion of serum/plasma, enabling more sensitive MS-based discovery of novel drug targets, and/or clinical biomarkers from complex clinical samples. Provided is a detailed protocol designed to guide the researcher through the preparation and immunoaffinity depletion of fresh frozen tissue homogenates for two-dimensional liquid chromatography, tandem mass spectrometry (2D-LC-MS/MS)-based molecular profiling of tissue specimens in the context of drug target and/or biomarker discovery.

A new insight into male fertility preservation for patients with completely immotile spermatozoa.

PubMed

Chen, Huanhua; Feng, Guixue; Zhang, Bo; Zhou, Hong; Wang, Caizhu; Shu, Jinhui; Gan, Xianyou; Lin, Ruoyun; Huang, Dongmei; Huang, Yingqin

2017-09-18

Sperm cryopreservation is the most effective method to preserve male fertility but this is normally used for motile spermatozoa. Thus, only motile spermatozoa are used for cryopreservation in most reproductive medicine centers worldwide. The immotile spermatozoa from some problematic patients are usually discarded, resulting in a missed opportunity of sterility cryopreservation for future assisted reproductive treatments. Many studies have shown that successful fertilization can be obtained after selection of viable sperm from the completely immotile spermatozoa before ICSI. Whether the completely immotile spermatozoa are worth of freezing has not been realized The aim of this study is to explore the clinical value of cryopreservation of immotile spermatozoa. Completely immotile spermatozoa were collected and frozen, and subsequently viable but immotile frozen-thawed spermatozoa were selected by laser plus for ICSI. Main outcomes included spermatozoa survival index, fertilization rate and good quality embryo rate. After identification by laser, the fresh samples of spermatozoa presented with a mean survival rate of 54.86% and 26.05%, and this was reduced to 44.13% and 18.13% in frozen-thawed spermatozoa samples, which showed a frozen-thawed spermatozoa survival index of 0.80 and 0.70 in the testicular and ejaculate sperm, respectively. There were no statistically differences in fertilization rate (80% vs80.51%, 75.00% vs 81.48%), cleavage rate (95.45% vs 98.95%, 100.00% vs 95.45%) and good quality embryo rate (40.48% vs 52.13%, 33.33%vs38.10%) between the frozen-thawed immotile spermatozoa group and the routine fresh immotile spermatozoa ICSI group in both testicular and ejaculate sperm, respectively. The results of the study show that completely immotile spermatozoa can be frozen in order to preserve male fertility as long as viable spermatozoa are present. This procedure provides a further possibility for fertility preservation for patients with completely immotile spermatozoa.

Freezing and calcium chloride marination effects on beef tenderness and calpastatin activity.

PubMed

Whipple, G; Koohmaraie, M

1992-10-01

Because freezing samples decreases calpastatin activity and the application of exogenous calcium activates the calpain proteolytic system, thereby improving tenderness, the objective of this study was to determine whether freezing would enhance the effects of CaCl2 marination on the tenderness of beef steaks. Longissimus steaks were obtained from 10 beef steers 6 d postmortem. One-half of the steaks were frozen at -30 degrees C for 6 wk. The remaining steaks were treated fresh; one-half were subjected to a 150 mM CaCl2 marinade for 48 h. Frozen steaks were thawed and subjected to the same treatment. Treatments consisted of 1) fresh control, 2) fresh marinated, 3) frozen control, and 4) frozen marinated. Samples were taken before and after treatment (6 and 8 d) for calpastatin activity determination and d 8 for SDS-PAGE. Warner-Bratzler shear force values were measured 8 d postmortem. Data were analyzed using a paired comparison t-test procedure. Results showed that freezing and marination significantly decreased calpastatin activity. A .35-kg improvement (P = .07) in Warner-Bratzler shear force was observed with freezing, whereas a .78-kg improvement (P less than .01) in tenderness was observed with marination. However, prior freezing enhanced the effects of marination. Therefore, the decrease in calpastatin activity seemed to allow greater proteolysis by the calpains with the application of Ca2+. The SDS-PAGE of myofibril preparations indicated that more small polypeptide fragments (28 to 32 kDa) appeared and a 95-kDa fragment was more intense in the marinated samples than in control samples, indicating that proteolysis was enhanced.(ABSTRACT TRUNCATED AT 250 WORDS)

Heating or freezing bone. Effects on angiogenesis induction and growth potential in mice.

PubMed

Leunig, M; Yuan, F; Berk, D A; Gerweck, L E; Jain, R K

1996-08-01

We have characterized the effect of bone graft treatment by heating or freezing (with or without dimethyl sulfoxide (DMSO)). Tissue culture and dorsal skin-fold chambers in mice were used as sites to quantify the effect on angiogenesis, growth and calcification of neonatal femora. Fresh femora increased in both length and cartilage diameter (calcification in vivo only), but cryopreservation or heating abolished the increase in femoral dimensions. In vivo, femora of all experimental groups elicited an angiogenic response from the host tissue, which was most pronounced for fresh femora, weaker for DMSO-preserved frozen bone and poor for unprotected frozen bone and boiled femora. Freezing in the presence of a cryopreservative (DMSO) was found to preserve the angiogenic potential of frozen bone, whereas unprotected heating or freezing significantly impaired angiogenesis induction and growth potential.

Fertility prediction of frozen boar sperm using novel and conventional analyses

USDA-ARS?s Scientific Manuscript database

Frozen-thawed boar sperm is seldom used for artificial insemination (AI) because fertility is lower than fresh or cooled semen. Despite the many advantages of AI including reduced pathogen exposure and ease of semen transport, cryo-induced damage to sperm usually results in decreased litter sizes a...

Antimüllerian hormone as a predictor of live birth following assisted reproduction: an analysis of 85,062 fresh and thawed cycles from the Society for Assisted Reproductive Technology Clinic Outcome Reporting System database for 2012-2013.

PubMed

Tal, Reshef; Seifer, David B; Wantman, Ethan; Baker, Valerie; Tal, Oded

2018-02-01

To determine if serum antimüllerian hormone (AMH) is associated with and/or predictive of live birth assisted reproductive technology (ART) outcomes. Retrospective analysis of Society for Assisted Reproductive Technology Clinic Outcome Reporting System database from 2012 to 2013. Not applicable. A total of 69,336 (81.8%) fresh and 15,458 (18.2%) frozen embryo transfer (FET) cycles with AMH values. None. Live birth. A total of 85,062 out of 259,499 (32.7%) fresh and frozen-thawed autologous non-preimplantation genetic diagnosis cycles had AMH reported for cycles over this 2-year period. Of those, 70,565 cycles which had embryo transfers were included in the analysis. Serum AMH was significantly associated with live birth outcome per transfer in both fresh and FET cycles. Multiple logistic regression demonstrated that AMH is an independent predictor of live birth in fresh transfer cycles and FET cycles when controlling for age, body mass index, race, day of transfer, and number of embryos transferred. Receiver operating characteristic (ROC) curves demonstrated that the areas under the curve (AUC) for AMH as predictors of live birth in fresh cycles and thawed cycles were 0.631 and 0.540, respectively, suggesting that AMH alone is a weak independent predictor of live birth after ART. Similar ROC curves were obtained also when elective single-embryo transfer (eSET) cycles were analyzed separately in either fresh (AUC 0.655) or FET (AUC 0.533) cycles, although AMH was not found to be an independent predictor in eSET cycles. AMH is a poor independent predictor of live birth outcome in either fresh or frozen embryo transfer for both eSET and non-SET transfers. Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

Assays for endogenous components of human milk: comparison of fresh and frozen samples and corresponding analytes in serum.

PubMed

Hines, Erin P; Rayner, Jennifer L; Barbee, Randy; Moreland, Rae Ann; Valcour, Andre; Schmid, Judith E; Fenton, Suzanne E

2007-05-01

Breast milk is a primary source of nutrition that contains many endogenous compounds that may affect infant development. The goals of this study were to develop reliable assays for selected endogenous breast milk components and to compare levels of those in milk and serum collected from the same mother twice during lactation (2-7 weeks and 3-4 months). Reliable assays were developed for glucose, secretory IgA, interleukin-6, tumor necrosis factor-a, triglycerides, prolactin, and estradiol from participants in a US EPA study called Methods Advancement in Milk Analysis (MAMA). Fresh and frozen (-20 degrees C) milk samples were assayed to determine effects of storage on endogenous analytes. The source effect (serum vs milk) seen in all 7 analytes indicates that serum should not be used as a surrogate for milk in children's health studies. The authors propose to use these assays in studies to examine relationships between the levels of milk components and children's health.

Outcome Analysis of Day-3 Frozen Embryo Transfer v/s Fresh Embryo Transfer in Infertility: A Prospective Therapeutic Study in Indian Scenario.

PubMed

Chandel, Neha Palo; Bhat, Vidya V; Bhat, B S; Chandel, Sidharth S

2016-10-01

Advanced fertilization techniques like frozen embryo transfer (FET) and assisted reproductive technology have become popular and commonly used methods to treat patients suffering from infertility. Incidences of infertility are on a rise due to increased representation of females in the work place, delay in marriages, stress, and ignorance. We performed this prospective therapeutic study to compare FET and fresh embryo transfer in the treatment of infertility in terms of conception rate, patient acceptance, complications, and patient's compliance. A prospective screening therapeutic study on 108 patients, from September 2013 to September 2014 in Karnataka, India, randomized the patients into 2 groups (n = 54), Group-I treated with day-3 FET while Group-II was treated with fresh embryo transfer, after performing ICSI. In 108 patients, 45 % patients were within 35 years of age, 35 % were in the age group 35-39. Significantly, 22 (40.75 %) patients treated with FET conceived (P = 0.022), whereas 16 (29.63 %) patients treated with fresh embryo transfer conceived (P = 0.59). There is limited published literature from the subcontinent, comparing techniques like FET and embryo transfers in the treatment of infertility. Awareness and economic reforms must be formulated in India to facilitate individuals facing infertility problems to conceive. FET has better and significant conception rates compared to fresh embryo transfers. FET shares an advantage of providing good quality embryos for future and subsequent implantations in cases of failure. Patient counseling and motivation play a pivotal role in the success of therapeutic procedure.

Validation of the Lung Subtyping Panel in Multiple Fresh-Frozen and Formalin-Fixed, Paraffin-Embedded Lung Tumor Gene Expression Data Sets.

PubMed

Faruki, Hawazin; Mayhew, Gregory M; Fan, Cheng; Wilkerson, Matthew D; Parker, Scott; Kam-Morgan, Lauren; Eisenberg, Marcia; Horten, Bruce; Hayes, D Neil; Perou, Charles M; Lai-Goldman, Myla

2016-06-01

Context .- A histologic classification of lung cancer subtypes is essential in guiding therapeutic management. Objective .- To complement morphology-based classification of lung tumors, a previously developed lung subtyping panel (LSP) of 57 genes was tested using multiple public fresh-frozen gene-expression data sets and a prospectively collected set of formalin-fixed, paraffin-embedded lung tumor samples. Design .- The LSP gene-expression signature was evaluated in multiple lung cancer gene-expression data sets totaling 2177 patients collected from 4 platforms: Illumina RNAseq (San Diego, California), Agilent (Santa Clara, California) and Affymetrix (Santa Clara) microarrays, and quantitative reverse transcription-polymerase chain reaction. Gene centroids were calculated for each of 3 genomic-defined subtypes: adenocarcinoma, squamous cell carcinoma, and neuroendocrine, the latter of which encompassed both small cell carcinoma and carcinoid. Classification by LSP into 3 subtypes was evaluated in both fresh-frozen and formalin-fixed, paraffin-embedded tumor samples, and agreement with the original morphology-based diagnosis was determined. Results .- The LSP-based classifications demonstrated overall agreement with the original clinical diagnosis ranging from 78% (251 of 322) to 91% (492 of 538 and 869 of 951) in the fresh-frozen public data sets and 84% (65 of 77) in the formalin-fixed, paraffin-embedded data set. The LSP performance was independent of tissue-preservation method and gene-expression platform. Secondary, blinded pathology review of formalin-fixed, paraffin-embedded samples demonstrated concordance of 82% (63 of 77) with the original morphology diagnosis. Conclusions .- The LSP gene-expression signature is a reproducible and objective method for classifying lung tumors and demonstrates good concordance with morphology-based classification across multiple data sets. The LSP panel can supplement morphologic assessment of lung cancers, particularly when classification by standard methods is challenging.

Hamstring autograft versus soft-tissue allograft in anterior cruciate ligament reconstruction: a systematic review and meta-analysis of randomized controlled trials.

PubMed

Cvetanovich, Gregory L; Mascarenhas, Randy; Saccomanno, Maristella F; Verma, Nikhil N; Cole, Brian J; Bush-Joseph, Charles A; Bach, Bernard R

2014-12-01

To compare outcomes of anterior cruciate ligament (ACL) reconstruction with hamstring autograft versus soft-tissue allograft by systematic review and meta-analysis. A systematic review of randomized controlled studies comparing hamstring autograft with soft-tissue allograft in ACL reconstruction was performed. Studies were identified by strict inclusion and exclusion criteria. Descriptive statistics were reported. Where possible, the data were pooled and a meta-analysis was performed using RevMan software (The Nordic Cochrane Centre, The Cochrane Collaboration, Copenhagen, Denmark). Dichotomous data were reported as risk ratios, whereas continuous data were reported as standardized mean differences and 95% confidence intervals. Heterogeneity was assessed by use of I(2) for each meta-analysis. Study methodologic quality was analyzed with the Modified Coleman Methodology Score and Jadad scale. Five studies with 504 combined patients (251 autograft and 253 allograft; 374 male and 130 female patients) with a mean age of 29.9 ± 2.2 years were included. The allografts used were fresh-frozen hamstring, irradiated hamstring, mixture of fresh-frozen and cryopreserved hamstring, fresh-frozen tibialis anterior, and fresh-frozen Achilles tendon grafts without bone blocks. The mean follow-up period was 47.4 ± 26.9 months, with a mean follow-up rate of 83.3% ± 8.6%. Two studies found a longer operative time with autograft than with allograft (77.1 ± 2.0 minutes v 59.9 ± 0.9 minutes, P = .008). Meta-analysis showed no statistically significant differences between autografts and allografts for any outcome measures (P > .05 for all tests). One study found significantly greater laxity for irradiated allograft than for autograft. The methodologic quality of the 5 studies was poor, with a mean Modified Coleman Methodology Score of 54.4 ± 6.9 and mean Jadad score of 1.6 ± 1.5. On the basis of this systematic review and meta-analysis of 5 randomized controlled trials, there is no statistically significant difference in outcome between patients undergoing ACL reconstruction with hamstring autograft and those undergoing ACL reconstruction with soft-tissue allograft. These results may not extrapolate to younger patient populations. The methodology of the available randomized controlled trials comparing hamstring autograft and soft-tissue allograft is poor. Level II, systematic review of Level I and II studies. Copyright © 2014 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.

[Proposed difficult airway teaching methodology. Presentation of an interactive fresh frozen cadaver model].

PubMed

Catalá Bauset, J C; de Andres Ibañez, J A; Valverde Navarro, A; Martinez Soriano, F

2014-04-01

The aim of this paper is to present a methodology based on the use of fresh-frozen cadavers for training in the management of the airway, and to evaluate the degree of satisfaction among learning physicians. About 6 fresh-frozen cadavers and 14 workstations were prepared where participants were trained in the different skills needed for airway management. The details of preparation of the cadavers are described. The level of satisfaction of the participant was determined using a Likert rating scale of 5 points, at each of the 14 stations, as well as the overall assessment and clinical usefulness of the course. The mean overall evaluation of the course and its usefulness was 4.75 and 4.9, out of 5, respectively. All parts of the course were rated above 4 out of 5. The high level of satisfaction of the course remained homogeneous in the 2 editions analysed. The overall satisfaction of the course was not finally and uniquely determined by any of its particular parts. The fresh cadaver model for training physicians in techniques of airway management is a proposal satisfactory to the participant, and with a realism that approaches the live patient. Copyright © 2013 Sociedad Española de Anestesiología, Reanimación y Terapéutica del Dolor. Published by Elsevier España. All rights reserved.

48 CFR 32.904 - Determining payment due dates.

Code of Federal Regulations, 2013 CFR

2013-10-01

... defined in Public Law 98-181, including any edible fresh or frozen poultry meat, any perishable poultry... or frozen fish. As defined in section 204(3) of the Fish and Seafood Promotion Act of 1986 (16 U.S.C... products, butter, yogurt, ice cream, mayonnaise, salad dressings, and other similar products fall within...

48 CFR 32.904 - Determining payment due dates.

Code of Federal Regulations, 2012 CFR

2012-10-01

... defined in Public Law 98-181, including any edible fresh or frozen poultry meat, any perishable poultry... or frozen fish. As defined in section 204(3) of the Fish and Seafood Promotion Act of 1986 (16 U.S.C... products, butter, yogurt, ice cream, mayonnaise, salad dressings, and other similar products fall within...

Cryopreservation of bull spermatozoa alters the perinuclear theca.

PubMed

Martínez, Carmen Omega; Juárez-Mosqueda, María de Lourdes; Hernández, Jorge; Valencia, Javier

2006-11-01

The perinuclear theca (PT) is involved in several important sperm functions leading to fertilization. The objective of this study was to investigate the effect of cryopreservation of bull spermatozoa on the integrity of the PT and the relationship between PT integrity and semen characteristics. Semen from seven bulls was evaluated before and after cryopreservation, comparing the integrity of the plasma membrane (hypo-osmotic test), percentage of live and dead spermatozoa (triple stain), acrosome integrity (triple stain) and the integrity of the PT (negative stain by electron microscopy). Cryopreservation of bull semen caused substantial damage to the PT; the proportion of spermatozoa with a damaged PT was 15.2% versus 52.5% (P<0.05) in fresh versus frozen-thawed spermatozoa, respectively. Furthermore, on average, 67.4% (range, 64-72%) of fresh spermatozoa were live, compared to 53.1% (range, 49-58%) for frozen-thawed spermatozoa; there was an inverse correlation between the percentage of live spermatozoa and the percentage with damage to the PT. Although 59.1% of frozen-thawed spermatozoa had an intact acrosome, only 43.7% of them still remained alive. In frozen-thawed semen, there was a high correlation (r=0.69) between live spermatozoa with an intact acrosome and spermatozoa that maintained an intact PT. In conclusion, freezing/thawing of bull spermatozoa altered the PT and maintaining PT integrity may be necessary to maintain acrosome integrity.

MicroRNA expression in melanocytic nevi: the usefulness of formalin-fixed, paraffin-embedded material for miRNA microarray profiling.

PubMed

Glud, Martin; Klausen, Mikkel; Gniadecki, Robert; Rossing, Maria; Hastrup, Nina; Nielsen, Finn C; Drzewiecki, Krzysztof T

2009-05-01

MicroRNAs (miRNAs) are small, noncoding RNA molecules that regulate cellular differentiation, proliferation, and apoptosis. MiRNAs are expressed in a developmentally regulated and tissue-specific manner. Aberrant expression may contribute to pathological processes such as cancer, and miRNA may therefore serve as biomarkers that may be useful in a clinical environment for diagnosis of various diseases. Most miRNA profiling studies have used fresh tissue samples. However, in some types of cancer, including malignant melanoma, fresh material is difficult to obtain from primary tumors, and most surgical specimens are formalin fixed and paraffin embedded (FFPE). To explore whether FFPE material would be suitable for miRNA profiling in melanocytic lesions, we compared miRNA expression patterns in FFPE versus fresh frozen samples, obtained from 15 human melanocytic nevi. Out of microarray data, we identified 84 miRNAs that were expressed in both types of samples and represented an miRNA profile of melanocytic nevi. Our results showed a high correlation in miRNA expression (Spearman r-value of 0.80) between paired FFPE and fresh frozen material. The data were further validated by quantitative RT-PCR. In conclusion, FFPE specimens of melanocytic lesions are suitable as a source for miRNA microarray profiling.

Environmental Factors Related to a Semiarid Climate Influence the Freezability of Sperm from Collared Peccaries (Pecari tajacu Linnaeus, 1758).

PubMed

Maia, Keilla M; Souza, Ana L P; Praxedes, Erica C G; Bezerra, Luana G P; Silva, Andreia M; Campos, Livia B; Moreira, Samara S J; Apolinário, Carlos A C; Souza, João B F; Silva, Alexandre R

2018-04-30

The influence of environmental factors in a semiarid climate on characteristics of fresh and frozen/thawed sperm collected from collared peccaries (Pecari tajacu) was assessed. Semen from 11 male collared peccaries was collected by electroejaculation during the peaks of the dry and rainy periods while rainfall indices, air temperatures, relative humidity levels, and wind speeds were measured. The number, motility, morphology, osmotic response, and membrane integrity of sperm in the collected ejaculates were assessed. Samples were then frozen in liquid nitrogen, thawed, and reassessed. The rainfall index of the rainy period (73.2 mm) was significantly higher than that of the dry period (13.6 mm) and the relative humidity was significantly higher during the rainy period (74.6%) than it was during the dry period (66.8%). Air temperature and wind speed did not differ between the two periods. Characteristics of sperm in the fresh samples were not affected by environmental parameters. In contrast, computerized analysis revealed that sperm in samples frozen during the rainy period exhibited better post-thaw membrane integrity (28.6 ± 6%), motility (29.5 ± 7.7%), and rapid sperm population (13.7 ± 6.2%) than did sperm in samples frozen during the dry period (23.4 ± 3% membrane integrity, 14.6 ± 4.1% motility, and 4.1 ± 1.2% rapid sperm; p < 0.05). Other characteristics of the frozen/thawed sperm did not differ depending on the period in which they were collected. We demonstrated that environmental parameters did not affect the quality of fresh sperm, but could influence the freezability of sperm collected from collared peccaries raised under a semiarid climate.

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

PubMed

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

2014-10-01

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

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

PubMed Central

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

2014-01-01

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

Assessment of sperm nuclear quality after in vitro maturation of fresh or frozen/thawed mouse pre-pubertal testes.

PubMed

Oblette, A; Rives, N; Dumont, L; Rives, A; Verhaeghe, F; Jumeau, F; Rondanino, C

2017-10-01

Is nuclear quality of in vitro generated spermatozoa from fresh or frozen/thawed pre-pubertal mouse testes similar to that of their in vivo counterparts? The production of spermatozoa with aneuploidy, DNA fragmentation or chromatin condensation defects was not significantly increased in organotypic cultures compared to in vivo controls. Although murine spermatozoa have been produced in vitro from pre-pubertal testes, their nuclear DNA integrity has never been investigated. Fresh and frozen/thawed testicular fragments from 6 to 7 days postpartum (dpp) mice were cultured for 30 days. Testicular tissues were frozen by controlled slow freezing (CSF) or solid surface vitrification (SSV). In total, 30 fresh, 30 CSF, 30 SSV testes were used for in vitro maturation and 6 testes from 36 to 37 dpp mice were used as in vivo controls. Murine spermatozoa were extracted from pooled in vitro cultured testicular fragments and from in vivo controls. Sperm aneuploidy was analyzed by fluorescence in situ hybridization (FISH), DNA fragmentation by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling, chromatin condensation by aniline blue staining, telomere length and number by quantitative FISH, DNA oxidation by immunocytochemical detection of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Because of the low spermatogenic yield in cultures, a hundred spermatozoa extracted from pooled tissues were examined and compared to their in vivo counterparts. Most of spermatozoa generated in vitro and in vivo were haploid, contained unfragmented DNA and normally condensed chromatin. A similar proportion of spermatozoa with aneuploidy, DNA fragmentation or chromatin condensation defects was found in cultures and in vivo. No significant difference in telomere length was found within the nuclei of in vitro and in vivo generated spermatozoa. However, the number of telomere spots was lower in gametes obtained from cultures of fresh, CSF and SSV testes than in their natural counterparts (P < 0.01). Moreover, the proportion of spermatozoa containing 8-OHdG was significantly increased in frozen/thawed tissues in comparison to fresh tissues and in vivo controls (P < 0.05). None. Further studies will be needed to enhance the production of spermatozoa in organotypic cultures while preserving their quality, to investigate epigenetic modifications and embryonic development. This is the first study comparing the nuclear quality of in vitro and in vivo generated murine spermatozoa. The organotypic culture system will have to be adapted for human tissue and extensive analyses of human gamete quality will have to be performed before potential clinical applications can be envisaged. This work was supported by Rouen University Hospital, Ligue contre le Cancer, Agence de la Biomédecine, Association Laurette Fugain, France Lymphome Espoir, and co-supported by European Union and Région Normandie. Europe gets involved in Normandie with European Régional Development Fund (ERDF). The authors declare that they have no conflict of interest. © The Author 2017. 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

Effect of refrigeration and frozen storage on the Campylobacter jejuni recovery from naturally contaminated broiler carcasses

PubMed Central

Maziero, Maike T.; de Oliveira, Tereza Cristina R. M.

2010-01-01

Campylobacter jejuni is the most common thermophilic Campylobacter associated with human enteritis in many countries. Broilers and their by-products are the main sources for human enteritis. Refrigeration and freezing are used to control bacterial growth in foods. The effect of these interventions on survival of Campylobacter jejuni is yet not quite understood. This study evaluated the effect of storage temperature on the survival of C. jejuni in chicken meat stored for seven days at 4°C and for 28 days at -20°C. The influence of selective enrichment on recovery of Campylobacter was also evaluated. Thirty fresh chicken meat samples were analyzed and 93.3% was contaminated with termotolerant Campylobacter spp. with average count of 3.08 Log10 CFU/g on direct plating. After refrigeration, 53.3% of the analyzed samples tested positive for Campylobacter and the average count was 1.19 Log10 CFU/g. After storage at -20°C, 36.6% of the samples were positive with a verage count of 0.75 Log10 CFU/g. C. jejuni was detected after enrichment, respectively, in 50% of the fresh, 36.7% of the refrigerated and 33.3% of the frozen meat samples analyzed. No difference was detected for the recovery of C. jejuni from fresh, refrigerated or frozen samples after selective enrichment, showing that this microorganism can survive under the tested storage conditions. PMID:24031523

Management of Sacral Tumors Requiring Spino-Pelvic Reconstruction with Different Histopathologic Diagnosis: Evaluation with Four Cases

PubMed Central

Togral, Guray; Hasturk, Askin Esen; Kekec, Fevzi; Parpucu, Murat; Gungor, Safak

2015-01-01

In this retrospective study, surgical results of four patients with sacral tumors having disparate pathologic diagnoses, who were treated with partial or total sacrectomy and lumbopelvic stabilization were abstracted. Two patients were treated with partial sacral resection and two patients were treated with total sacrectomy and spinopelvic fixation. Fixation methods included spinopelvic fixation with rods and screws in two cases, reconstruction plate in one case, and fresh frozen allografts in two cases. Fibular allografts used for reconstruction accelerated bony union and enhanced the stability in two cases. Addition of polymethyl methacrylate in the cavity in the case of a giant cell tumor had a positive stabilizing effect on fixation. As a result, we can conclude that mechanical instability after sacral resection can be stabilized securely with lumbopelvic fixation and polymethyl methacrylate application or addition of fresh frozen allografts between the rods can augment the stability of the reconstruction. PMID:26713133

Management of Sacral Tumors Requiring Spino-Pelvic Reconstruction with Different Histopathologic Diagnosis: Evaluation with Four Cases.

PubMed

Arıkan, Murat; Togral, Guray; Hasturk, Askin Esen; Kekec, Fevzi; Parpucu, Murat; Gungor, Safak

2015-12-01

In this retrospective study, surgical results of four patients with sacral tumors having disparate pathologic diagnoses, who were treated with partial or total sacrectomy and lumbopelvic stabilization were abstracted. Two patients were treated with partial sacral resection and two patients were treated with total sacrectomy and spinopelvic fixation. Fixation methods included spinopelvic fixation with rods and screws in two cases, reconstruction plate in one case, and fresh frozen allografts in two cases. Fibular allografts used for reconstruction accelerated bony union and enhanced the stability in two cases. Addition of polymethyl methacrylate in the cavity in the case of a giant cell tumor had a positive stabilizing effect on fixation. As a result, we can conclude that mechanical instability after sacral resection can be stabilized securely with lumbopelvic fixation and polymethyl methacrylate application or addition of fresh frozen allografts between the rods can augment the stability of the reconstruction.

Effects of sanitation, freezing and frozen storage on enteric viruses in berries and herbs.

PubMed

Butot, S; Putallaz, T; Sánchez, G

2008-08-15

Norovirus (NV) and hepatitis A virus (HAV) are foodborne enteric viruses associated with outbreaks of disease following consumption of fresh or frozen produce. Model experiments were performed to determine the effectiveness of certain commercial processes for the removal of enteric viruses that might be present in berries and herbs. The survival and persistence of HAV, NV, rotavirus (RV) and feline calicivirus (FCV), a surrogate for NV, in frozen produce over time were determined. Survival and inactivation of HAV, RV and FCV were assessed by viral culture and quantitative reverse transcription-PCR (RT-PCR), whereas NV persistence was determined by quantitative RT-PCR only. Freezing did not significantly reduce the viability of any of the viruses except the infectivity of FCV in strawberries. Frozen storage for 3 months had limited effects on HAV and RV survival in all tested food products, whereas in frozen raspberries and strawberries FCV infectivity showed the highest decay rate due to acid pH. To simulate postharvesting conditions, fresh berries and herbs were rinsed with tap, warm or chlorinated water or with a chlorine dioxide (ClO(2)) solution. Available chlorine at a concentration of 200 ppm and ClO(2) at 10 ppm reduced measurable enteric viruses in raspberry and parsley samples by less than 2 log(10) units.

Results of liver transplantation in patients with acute liver failure due to Amanita phalloides and paracetamol (acetaminophen) intoxication

PubMed Central

Grąt, Michał; Hołówko, Wacław; Masior, Łukasz; Wronka, Karolina M.; Grąt, Karolina; Stypułkowski, Jan; Patkowski, Waldemar; Krawczyk, Marek

2015-01-01

Introduction Amanita phalloides and paracetamol intoxications are responsible for the majority of acute liver failures. Aim To assess survival outcomes and to analyse risk factors affecting survival in the studied group. Material and methods Of 1369 liver transplantations performed in the Department of General, Transplant, and Liver Surgery, Medical University of Warsaw before December 2013, 20 (1.46%) patients with Amanita phalloides (n = 13, 0.95%) and paracetamol (n = 7, 0.51%) intoxication were selected for this retrospective study. Overall and graft survival at 5 years were set as primary outcome measures. Results Five-year overall survival after liver transplantation in the studied group was 53.57% and 53.85% in patients with paracetamol and Amanita phalloides poisoning, respectively (p = 0.816). Five-year graft survival was 26.79% for patients with paracetamol and 38.46% with Amanita phalloides intoxication (p = 0.737). Risk factors affecting patient survival were: pre-transplant bilirubin concentration (p = 0.023) and higher number of red blood cells (p = 0.013) and fresh frozen plasma (p = 0.004) transfused intraoperatively. Likewise, higher number of red blood cells (p = 0.012) and fresh frozen plasma (p = 0.007) transfused were risk factors affecting 5-year graft survival. Surprisingly, donor and recipient blood type incompatibility was neither the risk factor for 5-year overall survival (p = 0.939) nor the risk factor for 5-year graft survival (p = 0.189). Conclusions In selected intoxicated patients urgent liver transplantation is the only successful modality of treatment. Risk factors affecting survival are in correspondence with the patient's pre-transplant status (bilirubin level in serum) and intraoperative status (number of red blood cells and fresh frozen plasma transfused). PMID:27350835

Hormone Receptor Expression Analyses in Neoplastic and Non-Neoplastic Canine Mammary Tissue by a Bead Based Multiplex Branched DNA Assay: A Gene Expression Study in Fresh Frozen and Formalin-Fixed, Paraffin-Embedded Samples.

PubMed

Mohr, Annika; Lüder Ripoli, Florenza; Hammer, Susanne Conradine; Willenbrock, Saskia; Hewicker-Trautwein, Marion; Kiełbowicz, Zdzisław; Murua Escobar, Hugo; Nolte, Ingo

2016-01-01

Immunohistochemistry (IHC) is currently considered the method of choice for steroid hormone receptor status evaluation in human breast cancer and, therefore, it is commonly utilized for assessing canine mammary tumors. In case of low hormone receptor expression, IHC is limited and thus is complemented by molecular analyses. In the present study, a multiplex bDNA assay was evaluated as a method for hormone receptor gene expression detection in canine mammary tissues. Estrogen receptor (ESR1), progesterone receptor (PGR), prolactin receptor (PRLR) and growth hormone receptor (GHR) gene expressions were evaluated in neoplastic and non-neoplastic canine mammary tissues. A set of 119 fresh frozen and 180 formalin-fixed, paraffin-embedded (FFPE) was comparatively analyzed and used for assay evaluation. Furthermore, a possible association between the hormone receptor expression in different histological subtypes of canine malignant mammary tumors and the castration status, breed and invasive growth of the tumor were analyzed. The multiplex bDNA assay proved to be more sensitive for fresh frozen specimens. Hormone receptor expression found was significantly decreased in malignant mammary tumors in comparison to non-neoplastic tissue and benign mammary tumors. Among the histological subtypes the lowest gene expression levels of ESR1, PGR and PRLR were found in solid, anaplastic and ductal carcinomas. In summary, the evaluation showed that the measurement of hormone receptors with the multiplex bDNA assay represents a practicable method for obtaining detailed quantitative information about gene expression in canine mammary tissue for future studies. Still, comparison with IHC or quantitative real-time PCR is needed for further validation of the present method.

Sterilization with electron beam irradiation influences the biomechanical properties and the early remodeling of tendon allografts for reconstruction of the anterior cruciate ligament (ACL).

PubMed

Schmidt, Tanja; Hoburg, Arnd; Broziat, Christine; Smith, Mark D; Gohs, Uwe; Pruss, Axel; Scheffler, Sven

2012-08-01

Although allografts for anterior cruciate ligament (ACL) replacement have shown advantages compared to autografts, their use is limited due to the risk of disease transmission and the limitations of available sterilization methods. Gamma sterilization has shown detrimental effects on graft properties at the high doses required for sufficient pathogen inactivation. In our previous in vitro study on human patellar tendon allografts, Electron beam (Ebeam) irradiation showed less detrimental effects compared to gamma sterilization (Hoburg et al. in Am J Sports Med 38(6):1134-1140, 2010). To investigate the biological healing and restoration of the mechanical properties of a 34 kGy Ebeam treated tendon allograft twenty-four sheep underwent ACL replacement with either a 34 kGy Ebeam treated allograft or a non-sterilized fresh frozen allograft. Biomechanical testing of stiffness, ultimate failure load and AP-laxity as well as histological analysis to investigate cell, vessel and myofibroblast-density were performed after 6 and 12 weeks. Native sheep ACL and hamstring tendons (HAT, each n = 9) served as controls. The results of a previous study analyzing the remodeling of fresh frozen allografts (n = 12) and autografts (Auto, n = 18) with the same study design were also included in the analysis. Statistics were performed using Mann-Whitney U test followed by Bonferroni-Holm correction. Results showed significantly decreased biomechanical properties during the early remodeling period in Ebeam treated grafts and this was accompanied with an increased remodeling activity. There was no recovery of biomechanical function from 6 to 12 weeks in this group in contrast to the results observed in fresh frozen allografts and autografts. Therefore, high dose Ebeam irradiation investigated in this paper cannot be recommended for soft tissue allograft sterilization.

Horizontal Resorption of Fresh-Frozen Corticocancellous Bone Blocks in the Reconstruction of the Atrophic Maxilla at 5 Months

PubMed Central

Pereira, Eugénio; Messias, Ana; Dias, Ricardo; Judas, Fernando; Salvoni, Alexander; Guerra, Fernando

2015-01-01

Background Reliable implant-supported rehabilitation of an alveolar ridge needs sufficient volume of bone. In order to achieve a prosthetic-driven positioning, bone graft techniques may be required. Purpose This prospective cohort study aims to clinically evaluate the amount of resorption of corticocancellous fresh-frozen allografts bone blocks used in the reconstruction of the severe atrophic maxilla. Materials and Methods Twenty-two partial and totally edentulous patients underwent bone augmentation procedures with fresh-frozen allogenous blocks from the iliac crest under local anesthesia. Implants were inserted into the grafted sites after a healing period of 5 months. Final fixed prosthesis was delivered ± 4 months later. Ridge width analysis and measurements were performed with a caliper before and after grafting and at implant insertion. Bone biopsies were performed in 16 patients. Results A total of 98 onlay block allografts were used in 22 patients with an initial mean alveolar ridge width of 3.41 ± 1.36 mm. Early exposure of blocks was observed in four situations and one of these completely resorbed. Mean horizontal bone gain was 3.63 ± 1.28 mm (p < .01). Mean buccal bone resorption between allograph placement and the reopening stage was 0.49 ± 0.54 mm, meaning approximately 7.1% (95% confidence interval: [5.6%, 8.6%]) of total ridge width loss during the integration period. One hundred thirty dental implants were placed with good primary stability (≥ 30 Ncm). Four implants presented early failure before the prosthetic delivery (96.7% implant survival). All patients were successfully rehabilitated. Histomorphometric analysis revealed 20.9 ± 5.8% of vital bone in close contact to the remaining grafted bone. A positive strong correlation (adjusted R2 = 0.44, p = .003) was found between healing time and vital bone percentage. Conclusions Augmentation procedures performed using fresh-frozen allografts from the iliac crest are a suitable alternative in the reconstruction of the atrophic maxilla with low resorption rate at 5 months, allowing proper stability of dental implants followed by fixed prosthetic rehabilitation. PMID:25346211

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

PubMed

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

2012-12-01

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

9 CFR 319.281 - Bockwurst.

Code of Federal Regulations, 2012 CFR

2012-01-01

... Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE AGENCY... Loaves § 319.281 Bockwurst. (a) Bockwurst is an uncured, comminuted meat food product which may or may... and veal, pork and beef, or pork, veal, and beef. Such meat shall be fresh or fresh frozen meat. Pork...

9 CFR 319.281 - Bockwurst.

Code of Federal Regulations, 2013 CFR

2013-01-01

... Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE AGENCY... Loaves § 319.281 Bockwurst. (a) Bockwurst is an uncured, comminuted meat food product which may or may... and veal, pork and beef, or pork, veal, and beef. Such meat shall be fresh or fresh frozen meat. Pork...

9 CFR 319.281 - Bockwurst.

Code of Federal Regulations, 2014 CFR

2014-01-01

... Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE AGENCY... Loaves § 319.281 Bockwurst. (a) Bockwurst is an uncured, comminuted meat food product which may or may... and veal, pork and beef, or pork, veal, and beef. Such meat shall be fresh or fresh frozen meat. Pork...

9 CFR 319.281 - Bockwurst.

Code of Federal Regulations, 2011 CFR

2011-01-01

... Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE AGENCY... Loaves § 319.281 Bockwurst. (a) Bockwurst is an uncured, comminuted meat food product which may or may... and veal, pork and beef, or pork, veal, and beef. Such meat shall be fresh or fresh frozen meat. Pork...

Effect of Storage Temperature on Quality of Frozen Horse-mackerel

NASA Astrophysics Data System (ADS)

Kozima, Tsuneo; Ohtaka, Tateo

Quality change of frozen horse-mackerel were studied under storage temperature at -18, -23, -30 and -40°C for 12 months. Quality were measured with K value (Freshness index of muscle, degradation ratio of ATP), amount of drip (free and expressible drip), water-holdiog capacity, weight ratio of cooking loss, organoleptic test, and histological feature of muscle. K value, a mount of free drip, w eight ratio of cooking loss, histological feature of muscle, and organoleptic test in color, form and flavor were not detected any changes during frozen storage for 12 months at various temperature. However expressible drip, water-holding capacity and score of taste in organoleptic test showed some changes after 8 or 12 months at -18 and/or -23°C, it was not serious change to-loss quality as food. Frozen horse-mackerel can store under below ~ 18°C for 12 months.

78 FR 35643 - Frozen Warmwater Shrimp from China, Ecuador, India, Indonesia, Malaysia, Thailand, and Vietnam...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-13

...; (5) dried shrimp and prawns; (6) canned warmwater shrimp and prawns; and (7) certain `battered shrimp.' ```Battered shrimp' is a shrimp-based product: (1) That is produced from fresh (or thawed-from-frozen) and... accordance with the definition of dusting above, the battered shrimp product is also coated with a wet...

76 FR 64307 - Certain Frozen Warmwater Shrimp From the Socialist Republic of Vietnam: Amended Final Results and...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-10-18

... prawns; (6) canned warmwater shrimp and prawns (HTS subheading 1605.20.1040); and (7) certain battered shrimp. Battered shrimp is a shrimp-based product: (1) That is produced from fresh (or thawed-from-frozen.... When dusted in accordance with the definition of dusting above, the battered shrimp product is also...

Texture and microstructure properties of frozen chicken breasts pretreated with salt and phosphate solutions.

PubMed

Yoon, K S

2002-12-01

This study investigated the effects of 10% NaCl, trisodium phosphate (TSP), sodium tripolyphosphate (STPP), and tetrapotassium pyrophosphate (TKPP) treatments on textural and microstructural properties of chicken breasts during 10 mo of frozen storage at -20 C. Fresh chicken breasts were treated for 10 min with 10% NaCl and various phosphate solutions, including TSP, STPP, and TKPP, and stored in a -20 C freezer for 10 mo. Frozen chicken breasts were completely thawed at 4 C and oven-baked at 177 C for 20 min. Shear force, drip loss, and cooking loss were measured. In addition, ice crystal formation and structure changes of frozen chicken breasts during storage were evaluated using transmission electron microscopy (TEM). Treating chicken breasts with 10% TSP and STPP solution significantly reduced drip and cooking losses as well as minimized ice crystal formation and freeze-induced shrinkage of myofibrils. No significant texture toughening was observed in frozen chicken breasts regardless of treatments. These results suggest that the perceived quality losses of frozen chicken breast were not associated with texture toughening. The water-binding ability of chicken meat was the most important factor in maintaining the quality of chicken breast during extended frozen storage, which can be accomplished by treating chicken breasts with 10% TSP and STPP solutions before frozen storage.

Differences in semen freezability and intracellular ATP content between the rooster (Gallus gallus domesticus) and the Barbary partridge (Alectoris barbara).

PubMed

Madeddu, M; Berlinguer, F; Pasciu, V; Succu, S; Satta, V; Leoni, G G; Zinellu, A; Muzzeddu, M; Carru, C; Naitana, S

2010-10-01

This study aimed to compare viability, ATP content, and DNA integrity of rooster (Gallus gallus domesticus) and Barbary partridge (Alectoris barbara) fresh and frozen spermatozoa in order to identify factors possibly related to differences in semen freezability. Ejaculates were obtained from March to May by the abdominal massage method from 3 adult roosters and 12 adult Barbary partridges. Semen was frozen with different cryoprotectants using Lake's diluents as a base medium: 1) glycerol 11%; 2) glycerol 11% and trehalose 70 mmol/L; 3) dimethylacetamide (DMA) 6%; 4) DMA 6% and trehalose 70 mmol/L. Both fresh and frozen semen showed a lower viability and higher intracellular ATP concentrations in the Barbary partridge compared with the rooster (P < 0.05). In the Barbary partridge, semen viability after thawing did not differ among the 4 media used, but glycerol showed positive effects in avoiding a significant loss of ATP after thawing, compared with DMA containing media (P < 0.05). On the other hand, in the rooster a higher viability was recorded when semen was frozen in glycerol containing media compared to DMA (P < 0.0001), while ATP values significantly decreased after thawing (P < 0.05) without showing any differences among the semen frozen in the 4 different media. DNA integrity, as evaluated by the comet assay, was assessed only in frozen semen. In the Barbary partridge, mean scored parameter did not differ significantly among semen frozen in the 4 different media. In the rooster DNA fragmentation was higher in DMA ctr medium compared with the other media and with values found in Barbary partridge semen frozen in the same medium (P < 0.001). In both species, the addition of trehalose did not show any positive effects on viability, ATP levels and DNA integrity after thawing. In conclusion, species-related differences in semen features exist between the rooster and the Barbary partridge and the wide variation observed in ATP levels may account for differences in semen freezability between the two species. (c) 2010 Elsevier Inc. All rights reserved.

Development of frozen-thawed demi-embryos and production of identical twin calves of different ages.

PubMed

Seike, N; Sakai, M; Kanagawa, H

1991-02-01

The percentages of morphologically transferable embryos obtained from frozen-thawed demi-embryos which were embedded with or without agar, and from those with or without zonae pellucidae were 26.3% (5/19), 36.4% (8/22), 39.5% (15/38) and 40.0% (22/55), respectively. No significant differences were observed between these groups. Development to calves of frozen-thawed demi-embryos with or without zonae pellucidae was 25.0% (3/12) and 26.7% (4/15), respectively. There was also no significant difference between them. On the trial for production of identical twin calves of different ages, the pregnancy rates of fresh and frozen demi-embryos after transfer were 69.2% (9/13) and 11.1% (1/9), respectively. Out of 13 fresh demi-embryos and 9 frozen demi-embryos transferred, only one pair of identical twin male calves of different ages were produced. This frozen-thawed demi-embryo was stored for 43 days in liquid nitrogen before thawing and transfer. These twin calves were confirmed to be identical by blood typing. Although these calves had different birth dates, their growth rates indicated similar developmental patterns. We suggest that it is possible to produce identical twin calves of different ages. This possibility would be useful for predicting the sex, milk producing ability and progeny test of a pair of demi-embryos before a decision to transfer the other half of a pair is made.

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

PubMed

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

2018-01-01

Initial observational studies and a systematic review published 5 years ago have suggested that obstetric and perinatal outcomes are better in offspring conceived following frozen rather than fresh embryo transfers, with reduced risks of preterm birth, small for gestational age, low birth weight and pre-eclampsia. More recent primary studies are beginning to challenge some of these findings. We therefore conducted an updated systematic review and cumulative meta-analysis to examine if these results have remained consistent over time. The aim of this study was to perform a systematic review and cumulative meta-analysis (trend with time) of obstetric and perinatal complications in singleton pregnancies following the transfer of frozen thawed and fresh embryos generated through in-vitro fertilisation. Data Sources from Medline, EMBASE, Cochrane Central Register of Clinical Trials DARE and CINAHL (1984-2016) were searched using appropriate key words. Observational and randomised studies comparing obstetric and perinatal outcomes in singleton pregnancies conceived through IVF using either fresh or frozen thawed embryos. Two independent reviewers extracted data in 2 × 2 tables and assessed the methodological quality of the relevant studies using CASP scoring. Both aggregated as well as cumulative meta-analysis was done using STATA. Twenty-six studies met the inclusion criteria. Singleton babies conceived from frozen thawed embryos were at lower relative risk (RR) of preterm delivery (0.90; 95% CI 0.84-0.97) low birth weight (0.72; 95% CI 0.67-0.77) and small for gestational age (0.61; 95% CI 0.56-0.67) compared to those conceived from fresh embryo transfers, but faced an increased risk (RR) of hypertensive disorders of pregnancy (1.29; 95% CI 1.07-1.56) large for gestational age (1.54; 95% CI 1.48-1.61) and high birth weight (1.85; 95% CI 1.46-2.33). There was no difference in the risk of congenital anomalies and perinatal mortality between the two groups. The direction and magnitude of effect for these outcomes have remained virtually unchanged over time while the degree of precision has improved with the addition of data from newer studies. The results of this cumulative meta-analysis confirm that the decreased risks of small for gestational age, low birth weight and preterm delivery and increased risks of large for gestational age and high birth weight associated with pregnancies conceived from frozen embryos have been consistent in terms of direction and magnitude of effect over several years, with increasing precision around the point estimates. Replication in a number of different populations has provided external validity for the results, for outcomes of birth weight and preterm delivery. Meanwhile, caution should be exercised about embarking on a policy of electively freezing all embryos in IVF as there are increased risks for large for gestational age babies and hypertensive disorders of pregnancy. Therefore, elective freezing should ideally be undertaken in specific cases such as ovarian hyperstimulation syndrome, fertility preservation or in the context of randomised trials. © The Author 2017. 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

Assisted reproductive technology in Japan: a summary report for 2015 by The Ethics Committee of The Japan Society of Obstetrics and Gynecology.

PubMed

Saito, Hidekazu; Jwa, Seung Chik; Kuwahara, Akira; Saito, Kazuki; Ishikawa, Tomonori; Ishihara, Osamu; Kugu, Koji; Sawa, Rintaro; Banno, Kouji; Irahara, Minoru

2018-01-01

The Japan Society of Obstetrics and Gynecology (JSOG) implemented an assisted reproductive technology (ART) registry system in 1986. Here are reported the characteristics and treatment outcomes of ART cycles that were registered in 2015. JSOG has requested all participating ART facilities to register cycle-specific information for all ART cycles since 2007. A descriptive analysis was performed by using the registry database for 2015. In total, 424 151 cycles and 51 001 neonates (1 in 19.7 neonates born in Japan) were registered in 2015. The patients' mean age was 38.2 years (standard deviation = 4.5). Among the fresh cycles, 94 158 of 244 718 (38.5%) egg retrieval cycles were cycles with freeze-all embryos or oocytes, while fresh embryo transfer (ET) was performed in 70 254 cycles, signaling a decrease from 2014. There were 169 898 frozen-thawed ET cycles, resulting in 56 355 pregnancies and 40 599 neonates. Single ET was performed at a rate of 79.7% for fresh and 81.8% for frozen cycles and the singleton pregnancy/live birth rates were 96.9%/96.5% and 96.8%/96.4% for the respective cycles. The total ART cycles and live births resulting from ART has been increasing in Japan. Single ET was performed at a rate of almost 80% and ET cycles have shifted from fresh to frozen cycles.

Choice of the replacement fluid during large volume plasma-exchange.

PubMed

Nydegger, U E

1983-01-01

The replacement fluid used during therapeutic large volume plasma-exchange can be seen as an important factor influencing the result of such treatment. The choice includes fluids such as electrolyte solutions, gelatin, hydroxyethyl-starch, albumin and fresh frozen plasma. By evaluating the pathophysiology of the underlying disease, it is possible to choose between merely replacing the removed volume by non-protein fluids or rather to introduce plasma protein components into the patient's circulation by substituting with purified or enriched proteins such as albumin, clotting factors, antithrombin III or fresh frozen plasma. This paper analyzes the rationale for the choice of the appropriate replacement fluid taking into account pathophysiologic, pharmacologic and logistic criteria.

In vitro fertilization and sperm cryopreservation in the black-footed cat (Felis nigripes) and sand cat (Felis margarita).

PubMed

Herrick, J R; Campbell, M; Levens, G; Moore, T; Benson, K; D'Agostino, J; West, G; Okeson, D M; Coke, R; Portacio, S C; Leiske, K; Kreider, C; Polumbo, P J; Swanson, W F

2010-03-01

Studies of in vitro fertilization (IVF) and sperm cryopreservation have been conducted in several small cat species, but virtually no data exist for black-footed cats (Felis nigripes) (BFCs) or sand cats (Felis margarita) (SCs). The objectives of this study were 1) to compare in vitro motility and acrosome status of fresh and cryopreserved (frozen in pellets on dry ice or in straws in liquid nitrogen vapor) BFC and SC spermatozoa cultured in feline-optimized culture medium (FOCM) or Ham F-10, 2) to assess ovarian responsiveness in BFCs and SCs following exogenous gonadotropin treatment and laparoscopic oocyte recovery, and 3) to evaluate the fertility of fresh and frozen-thawed spermatozoa from both species using homologous and heterologous (domestic cat oocytes) IVF in the two culture media. Motility and acrosomal integrity of fresh and frozen-thawed spermatozoa from BFCs and SCs were similar (P > 0.05) in both media during 6 h of culture. Although effects were more pronounced in SCs, cryopreservation in straws was superior (P < 0.05) to cryopreservation in pellets for both species. Gonadotropin stimulation produced approximately 16 ovarian follicles per female, and >80% of recovered oocytes were of optimal (grade 1) quality. The BFC and SC spermatozoa fertilized 60.0%-79.4% of homologous and 37.7%-42.7% of heterologous oocytes in both culture media, with increased (P < 0.05) cleavage of homologous (SC) and heterologous (BFC and SC) oocytes in FOCM. These results provide the first information to date on the gamete biology of two imperiled cat species and further our capacity to apply reproductive technologies for their conservation.

City Level of Income and Urbanization and Availability of Food Stores and Food Service Places in China.

PubMed

Liao, Chunxiao; Tan, Yayun; Wu, Chaoqun; Wang, Shengfeng; Yu, Canqing; Cao, Weihua; Gao, Wenjing; Lv, Jun; Li, Liming

2016-01-01

The contribution of unhealthy dietary patterns to the epidemic of obesity has been well recognized. Differences in availability of foods may have an important influence on individual eating behaviors and health disparities. This study examined the availability of food stores and food service places by city characteristics on city level of income and urbanization. The cross-sectional survey was comprised of two parts: (1) an on-site observation to measure availability of food stores and food service places in 12 cities of China; (2) an in-store survey to determine the presence of fresh/frozen vegetables or fruits in all food stores. Trained investigators walked all the streets/roads within study tracts to identify all the food outlets. An observational survey questionnaire was used in all food stores to determine the presence of fresh/frozen vegetables or fruits. Urbanization index was determined for each city using a principal components factor analysis. City level of income and urbanization and numbers of each type of food stores and food service places were examined using negative binomial regression models. Large-sized supermarkets and specialty retailers had higher number of fresh/frozen vegetables or fruits sold compared to small/medium-sized markets. High-income versus low-income, high urbanized versus low urbanized areas had significantly more large-sized supermarkets and fewer small/medium-sized markets. In terms of restaurants, high urbanized cities had more western fast food restaurants and no statistically significant difference in the relative availability of any type of restaurants was found between high- and low-income areas. The findings suggested food environment disparities did exist in different cities of China.

City Level of Income and Urbanization and Availability of Food Stores and Food Service Places in China

PubMed Central

Liao, Chunxiao; Tan, Yayun; Wu, Chaoqun; Wang, Shengfeng; Yu, Canqing; Cao, Weihua; Gao, Wenjing; Lv, Jun; Li, Liming

2016-01-01

Objective The contribution of unhealthy dietary patterns to the epidemic of obesity has been well recognized. Differences in availability of foods may have an important influence on individual eating behaviors and health disparities. This study examined the availability of food stores and food service places by city characteristics on city level of income and urbanization. Methods The cross-sectional survey was comprised of two parts: (1) an on-site observation to measure availability of food stores and food service places in 12 cities of China; (2) an in-store survey to determine the presence of fresh/frozen vegetables or fruits in all food stores. Trained investigators walked all the streets/roads within study tracts to identify all the food outlets. An observational survey questionnaire was used in all food stores to determine the presence of fresh/frozen vegetables or fruits. Urbanization index was determined for each city using a principal components factor analysis. City level of income and urbanization and numbers of each type of food stores and food service places were examined using negative binomial regression models. Results Large-sized supermarkets and specialty retailers had higher number of fresh/frozen vegetables or fruits sold compared to small/medium-sized markets. High-income versus low-income, high urbanized versus low urbanized areas had significantly more large-sized supermarkets and fewer small/medium-sized markets. In terms of restaurants, high urbanized cities had more western fast food restaurants and no statistically significant difference in the relative availability of any type of restaurants was found between high- and low-income areas. Conclusions The findings suggested food environment disparities did exist in different cities of China. PMID:26938866

Comparison of fresh-frozen cadaver and high-fidelity virtual reality simulator as methods of laparoscopic training.

PubMed

Sharma, Mitesh; Horgan, Alan

2012-08-01

The aim of this study was to compare fresh-frozen cadavers (FFC) with a high-fidelity virtual reality simulator (VRS) as training tools in minimal access surgery for complex and relatively simple procedures. A prospective comparative face validity study between FFC and VRS (LAP Mentor(™)) was performed. Surgeons were recruited to perform tasks on both FFC and VRS appropriately paired to their experience level. Group A (senior) performed a laparoscopic sigmoid colectomy, Group B (intermediate) performed a laparoscopic incisional hernia repair, and Group C (junior) performed basic laparoscopic tasks (BLT) (camera manipulation, hand-eye coordination, tissue dissection and hand-transferring skills). Each subject completed a 5-point Likert-type questionnaire rating the training modalities in nine domains. Data were analysed using nonparametric tests. Forty-five surgeons were recruited to participate (15 per skill group). Median scores for subjects in Group A were significantly higher for evaluation of FFC in all nine domains compared to VRS (p < 0.01). Group B scored FFC significantly better (p < 0.05) in all domains except task replication (p = 0.06). Group C scored FFC significantly better (p < 0.01) in eight domains but not on performance feedback (p = 0.09). When compared across groups, juniors accepted VRS as a training model more than did intermediate and senior groups on most domains (p < 0.01) except team work. Fresh-frozen cadaver is perceived as a significantly overall better model for laparoscopic training than the high-fidelity VRS by all training grades, irrespective of the complexity of the operative procedure performed. VRS is still useful when training junior trainees in BLT.

[Adaptability of sweet corn ears to a frozen process].

PubMed

Ramírez Matheus, Alejandra O; Martínez, Norelkys Maribel; de Bertorelli, Ligia O; De Venanzi, Frank

2004-12-01

The effects of frozen condition on the quality of three sweet corn ears (2038, 2010, 2004) and the pattern (Bonanza), were evaluated. Biometrics characteristics like ear size, ear diameter, row and kernel deep were measured as well as chemical and physical measurement in fresh and frozen states. The corn ears were frozen at -95 degrees C by 7 minutes. The yield and stability of the frozen ears were evaluated at 45 and 90 days of frozen storage (-18 degrees C). The average commercial yield as frozen corn ear for all the hybrids was 54.2%. The industry has a similar value range of 48% to 54%. The ear size average was 21.57 cm, row number was 15, ear diameter 45.54 mm and the kernel corn deep was 8.57 mm. All these measurements were found not different from commercial values found for the industry. All corn samples evaluated showed good stability despites the frozen processing and storage. Hybrid 2038 ranked higher in quality.

The DNA isolation method has effect on allele drop-out and on the results of fluorescent PCR and DNA fragment analysis.

PubMed

Nagy, Bálint; Bán, Zoltán; Papp, Zoltán

2005-10-01

The quality and the quantity of isolated DNA have an effect on PCR amplifications. The authors studied three DNA isolation protocols (resin binding method using fresh and frozen amniotic fluid samples, and silica adsorption method using fresh samples) on the quantity and on the quality of the isolated DNA. Amniotic fluid samples were obtained from 20 pregnant women. The isolated DNA concentrations were determined by real-time fluorimeter using SYBRGreen I method. Each sample was studied for the presence of 8 STR markers. The authors compared the number of the detected alleles, electrophoretograms and peak areas. There was a significant difference between the concentration of the obtained DNA and in the peak areas between the three isolation protocols. The numbers of detected alleles were different, we observed the most allele drop outs in the resin type DNA isolation protocol from the fresh sample (detected allele numbers 182), followed by resin binding protocol from the frozen samples (detected allele number 243) and by the silica adsorption method (detected allele number 264). The authors demonstrated that the DNA isolation method has an effect on the quantity and quality of the isolated DNA, and on further PCR amplifications.

9 CFR 94.2 - Fresh (chilled or frozen) products (other than meat), and milk and milk products of ruminants and...

Code of Federal Regulations, 2014 CFR

2014-01-01

... (other than meat), and milk and milk products of ruminants and swine. 94.2 Section 94.2 Animals and... DISEASE, HIGHLY PATHOGENIC AVIAN INFLUENZA, AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR... (chilled or frozen) products (other than meat), and milk and milk products of ruminants and swine. (a) The...

77 FR 53856 - Administrative Review of Certain Frozen Warmwater Shrimp From the People's Republic of China...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-09-04

... include, but are not limited to, white-leg shrimp (Penaeus vannemei), banana prawn (Penaeus merguiensis... margin and determine an appropriate margin. For example, in Fresh Cut Flowers from Mexico,\\42\\ the...\\ See Fresh Cut Flowers from Mexico; Final Results of Antidumping Administrative Review, 61 FR 6812...

Potential of DNA methylation in rectal cancer as diagnostic and prognostic biomarkers

PubMed Central

Exner, Ruth; Pulverer, Walter; Diem, Martina; Spaller, Lisa; Woltering, Laura; Schreiber, Martin; Wolf, Brigitte; Sonntagbauer, Markus; Schröder, Fabian; Stift, Judith; Wrba, Fritz; Bergmann, Michael; Weinhäusel, Andreas; Egger, Gerda

2015-01-01

Background: Aberrant DNA methylation is more prominent in proximal compared with distal colorectal cancers. Although a number of methylation markers were identified for colon cancer, yet few are available for rectal cancer. Methods: DNA methylation differences were assessed by a targeted DNA microarray for 360 marker candidates between 22 fresh frozen rectal tumour samples and 8 controls and validated by microfluidic high-throughput and methylation-sensitive qPCR in fresh frozen and formalin-fixed paraffin-embedded (FFPE) samples, respectively. The CpG island methylator phenotype (CIMP) was assessed by MethyLight in FFPE material from 78 patients with pT2 and pT3 rectal adenocarcinoma. Results: We identified and confirmed two novel three-gene signatures in fresh frozen samples that can distinguish tumours from adjacent tissue as well as from blood with a high sensitivity and specificity of up to 1 and an AUC of 1. In addition, methylation of individual CIMP markers was associated with specific clinical parameters such as tumour stage, therapy or patients' age. Methylation of CDKN2A was a negative prognostic factor for overall survival of patients. Conclusions: The newly defined methylation markers will be suitable for early disease detection and monitoring of rectal cancer. PMID:26335606

Alterations in the host defense properties of human milk following prolonged storage or pasteurization.

PubMed

Akinbi, Henry; Meinzen-Derr, Jareen; Auer, Christine; Ma, Yan; Pullum, Derek; Kusano, Ryosuke; Reszka, Krzysztof J; Zimmerly, Kira

2010-09-01

Preterm infants are often fed pasteurized donor milk or mother's milk that has been stored frozen for up to 4 weeks. Our objectives were to assess the impact of pasteurization or prolonged storage at -20 degrees C on the immunologic components of human milk and the capability of the different forms of human milk to support bacterial proliferation. The concentrations and activities of major host defense proteins in the whey fractions of mother's milk stored for 4 weeks at -20 degrees C or pasteurized human donor milk were compared with freshly expressed human milk. Proliferation of bacteria incubated in the 3 forms of human milk was assessed. Relative to freshly expressed human milk, the concentrations of lysozyme, lactoferrin, lactoperoxidase, and secretory immunoglobulin A were reduced 50% to 82% in pasteurized donor milk and the activities of lysozyme and lactoperoxidase were 74% to 88% lower (P < 0.01). Proliferation of bacterial pathogens in pasteurized donor milk was enhanced 1.8- to 4.6-fold compared with fresh or frozen human milk (P < 0.01). The immunomodulatory proteins in human milk are reduced by pasteurization and, to a lesser extent, by frozen storage, resulting in decreased antibacterial capability. Stringent procedure to minimize bacterial contamination is essential during handling of pasteurized milk.

Establishment and Characterization of Primary Glioblastoma Cell Lines from Fresh and Frozen Material: A Detailed Comparison

PubMed Central

Mullins, Christina Susanne; Schneider, Björn; Stockhammer, Florian; Krohn, Mathias; Classen, Carl Friedrich; Linnebacher, Michael

2013-01-01

Background Development of clinically relevant tumor model systems for glioblastoma multiforme (GBM) is important for advancement of basic and translational biology. High molecular heterogeneity of GBM tumors is well recognized, forming the rationale for molecular tests required before administration of several of the novel therapeutics rapidly entering the clinics. One model that has gained wide acceptance is the primary cell culture model. The laborious and time consuming process is rewarded with a relative high success rate (about 60%). We here describe and evaluate a very simple cryopreservation procedure for GBM tissue prior to model establishment that will considerably reduce the logistic complexity. Methods Twenty-seven GBM samples collected ad hoc were prepared for primary cell culture freshly from surgery (#1) and after cryopreservation (#2). Results Take rates after cryopreservation (59%) were as satisfactory as from fresh tissue (63%; p = 1.000). We did not observe any relevant molecular or phenotypic differences between cell lines established from fresh or vitally frozen tissue. Further, sensitivity both towards standard chemotherapeutic agents (Temozolomide, BCNU and Vincristine) and novel agents like the receptor tyrosine kinase inhibitor Imatinib did not differ. Conclusions Our simple cryopreservation procedure facilitates collection, long-time storage and propagation (modeling) of clinical GBM specimens (potentially also from distant centers) for basic research, (pre-) clinical studies of novel therapies and individual response prediction. PMID:23951083

FROZEN THIN SECTIONS OF FRESH TISSUE FOR ELECTRON MICROSCOPY, WITH A DESCRIPTION OF PANCREAS AND LIVER

PubMed Central

Christensen, A. Kent

1971-01-01

A simple method has been developed that allows frozen thin sections of fresh-frozen tissue to be cut on a virtually unmodified ultramicrotome kept at room temperature. A bowl-shaped Dewar flask with a knifeholder in its depths replaces the stage of the microtome; a bar extends down into the bowl from the microtome's cutting arm and bears the frozen tissue near its lower end. When the microtome is operated, the tissue passes a glass or diamond knife in the depths of the bowl as in normal cutting. The cutting temperature is maintained by flushing the bowl with cold nitrogen gas, and can be set anywhere from about -160°C up to about -30°C. The microtome is set for a cutting thickness of 540–1000 A. Sections are picked up from the dry knife edge, and are placed on membrane-coated grids, flattened with the polished end of a copper rod, and either dried in nitrogen gas or freeze-dried. Throughout the entire process the tissue is kept cold and does not come in contact with any solvent. The morphology seen in frozen thin sections of rat pancreas and liver generally resembles that in conventional preparations, although freezing damage and low contrast limit the detail that can be discerned. Among unusual findings is a frequent abundance of mitochondrial granules in material prepared by this method. PMID:4942776

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

PubMed

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

2013-01-01

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

HP-HMG versus rFSH in treatments combining fresh and frozen IVF cycles: success rates and economic evaluation.

PubMed

Wex-Wechowski, Jaro; Abou-Setta, Ahmed M; Kildegaard Nielsen, Sandy; Kennedy, Richard

2010-08-01

The economic implications of the choice of gonadotrophin influence decision making but their cost-effectiveness in frozen-embryo transfer cycles has not been adequately studied. An economic evaluation was performed comparing highly purified human menopausal gonadotrophin (HP-HMG) and recombinant FSH (rFSH) using individual patient data (n=986) from two large randomized controlled trials using a long agonist IVF protocol. The simulation model incorporated live birth data and published UK costs of IVF-related medical resources. After treatment for up-to-three cycles (one fresh and up to two subsequent fresh or frozen cycles conditional on availability of cryopreserved embryos), the cumulative live birth rate was 53.7% (95% CI 49.3-58.1%) for HP-HMG and 44.6% (40.2-49.0%) for rFSH (OR 1.44, 95% CI 1.12-1.85; P<0.005). The mean costs per IVF treatment for HP-HMG and rFSH were pound5393 ( pound5341-5449) and pound6269 ( pound6210-6324), respectively (number needed to treat to fund one additional treatment was seven; P<0.001). With maternal and neonatal costs applied, the median cost per IVF baby delivered with HP-HMG was pound11,157 ( pound11,089-11,129) and pound14,227 ( pound14,183-14,222) with rFSH (P<0.001). The cost saving using HP-HMG remained after varying model parameters in a probabilistic sensitivity analysis. 2010 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

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

PubMed

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

2015-01-15

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

Preservation Methods Alter Carbon and Nitrogen Stable Isotope Values in Crickets (Orthoptera: Grylloidea)

PubMed Central

Jesus, Fabiene Maria; Pereira, Marcelo Ribeiro; Rosa, Cassiano Sousa; Moreira, Marcelo Zacharias; Sperber, Carlos Frankl

2015-01-01

Stable isotope analysis (SIA) is an important tool for investigation of animal dietary habits for determination of feeding niche. Ideally, fresh samples should be used for isotopic analysis, but logistics frequently demands preservation of organisms for analysis at a later time. The goal of this study was to establish the best methodology for preserving forest litter-dwelling crickets for later SIA analysis without altering results. We collected two cricket species, Phoremia sp. and Mellopsis doucasae, from which we prepared 70 samples per species, divided among seven treatments: (i) freshly processed (control); preserved in fuel ethanol for (ii) 15 and (iii) 60 days; preserved in commercial ethanol for (iv) 15 and (v) 60 days; fresh material frozen for (vi) 15 and (vii) 60 days. After oven drying, samples were analyzed for δ 15N, δ 13C values, N(%), C(%) and C/N atomic values using continuous flow isotope ratio mass spectrometry. All preservation methods tested, significantly impacted δ 13C and δ 15N and C/N atomic values. Chemical preservatives caused δ 13C enrichment as great as 1.5‰, and δ 15N enrichment as great as 0.9‰; the one exception was M. doucasae stored in ethanol for 15 days, which had δ 15N depletion up to 1.8‰. Freezing depleted δ 13C and δ 15N by up to 0.7 and 2.2‰, respectively. C/N atomic values decreased when stored in ethanol, and increased when frozen for 60 days for both cricket species. Our results indicate that all preservation methods tested in this study altered at least one of the tested isotope values when compared to fresh material (controls). We conclude that only freshly processed material provides adequate SIA results for litter-dwelling crickets. PMID:26390400

Preservation Methods Alter Carbon and Nitrogen Stable Isotope Values in Crickets (Orthoptera: Grylloidea).

PubMed

Jesus, Fabiene Maria; Pereira, Marcelo Ribeiro; Rosa, Cassiano Sousa; Moreira, Marcelo Zacharias; Sperber, Carlos Frankl

2015-01-01

Stable isotope analysis (SIA) is an important tool for investigation of animal dietary habits for determination of feeding niche. Ideally, fresh samples should be used for isotopic analysis, but logistics frequently demands preservation of organisms for analysis at a later time. The goal of this study was to establish the best methodology for preserving forest litter-dwelling crickets for later SIA analysis without altering results. We collected two cricket species, Phoremia sp. and Mellopsis doucasae, from which we prepared 70 samples per species, divided among seven treatments: (i) freshly processed (control); preserved in fuel ethanol for (ii) 15 and (iii) 60 days; preserved in commercial ethanol for (iv) 15 and (v) 60 days; fresh material frozen for (vi) 15 and (vii) 60 days. After oven drying, samples were analyzed for δ15N, δ13C values, N(%), C(%) and C/N atomic values using continuous flow isotope ratio mass spectrometry. All preservation methods tested, significantly impacted δ13C and δ15N and C/N atomic values. Chemical preservatives caused δ13C enrichment as great as 1.5‰, and δ15N enrichment as great as 0.9‰; the one exception was M. doucasae stored in ethanol for 15 days, which had δ15N depletion up to 1.8‰. Freezing depleted δ13C and δ15N by up to 0.7 and 2.2‰, respectively. C/N atomic values decreased when stored in ethanol, and increased when frozen for 60 days for both cricket species. Our results indicate that all preservation methods tested in this study altered at least one of the tested isotope values when compared to fresh material (controls). We conclude that only freshly processed material provides adequate SIA results for litter-dwelling crickets.

Decellularized Versus Fresh-Frozen Allografts in Anterior Cruciate Ligament Reconstruction: An In Vitro Study in a Rabbit Model.

PubMed

Dong, Shikui; Huangfu, Xiaoqiao; Xie, Guoming; Zhang, Yang; Shen, Peng; Li, Xiaoxi; Qi, Jin; Zhao, Jinzhong

2015-08-01

The common fresh-frozen allografts that are used for anterior cruciate ligament (ACL) reconstructions behave slower during the remodeling process and produce weaker tendon-bone integrations than do autografts. Decellularization of allogenic tendons results in a clean and porous collagen scaffold with low antigenicity and high compatibility, which may be more suitable for ACL reconstructions. Allograft decellularization will result in a tissue structure with suitable mechanical characteristics for ACL reconstruction, thereby promoting graft remodeling and enhancing tendon-bone healing. Controlled laboratory study. Decellularized allograft tissues were prepared with a pH-modified decellularization process and evaluated for their biocompatibility and biomechanical character in vitro. Eighty New Zealand White rabbits were divided into 2 groups, with 40 in each group, to receive ACL reconstruction with either fresh-frozen (common) allografts or decellularized allografts on both knees. At 2, 4, 8, and 12 weeks postoperatively, the rabbits were euthanized for biomechanical testing, micro-computed tomography analysis, and histologic analysis. The pH-modified decellularized allograft tissues kept excellent biocompatibility and biomechanical character during the in vitro study. Biomechanical testing indicated that the decellularized allograft had significantly higher ultimate load (P = .02) and stiffness (P = .01) levels than the common allograft at 12 weeks, and there was no significant difference between the 2 groups at any other time point. The micro-CT evaluation determined significantly higher bone mineral density (P < .01) in the decellularized allograft group than that in the common allograft group at 12 weeks, but no difference between the 2 groups was observed at any other time point. Regarding bone volume/total volume, there was no difference between the 2 groups at any time point. Fibroblast ingrowths, vascular formation, and connective tissue formation in the tendon-bone interface were better in the decellularized group within 8 weeks. New bone formation was more common in the decellularized allograft group. The collagen birefringence was restored more quickly in the decellularized allograft group than in the common allograft group at all time points. The use of pH-modified decellularized allografts compared with the common allografts resulted in better cellularity, vascularity, collagen matrix remolding, new bone formation around the graft, enhanced tendon-bone healing, and higher ultimate failure load and stiffness of the graft after ACL reconstruction in the rabbit model. The pH-modified decellularized allograft may be a better graft option than the common fresh-frozen allograft for knee ligament reconstructions. © 2015 The Author(s).

The Effect of Supraphysiological Estradiol on Pregnancy Outcomes Differs between Women with PCOS and Ovulatory Women.

PubMed

Wei, Daimin; Yu, Yunhai; Sun, Mei; Shi, Yuhua; Sun, Yun; Deng, Xiaohui; Li, Jing; Wang, Ze; Zhao, Shigang; Zhang, Heping; Legro, Richard S; Chen, Zi-Jiang

2018-04-27

Supra-physiological estradiol exposure after ovarian stimulation may disrupt embryo implantation after fresh embryo transfer, compared with physiological estradiol exposure during frozen embryo transfer(FET). Women with polycystic ovary syndrome (PCOS) who usually over-respond to ovarian stimulation have a better live birth rate after elective FET than fresh embryo transfer; however ovulatory women don't. To evaluate whether the discrepancy in live birth rate after fresh versus FET between women with PCOS and ovulatory women was due to the variation in ovarian response, i.e. peak estradiol level or oocyte number. This was a secondary analysis of data from two multicenter randomized trials with similar study designs. A total of 1508 women with PCOS in the first trial and 2157 ovulatory women in the second trial were randomized to undergo either fresh or frozen embryo transfer. The primary outcome was live birth. Compared with fresh embryo transfer, FET resulted in a higher live birth rate(51.9% vs. 40.7%, OR: 1.57, 95%CI: 1.22-2.03) in PCOS women with peak estradiol level >3000pg/ml but not in those with estradiol level ≤3000pg/ml. In PCOS women with oocyte number ≥16, FET yielded a higher live birth rate(54.8% vs. 42.1%, OR: 1.67, 95%CI: 1.20-2.31), but not in those with oocyte number <16. However, in ovulatory women, the pregnancy outcomes were comparable after fresh and FET in all subgroups. Supra-physiological level of estradiol after ovarian stimulation may adversely affect pregnancy outcomes in women with PCOS; but not in ovulatory women.

Effects of Storage Temperatures on the Quality of Frozen Sardine, Mackerel, and Saury

NASA Astrophysics Data System (ADS)

Kozima, Tsuneo; Ohataka, Tateo

The three Japanese coastal fish species, sardine (Sardinops melanosticta), mackerel (Scomber japonicus) and saury (Cololabis saira) was frozen under commercial condition and stored at -18, -23, -30 and -40°C for 12 months. During frozen storage the quality was measured by determining the K value (freshness index), peroxide value (POV) of fat, a mount of free drip, water-holding capacity of muscle, weight ratio of cooking loss, histological feature of frozen and thawed muscle, and organoleptic test at regular intervals (each 2 months). Storage life of frozen sardine was 6 months at -18°C and 12 months at below -23°C. On frozen mackerel it was 6 months at -18°C. 8 months at -23°C and 12 months at below -30°C. On saury it was 6 months at 18°C and 12 months at below -23°C.

Sample storage-induced changes in the quantity and quality of soil labile organic carbon

PubMed Central

Sun, Shou-Qin; Cai, Hui-Ying; Chang, Scott X.; Bhatti, Jagtar S.

2015-01-01

Effects of sample storage methods on the quantity and quality of labile soil organic carbon are not fully understood even though their effects on basic soil properties have been extensively studied. We studied the effects of air-drying and frozen storage on cold and hot water soluble organic carbon (WSOC). Cold- and hot-WSOC in air-dried and frozen-stored soils were linearly correlated with those in fresh soils, indicating that storage proportionally altered the extractability of soil organic carbon. Air-drying but not frozen storage increased the concentrations of cold-WSOC and carbohydrate in cold-WSOC, while both increased polyphenol concentrations. In contrast, only polyphenol concentration in hot-WSOC was increased by air-drying and frozen storage, suggesting that hot-WSOC was less affected by sample storage. The biodegradability of cold- but not hot-WSOC was increased by air-drying, while both air-drying and frozen storage increased humification index and changed specific UV absorbance of both cold- and hot-WSOC, indicating shifts in the quality of soil WSOC. Our results suggest that storage methods affect the quantity and quality of WSOC but not comparisons between samples, frozen storage is better than air-drying if samples have to be stored, and storage should be avoided whenever possible when studying the quantity and quality of both cold- and hot-WSOC. PMID:26617054

Cryogenic preservation of semen from the Aleutian Canada goose (Branta canadensis leucopareia)

USGS Publications Warehouse

Gee, G.F.; Sexton, T.J.

1990-01-01

Aleutian Canada geese (Branta canadensis leucopareia) were inseminated with frozen-thawed semen containing 6% or 7% dimethylsulfoxide (DMSO) resulting in 32 fertile eggs and 17 goslings; with 7% DMSO, 19 of 31 eggs were fertile. Beltsville Poultry Semen Extender (BPSE), adjusted to 270 ? 30 mOs and 7.5 ? 0.4 pH, was used to dilute semen samples and the DMSO before cryopreservation. About half of the live spermatozoa in the fresh semen (92.9 ? 2.5% live cells, laboratory studies; 87.3 ? 7.3%, insemination trials) survived the freeze-thaw process (46.7 ? 7.8%, laboratory; 33.3 ? 17.8%, insemination trials). Samples of frozen-thawed semen contained a greater percentage of bent spermatozoa (27.1 ? 8.4% of live cells) than fresh semen (14.4 ? 3.0% of live cells). Fecal- and urate-contaminated semen (a common problem when collecting goose semen) reduced the sperm motility score from 3.2 ? 0.6 to 2.7? 0.7 and number of live spermatozoa in frozen-thawed semen from 49 ? 9% to 24 ?18%. Other variables examined that had less of an effect on semen quality included semen extenders, semen holding temperature, dilution and equilibration, relationship between hour of semen collection and level of semen contamination, and the relationship between season and sperm concentration.

Cryopreservation of epididymal stallion sperm.

PubMed

Olaciregui, M; Gil, L; Montón, A; Luño, V; Jerez, R A; Martí, J I

2014-02-01

Any event that makes semen collection or mating impossible, such as death, castration, or injury, may terminate a stallion's breeding career. Fortunately, stallion sperm which are capable of fertilization can be harvested from the epididymis, and frozen for future use. However, the fertility of frozen-thawed epididymal sperm has been found to be lower than that of ejaculated sperm. Therefore, this study aimed to optimize the fertility of frozen epididymal stallion sperm by investigating the effects of different cryoprotectants and freezing protocols on sperm quality. Dimethylformamide was tested alone or combination with pasteurized egg yolk as substitute of fresh egg yolk. In addition, the effect of the pre-freeze stabilization on sperm quality was analyzed. Heterospermic samples obtained from stallion epididymis were collected and cryopreserved in lactose-egg-yolk extender or in the same extender with varying content of cryoprotectant and content of egg yolk, stabilized and no-stabilized. Sperm motility, viability, hypoosmotic swelling test (HOST) and acrosome integrity were evaluated post-thawing. No improvement was observed on the replacement of fresh yolk by pasteurized egg yolk, whereas the results suggest that dimethylformamide is a cryoprotectant suitable for cryopreservation of equine epididymal semen, even better than glycerol. In addition, we found that the stabilization before freezing on epididymal stallion sperm, can improve sperm quality parameters. Copyright © 2014 Elsevier Inc. All rights reserved.

9 CFR 319.182 - Braunschweiger and liver sausage or liverwurst.

Code of Federal Regulations, 2013 CFR

2013-01-01

... liverwurst. 319.182 Section 319.182 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE... made from fresh, cured, and/or frozen pork, beef, and/or veal and at least 30 percent pork, beef, and/or veal livers computed on the weight of the fresh livers. It may also contain pork and/or beef fat...

9 CFR 319.182 - Braunschweiger and liver sausage or liverwurst.

Code of Federal Regulations, 2012 CFR

2012-01-01

... liverwurst. 319.182 Section 319.182 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE... made from fresh, cured, and/or frozen pork, beef, and/or veal and at least 30 percent pork, beef, and/or veal livers computed on the weight of the fresh livers. It may also contain pork and/or beef fat...

9 CFR 319.182 - Braunschweiger and liver sausage or liverwurst.

Code of Federal Regulations, 2014 CFR

2014-01-01

... liverwurst. 319.182 Section 319.182 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE... made from fresh, cured, and/or frozen pork, beef, and/or veal and at least 30 percent pork, beef, and/or veal livers computed on the weight of the fresh livers. It may also contain pork and/or beef fat...

9 CFR 319.182 - Braunschweiger and liver sausage or liverwurst.

Code of Federal Regulations, 2010 CFR

2010-01-01

... liverwurst. 319.182 Section 319.182 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE... made from fresh, cured, and/or frozen pork, beef, and/or veal and at least 30 percent pork, beef, and/or veal livers computed on the weight of the fresh livers. It may also contain pork and/or beef fat...

9 CFR 319.182 - Braunschweiger and liver sausage or liverwurst.

Code of Federal Regulations, 2011 CFR

2011-01-01

... liverwurst. 319.182 Section 319.182 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE... made from fresh, cured, and/or frozen pork, beef, and/or veal and at least 30 percent pork, beef, and/or veal livers computed on the weight of the fresh livers. It may also contain pork and/or beef fat...

Increased coagulation and fibrinolytic potential of solvent-detergent plasma: a comparative study between Omniplasma and fresh frozen plasma.

PubMed

van Beers, J J B C; van Egmond, L T; Wetzels, R J H; Verhezen, P W M; Beckers, E A M; van Oerle, R; Spronk, H M H; Straat, R J M H E; Henskens, Y M C

2016-07-01

In this study, differences in levels of proteins involved in coagulation and fibrinolysis were compared between fresh frozen (quarantine plasma) and Omniplasma. Furthermore, thawing conditions and plasma stability after thawing were studied. 10 Omniplasma and 10 quarantine plasma units were used to study different procoagulation, anticoagulation and fibrinolytic parameters. Analysis took place at different time-points during plasma storage at 2-6°C. At baseline, significant reduced levels of factor V, free protein S, α2-antiplasmin and tPA-induced ROTEM lysis time were observed in Omniplasma as compared to quarantine plasma. Moreover, thrombin generation, IXa-AT complex levels and factor XIa were significantly increased in Omniplasma. The majority of the parameters studied remained stable in Omniplasma 48 h after thawing, with the exception of factor VIII (decrease) and IXa-AT (increase). Our results suggest an increased coagulation potential, presumingly as a result of contact activation during the production process and also, an increased fibrinolytic potential in Omniplasma. The stability of Omniplasma, based upon the different parameters studied, is comparable to Q-plasma. A maximum post-thawing time of 48 hfor Omniplasma can be suggested. © 2016 International Society of Blood Transfusion.

TVT-S in the U position--anatomical study.

PubMed

Hubka, Petr; Nanka, Ondrej; Martan, Alois; Grim, Milos; Zvarova, Jana; Masata, Jaromir

2011-02-01

The objective is to describe the anatomical position of tension-free vaginal tape Secur (TVT-S) in the U position regarding possible injury and fixation site. We placed TVT-S inserters bilaterally in 13 embalmed and five fresh frozen female bodies. After dissection, we measured distances from the obturator bundle. In embalmed bodies, the mean distance of TVT-S from the obturator bundle was 2.83 cm (standard deviation (SD) 0.87 cm) on the left, 2.92 cm (SD 1.24 cm) on the right. Perforation of the fascia of obturator internus muscle occurred in 38.5%. In fresh frozen bodies, results were fundamentally similar (p > 0.05). There is a risk of injury to the obturator bundle and urinary bladder during TVT-S; however, there is a significant risk of inserting the TVT-S inserter outside the obturator internus muscle (into the lesser pelvis). The position of TVT-S does not change significantly after legs mal-positioning.

A study of the spread of Campylobacter jejuni in four large kitchens.

PubMed Central

Dawkins, H. C.; Bolton, F. J.; Hutchinson, D. N.

1984-01-01

Campylobacters were sought in swabs taken from work surfaces, sinks and floors of four kitchens-i.e. hospital, university, cook-freeze and commercial, processing frozen or fresh chickens. Each kitchen was visited on four occasions. In the large commercial kitchen environmental contamination was found on each visit, whereas campylobacters were isolated on six of the twelve visits to the other kitchens. The hands of operatives were contaminated with campylobacters on only two of the 45 swabs taken during processing. Cleaning with detergent and hot water (or steam) and drying appears to be sufficient to remove the organism from the environment. Evidence of carriage of campylobacters by the birds was obtained on all 16 visits. In the three kitchens where only frozen birds were used the organism was isolated from 30% and 9.8% of swabs taken from the internal and external surfaces respectively, while 41% of giblets and 22.2% of thawed juices yielded campylobacters. The external surface of 30 (88%) of 34 fresh birds grew campylobacters. PMID:6736643

Effects of Extended Freezer Storage on the Integrity of Human Milk.

PubMed

Ahrabi, Ali Faraghi; Handa, Deepali; Codipilly, Champa N; Shah, Syed; Williams, Janet E; McGuire, Mark A; Potak, Debra; Aharon, Grace Golda; Schanler, Richard J

2016-10-01

To examine the integrity (pH, bacterial counts, host defense factors, nutrient contents, and osmolality) of freshly expressed and previously refrigerated human milk subjected to long-term freezer storage. Mothers donated 100 mL of freshly expressed milk. Samples were divided into baseline, storage at -20°C (fresh frozen) for 1, 3, 6, and 9 months, and prior storage at +4°C for 72 hours (refrigerated frozen) before storage at -20°C for 1 to 9 months. Samples were analyzed for pH, total bacterial colony count, gram-positive and gram-negative colony counts, and concentrations of total protein, fat, nonesterified fatty acids, lactoferrin, secretory IgA, and osmolality. Milk pH, total bacterial colony count, and Gram-positive colony counts decreased significantly with freezer storage (P < .001); bacterial counts decreased most rapidly in the refrigerated frozen group. The gram-negative colony count decreased significantly over time (P < .001). Nonesterified fatty acid concentrations increased significantly with time in storage (P < .001). Freezing for up to 9 months did not affect total protein, fat, lactoferrin, secretory IgA, or osmolality in either group. Freezer storage of human milk for 9 months at -20°C is associated with decreasing pH and bacterial counts, but preservation of key macronutrients and immunoactive components, with or without prior refrigeration for 72 hours. These data support current guidelines for freezer storage of human milk for up to 9 months for both freshly expressed and refrigerated milk. Copyright © 2016 Elsevier Inc. All rights reserved.

In vitro embryo production in buffalo (Bubalus bubalis) using sexed sperm and oocytes from ovum pick up.

PubMed

Liang, X W; Lu, Y Q; Chen, M T; Zhang, X F; Lu, S S; Zhang, M; Pang, C Y; Huang, F X; Lu, K H

2008-04-15

The objective was to explore the use of sexed sperm and OPU-derived oocytes in an IVP system to produce sex-preselected bubaline embryos. Oocytes were recovered from 20 fertile Murrah and Nili-Ravi buffalo cows by repeated (twice weekly) ultrasound-guided transvaginal ovum pick up (OPU), or by aspiration of abbatoir-derived bubaline ovaries, and subjected to IVF, using frozen-thawed sexed or unsexed bubaline semen. On average, 4.6 oocytes were retrieved per buffalo per session (70.9% were Grades A or B). Following IVF with sexed sperm, oocytes derived from OPU had similar developmental competence as those from abattoir-derived ovaries, in terms of cleavage rate (57.6 vs. 50.4%, P=0.357) and blastocyst development rate (16.0 vs. 23.9%, P=0.237). Furthermore, using frozen-thawed sexed versus unsexed semen did not affect rates of cleavage (50.5 vs. 50.9%, P=0.978) or blastocyst development (15.3 vs. 19.1%, P=0.291) after IVF using OPU-derived oocytes. Of the embryos produced in an OPU-IVP system, 9 of 34 sexed fresh embryos (26.5%) and 5 of 43 sexed frozen embryos (11.6%) transferred to recipients established pregnancies, whereas 7 of 26 unsexed fresh embryos (26.9%) and 6 out of 39 unsexed frozen embryos (15.4%) transferred to recipients established pregnancies. Eleven sex-preselected buffalo calves (10 females and one male) and 10 sexed buffalo calves (six females and four males) were born following embryo transfer. In the present study, OPU, sperm sexing technology, IVP, and embryo transfer, were used to produce sex-preselected buffalo calves. This study provided proof of concept for further research and wider field application of these technologies in buffalo.

A randomized controlled trial of fresh frozen plasma for coagulopathy in Russell's viper (Daboia russelii) envenoming.

PubMed

Isbister, G K; Jayamanne, S; Mohamed, F; Dawson, A H; Maduwage, K; Gawarammana, I; Lalloo, D G; de Silva, H J; Scorgie, F E; Lincz, L F; Buckley, N A

2017-04-01

Essentials Russell's viper envenoming is a major health issue in South Asia and causes coagulopathy. We studied the effect of fresh frozen plasma and two antivenom doses on correcting coagulopathy. Fresh frozen plasma did not hasten recovery of coagulopathy. Low-dose antivenom did not worsen coagulopathy. Background Russell's viper (Daboia russelii) envenoming is a major health issue in South Asia and causes venom-induced consumption coagulopathy (VICC). Objectives To investigate the effects of fresh frozen plasma (FFP) and two antivenom doses in correcting VICC. Methods We undertook an open-label randomized controlled trial in patients with VICC at two Sri Lankan hospitals. Patients with suspected Russell's viper bites and coagulopathy were randomly allocated (1 : 1) to high-dose antivenom (20 vials) or low-dose antivenom (10 vials) plus 4 U of FFP. The primary outcome was the proportion of patients with an International Normalized Ratio (INR) of < 2 at 6 h after antivenom administration. Secondary outcomes included anaphylaxis, major hemorrhage, death, and clotting factor recovery. Results From 214 eligible patients, 141 were randomized: 71 to high-dose antivenom, and 70 to low-dose antivenom/FFP; five had no post-antivenom blood tests. The groups were similar except for a delay of 1 h in antivenom administration for FFP patients. Six hours after antivenom administration, 23 of 69 (33%) patients allocated to high-dose antivenom had an INR of < 2, as compared with 28 of 67 (42%) allocated to low-dose antivenom/FFP (absolute difference 8%; 95% confidence interval - 8% to 25%). Fifteen patients allocated to FFP did not receive it. Severe anaphylaxis occurred equally frequently in each group. One patient given FFP developed transfusion-related acute lung injury. Three deaths occurred in low-dose antivenom/FFP patients, including one intracranial hemorrhage. There was no difference in recovery rates of INR or fibrinogen, but there was more rapid initial recovery of factor V and FX in FFP patients. Conclusion FFP after antivenom administration in patients with Russell's viper bites did not hasten recovery of coagulopathy. Low-dose antivenom/FFP did not worsen VICC, suggesting that low-dose antivenom is sufficient. © 2017 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals, Inc. on behalf of International Society on Thrombosis and Haemostasis.

ASSAYS FOR ENDOGENOUS COMPONENTS OF HUMAN MILK: COMPARISON OF FRESH AND FROZEN SAMPLES AND CORRESPONDING ANALYTES IN SERUM

EPA Science Inventory

Breast milk is a primary source of nutrition that contains many endogenous compounds that may affect infant development. The goals of this study were to develop reliable assays for selected endogenous breast milk components and to compare levels of those in milk and serum collect...

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) ...

Leakage of fluid around endotracheal tube cuffs: a cadaver study

PubMed Central

Lucius, Ralph; Ewald, Kristian

2013-01-01

Background The aim of the study was to evaluate the leakage of liquid past the cuffs of tracheal tubes in fresh frozen human heads. Methods Six truncated fresh frozen heads were used and intubated with 8.0 mm endotracheal tubes. The intracuff pressures tested were 30 and 100 cmH2O. Subsequently, 20 ml of each of two oral antiseptic rinses (0.2% chlorhexidine and octenidine [octenidol®, Schülke & Mayr GmbH, Norderstedt, Germany]) was applied for thirty seconds in the mouth. During the trial, leakage of the cuffs was examined. Results The sealing between the tracheal cuff and tracheal wall was leakage-proof for all tested intracuff pressures and all tested antiseptic rinses. However, approximately 5.6 ml and 1.8 ml leaked into the esophagus and remained as a cuff-puddle, respectively. Conclusions The sealing between an endotracheal tube cuff with an intracuff pressure of 30 cmH2O and the tracheal wall is leakage-proof during oral care with antiseptic rinsing. An increase of intracuff pressure to 100 cmH2O does not appear to be required. PMID:24363847

Targeted or whole genome sequencing of formalin fixed tissue samples: potential applications in cancer genomics.

PubMed

Munchel, Sarah; Hoang, Yen; Zhao, Yue; Cottrell, Joseph; Klotzle, Brandy; Godwin, Andrew K; Koestler, Devin; Beyerlein, Peter; Fan, Jian-Bing; Bibikova, Marina; Chien, Jeremy

2015-09-22

Current genomic studies are limited by the poor availability of fresh-frozen tissue samples. Although formalin-fixed diagnostic samples are in abundance, they are seldom used in current genomic studies because of the concern of formalin-fixation artifacts. Better characterization of these artifacts will allow the use of archived clinical specimens in translational and clinical research studies. To provide a systematic analysis of formalin-fixation artifacts on Illumina sequencing, we generated 26 DNA sequencing data sets from 13 pairs of matched formalin-fixed paraffin-embedded (FFPE) and fresh-frozen (FF) tissue samples. The results indicate high rate of concordant calls between matched FF/FFPE pairs at reference and variant positions in three commonly used sequencing approaches (whole genome, whole exome, and targeted exon sequencing). Global mismatch rates and C · G > T · A substitutions were comparable between matched FF/FFPE samples, and discordant rates were low (<0.26%) in all samples. Finally, low-pass whole genome sequencing produces similar pattern of copy number alterations between FF/FFPE pairs. The results from our studies suggest the potential use of diagnostic FFPE samples for cancer genomic studies to characterize and catalog variations in cancer genomes.

Requirements for 25,000 Servings of Items Selected From the 42-Day Menu

DTIC Science & Technology

1975-04-01

1.1 Eggs, whole, frozen 563 563 640 37.0 Cake, Coffee, Glazed Nut Mix, sweet dough 3,750 3,750 4,500 137.5 Eggs, whole, frozen 125 125 142 8.2...Twist Mix, sweet dough 3,750 3,750 4,500 137.5 Butter 250 250 294 4.8 Cinnamon, ground 20 20 28 0.9 Sugar, brown 500 500 525 8.8 Raisins 500...1,688 1,923 111.2 Lemons, fresh 1,688 1,688 1,928 60.7 \\ Cookies , Butternut Refrigerator i Eggs, whole, frozen 188 188 213 12.3 \\ Shortening 656

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

PubMed

Seong, Pil Nam; Seo, Hyun Woo; Kim, Jin-Hyoung; Kang, Geun Ho; Cho, Soo-Hyun; Chae, Hyun Seok; Park, Beom Young; Van Ba, Hoa

2017-12-01

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

Modifying the processing and handling of frozen broccoli for increased sulforaphane formation.

PubMed

Dosz, Edward B; Jeffery, Elizabeth H

2013-09-01

Frozen broccoli can provide a cheaper product, with a longer shelf life and less preparation time than fresh broccoli. We previously showed that several commercially available frozen broccoli products do not retain the ability to generate the cancer-preventative agent sulforaphane. We hypothesized that this was because the necessary hydrolyzing enzyme myrosinase was destroyed during blanching, as part of the processing that frozen broccoli undergoes. This study was carried out to determine a way to overcome loss of hydrolyzing activity. Industrial blanching usually aims to inactivate peroxidase, although lipoxygenase plays a greater role in product degradation during frozen storage of broccoli. Blanching at 86 °C or higher inactivated peroxidase, lipoxygenase, and myrosinase. Blanching at 76 °C inactivated 92% of lipoxygenase activity, whereas there was only an 18% loss in myrosinase-dependent sulforaphane formation. We considered that thawing frozen broccoli might disrupt membrane integrity, allowing myrosinase and glucoraphanin to come into contact. Thawing frozen broccoli for 9 h did not support sulforaphane formation unless an exogenous source of myrosinase was added. Thermal stability studies showed that broccoli root, as a source of myrosinase, was not more heat stable than broccoli floret. Daikon radish root supported some sulforaphane formation even when heated at 125 °C for 10 min, a time and temperature comparable to or greater than microwave cooking. Daikon radish (0.25%) added to frozen broccoli that was then allowed to thaw supported sulforaphane formation without any visual alteration to that of untreated broccoli. © 2013 Institute of Food Technologists®

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

PubMed

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

2016-10-01

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

9 CFR 94.2 - Fresh (chilled or frozen) products (other than meat), and milk and milk products of ruminants and...

Code of Federal Regulations, 2013 CFR

2013-01-01

... (other than meat), and milk and milk products of ruminants and swine. 94.2 Section 94.2 Animals and... NEWCASTLE DISEASE, AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR DISEASE, AND BOVINE... (other than meat), and milk and milk products of ruminants and swine. (a) The importation of fresh...

9 CFR 94.2 - Fresh (chilled or frozen) products (other than meat), and milk and milk products of ruminants and...

Code of Federal Regulations, 2012 CFR

2012-01-01

... (other than meat), and milk and milk products of ruminants and swine. 94.2 Section 94.2 Animals and... NEWCASTLE DISEASE, AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR DISEASE, AND BOVINE... (other than meat), and milk and milk products of ruminants and swine. (a) The importation of fresh...

9 CFR 94.2 - Fresh (chilled or frozen) products (other than meat), and milk and milk products of ruminants and...

Code of Federal Regulations, 2011 CFR

2011-01-01

... (other than meat), and milk and milk products of ruminants and swine. 94.2 Section 94.2 Animals and... NEWCASTLE DISEASE, AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR DISEASE, AND BOVINE... (other than meat), and milk and milk products of ruminants and swine. (a) The importation of fresh...

Perinatal outcomes after gestational surrogacy versus autologous IVF: analysis of national data.

PubMed

Sunkara, Sesh Kamal; Antonisamy, Belavendra; Selliah, Hepsy Y; Kamath, Mohan S

2017-12-01

Anonymized data were obtained from the Human Fertilization and Embryology Authority to determine whether gestational surrogacy influences perinatal outcomes compared with pregnancies after autologous IVF. A total of 103,160 singleton live births, including 244 after gestational surrogacy, 87,571 after autologous fresh IVF and intractyoplasmic sperm injection (ICSI) and 15,345 after autologous frozen embryo transfers were analysed. Perinatal outcomes of pretern birth (PTB), low birth weight (LBW) and high birth weight (HBW) were compared. No difference was found in the risk of PTB and LBW after gestational surrogacy compared with autologous fresh IVF-ICSI: PTB (adjusted OR 0.90, 95% CI 0.56 to 1.42), LBW (adjusted OR 0.90, 95% CI 0.57 to 1.43) and gestational surrogacy compared with autologous frozen embryo transfers: PTB (adjusted OR 0.96, 95% CI 0.58 to 1.60), LBW (adjusted OR 1.16, 95% CI 0.69 to 1.96). The incidence of HBW was significantly higher after gestational surrogacy compared with fresh IVF-ICSI (adjusted OR 1.94, 95% CI 1.38 to 2.75); no difference was found in HBW between gestational surrogacy and autologous frozen embryo transfers. The dataset is limited by lack of information on confounders, i.e. ethnicity, body mass index, underlying medical history, which could result in residual confounding. Copyright © 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Penetration and decontamination of americium-241 ex vivo using fresh and frozen pig skin.

PubMed

Tazrart, A; Bolzinger, M A; Moureau, A; Molina, T; Coudert, S; Angulo, J F; Briancon, S; Griffiths, N M

2017-04-01

Skin contamination is one of the most probable risks following major nuclear or radiological incidents. However, accidents involving skin contamination with radionuclides may occur in the nuclear industry, in research laboratories and in nuclear medicine departments. This work aims to measure the penetration of the radiological contaminant Americium ( 241 Am) in fresh and frozen skin and to evaluate the distribution of the contamination in the skin. Decontamination tests were performed using water, Fuller's earth and diethylene triamine pentaacetic acid (DTPA), which is the recommended treatment in case of skin contamination with actinides such as plutonium or americium. To assess these parameters, we used the Franz cell diffusion system with full-thickness skin obtained from pigs' ears, representative of human skin. Solutions of 241 Am were deposited on the skin samples. The radioactivity content in each compartment and skin layers was measured after 24 h by liquid scintillation counting and alpha spectrophotometry. The Am cutaneous penetration to the receiver compartment is almost negligible in fresh and frozen skin. Multiple washings with water and DTPA recovered about 90% of the initial activity. The rest remains fixed mainly in the stratum corneum. Traces of activity were detected within the epidermis and dermis which is fixed and not accessible to the decontamination. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

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

PubMed Central

Cavallo, Carola; Cenacchi, Annarita; Facchini, Andrea; Grigolo, Brunella; Kon, Elizaveta; Mariani, Erminia; Pratelli, Loredana; Marcacci, Maurilio

2014-01-01

PRP cryopreservation remains a controversial point. Our purpose was to investigate the effect of freezing/thawing on PRP molecule release, and its effects on the metabolism of chondrocytes and synoviocytes. PRP was prepared from 10 volunteers, and a half volume underwent one freezing/thawing cycle. IL-1β, HGF, PDGF AB/BB, TGF-β1, and VEGF were assayed 1 hour and 7 days after activation. Culture media of chondrocytes and synoviocytes were supplemented with fresh or frozen PRP, and, at 7 days, proliferation, gene expression, and secreted proteins levels were evaluated. Results showed that in the freeze-thawed PRP the immediate and delayed molecule releases were similar or slightly lower than those in fresh PRP. TGF-β1 and PDGF AB/BB concentrations were significantly reduced after freezing both at 1 hour and at 7 days, whereas HGF concentration was significantly lower in frozen PRP at 7 days. In fresh PRP IL-1β and HGF concentrations underwent a significant further increase after 7 days. Similar gene expression was found in chondrocytes cultured with both PRPs, whereas in synoviocytes HGF gene expression was higher in frozen PRP. PRP cryopreservation is a safe procedure, which sufficiently preserves PRP quality and its ability to induce proliferation and the production of ECM components in chondrocytes and synoviocytes. PMID:25136613

Comparison of biochemical values in serum and plasma, fresh and frozen plasma, and hemolyzed samples from orange-winged Amazon parrots (Amazona amazonica).

PubMed

Hawkins, Michelle G; Kass, Philip H; Zinkl, Joseph G; Tell, Lisa A

2006-06-01

To the authors' knowledge, on the basis of sample type, storage condition, or hemolysis, differences in serum and plasma biochemical values have not been evaluated in orange-winged Amazon parrots (Amazona amazonica). The purpose of this study was to compare values for biochemical analytes in serum vs plasma, fresh vs frozen plasma, and nonhemolyzed vs hemolyzed samples in orange-winged Amazon parrots. We also compared differences in serum and plasma yield from whole-blood aliquots. Fifteen biochemical analytes were evaluated in paired serum and plasma, fresh and frozen plasma, nonhemolyzed and hemolyzed serum and plasma samples from orange-winged Amazon parrots (n = 10) using a wet reagent analyzer. Hemolysis was assessed qualitatively (visually) and quantitatively (hemoglobin [Hgb] measured spectrophotometrically). Serum and plasma yields from 500-microl whole-blood aliquots were determined from centrifuged samples. Analyte values significantly differed among sample groups, but were still within published reference intervals, with the exception of increases in potassium concentration in markedly hemolyzed serum and plasma samples. Clinically important changes in hemolyzed serum and plasma samples included increases in potassium, phosphorus, and albumin concentrations and lactate dehydrogenase activity. The degree of hemolysis assigned qualitatively did not correlate with quantitative Hgb concentration. A significantly greater yield of plasma (288 +/- 13 microL) than serum (241 +/- 44 microL) was obtained. Significant differences may occur in different sample types, however, only changes in potassium, phosphorus, albumin, and lactate dehydrogenase values in hemolyzed samples were considered clinically relevant. Lack of agreement between qualitative and quantitative Hgb concentration indicates the unreliability of visual estimation. Based on higher sample yield, and lack of clinically relevant differences from serum, plasma is a better sample choice for clinical chemistry analysis in birds.

Impact of the revised Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) food package policy on fruit and vegetable prices.

PubMed

Zenk, Shannon N; Powell, Lisa M; Odoms-Young, Angela M; Krauss, Ramona; Fitzgibbon, Marian L; Block, Daniel; Campbell, Richard T

2014-02-01

Obesity is generally inversely related to income among women in the United States. Less access to healthy foods is one way lower income can influence dietary behaviors and body weight. Federal food assistance programs, such as the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), are an important source of healthy food for low-income populations. In 2009, as part of a nationwide policy revision, WIC added a fruit and vegetable (F/V) voucher to WIC food packages. This quasi-experimental study determined whether F/V prices at stores authorized to accept WIC (ie, WIC vendors) decreased after the policy revision in seven Illinois counties. It also examined cross-sectional F/V price variations by store type and neighborhood characteristics. Two pre-policy observations were conducted in 2008 and 2009; one post-policy observation was conducted in 2010. Small pre- to post-policy reductions in some F/V prices were found, particularly for canned fruit and frozen vegetables at small stores. Compared with chain supermarkets, mass merchandise stores had lower prices for fresh F/V and frozen F/V and small stores and non-chain supermarkets had higher canned and frozen F/V prices, but lower fresh F/V prices. Limited price differences were found across neighborhoods, although canned vegetables were more expensive in neighborhoods with higher concentrations of either Hispanics or blacks and fresh F/V prices were lower in neighborhoods with more Hispanics. Results suggest the WIC policy revision contributed to modest reductions in F/V prices. WIC participants' purchasing power can differ depending on the type and neighborhood of the WIC vendor used. Copyright © 2014 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Impact of the Revised Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) Food Package Policy on Fruit and Vegetable Prices

PubMed Central

Zenk, Shannon N.; Powell, Lisa M.; Odoms-Young, Angela M.; Krauss, Ramona; Fitzgibbon, Marian L.; Block, Daniel; Campbell, Richard T.

2014-01-01

Obesity is generally inversely related to income among women in the United States. Less access to healthy foods is one way lower income can influence dietary behaviors and body weight. Federal food assistance programs, such as the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC), are an important source of healthy food for low-income populations. In 2009, as part of a nationwide policy revision, WIC added a fruit and vegetable (F/V) voucher to WIC food packages. This quasi-experimental study determined whether F/V prices at stores authorized to accept WIC (ie, WIC vendors) decreased after the policy revision in seven Illinois counties. It also examined cross-sectional F/V price variations by store type and neighborhood characteristics. Two pre-policy observations were conducted in 2008 and 2009; one post-policy observation was conducted in 2010. Small pre- to post-policy reductions in some F/V prices were found, particularly for canned fruit and frozen vegetables at small stores. Compared with chain supermarkets, mass merchandise stores had lower prices for fresh F/V and frozen F/V and small stores and non-chain supermarkets had higher canned and frozen F/V prices, but lower fresh F/V prices. Limited price differences were found across neighborhoods, although canned vegetables were more expensive in neighborhoods with higher concentrations of either Hispanics or blacks and fresh F/V prices were lower in neighborhoods with more Hispanics. Results suggest the WIC policy revision contributed to modest reductions in F/V prices. WIC participants’ purchasing power can differ depending on the type and neighborhood of the WIC vendor used. PMID:24183996

21 CFR 1250.3 - Definitions.

Code of Federal Regulations, 2010 CFR

2010-04-01

... Rico and the Virgin Islands. (l) Utensil. Includes any kitchenware, tableware, glassware, cutlery... shell, and any fresh, frozen, or incompletely cooked edible products thereof. [40 FR 5624, Feb. 6, 1975...

Evaluation and utilization of preassembled frozen commercial fast real-time qPCR master mixes for detection of cytomegalovirus and BK virus.

PubMed

Glover, William A; Atienza, Ederlyn E; Nesbitt, Shannon; Kim, Woo J; Castor, Jared; Cook, Linda; Jerome, Keith R

2016-01-01

Quantitative DNA detection of cytomegalovirus (CMV) and BK virus (BKV) is critical in the management of transplant patients. Quantitative laboratory-developed procedures for CMV and BKV have been described in which much of the processing is automated, resulting in rapid, reproducible, and high-throughput testing of transplant patients. To increase the efficiency of such assays, the performance and stability of four commercial preassembled frozen fast qPCR master mixes (Roche FastStart Universal Probe Master Mix with Rox, Bio-Rad SsoFast Probes Supermix with Rox, Life Technologies TaqMan FastAdvanced Master Mix, and Life Technologies Fast Universal PCR Master Mix), in combination with in-house designed primers and probes, was evaluated using controls and standards from standard CMV and BK assays. A subsequent parallel evaluation using patient samples was performed comparing the performance of freshly prepared assay mixes versus aliquoted frozen master mixes made with two of the fast qPCR mixes (Life Technologies TaqMan FastAdvanced Master Mix, and Bio-Rad SsoFast Probes Supermix with Rox), chosen based on their performance and compatibility with existing PCR cycling conditions. The data demonstrate that the frozen master mixes retain excellent performance over a period of at least 10 weeks. During the parallel testing using clinical specimens, no difference in quantitative results was observed between the preassembled frozen master mixes and freshly prepared master mixes. Preassembled fast real-time qPCR frozen master mixes perform well and represent an additional strategy laboratories can implement to reduce assay preparation times, and to minimize technical errors and effort necessary to perform clinical PCR. © 2015 Wiley Periodicals, Inc.

Endometrial preparation for women undergoing embryo transfer with frozen embryos or embryos derived from donor oocytes.

PubMed

Glujovsky, Demián; Pesce, Romina; Fiszbajn, Gabriel; Sueldo, Carlos; Hart, Roger J; Ciapponi, Agustín

2010-01-20

If a fresh embryo, assisted reproductive technology procedure cycle is unsuccessful and there are frozen embryos available, a frozen-thawed embryo transfer is performed. In some specific cases women may undergo oocyte donation treatment. In both situations the endometrium is primed by the administration of estrogen and progesterone. To prevent the possibility of spontaneous ovulation, gonadotropin-releasing hormone (GnRH) agonists are frequently used. To evaluate the most effective endometrial preparation for women undergoing transfer with frozen embryos or embryos from donor oocytes with regard to the subsequent live birth rate. We searched the Cochrane Menstrual Disorders and Subfertility Group Trials Register, Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library), MEDLINE, EMBASE, LILACS, and abstracts of reproductive societies' meetings (from inception). No language restrictions were applied. Experts in the field were contacted. Randomised controlled trials evaluating endometrial preparation in women undergoing fresh donor cycles and frozen embryo transfers. Two review authors independently applied the inclusion criteria, assessed trial risk of bias, and extracted data. Twenty two randomised controlled trials were included. Five studies analysed the use of a GnRH agonist versus control. No significant benefit was demonstrated when using GnRH agonists. No evidence of statistically significant benefit was found for one GnRH agonist over another, or vaginal over intramuscular progesterone administration. No difference in pregnancy rate was demonstrated when no treatment was compared to aspirin, steroids, ovarian stimulation, or human chorionic gonadotropin (hCG) prior to embryo transfer, although using hCG several times before the oocyte retrieval decreases the pregnancy rate. Finally, when oocyte recipients were studied further, starting progesterone on the day of oocyte pick-up (OPU) or the day after OPU produced a significantly higher pregnancy rate (OR 1.87, 95% CI 1.13 to 3.08) than when recipients started progesterone the day prior to OPU. There is insufficient evidence to recommend any one particular protocol for endometrial preparation over another with regard to pregnancy rates after embryo transfers. These were either frozen embryos or embryos derived from donor oocytes. However, there is evidence of a lower pregnancy rate and a higher cycle cancellation rate when the progesterone supplementation is commenced prior to oocyte retrieval in oocyte donation cycles. Adequately powered studies are needed to evaluate each treatment more accurately.

Halyomorpha halys (Heteroptera: Pentatomidae) egg surface chemicals inhibit North American Telenomus and Trissolcus (Hymenoptera: Scelionidae) parasitism

USDA-ARS?s Scientific Manuscript database

Halyomorpha halys (Stål) (Heteroptera: Pentatomidae), the brown marmorated stink bug (BMSB), is an invasive pest in the U.S., where it has been largely unaffected by natural enemies. Using rinsed and unrinsed, frozen and fresh egg masses in laboratory bioassays, we studied how surface chemicals on t...

Subpopulations of bovine T lymphocytes collected during foot-and-mouth disease virus infection are affected by freezing, but are subsequently stable in frozen samples

USDA-ARS?s Scientific Manuscript database

Immunophenotyping of peripheral-blood lymphocytes by flow cytometry is an important tool for infectious disease research. In many live-animal experiments and other longitudinal studies, the processing, prompt staining, and analysis of fresh samples is a logistical challenge and daily variation can c...

Disappearance of six pesticides in fresh and processed zucchini, bioavailability and health risk assessment.

PubMed

Oliva, J; Cermeño, S; Cámara, M A; Martínez, G; Barba, A

2017-08-15

A field study was carried out on the dissipation of three insecticides and three fungicides during the freezing of zucchini. A simultaneous residue analysis method is validated using QuEChERS extraction with acetonitrile and CG-MS and LC-MS analysis. The residues detected after field application never exceeded the established maximum residue limits. The processing factors calculated (fresh product/frozen product) are lower than 1, indicating a clear influence of the stages of the freezing process, especially the washing and blanching. The in vitro study of bioavailability establishes a low percentage of stomach absorption capacity. The level of residues detected in fresh zucchini and the Estimated Daily Intake calculated for Spain suggest that there is no risk of acute toxicity due to dietary exposure. Copyright © 2017 Elsevier Ltd. All rights reserved.

Foods - fresh vs. frozen or canned

MedlinePlus

... and U.S. Department of Agriculture website. 2015-2020 Dietary Guidelines for Americans. 8th Edition. December 2015. health.gov/dietaryguidelines/2015/resources/2015-2020_Dietary_Guidelines.pdf . Accessed July 13,2017.

Does premature elevated progesterone on the day of trigger increase spontaneous abortion rates in fresh and subsequent frozen embryo transfers?

PubMed

Healy, Mae; Patounakis, George; Zanelotti, Austin; Devine, Kate; DeCherney, Alan; Levy, Michael; Hill, Micah J

2017-06-01

Recent evidence has shown elevated progesterone (P) advances the endometrium in fresh ART cycles, creating asynchrony with the embryo and thus implantation failure and decreased live birth rates. If the window of implantation is closing as the embryo attempts to implant, there may be difficulty with trophoblastic invasion, leading to failure of early pregnancies. Our objective was to evaluate if P on the day of trigger was associated with spontaneous abortion (SAB) rates in fresh ART transfers. This was a retrospective cohort study involving fresh autologous and FET cycles from 2011 to 2013. The main outcome was spontaneous abortion rates. About 4123 fresh and FET transfer cycles were included which resulted in 1547 fresh and 491 FET pregnancies. The overall SAB rate was 20% among fresh cycles and 19% in FET cycles. P on the day of trigger, as a continuous variable or when > 2 ng/mL, was not associated with SAB in fresh cycles. Similar results were found after adjusting for age, embryo quality, and embryo stage. Despite elevated P likely advancing the window of implantation, once implantation occurs, pregnancies were no longer negatively impacted by progesterone.

Evaluation of cholesterol- treated dromedary camel sperm function by heterologous IVF and AI.

PubMed

Crichton, Elizabeth G; Malo, Clara; Pukazhenthi, Budhan S; Nagy, Peter; Skidmore, Julian A

2016-11-01

Cholesterol (cholesterol-loaded cyclodextrins: CLC) treatment of dromedary camel sperm prior to freezing enhances cryosurvival. The present study first validated the efficacy of a heterologous zona-free goat oocyte assay (n=115 oocytes) to evaluate camel sperm function in vitro (Experiment 1: n=6 bulls), then examined the effects of CLC treatment (1.5mg/mL CLC; CLC+) versus no treatment (0 CLC) of fresh (Experiment 2: n=4 bulls) and frozen-thawed (Experiment 3: n=5 bulls) camel sperm to penetrate, de-condense and form pro-nuclei in in vitro-matured goat oocytes. Finally, the ability of fresh 0 CLC and CLC+ sperm to fertilize in vivo was studied by artificially inseminating super-ovulated females (n=7-9 per treatment) and examining embryo production (Experiment 4: n=4-5 bulls/treatment). Camel spermatozoa penetrated (60%) and formed pro-nuclei (33%) in goat oocytes demonstrating the utility of this heterologous system for assessing sperm function in vitro. For fresh spermatozoa, 0 CLC-treated sperm performed better than their CLC+ counterparts for all parameters measured (P<0.05). In contrast, cryopreservation resulted in a sharp decline in sperm-oocyte interaction in 0 CLC aliquots but remained unaltered in CLC+ aliquots demonstrating a protective effect of cholesterol treatment. There was no difference between treatments in the in vitro fertilizing ability of frozen-thawed sperm or in the numbers of embryos retrieved following AI with fresh 0 CLC or CLC+ sperm. We conclude that although CLC treatment of dromedary camel sperm improves sperm motility it fails to confer an advantage to them in terms of improved in vitro sperm-oocyte interaction or in vivo fertilization under the conditions tested. Copyright © 2016 Elsevier B.V. All rights reserved.

All about Carbohydrate Counting

MedlinePlus

... barley, bulgur, couscous, • fruit (canned, dried, grits, kasha, pasta, and fresh, and frozen) and rice fruit juice • ... ½ cup banana: 1 extra small (4 ounces) pasta or rice (cooked): ⅓ cup bread: 1 slice ( ...

Microbial characterization of bee pollen from the Vesuvius area collected by using three different traps.

PubMed

Mauriello, Gianluigi; De Prisco, Annachiara; Di Prisco, Gennaro; La Storia, Antonietta; Caprio, Emilio

2017-01-01

Flower pollen is collected by honeybee foragers, adhered on their rear legs and transported into the hives in the form of pellets. Once in the hives, bee pollen is moisturised with nectar and bee mouth secretions and due to enzymatically modifications it becomes the so-called bee-bread, the protein reservoir of young bees. Bee pollen can be artificially removed from bee legs and collected by using specific systems, the bee pollen traps. Bee pollen is commercialized for human consumption as fresh product and after freezing or drying. Although bee pollen is nowadays largely consumed in developed countries, as food or food supplement according to local legislation, little is known on its safety related to microbiological hazards. In this work, we aimed to characterize for the first time the microbiological profile of Italian bee pollen in fresh, frozen and dried form collected along an entire harvesting season. Moreover, monthly microbiological analyses were performed on frozen (storage at -18°C) and dried (storage at room temperature) bee pollen over a 4 months period. Further aim of this work was the evaluation of the possible impact on production level of three different traps used for pollen collection. Our results on microbial contamination of fresh and frozen bee pollen show that a more comprehensive microbiological risk assessment of bee pollen is required. On the other side, dried pollen showed very low microbial contamination and no pathogen survived after the drying process and during storage.

Microbial characterization of bee pollen from the Vesuvius area collected by using three different traps

PubMed Central

De Prisco, Annachiara; Di Prisco, Gennaro; La Storia, Antonietta; Caprio, Emilio

2017-01-01

Flower pollen is collected by honeybee foragers, adhered on their rear legs and transported into the hives in the form of pellets. Once in the hives, bee pollen is moisturised with nectar and bee mouth secretions and due to enzymatically modifications it becomes the so-called bee-bread, the protein reservoir of young bees. Bee pollen can be artificially removed from bee legs and collected by using specific systems, the bee pollen traps. Bee pollen is commercialized for human consumption as fresh product and after freezing or drying. Although bee pollen is nowadays largely consumed in developed countries, as food or food supplement according to local legislation, little is known on its safety related to microbiological hazards. In this work, we aimed to characterize for the first time the microbiological profile of Italian bee pollen in fresh, frozen and dried form collected along an entire harvesting season. Moreover, monthly microbiological analyses were performed on frozen (storage at -18°C) and dried (storage at room temperature) bee pollen over a 4 months period. Further aim of this work was the evaluation of the possible impact on production level of three different traps used for pollen collection. Our results on microbial contamination of fresh and frozen bee pollen show that a more comprehensive microbiological risk assessment of bee pollen is required. On the other side, dried pollen showed very low microbial contamination and no pathogen survived after the drying process and during storage. PMID:28934240

Use of recombinant human thyroid-stimulating hormone for thyrotropin stimulation test in healthy, hypothyroid and euthyroid sick dogs.

PubMed

Daminet, Sylvie; Fifle, Lyanne; Paradis, Manon; Duchateau, Luc; Moreau, Maxim

2007-12-01

Recombinant human thyroid-stimulating hormone (rhTSH) was evaluated for the diagnosis of canine hypothyroidism, using TSH response tests. Phase I stimulation tests were performed in 6 healthy dogs weighing over 20 kg, using 50 and then 100 microg of freshly reconstituted rhTSH administered intravenously. In phase II, the same dogs were stimulated by using 100 microg of rhTSH frozen for 3 months at -20 degrees C. Phase III stimulation tests were performed by using 50 or 100 microg of freshly reconstituted or frozen rhTSH in healthy (n = 14), euthyroid sick (n = 11) and hypothyroid dogs (n = 9). A dose of 100 microg of rhTSH was judged more appropriate for dogs weighing more than 20 kg. Biological activity of rhTSH after freezing at -20 degrees C for up to 12 weeks was maintained. When stimulated, significant (P < 0.05) increases in total thyroxine concentration were observed only in healthy and euthyroid sick dogs. Results of this study show that the rhTSH stimulation test is able to differentiate euthyroidism from hypothyroidism in dogs.

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

PubMed

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

2015-04-01

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

Survival and growth of fresh and stored planting stock.

Treesearch

Robert H. Ruth

1953-01-01

Does planting stock that has been kept in storage survive and grow as well as freshly dug stock? This question is important because the ground at a forest nursery may still be frozen when spring planting time arrives in the warmer parts of the region. This means that seedlings for spring planting need to be dug in the fall, kept in cold storage over winter, and shipped...

Evaluating the effect of sample type on American alligator ( Alligator mississippiensis) analyte values in a point-of-care blood analyser

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

Hamilton, Matthew T.; Finger, John W.; Winzeler, Megan E.

The assessment of wildlife health has been enhanced by the ability of point-of-care (POC) blood analysers to provide biochemical analyses of non-domesticated animals in the field. However, environmental limitations (e.g. temperature, atmospheric humidity and rain) and lack of reference values may inhibit researchers from using such a device with certain wildlife species. Evaluating the use of alternative sample types, such as plasma, in a POC device may afford researchers the opportunity to delay sample analysis and the ability to use banked samples. In this study, we examined fresh whole blood, fresh plasma and frozen plasma (sample type) pH, partial pressuremore » of carbon dioxide (PCO 2), bicarbonate (HCO 3₋), total carbon dioxide (TCO 2), base excess (BE), partial pressure of oxygen (PO 2), oxygen saturation (sO 2) and lactate concentrations in 23 juvenile American alligators (Alligator mississippiensis) using an i-STAT CG4+ cartridge. Our results indicate that sample type had no effect on lactate concentration values (F 2,65 = 0.37, P = 0.963), suggesting that the i-STAT analyser can be used reliably to quantify lactate concentrations in fresh and frozen plasma samples. In contrast, the other seven blood parameters measured by the CG4+ cartridge were significantly affected by sample type. In conclusion, we were able to collect blood samples from all alligators within 2 min of capture to establish preliminary reference ranges for juvenile alligators based on values obtained using fresh whole blood.« less

Evaluating the effect of sample type on American alligator (Alligator mississippiensis) analyte values in a point-of-care blood analyser

PubMed Central

Hamilton, Matthew T.; Finger, John W.; Winzeler, Megan E.; Tuberville, Tracey D.

2016-01-01

The assessment of wildlife health has been enhanced by the ability of point-of-care (POC) blood analysers to provide biochemical analyses of non-domesticated animals in the field. However, environmental limitations (e.g. temperature, atmospheric humidity and rain) and lack of reference values may inhibit researchers from using such a device with certain wildlife species. Evaluating the use of alternative sample types, such as plasma, in a POC device may afford researchers the opportunity to delay sample analysis and the ability to use banked samples. In this study, we examined fresh whole blood, fresh plasma and frozen plasma (sample type) pH, partial pressure of carbon dioxide (PCO2), bicarbonate (HCO3−), total carbon dioxide (TCO2), base excess (BE), partial pressure of oxygen (PO2), oxygen saturation (sO2) and lactate concentrations in 23 juvenile American alligators (Alligator mississippiensis) using an i-STAT CG4+ cartridge. Our results indicate that sample type had no effect on lactate concentration values (F2,65 = 0.37, P = 0.963), suggesting that the i-STAT analyser can be used reliably to quantify lactate concentrations in fresh and frozen plasma samples. In contrast, the other seven blood parameters measured by the CG4+ cartridge were significantly affected by sample type. Lastly, we were able to collect blood samples from all alligators within 2 min of capture to establish preliminary reference ranges for juvenile alligators based on values obtained using fresh whole blood. PMID:27382469

Strategies for the management of OHSS: Results from freezing-all cycles.

PubMed

Borges, Edson; Braga, Daniela Paes Almeida Ferreira; Setti, Amanda S; Vingris, Livia S; Figueira, Rita Cássia S; Iaconelli, Assumpto

2016-03-01

To compare the use of GnRH agonist (GnRHa) or hCG trigger in potential OHSS patients undergoing freeze-all programs. We also compared the clinical outcomes when fresh versus freeze-thawed embryo transfers were performed in cycles with a high number of retrieved oocytes. The study included potential OHSS patients who received GnRHa (n=74) or hCG (n=49) trigger. The protocols were compared with respect to the clinical outcomes. We also compared the clinical outcomes of cycles in which hCG trigger was used and more than 20 MII oocytes were retrieved when: fresh embryo transfer protocol (n=153) or freeze-all protocol (n=123) were performed. A decreased serum estradiol level, a decreased number of retrieved oocytes, an increased MII retrieved rate, and decreased fertilization rate was observed in the hCG when compared with the GnRHa group. No significant differences were noted concerning clinical outcomes. When fresh cycles were compared with frozen-thawed cycles, the estradiol serum level and the number of cryopreserved embryos were higher in the frozen-thawed cycles. The clinical pregnancy rate was higher among freeze-all cycles, as well as the implantation and cumulative pregnancy rates, when compared with fresh embryo transfer cycles. The use of GnRHa trigger may be a good alternative to prevent the OHSS in patients presenting an extreme ovarian response to COS, leading to similar clinical outcomes, when compared with the traditional hCG trigger. Moreover, our findings demonstrated that the strategy of freezing-all embryos not only decreases the risk of OHSS but also leads to a better pregnancy rate.

Evaluating the effect of sample type on American alligator ( Alligator mississippiensis) analyte values in a point-of-care blood analyser

DOE PAGES

Hamilton, Matthew T.; Finger, John W.; Winzeler, Megan E.; ...

2016-01-01

The assessment of wildlife health has been enhanced by the ability of point-of-care (POC) blood analysers to provide biochemical analyses of non-domesticated animals in the field. However, environmental limitations (e.g. temperature, atmospheric humidity and rain) and lack of reference values may inhibit researchers from using such a device with certain wildlife species. Evaluating the use of alternative sample types, such as plasma, in a POC device may afford researchers the opportunity to delay sample analysis and the ability to use banked samples. In this study, we examined fresh whole blood, fresh plasma and frozen plasma (sample type) pH, partial pressuremore » of carbon dioxide (PCO 2), bicarbonate (HCO 3₋), total carbon dioxide (TCO 2), base excess (BE), partial pressure of oxygen (PO 2), oxygen saturation (sO 2) and lactate concentrations in 23 juvenile American alligators (Alligator mississippiensis) using an i-STAT CG4+ cartridge. Our results indicate that sample type had no effect on lactate concentration values (F 2,65 = 0.37, P = 0.963), suggesting that the i-STAT analyser can be used reliably to quantify lactate concentrations in fresh and frozen plasma samples. In contrast, the other seven blood parameters measured by the CG4+ cartridge were significantly affected by sample type. In conclusion, we were able to collect blood samples from all alligators within 2 min of capture to establish preliminary reference ranges for juvenile alligators based on values obtained using fresh whole blood.« less

Isolation and molecular characterization of Vibrio parahaemolyticus from fresh, low-temperature preserved, dried, and salted seafood products in two coastal areas of eastern China.

PubMed

Yang, Zhen-Quan; Jiao, Xin-An; Zhou, Xiao-Hui; Cao, Guo-Xiang; Fang, Wei-Ming; Gu, Rui-Xia

2008-07-31

A total of 1293 seafood samples from fishing farm, retail markets, restaurants and cooking rooms of hotels in Jiangsu province and Shanghai city of China were collected and analyzed for the prevalence of Vibrio parahaemolyticus during July to October in 2007. Two hundred and fifty one isolates of V. parahaemolyticus were identified, of which 8 isolates were positive for tdh and 2 were positive for trh gene. Three tdh positive isolates were identified from low-temperature preserved seafood samples and 5 isolates from fresh seafood samples, of these tdh positive isolates, 3 were positive in ORF8-PCR test. The genetic diversity among V. parahaemolyticus isolates was assessed using random amplified polymorphic DNA (RAPD)-PCR and the results showed that there were 33 different genetic patterns that were clustered into nine groups (groups A to I) at 82% similarity level. About 31.9% of the isolates belong to type III9d that were widely distributed in fresh, iced, frozen, dried and salted seafood samples. Seven tdh positive isolates belonged to group A and one belonged to group C, 2 trh positive isolates were type I10d belonging to group F, which was identical to that of reference strains isolated from patients. This study demonstrated genetic variability within V. parahaemolyticus isolates from seafood in Chinese markets and confirmed the presence of toxigenic V. parahaemolyticus not only in fresh but also in iced and frozen seafood products indicating that low-temperature preserved seafood might be also a vehicle for transmitting pathogenic V. parahaemolyticus.

Slow Freezing, but Not Vitrification Supports Complete Spermatogenesis in Cryopreserved, Neonatal Sheep Testicular Xenografts

PubMed Central

Pukazhenthi, Budhan S.; Nagashima, Jennifer; Travis, Alexander J.; Costa, Guilherme M.; Escobar, Enrique N.; França, Luiz R.; Wildt, David E.

2015-01-01

The ability to spur growth of early stage gametic cells recovered from neonates could lead to significant advances in rescuing the genomes of rare genotypes or endangered species that die unexpectedly. The purpose of this study was to determine, for the first time, the ability of two substantially different cryopreservation approaches, slow freezing versus vitrification, to preserve testicular tissue of the neonatal sheep and subsequently allow initiation of spermatogenesis post-xenografting. Testis tissue from four lambs (3-5 wk old) was processed and then untreated or subjected to slow freezing or vitrification. Tissue pieces (fresh, n = 214; slow freezing, then thawing, n = 196; vitrification, then warming, n = 139) were placed subcutaneously under the dorsal skin of SCID mice and then grafts recovered and evaluated 17 wk later. Grafts from fresh and slow frozen tissue contained the most advanced stages of spermatogenesis, including normal tubule architecture with elongating spermatids in ~1% (fresh) and ~10% (slow frozen) of tubules. Fewer than 2% of seminiferous tubules advanced to the primary spermatocyte stage in xenografts derived from vitrified tissue. Results demonstrate that slow freezing of neonatal lamb testes was far superior to vitrification in preserving cellular integrity and function after xenografting, including allowing ~10% of tubules to retain the capacity to resume spermatogenesis and yield mature spermatozoa. Although a first for any ruminant species, findings also illustrate the importance of preemptive studies that examine cryo-sensitivity of testicular tissue before attempting this type of male fertility preservation on a large scale. PMID:25923660

Fresh, Frozen or Canned Fruits and Vegetables: All Can Be Healthy Choices!

MedlinePlus

... a Healthy Heart Healthy Kids Our Kids Programs Childhood Obesity What is childhood obesity Overweight in Children BMI in Children Is Childhood Obesity an Issue in Your Home? Addressing Your Child's ...

PARAMETERS OF TEXTURE CHANGE IN PROCESSED FISH: MYOSIN DENATURATION.

PubMed

Chu, George Hao; Sterling, Clarence

1970-03-01

The white muscle of the Sacramento blackfish (Orthodon microlepidotus) was processed by freezing, dehydration, and cooking. Myosin was extracted immediately afterwards or following a period of storage in order to examine evidence for denaturation. The tests used were the solubility of whole muscle protein and the intrinsic viscosity, isoelectric point, ATPase activity, ultra-violet absorption spectrum, and optical rotatory dispersion of purified myosin extract. Almost all measures used showed that denaturation increased in the order: fresh < frozen < frozen-stored < dehydrated < dehydrated-stored < cooked.

Frozen embryo transfer prevents the detrimental effect of high estrogen on endometrium receptivity.

PubMed

Adeviye Erşahin, Aynur; Acet, Mustafa; Erşahin, Suat Süphan; Dokuzeylül Güngör, Nur

2017-03-15

To investigate whether serum levels of estradiol affect reproductive outcomes of normoresponder women undergoing fresh embryo transfer (ET) versus frozen-thawed ET (FET). Two hundred fifty-five normoresponder women underwent fresh ET in their first or second in vitro fertilization cycle. Ninety-two women with negative pregnacy test results underwent FET. Clinical and ongoing pregnancy rates, implantation, and live birth rates of women undergoing fresh ET versus FET were compared. One hundred forty-seven (57.65%) out of the 255 normoresponder women receiving FET had positive beta-human chorionic gonadotrophin (hCG) results. The remaining 108 women had negative beta-hCG results. The clinical pregnancy rates of the fresh ET group were found as 55.69% (n=142). Ninety-two of the 108 women with failed pregnancies underwent FET; 72.83% had positive beta-hCG results (n=67), and 70.65% had clinical pregnancy (n=65). Both biochemical and clinical pregnancy rates of women undergoing FET increased significantly (p<0.012 and p<0.013, respectively). Ongoing pregnancy (60.87% vs. 52.94%) and live birth rates (59.87% vs. 48.63%) were similar in both fresh and FET groups. Serum E2 levels of women who failed to conceive were significantly higher than those women did conceive. Serum progesterone levels of women who conceived versus those that did not were similar. The detrimental effect of high serum estradiol levels on endometrial receptivity could be prevented by FET.

Frozen embryo transfer prevents the detrimental effect of high estrogen on endometrium receptivity

PubMed Central

Erşahin, Aynur Adeviye; Acet, Mustafa; Erşahin, Suat Süphan; Dokuzeylül Güngör, Nur

2017-01-01

Objective: To investigate whether serum levels of estradiol affect reproductive outcomes of normoresponder women undergoing fresh embryo transfer (ET) versus frozen-thawed ET (FET). Material and Methods: Two hundred fifty-five normoresponder women underwent fresh ET in their first or second in vitro fertilization cycle. Ninety-two women with negative pregnacy test results underwent FET. Clinical and ongoing pregnancy rates, implantation, and live birth rates of women undergoing fresh ET versus FET were compared. Results: One hundred forty-seven (57.65%) out of the 255 normoresponder women receiving FET had positive beta-human chorionic gonadotrophin (hCG) results. The remaining 108 women had negative beta-hCG results. The clinical pregnancy rates of the fresh ET group were found as 55.69% (n=142). Ninety-two of the 108 women with failed pregnancies underwent FET; 72.83% had positive beta-hCG results (n=67), and 70.65% had clinical pregnancy (n=65). Both biochemical and clinical pregnancy rates of women undergoing FET increased significantly (p<0.012 and p<0.013, respectively). Ongoing pregnancy (60.87% vs. 52.94%) and live birth rates (59.87% vs. 48.63%) were similar in both fresh and FET groups. Serum E2 levels of women who failed to conceive were significantly higher than those women did conceive. Serum progesterone levels of women who conceived versus those that did not were similar. Conclusion: The detrimental effect of high serum estradiol levels on endometrial receptivity could be prevented by FET. PMID:28506949

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.

Cryoprotective roles of trehalose and alginate oligosaccharides during frozen storage of peeled shrimp (Litopenaeus vannamei).

PubMed

Zhang, Bin; Wu, Hai-Xiao; Yang, Hui-Cheng; Xiang, Xing-Wei; Li, Hai-Bo; Deng, Shang-Gui

2017-08-01

Cryoprotective saccharides are widely accepted additives that reduce thawing loss, maintain texture, and retard protein denaturation in the frozen seafood. The present study aimed to investigate the roles of trehalose and alginate oligosaccharides on cryoprotection of frozen shrimp, primarily focusing on the interactions between myosin and saccharide molecules using a molecular dynamics (MD) simulation analysis. The results indicated that soaking in the trehalose and alginate oligosaccharides solutions markedly reduced thawing and cooking losses in frozen shrimp, with respective values decreasing to 6.02%, 8.14%, and 5.99%, 8.19% after 9weeks of storage, which were significantly lower than that of fresh water treatment (9.75% and 15.09%). Our assumption was that water replacement played a leading role in cryoprotection, as shown in previous experimental results and reports. Furthermore, homology modeling and MD simulations confirmed that trehalose and alginate oligosaccharides substituted the water molecules around the myosin surface by forming hydrogen bonds with polar residues of amino acids, thereby stabilizing the structures in the absence of water during frozen storage. These conditions affected the flexibility of particular amino acid residues, enhanced the residue cross correlations within the two chains of myosin, and also increased the total interaction energy between myosin and water/saccharide molecules, thereby leading to an increase in protein stability. Finally, by comparing the experimental results to that of MD simulation, significant positive correlation existed between saccharides and the stabilization of myosin in shrimp muscle. The findings of the present study may help better understand the cryoprotective mechanisms of saccharides in frozen shrimp, and the two saccharides may be potentially used as alternative additives in seafood to maintain better quality during frozen storage. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of home freezing and Italian style of cooking on antioxidant activity of edible vegetables.

PubMed

Danesi, F; Bordoni, A

2008-08-01

In this study, we analyzed the modifications of antioxidant activity consequent to 3 typical home cooking practices (steaming, boiling, and microwave cooking) in fresh and home frozen vegetables. Six different vegetable species were examined: carrots (Daucus carota L.), zucchini (Cucurbita pepo L.), tomatoes (Solanumn lycopersicum L.), green beans (Phaseolus vulgaris L.), peas (Pisum sativum L.), and yellow peppers (Capsicum annuum L.). All vegetables were conventional products and were analyzed in season to minimize differences due to agricultural practice and storage. Cooking and freezing are generally regarded as destructive to antioxidants, and this has fostered a belief among many consumers that raw vegetables are nutritionally superior to their frozen and/or cooked forms. In the current study, we provide evidence that this is not always the case.

Selection of euploid blastocysts for cryopreservation with array comparative genomic hybridization (aCGH) results in increased implantation rates in subsequent frozen and thawed embryo transfer cycles

PubMed Central

2013-01-01

Background In assisted reproductive treatments, embryos remaining after fresh embryo transfer are usually selected for cryopreservation based on traditional morphology assessment. Our previous report has demonstrated that array comparative genomic hybridization (aCGH) screening for IVF patients with good prognosis significantly improves clinical and ongoing pregnancy rates in fresh embryo transfer cycles. The current study further investigates the efficiency of applying aCGH in the selection of euploid embryos for cryopreservation as related to pregnancy and implantation outcomes in subsequent frozen embryo transfer (FET) cycles. Methods First-time IVF patients with good prognosis undergoing fresh single embryo transfer and having at least one remaining blastocyst for cryopreservation were prospectively randomized into two groups: 1) Group A patients had embryos assessed by morphology first and then by aCGH screening of trophectoderm cells and 2) Group B patients had embryos evaluated by morphology alone. All patients had at least one blastocyst available for cryopreservation after fresh embryo transfer. There were 15 patients in Group A and 23 patients in Group B who failed to conceive after fresh embryo transfer and completed the FET cycles. Blastocyst survival and implantation rates were compared between the two groups. Results There were no significant differences in blastocyst survival rates between Group A and Group B (90.9% vs. 91.3%, respectively; p >0.05). However, a significantly higher implantation rate was observed in the morphology assessment plus aCGH screening group compared to the morphology assessment alone group (65.0% vs. 33.3%, respectively; p = 0.038). There was no miscarriage observed in Group A while a 16.7% miscarriage rate was recorded in Group B (0% vs. 16.7%, respectively; p >0.05). Conclusions While aCGH screening has been recently applied to select euploid blastocysts for fresh transfer in young, low-risk IVF patients, this is the first prospective study on the impact of aCGH specifically on blastocyst survival and implantation outcomes in the subsequent FET cycles of IVF patients with good prognosis. The present study demonstrates that aCGH screening of blastocysts prior to cryopreservation significantly improves implantation rates and may reduce the risk of miscarriage in subsequent FET cycles. Further randomized clinical studies with a larger sample size are needed to validate these preliminary findings. PMID:23937723

Tissue Quality Assessment Using a Novel Direct Elasticity Assessment Device (The E-Finger): A Cadaveric Study of Prostatectomy Dissection

PubMed Central

Good, Daniel W.; Khan, Ashfaq; Hammer, Steven; Scanlan, Paul; Shu, Wenmiao; Phipps, Simon; Parson, Simon H.; Stewart, Grant D.; Reuben, Robert; McNeill, S. Alan

2014-01-01

Introduction Minimally invasive radical prostatectomy (RP) (robotic and laparoscopic), have brought improvements in the outcomes of RP due to improved views and increased degrees of freedom of surgical devices. Robotic and laparoscopic surgeries do not incorporate haptic feedback, which may result in complications secondary to inadequate tissue dissection (causing positive surgical margins, rhabdosphincter damage, etc). We developed a micro-engineered device (6 mm2 sized) [E-finger]) capable of quantitative elasticity assessment, with amplitude ratio, mean ratio and phase lag representing this. The aim was to assess the utility of the device in differentiating peri-prostatic tissue types in order to guide prostate dissection. Material and Methods Two embalmed and 2 fresh frozen cadavers were used in the study. Baseline elasticity values were assessed in bladder, prostate and rhabdosphincter of pre-dissected embalmed cadavers using the micro-engineered device. A measurement grid was created to span from the bladder, across the prostate and onto the rhabdosphincter of fresh frozen cadavers to enable a systematic quantitative elasticity assessment of the entire area by 2 independent assessors. Tissue was sectioned along each row of elasticity measurement points, and stained with haematoxylin and eosin (H&E). Image analysis was performed with Image Pro Premier to determine the histology at each measurement point. Results Statistically significant differences in elasticity were identified between bladder, prostate and sphincter in both embalmed and fresh frozen cadavers (p = <0.001). Intra-class correlation (ICC) reliability tests showed good reliability (average ICC = 0.851). Sensitivity and specificity for tissue identification was 77% and 70% respectively to a resolution of 6 mm2. Conclusions This cadaveric study has evaluated the ability of our elasticity assessment device to differentiate bladder, prostate and rhabdosphincter to a resolution of 6 mm2. The results provide useful data for which to continue to examine the use of elasticity assessment devices for tissue quality assessment with the aim of giving haptic feedback to surgeons performing complex surgery. PMID:25384014

Long-Term Frozen Storage of Urine Samples: A Trouble to Get PCR Results in Schistosoma spp. DNA Detection?

PubMed Central

Fernández-Soto, Pedro; Velasco Tirado, Virginia; Carranza Rodríguez, Cristina; Pérez-Arellano, José Luis; Muro, Antonio

2013-01-01

Background Human schistosomiasis remains a serious worldwide public health problem. At present, a sensitive and specific assay for routine diagnosis of schistosome infection is not yet available. The potential for detecting schistosome-derived DNA by PCR-based methods in human clinical samples is currently being investigated as a diagnostic tool with potential application in routine schistosomiasis diagnosis. Collection of diagnostic samples such as stool or blood is usually difficult in some populations. However, urine is a biological sample that can be collected in a non-invasive method, easy to get from people of all ages and easy in management, but as a sample for PCR diagnosis is still not widely used. This could be due to the high variability in the reported efficiency of detection as a result of the high variation in urine samples’ storage or conditions for handling and DNA preservation and extraction methods. Methodology/Principal Findings We evaluate different commercial DNA extraction methods from a series of long-term frozen storage human urine samples from patients with parasitological confirmed schistosomiasis in order to assess the PCR effectiveness for Schistosoma spp. detection. Patientś urine samples were frozen for 18 months up to 7 years until use. Results were compared with those obtained in PCR assays using fresh healthy human urine artificially contaminated with Schistosoma mansoni DNA and urine samples from mice experimentally infected with S. mansoni cercariae stored frozen for at least 12 months before use. PCR results in fresh human artificial urine samples using different DNA based extraction methods were much more effective than those obtained when long-term frozen human urine samples were used as the source of DNA template. Conclusions/Significance Long-term frozen human urine samples are probably not a good source for DNA extraction for use as a template in PCR detection of Schistosoma spp., regardless of the DNA method of extraction used. PMID:23613907

Long-term frozen storage of urine samples: a trouble to get PCR results in Schistosoma spp. DNA detection?

PubMed

Fernández-Soto, Pedro; Velasco Tirado, Virginia; Carranza Rodríguez, Cristina; Pérez-Arellano, José Luis; Muro, Antonio

2013-01-01

Human schistosomiasis remains a serious worldwide public health problem. At present, a sensitive and specific assay for routine diagnosis of schistosome infection is not yet available. The potential for detecting schistosome-derived DNA by PCR-based methods in human clinical samples is currently being investigated as a diagnostic tool with potential application in routine schistosomiasis diagnosis. Collection of diagnostic samples such as stool or blood is usually difficult in some populations. However, urine is a biological sample that can be collected in a non-invasive method, easy to get from people of all ages and easy in management, but as a sample for PCR diagnosis is still not widely used. This could be due to the high variability in the reported efficiency of detection as a result of the high variation in urine samples' storage or conditions for handling and DNA preservation and extraction methods. We evaluate different commercial DNA extraction methods from a series of long-term frozen storage human urine samples from patients with parasitological confirmed schistosomiasis in order to assess the PCR effectiveness for Schistosoma spp. detection. Patients urine samples were frozen for 18 months up to 7 years until use. Results were compared with those obtained in PCR assays using fresh healthy human urine artificially contaminated with Schistosoma mansoni DNA and urine samples from mice experimentally infected with S. mansoni cercariae stored frozen for at least 12 months before use. PCR results in fresh human artificial urine samples using different DNA based extraction methods were much more effective than those obtained when long-term frozen human urine samples were used as the source of DNA template. Long-term frozen human urine samples are probably not a good source for DNA extraction for use as a template in PCR detection of Schistosoma spp., regardless of the DNA method of extraction used.

In vitro survival of fresh and frozen/thawed bovine demi-embryos.

PubMed

Lucas-Hahn, A; Niemann, H

1991-10-01

Three experiments were conducted to investigate the effects of type of culture medium in freshly bisected bovine embryos and the effects of agar embedding and of 1.2 propanediol (PROH) as the cryoprotectant in frozen/thawed bisected bovine embryos on development in vitro. A total of 265 bovine embryos were used as controls or were microsurgically bisected and were cultured in vitro for 48 hours and development was determined 24 and 48 hours after the onset of culture. Whitten's medium supported more (P<0.05) intact and demi-embryos to grow to expanded blastocysts (92.9 and 73.1%, respectively) compared with Ham's F10 (43.8 and 26.3%, respectively) and PBS (53.8 and 12.5%, respectively). Embedding in agar and culture in Whitten's medium resulted in a higher (P<0.05) percentage of in vitro development of frozen/thawed demi-embryos after 24 hours than the freezing of nonembedded demi-embryos (44.1 versus 19.6%, respectively). This difference disappeared, however, after a 48 hours culture period (17.6 versus 11.8%, respectively). Following freezing in PROH, survival rates of 40 and 28%, respectively after 24 hours of culture were obtained for intact and demi-embryos. The respective percentages after 48 hours were 8.6 and 16%. Since neither embedding in agar nor the use of PROH as the cryoprotectant resulted in high survival rates of frozen/thawed demi-embryos in vitro, new freezing procedures are needed to overcome the sensitivity of demi-embryos to freezing and thawing.

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

PubMed

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

2016-03-01

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

Effects of retail style or food service style packaging type and storage time on sensory characteristics of bacon manufactured from commercially sourced pork bellies.

PubMed

Lowe, B K; Bohrer, B M; Holmer, S F; Boler, D D; Dilger, A C

2014-06-01

Objectives were to characterize differences in pork bellies that were stored frozen for different durations prior to processing and characterize sensory properties of the bacon derived from those bellies when stored in either retail or food service style packaging. Bellies (n = 102) were collected from 4 different time periods, fresh bellies (never frozen) and bellies frozen for 2, 5, or 7 mo, and manufactured into bacon under commercial conditions. Food service bacon was packaged in oxygen-permeable polyvinyl lined boxes layered on wax-covered lined paper and blast frozen (-33 °C) for 45 or 90 d after slicing. Retail bacon was vacuum-packaged in retail packages and refrigerated (2 °C) in the dark for 60 or 120 d after slicing. At the end of respective storage times after slicing, bacon was analyzed for sensory attributes and lipid oxidation. Off-flavor and oxidized odor of bacon increased (P < 0.01) with increasing storage time in both packaging types. Lipid oxidation increased (P < 0.01) as storage time increased from day 0 to day 45 in food service packaged bacon from frozen bellies, but was unchanged (P ≥ 0.07) with time in food service packaged bacon from fresh bellies. Lipid oxidation was also unchanged (P ≥ 0.21) over time in retail packaged bacon, with the exception of bellies frozen for 5 mo, which was increased from day 0 to day 90. Overall, off-flavor, oxidized odor, and lipid oxidation increased as storage time after processing increased. Freezing bellies before processing may exacerbate lipid oxidation as storage time after processing was extended. Bacon can be packaged and managed several different ways before it reaches the consumer. This research simulated food service (frozen) and retail packaged (refrigerated) bacon over a range of storage times after slicing. Off-flavor and oxidized odor increased as storage time after processing increased in both packaging types. Lipid oxidation increased as storage time after slicing increased to a greater extent in food service packaging. © 2014 Institute of Food Technologists®

Access to Fresh Fruits and Vegetables in School Lunches: A Policy Analysis.

PubMed

Schultz, Celeste; Thorlton, Janet

2018-01-01

Consumption of fresh fruits and vegetables helps to reduce childhood obesity and improves academic achievement and attendance. However, providing fresh fruits and vegetables is challenging for some schools due to cost, administrative burden, and concern for food waste. To address these challenges, the Fruit and Vegetable Access for Children Act proposes to allow federally funded programs to substitute fresh fruits and vegetables with canned, frozen, or pureed versions. In this policy analysis, we propose options for providing fresh fruits and vegetables to children enrolled in the National School Lunch Program. We recommend that school nurses actively facilitate the process of obtaining fresh fruits and vegetables by being appointed members of Team Nutrition giving them authority to collaborate with local famers, entrepreneurs, and land-grant universities in Farm to School Programs. This strategy empowers school nurses in promoting healthy eating habits, reducing obesity, and improving academic performance and school attendance.

DASH diet to lower high blood pressure

MedlinePlus

... breakfast. For dessert, choose fresh fruit or low-fat frozen yogurt instead of high-calorie sweets, such as cakes or pies. Choose healthy snacks, such as unsalted rice cakes or popcorn, raw vegetables, or yogurt. Dried fruits, seeds, and nuts ...

75 FR 68505 - National Organic Program; Proposed Amendments to the National List of Allowed and Prohibited...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-08

... including fresh and frozen baked goods, pastas, salad dressings, confectionery, processed cheese spreads... extraction or treatment is used in its manufacture. It functions to retain moisture in baked goods, pastas...

40 CFR 125.133 - What special definitions apply to this subpart?

Code of Federal Regulations, 2010 CFR

2010-07-01

... Subcategories of the Oil and Gas Extraction Point Source Category Effluent Guidelines in 40 CFR 435.10 or 40 CFR..., floating, mobile, facility engaged in the processing of fresh, frozen, canned, smoked, salted or pickled...

Dietary α-linolenic acid from flaxseed oil or eicosapentaenoic and docosahexaenoic acids from fish oil differentially alter fatty acid composition and characteristics of fresh and frozen-thawed bull semen.

PubMed

Moallem, Uzi; Neta, Noam; Zeron, Yoel; Zachut, Maya; Roth, Zvi

2015-04-15

Incorporation rates of dietary omega-3 (n-3) fatty acids (FAs) from different sources into bull plasma and sperm and the effects on physiological characteristics of fresh and frozen-thawed semen were determined. Fifteen fertile bulls were assigned to three treatment groups and supplemented for 13 weeks with encapsulated fat: (1) SFA-360 g/d per bull saturated FA; (2) FLX-450 g/d per bull providing 84.2 g/d C18:3n-3 (α-linolenic acid) from flaxseed oil; and (3) FO-450 g/d per bull providing 8.7 g/d C20:5n-3 (eicosapentaenoic acid) and 6.5 g/d C22:6n-3 (docosahexaenoic acid, DHA) from fish oil. Blood samples were taken every 2 weeks and semen was collected weekly. With respect to the FA supplements, the proportion of α-linolenic acid in plasma increased in the FLX bulls, whereas that of DHA was increased in the FO bulls, within 2 weeks. However, changes in the sperm FA fraction were first expressed in the sixth week of supplementation: in the FO and FLX bulls the DHA proportion increased (P < 0.001), whereas that of C22:5n-6 FAs (docosapentaenoic acid [DPA] n-6) decreased (P < 0.001). Sperm motility and progressive motility in fresh semen were higher (P < 0.05), and the fading rate tended to be lower in the FLX than in FO bulls (P < 0.06). Furthermore, sperm motility, progressive motility, and velocity in frozen-thawed semen were higher in FLX than in the other groups (P < 0.008). These findings indicate that the proportion of DHA in sperm can be increased at the expense of DPAn-6 by either FO or FLX supplementation, indicating de novo elongation and desaturation of short- into longer-chain n-3 FAs in testes. Furthermore, the moderate exchange of DHA and DPAn-6 in the FLX group's sperm was associated with changes in the characteristics of both fresh and frozen-thawed semen, suggesting the importance of the ratio between these two FAs for sperm structure and function. Copyright © 2015 Elsevier Inc. All rights reserved.

Evaluation of the BinaxNOW® Streptococcus pneumoniae antigen test on fresh, frozen and concentrated urine samples in elderly patients with and without community-acquired pneumonia.

PubMed

Saukkoriipi, Annika; Pascal, Thierry; Palmu, Arto A

2016-02-01

We evaluated the BinaxNOW® urine antigen test in elderly. For fresh un-concentrated urine samples, the sensitivity for pneumococcal pneumonia was 63% and specificity 97%. After freezing and concentration, the results comparable to positive control line in intensity at 60 min gave high sensitivity (81%) with no loss in specificity (96%). Copyright © 2015 Elsevier B.V. All rights reserved.

Waterborne Commerce of the United States, Calendar Year 1982. Part 3. Waterways and Harbors, Great Lakes.

DTIC Science & Technology

1984-07-01

34 2099 MISCELLANE1uS FOOD ONUUCT- 77,36 2 986 951 4,02:...........--.--.,...... lll TORACCn MANUFACT ,RES........... . ........... ... "..:.09...0102 Barley and rye 0103 Corn Group 20- Food and Kindred Products 0104 OatsS0105 Rice 2011 Meat, fresh, chilled, or frozen 0106 Sorghum Grains 2012 Meat...marine Products 2094 Groceries 2099 Miscellaneous food products 0911 Fresh fish, except shellfish 0912 Shellfish, except prepared or preserved Group

Lipaemic plasma induces haemolysis in resuspended red cell concentrate.

PubMed

Bashir, S; Wiltshire, M; Cardigan, R; Thomas, S

2013-04-01

We investigated whether haemolysis in red cells suspended in plasma was affected by the lipid content and/or methylene blue (MB) treatment of fresh-frozen plasma (FFP). We also investigated whether haemolysis was affected by the conditions under which lipaemic plasma was stored. Study 1: Visibly lipaemic (n = 22) or nonlipaemic FFP (n = 24) units were thawed, pooled and split into identical pairs, one of which was MB treated. These units were used to resuspend red cell concentrates (RCC) and tested for haemolysis immediately and after 24 and 48 h of storage at 2-6°C. Study 2: Fresh plasma was aliquoted into 15-ml tubes and stored in one of four ways as follows: room temperature; 2-6°C; frozen and thawed; or twice frozen and thawed. A sample of RCC was resuspended in each of these plasmas and haemolysis measured after 2 h. Study 3: Plasma was divided into 15-ml tubes and stored as in study 2 followed by storage left standing upright in a refrigerator (2-6°C) for 24 h (with the exception of the room temperature sample). Plasma was separated into top, middle and bottom fractions and used to resuspend RCC that were assessed for haemolysis after 2 h. The levels of haemolysis in RCC were immediately greater when suspended in lipaemic plasma (0·70 ± 0·53% v 0·05 ± 0·06% for nonlipaemic plasma), which increased further on subsequent storage for 48 h (1·22 ± 0·40% v 0·15 ± 0·14% for nonlipaemic plasma). This was irrespective of whether plasma was MB treated. Lipaemic plasma stored frozen and then thawed resulted in the greatest haemolysis. In lipaemic plasma stored at 2-6°C, the chylomicron-rich top fraction caused the highest level of haemolysis. Haemolysis in red cells is increased in those suspended in lipaemic plasma and is dependent upon the storage conditions of that plasma prior to suspension. These data are relevant to the choice of plasma used to suspend red cells for neonatal exchange transfusion. © 2012 The Author(s). Vox Sanguinis © 2012 International Society of Blood Transfusion.

Freezing of Rat Tibiae at -20°C Does Not Affect the Mechanical Properties of Intramedullary Bone/Implant-Interface: Brief Report

PubMed Central

Diefenbeck, Michael; Mückley, Thomas; Zankovych, Sergiy; Bossert, Jörg; Jandt, Klaus D; Schrader, Christian; Schmidt, Jürgen; Finger, Ulrich; Faucon, Mathilde

2011-01-01

Background: The effects of freezing-thawing cycles on intramedullary bone-implant interfaces have been studied in a rat model in mechanical pull-out tests. Implants: Twenty TiAl6V4 rods (Ø 0.8 mm, length 10 mm) implanted in rat tibiae Methods: 10 rats underwent bilateral tibial implantation of titanium rods. At eight weeks, the animals were sacrificed and tibiae harvested for biomechanical testing. Eight tibiae were frozen and stored at -20°C for 14 days, the remaining eight were evaluated immediately post-harvest. Pull-out tests were used to determine maximum force and interfacial shear strength. Results: There were no significant differences between fresh and those of the frozen-thawed group in maximum force or in interfacial shear strength. Conclusion: Frozen Storage of rat tibiae containing implants at -20° C has no effects on the biomechanical properties of Bone/ Implant interface. PMID:21760868

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

PubMed Central

Roberts, Diane

1972-01-01

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

Permeability of tritiated water through human cervical and vaginal tissue.

PubMed

Sassi, Alexandra B; McCullough, Kristy D; Cost, Marilyn R; Hillier, Sharon L; Rohan, Lisa Cencia

2004-08-01

The increased incidence of human immunodeficiency virus infection in women has identified an urgent need to develop a female-controlled method to prevent acquisition of human immunodeficiency virus and other sexually transmitted diseases. Women would apply the product intravaginally before intercourse. Development of such a product requires a better understanding of the permeability characteristics of the tissues with which such products would come into contact. However, limited studies have been performed in this area. In the present study, water permeability of fresh human cervical and vaginal tissue was evaluated. The average apparent permeability coefficient was found to be 8 x 10(-5) cm/s for fresh human cervical tissue and 7 x 10(-5) cm/s for fresh human vaginal tissue. Considering the lack of regularity in obtaining cervical and vaginal tissue from surgical specimens, additional tests were performed to evaluate the effect of freezing on tritiated water permeability. No statistically significant differences were observed in the permeability values obtained when comparing fresh versus frozen tissues. Copyright 2004 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 93:2009-2016, 2004

Update on meat irradiation

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

Olson, D.G.

1997-12-01

The irradiation of meat and poultry in the United States is intended to eliminate pathogenic bacteria from raw product, preferably after packaging to prevent recontamination. Irradiation will also increase the shelf life of raw meat and poultry products approximately two to three times the normal shelf life. Current clearances in the United States are for poultry (fresh or frozen) at doses from 1.5 to 3.0 kGy and for fresh pork at doses from 0.3 to 1.0 kGy. A petition for the clearance of all red meat was submitted to the Food and Drug Administration (FDA) in July 1994. The petitionmore » is for clearances of fresh meat at doses from 1.5 to 4.5 kGy and for frozen meat at {approximately}2.5 to 7.5 kGy. Clearance for red meat is expected before the end of 1997. There are 28 countries that have food irradiation clearances, of which 18 countries have clearances for meat or poultry. However, there are no uniform categories or approved doses for meat and poultry among the countries that could hamper international trade of irradiated meat and poultry.« less

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

PubMed

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

2017-09-01

With avian sperm cryopreservation protocols, the most widely used cryoprotectants (CPAs) are the glycerol (GLY; in gradual freezing: in-straw freezing method), and the dimethylacetamide (DMA; in pellets by plunging into liquid nitrogen: in-pellet rapid freezing method). Use of both methods results in a small portion of thawed live sperm with lesser fertilizing ability compared with the semen samples immediately after collection. This study was conducted to assess the pre-freezing damage occurring to the sperm due to the interaction with the cryoprotectants (CPAs) GLY (8%) and DMA (5%), as well as the post-freezing damage resulting from both freezing methods Data for each treatment, in fresh and frozen-thawed samples, were compared for sperm motility, fertilizing capacity and sperm-egg penetration holes/germinal disc (SP holes/GD). Hens (n=50) were artificially inseminated (10 hens/treatment) six times with 3day intervals between inseminations. The treatment of fresh sperm with DMA led to a reduction (P<0.05) in the count of SP holes/GD (21.4) and the fertility rate (66.7%). The addition and elimination of GLY in fresh samples resulted in a lesser (P<0.05) number of SP holes/GD (11.8) and the fertility rate (i.e., 50.0%). The number of SP-holes/GD was least in frozen-thawed samples using both DMA and GLY (14.2 and 9.2, respectively). The fertility rate when using semen frozen with DMA in- pellets was greater (P<0.05) than with use of semen that had been frozen using GLY in straws (46.4% compared with 31.3%). The reduction in fertility compared with the control when semen was cryopreserved using GLY was 64.1%; the GLY addition and elimination was responsible for two thirds of this reduction. The reduction in fertility when using semen cryopreserved with DMA was 46.7%; half of the reduction was attributed to the treatment with DMA. In conclusion, the mechanical damage attributed to the process for reducing GLY concentrations was more harmful to sperm fertilizing capacity than the toxicity of DMA and freeze/thaw process. For both freezing methods, the amount of sperm cryo-damage was similar, when the damage attributed to the CPA addition and elimination process was excluded. Copyright © 2017 Elsevier B.V. All rights reserved.

Use of recombinant human thyroid-stimulating hormone for thyrotropin stimulation test in healthy, hypothyroid and euthyroid sick dogs

PubMed Central

Daminet, Sylvie; Fifle, Lyanne; Paradis, Manon; Duchateau, Luc; Moreau, Maxim

2007-01-01

Recombinant human thyroid-stimulating hormone (rhTSH) was evaluated for the diagnosis of canine hypothyroidism, using TSH response tests. Phase I stimulation tests were performed in 6 healthy dogs weighing over 20 kg, using 50 and then 100 μg of freshly reconstituted rhTSH administered intravenously. In phase II, the same dogs were stimulated by using 100 μg of rhTSH frozen for 3 months at −20°C. Phase III stimulation tests were performed by using 50 or 100 μg of freshly reconstituted or frozen rhTSH in healthy (n = 14), euthyroid sick (n = 11) and hypothyroid dogs (n = 9). A dose of 100 μg of rhTSH was judged more appropriate for dogs weighing more than 20 kg. Biological activity of rhTSH after freezing at −20°C for up to 12 weeks was maintained. When stimulated, significant (P < 0.05) increases in total thyroxine concentration were observed only in healthy and euthyroid sick dogs. Results of this study show that the rhTSH stimulation test is able to differentiate euthyroidism from hypothyroidism in dogs. PMID:18189051

Effect of chilling and cryopreservation on expression of Pax genes in zebrafish (Danio rerio) embryos and blastomeres.

PubMed

Lin, C; Spikings, E; Zhang, T; Rawson, D M

2009-08-01

Cryopreservation is now common practice in the fields of aquaculture, conservation and biomedicine. However, there is a lack of information on the effect of chilling and cryopreservation at the molecular level. In the present study, we used real-time RT-PCR analysis to determine the effect of chilling and cryopreservation on expression of Pax2a, Pax2b, Pax5 and Pax8 which constitute one subgroup of the Pax gene family. As intact embryos of zebrafish have not yet been successfully cryopreserved, we have used two alternatives: chilling of intact embryos and cryopreservation of isolated blastomeres. Cryopreservation was found to affect the normal pattern of gene expression in zebrafish embryonic blastomeres. The trends, profile changes, in expression of Pax2a and Pax5 occurred to a lesser extent in frozen-thawed blastomeres than in fresh blastomeres whilst the opposite was true for Pax8. The trends in expression of Pax2b were delayed in frozen-thawed blastomeres compared to fresh blastomeres. Cryopreservation can therefore disrupt normal gene expression patterns in zebrafish embryonic blastomeres which could have a detrimental effect on embryo development.

A Comparison of RNA-Seq Results from Paired Formalin-Fixed Paraffin-Embedded and Fresh-Frozen Glioblastoma Tissue Samples

PubMed Central

Esteve-Codina, Anna; Arpi, Oriol; Martinez-García, Maria; Pineda, Estela; Mallo, Mar; Gut, Marta; Carrato, Cristina; Rovira, Anna; Lopez, Raquel; Tortosa, Avelina; Dabad, Marc; Del Barco, Sonia; Heath, Simon; Bagué, Silvia; Ribalta, Teresa; Alameda, Francesc; de la Iglesia, Nuria

2017-01-01

The molecular classification of glioblastoma (GBM) based on gene expression might better explain outcome and response to treatment than clinical factors. Whole transcriptome sequencing using next-generation sequencing platforms is rapidly becoming accepted as a tool for measuring gene expression for both research and clinical use. Fresh frozen (FF) tissue specimens of GBM are difficult to obtain since tumor tissue obtained at surgery is often scarce and necrotic and diagnosis is prioritized over freezing. After diagnosis, leftover tissue is usually stored as formalin-fixed paraffin-embedded (FFPE) tissue. However, RNA from FFPE tissues is usually degraded, which could hamper gene expression analysis. We compared RNA-Seq data obtained from matched pairs of FF and FFPE GBM specimens. Only three FFPE out of eleven FFPE-FF matched samples yielded informative results. Several quality-control measurements showed that RNA from FFPE samples was highly degraded but maintained transcriptomic similarities to RNA from FF samples. Certain issues regarding mutation analysis and subtype prediction were detected. Nevertheless, our results suggest that RNA-Seq of FFPE GBM specimens provides reliable gene expression data that can be used in molecular studies of GBM if the RNA is sufficiently preserved. PMID:28122052

Pseudocatalytic scavenging of the nerve agent VX with human blood components and the oximes obidoxime and HI-6.

PubMed

Wille, Timo; von der Wellen, Jens; Thiermann, Horst; Worek, Franz

2017-03-01

Despite six decades of extensive research in medical countermeasures against nerve agent poisoning, a broad spectrum acetylcholinesterase (AChE) reactivator is not yet available. One current approach is directed toward synthesizing oximes with high affinity and reactivatability toward butyrylcholinesterase (BChE) in plasma to generate an effective pseudocatalytic scavenger. An interim solution could be the administration of external AChE or BChE from blood products to augment pseudocatalytic scavenging with slower but clinically approved oximes to decrease nerve agent concentrations in the body. We here semiquantitatively investigate the ability of obidoxime and HI-6 to decrease the inhibitory activity of VX with human AChE and BChE from whole blood, erythrocyte membranes, erythrocytes, plasma, clinically available fresh frozen plasma and packed red blood cells. The main findings are that whole blood showed a VX concentration-dependent decrease in inhibitory activity with HI-6 being more potent than obidoxime. Using erythrocytes and erythrocyte membranes again, HI-6 was more potent compared to obidoxime. With freshly prepared plasma, obidoxime and HI-6 showed comparable results for the decrease in VX. The use of the clinically available blood products revealed that packed red blood cells showed similar kinetics as fresh erythrocytes. Fresh frozen plasma resulted in a slower and incomplete decrease in inhibitory plasma compared to freshly prepared plasma. In conclusion, the administration of blood products in combination with available oximes augments pseudocatalytic scavenging and might be useful to decrease the body load of persistent, highly toxic nerve agents.

75 FR 74684 - Notice of Final Results of Antidumping Duty Changed Circumstances Review: Certain Frozen...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-12-01

... battered shrimp. Dusted shrimp is a shrimp-based product: (1) That is produced from fresh (or thawed-from... subjected to IQF freezing immediately after application of the dusting layer. Battered shrimp is a shrimp...

Identification of protein markers for the occurrence of defrosted material in milk through a MALDI-TOF-MS profiling approach.

PubMed

Arena, Simona; Salzano, Anna Maria; Scaloni, Andrea

2016-09-16

Mozzarella di Bufala Campana is a soft, stretched curd Italian cheese made from fresh buffalo milk that obtained the Protected Designation of Origin (PDO) registration in EU legislation. Seasonality of buffalo milk production, rapid cheese decay and transport of its preserving liquid have relevant practical/economic consequences for mozzarella production; consequently, a progressive diffusion of cheese products realized with frozen curd or frozen milk has recently been observed. In order to meet the demand of the dairy producers and consumers for a reduction of starting material adulterations and for the certification of the raw milk used for cheese manufacturing, we have developed a rapid/robust MALDI-TOF-MS polypeptide profiling procedure that assays material quality through the identification of specific markers of its freshness. Massive analysis of fresh and frozen buffalo milks (stored for different times) was realized to this purpose; a tough statistical evaluation of the resulting data ultimately permitted the typing of milk samples. We identified 28 polypeptide markers of the milk freezing storage, among which 13 and 15 showed down- and over-representation, respectively. Quantitative data were confirmed by an independent analytical approach on selected markers. GLYCAM1-derived phosphopeptides (1-53), β-casein-derived phosphopeptides (1-68), β-casein-derived γ2-, γ3- and γ4-fragments, α-lactalbumin and β-lactoglobulin were components showing the highest significance. The occurrence of the first compounds in buffalo milk is here described for the first time; their formation in the frozen material was ascribed to the activity of plasmin or of unknown bacterial proteases/peptidases stable at low temperatures. In conclusion, data reported here suggest the application of this MALDI-TOF-MS polypeptide profiling platform to other high-quality dairy productions, in which milk freshness has important consequences on final product organoleptic properties. In the last decades, several studies have provided the molecular basis underlying the relation between food quality and human wellness/health. In this context, Foodomics emerged as a novel scientific discipline studying food and nutrition domains through the application of advanced omics technologies, including genomics, transcriptomics, proteomics and/or metabolomics. Above-mentioned technologies have been used in an integrated, holistic way to study foods for: i) compound profiling, authenticity, and/or biomarker-detection related to product quality or safety; ii) contaminants and their whole toxicity; iii) bioactivity and general effects on human health; iv) their digestion and assumption in human body; v) development of new transgenic products; and vi) evaluation of their modifications within the digestive tract. In the first context, a highly reproducible MALDI-TOF-MS polypeptide profiling procedure is here presented, which provides information on buffalo milk quality through the identification of specific markers of its freshness. Among identified markers, some were indicative of the action of various proteolytic enzymes and the resulting occurrence of specific defense components in buffalo milk having the physiological role to limit bacterial/virus content in this biological fluid. Data suggest the possible application of similar MALDI-TOF-based platforms to other high-quality food productions, where storage conditions of the starting materials may have important consequences on final product characteristics. Copyright © 2016 Elsevier B.V. All rights reserved.

Rapid detection of frozen-then-thawed minced beef using multispectral imaging and Fourier transform infrared spectroscopy.

PubMed

Ropodi, Athina I; Panagou, Efstathios Z; Nychas, George-John E

2018-01-01

In recent years, fraud detection has become a major priority for food authorities, as fraudulent practices can have various economic and safety consequences. This work explores ways of identifying frozen-then-thawed minced beef labeled as fresh in a rapid, large-scale and cost-effective way. For this reason, freshly-ground beef was purchased from seven separate shops at different times, divided in fifteen portions and placed in Petri dishes. Multi-spectral images and FTIR spectra of the first five were immediately acquired while the remaining were frozen (-20°C) and stored for 7 and 32days (5 samples for each time interval). Samples were thawed and subsequently subjected to similar data acquisition. In total, 105 multispectral images and FTIR spectra were collected which were further analyzed using partial least-squares discriminant analysis and support vector machines. Two meat batches (30 samples) were reserved for independent validation and the remaining five batches were divided in training and test set (75 samples). Results showed 100% overall correct classification for test and external validation MSI data, while FTIR data yielded 93.3 and 96.7% overall correct classification for FTIR test set and external validation set respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

ECONOMIC FEASIBILITY OF RADIATION-PASTEURIZING FRESH STRAWBERRIES, PEACHES, TOMATOES, GRAPES, ORANGES, AND GRAPEFRUIT

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

Droge, J.H.

1963-08-01

Results are reported from a survey of the produce industry on the feasibility of using radiation processing to extend the storage life of fresh strawberries, peaches, tomatoes, grapes, oranges, and grapefruit. A majority of the respondents thought that radiation processing would increase the production and market volume of the selected fruits and vegetables surveyed but would not change output and sales volume of camned, frozen, and other processed forms. Cost estimates of radiation processing are discussed. (C.H.)

An Array-Based Analysis of MicroRNA Expression Comparing Matched Frozen and Formalin-Fixed Paraffin-Embedded Human Tissue Samples

PubMed Central

Zhang, Xiao; Chen, Jiamin; Radcliffe, Tom; LeBrun, Dave P.; Tron, Victor A.; Feilotter, Harriet

2008-01-01

MicroRNAs (miRNAs) are small, noncoding RNAs that suppress gene expression at the posttranscriptional level via an antisense RNA-RNA interaction. miRNAs used for array-based profiling are generally purified from either snap-frozen or fresh samples. Because tissues found in most pathology departments are available only in formalin-fixed and paraffin-embedded (FFPE) states, we sought to evaluate miRNA derived from FFPE samples for microarray analysis. In this study, miRNAs extracted from matched snap-frozen and FFPE samples were profiled using the Agilent miRNA array platform (Agilent, Santa Clara, CA). Each miRNA sample was hybridized to arrays containing probes interrogating 470 human miRNAs. Seven cases were compared in either duplicate or triplicate. Intrachip and interchip analyses demonstrated that the processes of miRNA extraction, labeling, and hybridization from both frozen and FFPE samples are highly reproducible and add little variation to the results; technical replicates showed high correlations with one another (Kendall tau, 0.722 to 0.853; Spearman rank correlation coefficient, 0.891 to 0.954). Our results showed consistent high correlations between matched frozen and FFPE samples (Kendall tau, 0.669 to 0.815; Spearman rank correlation coefficient, 0.847 to 0.948), supporting the use of FFPE-derived miRNAs for array-based, gene expression profiling. PMID:18832457

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

PubMed

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

2008-06-01

The effects of residual salt in surimi on physicochemical properties as affected by various freeze and thaw (FT) cycles were examined. Fresh Alaska pollock surimi was mixed with 4.0% sugar and 5.0% sorbitol, along with 8 combinations of salt (0.4%, 0.6%, 0.8%, and 1.0% NaCl) and sodium polyphosphate (0.25% and 0.5%), vacuum-packed, and stored at -18 degrees C until used. FT cycles (0, 6, and 9) were used to mimic long-term frozen storage. At the time of gel preparation, each treatment was appropriately adjusted to maintain 2% salt and 78% moisture. The pH decreased as residual salt increased during frozen storage. Salt extractable protein (SEP) decreased (P < 0.05) as FT cycles extended from 0 to 9. Regardless of residual salt and phosphate concentration during frozen storage, whiteness value (L*- 3b*) decreased (P < 0.05) as FT cycles extended, except for samples with 0.4% salt/0.5% phosphate and 0.6% salt/0.25% phosphate. Water retention ability (WRA) and texture significantly (P < 0.05) decreased at higher salt content (0.8% and 1.0%) after 9 FT cycles, indicating higher residual salt concentration can shorten the shelf life of frozen surimi. Our study revealed lower residual salt concentration and higher phosphate concentration are likely to extend the shelf life of frozen surimi.

Mango Shake

MedlinePlus

... this page: https://medlineplus.gov/recipe/mangoshake.html Mango Shake To use the sharing features on this page, please enable JavaScript. Prep time: 5 minutes Cook time: 0 minutes ... cup low-fat (1 percent) milk 4 Tbsp frozen mango juice (or 1 fresh pitted mango) 1 small ...

Determination of toxic elements (mercury, cadmium, lead, tin and arsenic) in fish and shellfish samples. Risk assessment for the consumers.

PubMed

Olmedo, P; Pla, A; Hernández, A F; Barbier, F; Ayouni, L; Gil, F

2013-09-01

Although fish intake has potential health benefits, the presence of metal contamination in seafood has raised public health concerns. In this study, levels of mercury, cadmium, lead, tin and arsenic have been determined in fresh, canned and frozen fish and shellfish products and compared with the maximum levels currently in force. In a further step, potential human health risks for the consumers were assessed. A total of 485 samples of the 43 most frequently consumed fish and shellfish species in Andalusia (Southern Spain) were analyzed for their toxic elements content. High mercury concentrations were found in some predatory species (blue shark, cat shark, swordfish and tuna), although they were below the regulatory maximum levels. In the case of cadmium, bivalve mollusks such as canned clams and mussels presented higher concentrations than fish, but almost none of the samples analyzed exceeded the maximum levels. Lead concentrations were almost negligible with the exception of frozen common sole, which showed median levels above the legal limit. Tin levels in canned products were far below the maximum regulatory limit, indicating that no significant tin was transferred from the can. Arsenic concentrations were higher in crustaceans such as fresh and frozen shrimps. The risk assessment performed indicated that fish and shellfish products were safe for the average consumer, although a potential risk cannot be dismissed for regular or excessive consumers of particular fish species, such as tuna, swordfish, blue shark and cat shark (for mercury) and common sole (for lead). Copyright © 2013 Elsevier Ltd. All rights reserved.

Ex vivo fluorescence confocal microscopy for fast evaluation of tumour margins during Mohs surgery.

PubMed

Bennàssar, A; Vilata, A; Puig, S; Malvehy, J

2014-02-01

Ex vivo fluorescence confocal microscopy (FCM) enables real-time imaging of skin morphology directly in freshly excised tissue. FCM displays wide field-of-view mosaics with cellular resolution, thus enabling a rapid bedside pathology. An application of interest is rapid detection of residual basal cell carcinoma (BCC) in skin excisions during Mohs surgery. We sought to evaluate the sensitivity and specificity of ex vivo imaging with FCM for the detection of residual BCC in Mohs tissue excisions, and to calculate the time invested up to the diagnosis for both FCM and frozen sections. Eighty consecutive BCCs were prospectively collected and the margins scanned with ex vivo FCM, including excisions with and without residual BCC of all major subtypes. Each mosaic was divided into two or four, resulting in 480 submosaics for study. Every confocal submosaic was assessed for the presence or absence of BCC and compared with standard frozen sections as the gold standard. Furthermore, the time spent for each technique was calculated and compared. The overall sensitivity and specificity of detecting residual BCC were 88% and 99%, respectively. Moreover, the new technique reduced by almost two-thirds the time invested when compared with the processing of a frozen section (P < 0·001). The results demonstrate the feasibility of confocal mosaicing microscopy in fresh tissue for rapid surgical pathology, potentially to expedite and guide Mohs surgery with high accuracy. This observation is an important step towards the goal of using real-time surgical pathology for skin tumours. © 2013 British Association of Dermatologists.

The effect of Arabic gum on frozen dough properties and the sensory assessments of the bread produced.

PubMed

Tavakoli, Hamid Reza; Jonaidi Jafari, Nematollah; Hamedi, Hassan

2017-04-01

The use of hydrocolloids in frozen dough has become frequent as bread improvers due to their anti-staling effect. Nevertheless, the impact of both different frozen storage and Arabic gum level in non-prefermented flat dough with following thawing procedure have not been studied. This work intended to study the effect of three different ratio of Arabic gum on rheological properties of 1, 7, and 30 days of frozen storage and the quality of the bread made from. In order to gain the least detrimental effects on gluten network, we used rapid rate freezing and microwave heating in thawing stage. Rheological results showed that the unfrozen samples to which Arabic gum had been added rendered the highest resistance to extension. The resistance of gum fortified samples were less than fresh dough, however the decline was not significant in 3.0% Arabic gum dough kept in a month storage (p > .05). The similar findings were obtained for extensibility and adhesiveness; in which the maximum incorporation of Arabic gum lessen the destructive impact of long freezing storage. Addition of 3% gum could be able to retard staling through an increment in hydrophilic bonds between water molecules and amylose during thawing (p < .05). The overall rating of Arabic gum enriched samples was similar with bread made from non-frozen dough, even after 30 days of storage as indicated by the sensory evaluation of breads. Producing a chapatti-like fermented bread without long fermentation period. Formulation a frozen dough without using chemical additives. Introducing a proper use of a new defrosting method with the aim of achieving a better texture. Improvement in retarding staling by the use of Gum Arabic after 7 days. © 2016 Wiley Periodicals, Inc.

Availability of Foods and Beverages in Supplemental Nutrition Assistance Program-Authorized Dollar Stores in a Region of North Carolina.

PubMed

Racine, Elizabeth F; Batada, Ameena; Solomon, Corliss A; Story, Mary

2016-10-01

There are >25,000 Supplemental Nutrition Assistance Program (SNAP)-authorized dollar stores throughout the United States; many are located in lower-income neighborhoods and provide an accessible food and beverage source for area residents. The purpose of this research was to determine the percent of food deserts within 16 counties in North Carolina that include a SNAP dollar store; examine the types of foods and beverages at SNAP dollar stores in these counties; test whether the foods and beverages offered vary by SNAP dollar store chain; and test whether the foods and beverages available differ by rural and urban location. This cross-sectional study used a combination of publicly available data and primary data to investigate the research questions. Secondary data sources were obtained from the US Department of Agriculture's SNAP retailer locator, the US Census, and the US Department of Agriculture's Food Access Research Atlas. Availability of foods and beverages was assessed among a sample of 90 SNAP dollar stores in 16 counties in southern and western sections of North Carolina. Data were collected in June 2014. About half (52%) of the food deserts in the research area included a SNAP dollar store. Most of the sampled stores sold healthier food staples, such as frozen meats, brown rice, 100% whole-wheat bread, and dried beans. None of the stores sold fresh fruits or vegetables. Some of the foods and beverages offered (eg, frozen fruit, frozen unseasoned vegetables, nonfat or low-fat milk, frozen ground beef) varied by SNAP dollar store chain. The foods and beverages offered did not differ by rural or urban county location. SNAP dollar stores offer a number of healthy food staples; however, they do not sell fresh fruits or vegetables. Further food environment research should include dollar stores. Copyright © 2016 Academy of Nutrition and Dietetics. Published by Elsevier Inc. All rights reserved.

Sage (Salvia officinalis) and fennel (Foeniculum vulgare) improve cryopreserved boar epididymal semen quality study.

PubMed

Monton, A; Gil, L; Malo, C; Olaciregui, M; Gonzalez, N; de Blas, I

2015-01-01

The aim of this study was to evaluate the effect of fennel and sage extracts and the influence of the egg yolk source (fresh or pasteurized) on the success of freezing boar epididymal spermatozoa. In experiment 1, epididymal sperm was recovered by flushing and cryopreserved in a lactose-egg yolk solution supplemented with various concentrations (10, 5 and 2.5 g/L) of sage or fennel. Sperm quality was evaluated (motility, viability, HOST and acrosome integrity) at 0 h and 2 h after thawing. Fennel 10 g/L and sage 5 g/L and control (no extracts) were selected for experiment 2 which also compared fresh or pasteurized egg yolk in the freezing extender and measured DNA integrity of the frozen sperm. Results showed that the interaction between fennel and sage antioxidants with fresh egg yolk significantly improved post thaw sperm quality and protected boar epididymal spermatozoa from cryopreservation damage as a result of oxidative stress.

Hope for Restoration of Dead Valuable Bulls through Cloning Using Donor Somatic Cells Isolated from Cryopreserved Semen

PubMed Central

Selokar, Naresh L.; Saini, Monika; Palta, Prabhat; Chauhan, Manmohan S.; Manik, Radheysham; Singla, Suresh K.

2014-01-01

Somatic cells were isolated from cryopreserved semen of 4 buffalo bulls, 3 of which had died over 10 years earlier, and were established in culture. The cells expressed cytokeratin-18, keratin and vimentin indicating that they were of epithelial origin. The cells were used as nuclear donors for hand-made cloning for producing buffalo embryos. The blastocyst rate and quality, as indicated by apoptotic index, were comparable among embryos produced using cells obtained from fresh or frozen-thawed semen or those obtained from conventional cell sources such as skin. Examination of the epigenetic status revealed that the global level of H3K27me3 but not that of H3K9/14ac and H4K5ac differed significantly (P<0.05) among cloned embryos from different bulls. The relative mRNA abundance of HDAC1, DNMT1, P53 and CASPASE 3 but not that of DNMT3a differed in cells and in cloned embryos. Following transfer of 24 cloned embryos produced from fresh semen-derived cells to 12 recipients, one calf weighing 55 kg, which is now 6 months of age and is normal, was born through normal parturition. Following transfer of 20 embryos produced from frozen-thawed semen-derived cells to 10 recipients, 2 became pregnant, one of which aborted in the first trimester; the calf born was severely underweight (17 kg), and died 12 h after birth. The ability of cells derived from fresh and frozen-thawed semen to produce live offspring confirms the ability of these cells to be reprogrammed. Our findings pave the way for restoration of highly precious progeny-tested bulls, which has immense economic importance, and can also be used for restoration of endangered species. PMID:24614586

Hope for restoration of dead valuable bulls through cloning using donor somatic cells isolated from cryopreserved semen.

PubMed

Selokar, Naresh L; Saini, Monika; Palta, Prabhat; Chauhan, Manmohan S; Manik, Radheysham; Singla, Suresh K

2014-01-01

Somatic cells were isolated from cryopreserved semen of 4 buffalo bulls, 3 of which had died over 10 years earlier, and were established in culture. The cells expressed cytokeratin-18, keratin and vimentin indicating that they were of epithelial origin. The cells were used as nuclear donors for hand-made cloning for producing buffalo embryos. The blastocyst rate and quality, as indicated by apoptotic index, were comparable among embryos produced using cells obtained from fresh or frozen-thawed semen or those obtained from conventional cell sources such as skin. Examination of the epigenetic status revealed that the global level of H3K27me3 but not that of H3K9/14ac and H4K5ac differed significantly (P<0.05) among cloned embryos from different bulls. The relative mRNA abundance of HDAC1, DNMT1, P53 and CASPASE 3 but not that of DNMT3a differed in cells and in cloned embryos. Following transfer of 24 cloned embryos produced from fresh semen-derived cells to 12 recipients, one calf weighing 55 kg, which is now 6 months of age and is normal, was born through normal parturition. Following transfer of 20 embryos produced from frozen-thawed semen-derived cells to 10 recipients, 2 became pregnant, one of which aborted in the first trimester; the calf born was severely underweight (17 kg), and died 12 h after birth. The ability of cells derived from fresh and frozen-thawed semen to produce live offspring confirms the ability of these cells to be reprogrammed. Our findings pave the way for restoration of highly precious progeny-tested bulls, which has immense economic importance, and can also be used for restoration of endangered species.

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

PubMed

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

2015-05-01

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

Short communication: Preliminary investigation into the effect of freezing and a cryopreservant on the recovery of mastitis pathogens from ewe milk.

PubMed

Smith, E M; Monaghan, E M; Huntley, S J; Green, L E

2011-10-01

The objective of this study was to investigate the recovery of bacteria from ewe milk after freezing for 4 or 8 wk with and without the addition of glycerol as a cryopreservant. A total of 50 udder-half milk samples with a known range of bacterial species were selected, stored, and analyzed in 5 treatment groups: time zero; frozen for 4 wk with, and without, glycerol; and frozen for 8 wk with, and without, glycerol. A lower recovery was observed in all bacterial species studied after freezing. Samples containing fewer than 100 cfu/mL came from ewes with a lower somatic cell count and were more likely to be bacteriologically negative after freezing than those above this threshold. The addition of glycerol increased recovery of gram-negative bacteria after freezing, although this requires further study to draw strong conclusions. The effects on gram-positive species were inconsistent. We conclude that although the addition of glycerol had a small beneficial effect on the sensitivity of detection of bacteria from frozen sheep milk, sensitivity was highest in cultures from fresh milk. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

The effect of refrigerated and frozen storage on butter flavor and texture.

PubMed

Krause, A J; Miracle, R E; Sanders, T H; Dean, L L; Drake, M A

2008-02-01

Butter is often stored for extended periods of time; therefore, it is important for manufacturers to know the refrigerated and frozen shelf life. The objectives of this study were to characterize the effect of refrigerated and frozen storage on the sensory and physical characteristics of butter. Fresh butter was obtained on 2 occasions from 2 facilities in 113-g sticks and 4-kg bulk blocks (2 facilities, 2 package forms). Butters were placed into both frozen (-20 degrees C) and refrigerated storage (5 degrees C). Frozen butters were sampled after 0, 6, 12, 15, and 24 mo; refrigerated butters were sampled after 0, 3, 6, 9, 12, 15, and 18 mo. Every 3 mo, oxidative stability index (OSI) and descriptive sensory analysis (texture, flavor, and color) were conducted. Every 6 mo, peroxide value (PV), free fatty acid value (FFV), fatty acid profiling, vane, instrumental color, and oil turbidity were examined. A mixed-model ANOVA was conducted to characterize the effects of storage time, temperature, and package type. Storage time, temperature, and package type affected butter flavor, OSI, PV, and FFV. Refrigerated butter quarters exhibited refrigerator/stale off-flavors concurrent with increased levels of oxidation (lower oxidative stability and higher PV and FFV) within 6 mo of refrigerated storage, and similar trends were observed for refrigerated bulk butter after 9 mo. Off-flavors were not evident in frozen butters until 12 or 18 mo for quarters and bulk butters, respectively. Off-flavors in frozen butters were not correlated with instrumental oxidation measurements. Because butter is such a desirable fat source in terms of flavor and textural properties, it is important that manufacturers understand how long their product can be stored before negative attributes develop.

Safety of lumbar spine radiofrequency procedures in the presence of posterior pedicle screws: technical report of a cadaver study.

PubMed

Gazelka, Halena M; Welch, Tasha L; Nassr, Ahmad; Lamer, Tim J

2015-05-01

To determine whether the thermal energy associated with lumbar spine radiofrequency neurotomy (RFN) performed near titanium and stainless steel pedicle screws is conducted to the pedicle screws or adjacent tissues, or both, thus introducing potential for thermal damage to those tissues. Cadaver study. Cadaver laboratory equipped with fluoroscopy, surgical spine implements, and radiofrequency generator. No live human subject; a fresh frozen (and thawed) cadaver torso was used for the study. Titanium and stainless steel pedicle screws were placed in the lumbar spine of a fresh frozen cadaver torso with real-time fluoroscopic guidance. Conventional RFN cannula placement was performed at the level of pedicle screws and a control (nonsurgically altered) lumbar level. Neurotomy was performed with conventional radiofrequency lesioning parameters. Temperatures were recorded at multiple sites through thermistor probes. Direct contact of the radiofrequency cannula with the pedicle screws during conventional RFN produced a substantial increase in temperature in the surrounding soft tissues. A small increase in temperature occurred at the same sites at the control level. Titanium and stainless steel pedicle screws are capable of sustaining large increases in temperature when the radiofrequency probe comes in contact with the screw. These results are suggestive that pedicle screws could serve as a possible source of tissue heating and thermal injury during RFN. Wiley Periodicals, Inc.

Time-Dependent Effects of Chlorhexidine Soaks on Grossly Contaminated Bone

DTIC Science & Technology

2012-10-01

decontaminate . Methods: Fifty four bone segments were harvested from fresh frozen porcine legs. Each specimen was dropped onto a Mueller Hinton medium that was...P, 0.0001) and no difference between the 2%CHLand 4%CHLgroups. Conclusions: This study provides new data supporting the use of CHL to decontaminate ...bone may expedite the process of biofilm formation.1,6,7 Therefore, grossly contaminated bone seg- ments should be decontaminated with effective

Benign Breast Disease: Toward Molecular Prediction of Breast Cancer Risk

DTIC Science & Technology

2006-06-01

with benign breast disease ( BBD ) (1967-1991); 2) the application of potential biomarkers of risk to this archival tissue set; and 3) the discovery...of new, potentially relevant biomarkers of risk in fresh and frozen specimens of BBD . The Center includes a multi-institutional team of basic...State). I. Task 1: Establish Retrospective Cohort of BBD and Nested Case-Control Study A. Complete cohort follow-up We provide here an update of

Antigen detection systems

USDA-ARS?s Scientific Manuscript database

Infectious agents or their constituent parts (antigens or nucleic acids) can be detected in fresh, frozen, or fixed tissues or other specimens, using a variety of direct or indirect assays. The assays can be modified to yield the greatest sensitivity and specificity but in most cases a particular m...

Tumor Cold Ischemia | Office of Cancer Clinical Proteomics Research

Cancer.gov

In a recently published manuscript in the journal of Molecular and Cellular Proteomics, researchers from the National Cancer Institutes (NCI) Clinical Proteomic Tumor Analysis Consortium (CPTAC) investigated the effect of cold ischemia on the proteome of fresh frozen tumors.

Eat for Health

MedlinePlus

... pasta, or cereal Vegetables 4-5 1 cup raw leafy vegetables ½ cup cooked vegetables 6 oz vegetable juice Fruits 4-5 1 medium fruit ¼ cup dried fruit ½ cup fresh, frozen, or canned fruit ½ cup fruit juice Low-fat or fat-free milk and milk products 2- ...

7 CFR 926.13 - Processor.

Code of Federal Regulations, 2014 CFR

2014-01-01

... RECORDKEEPING REQUIREMENTS APPLICABLE TO CRANBERRIES NOT SUBJECT TO THE CRANBERRY MARKETING ORDER § 926.13 Processor. Processor means any person who receives or acquires fresh or frozen cranberries or cranberries in... uses such cranberries or concentrate, with or without other ingredients, in the production of a product...

7 CFR 926.13 - Processor.

Code of Federal Regulations, 2013 CFR

2013-01-01

... RECORDKEEPING REQUIREMENTS APPLICABLE TO CRANBERRIES NOT SUBJECT TO THE CRANBERRY MARKETING ORDER § 926.13 Processor. Processor means any person who receives or acquires fresh or frozen cranberries or cranberries in... uses such cranberries or concentrate, with or without other ingredients, in the production of a product...

7 CFR 926.13 - Processor.

Code of Federal Regulations, 2011 CFR

2011-01-01

... RECORDKEEPING REQUIREMENTS APPLICABLE TO CRANBERRIES NOT SUBJECT TO THE CRANBERRY MARKETING ORDER § 926.13 Processor. Processor means any person who receives or acquires fresh or frozen cranberries or cranberries in... uses such cranberries or concentrate, with or without other ingredients, in the production of a product...

7 CFR 926.13 - Processor.

Code of Federal Regulations, 2012 CFR

2012-01-01

... RECORDKEEPING REQUIREMENTS APPLICABLE TO CRANBERRIES NOT SUBJECT TO THE CRANBERRY MARKETING ORDER § 926.13 Processor. Processor means any person who receives or acquires fresh or frozen cranberries or cranberries in... uses such cranberries or concentrate, with or without other ingredients, in the production of a product...

Architecture of baked breads depicted by a magnetic resonance imaging.

PubMed

Ishida, N; Takano, H; Naito, S; Isobe, S; Uemura, K; Haishi, T; Kose, K; Koizumi, M; Kano, H

2001-07-01

The architecture of baked breads made of fresh dough and frozen dough was depicted by magnetic resonance imaging (MRI). Pieces of bread (16 mm cubic cakes) were soaked in organic solvents containing various concentrations of heavy metals (Cu(2+), Co(2+) and Fe(3+)) and images of the grain structure of the breads were obtained. Of the organic solvents tested, acetone was preferable because of its single peak that prevents chemical shift effects on images, the retention of the bread structure, and the solubility of heavy metals. The heavy metals, especially Fe(3+), shortened the overly long relaxation times of acetone to practical lengths for imaging and stained the materials to provide high contrasts. The images obtained in acetone with 8 mM Fe(3+) were suitable for analyzing crumb grain structures. The bread of fresh dough showed a uniform distribution of pores of various sizes made of thin gluten sheets, whereas the pores in the bread of frozen dough were less, prominently large, non-uniformly distributed, and made of thick gluten sheets.

Transfusion audit of fresh-frozen plasma in southern Taiwan.

PubMed

Yeh, C-J; Wu, C-F; Hsu, W-T; Hsieh, L-L; Lin, S-F; Liu, T-C

2006-10-01

The demand for transfusions has increased rapidly in southern Taiwan. Between 1993 and 2003, requests for fresh-frozen plasma (FFP) in particular rose dramatically at Kaohsiung Medical University Hospital (KMUH). Transfusion orders were not tightly regulated, and inappropriate use of blood products was common. We carried out a prospective analysis of transfusion requests from October 2003 to January 2004 at KMUH, and then repeated the audit for another 3-month period after the clinical faculty had undergone five sessions of education on transfusion guidelines. Later, our consultant haematologist applied computerized guidelines to periodic audits. A 5.2% decrease in inappropriate FFP usage followed the educational programme and a further 30% reduction took place after the application of computerized transfusion guidelines. With the guidelines and periodic audits, FFP transfusions decreased by 74.6% and inappropriate requests from 65.2% to 30%. Hospital policy, computerized transfusion guidelines and periodic audits greatly reduced inappropriate FFP transfusions. An educational campaign had a more limited effect.

Reverse Stability Kinetics of Meat Pigment Oxidation in Aqueous Extract from Fresh Beef.

PubMed

Frelka, John C; Phinney, David M; Wick, Macdonald P; Heldman, Dennis R

2017-12-01

The use of kinetic models is an evolving approach to describing quality changes in foods during processes, including storage. Previous studies indicate that the oxidation rate of myoglobin is accelerated under frozen storage conditions, a phenomenon termed reverse stability. The goal of this study was to develop a model for meat pigment oxidation to incorporate the phenomenon of reverse stability. In this investigation, the model system was an aqueous extract from beef which was stored under a range of temperatures, both unfrozen and frozen. The kinetic analysis showed that in unfrozen solutions, the temperature dependence of oxidation rate followed Arrhenius kinetics. However, under in frozen solutions the rate of oxidation increased with decreasing temperature until reaching a local maximum around -20 °C. The addition of NaCl to the model system increased oxidation rates at all temperatures, even above the initial freezing temperature. This observation suggests that this reaction is dependent on the ionic strength of the solution as well as temperature. The mechanism of this deviant kinetic behavior is not fully understood, but this study shows that the interplay of temperature and composition on the rate of oxidation of meat pigments is complicated and may involve multiple mechanisms. A better understanding of the kinetics of quality loss in a meat system allows for a re-examination of the current recommendations for frozen storage. The deviant kinetic behavior observed in this study indicates that the relationship between quality loss and temperature in a frozen food is not as simple as once thought. Product-specific recommendations could be implemented in the future that would allow for a decrease in energy consumption without a significant loss of quality. © 2017 Institute of Food Technologists®.

Spoilage and shelf-life extension of fresh fish and shellfish.

PubMed

Ashie, I N; Smith, J P; Simpson, B K

1996-01-01

Fresh fish and shellfish are highly perishable products due to their biological composition. Under normal refrigerated storage conditions, the shelf life of these products is limited by enzymatic and microbiological spoilage. However, with increasing consumer demands for fresh products with extended shelf life and increasing energy costs associated with freezing and frozen storage, the fish-processing industry is actively seeking alternative methods of shelf life preservation and marketability of fresh, refrigerated fish and at the same time economizing on energy costs. Additional methods that could fulfill these objectives include chemical decontamination, low-dose irradiation, ultra-high pressure, and modified atmosphere packaging (MAP). This review focuses on the biochemical and microbiological composition of fresh fish/shellfish, the spoilage patterns in these products, factors influencing spoilage, and the combination treatments that can be used in conjunction with refrigeration to extend the shelf life and keeping quality of fresh fish/shellfish. The safety concerns of minimally processed/MAP fish, specifically with respect to the growth of Clostridium botulinum type E, is also addressed.

High oestradiol concentration after ovarian stimulation is associated with lower maternal serum beta-HCG concentration and neonatal birth weight.

PubMed

Liu, Suying; Kuang, Yanping; Wu, Yu; Feng, Yun; Lyu, Qifeng; Wang, Li; Sun, Yijuan; Sun, Xiaoxi

2017-08-01

In this retrospective study, the relationship between maternal serum oestradiol and progesterone levels after fresh embryo transfer or frozen embryo transfer (FET), and serum beta-HCG levels in early pregnancy and neonatal birth weight was examined. Included for analysis were 5643 conceived singletons: 2610 after FET and 3033 after fresh embryo transfer. Outcome measures included maternal serum oestradiol, progesterone, beta-HCG levels during the peri-implantation period, birth weight and small-for-gestational-age (SGA). Results at 4, 5 and 6 weeks' gestation were as follows: serum oestradiol and progesterone levels were significantly higher in women who underwent fresh embryo transfer compared with FET (all P < 0.0001 except progesterone at 6 weeks; P = 0.009); for fresh embryo transfers, serum beta-HCG levels were significantly lower than in women who underwent FET (P < 0.0001); beta-HCG levels were negatively correlated with serum oestradiol; and birth weight was negatively correlated with serum oestradiol. Incidence of SGA in fresh embryo transfer was increased significantly compared with FET (P < 0.001). Higher maternal oestradiol levels after fresh embryo transfer was correlated with lower beta-HCG in early pregnancy, lower birth weight and higher incidence of SGA. Copyright © 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

A new use for long-term frozen brain tissue: Golgi impregnation

PubMed Central

Melendez-Ferro, Miguel; Perez-Costas, Emma; Roberts, Rosalinda C.

2009-01-01

The study of dendritic spine shape and number has become a standard in the analysis of synaptic transmission anomalies since a considerable number of neuropsychiatric and neurological diseases have their foundation in alterations in these structures. One of the best ways to study possible alterations of dendritic spines is the use of Golgi impregnation. Although usually the Golgi method implies the use of fresh or fixed tissue, here we report the use of Golgi-Cox for the staining of human and animal brain tissue kept frozen for long periods of time. We successfully applied the Golgi-Cox method to human brain tissue stored for up to 15 years in a freezer. The technique produced reliable and reproducible impregnation of dendrites and dendritic spines in different cortical areas. We also applied the same technique to rat brain frozen for up to one year, obtaining the same satisfactory results. The fact that Golgi-Cox can be successfully applied to this type of tissue adds a new value for hundreds of frozen human or animal brains kept in the freezers of the laboratories, that otherwise would not be useful for anything else. Researchers other than neuroanatomists, i.e. in fields such as biochemistry and molecular biology can also benefit from a simple and reliable technique that can be applied to tissue left from their primary experiments. PMID:18789970

Rapid demonstration of Legionella pneumophila in unembedded tissue. An adaptation of the Giménez stain.

PubMed

Greer, P W; Chandler, F W; Hicklin, M D

1980-06-01

The Giménez stain, originally developed for demonstrating rickettsiae, readily stained the Legionnaires' disease bacterium (Legionella pneumophila) in frozen tissue sections and smears of fresh or formalin-fixed lung tissue from patients who had confirmed Legionnaires' disease. With the Giménez procedure, the bacterium stained bright red against a blue-green background. The tissue Gram procedures also stained L. pneumophila in frozen sections and smears, but the staining reaction was weak, and these stains were neither as sensitive nor a consistent as the Giménez procedure.

Home Food Environment Factors Associated with the Presence of Fruit and Vegetables at Dinner: A Direct Observational Study

PubMed Central

Trofholz, Amanda C.; Tate, Allan D.; Draxten, Michelle L.; Neumark-Sztainer, Dianne; Berge, Jerica M.

2015-01-01

Background Little research exists about the factors influencing the foods available at family meals. This study examines the home food environment factors contributing to the presence of fruit and vegetables at family meals. Methods Home food inventory (HFI) and survey data were collected from low-income, minority families (n=120) with children 6-12 years old. Observations from video-recorded family dinner meals, totaling 800 videos, were used to measure the frequency at which fruit and vegetables were served. Multiple regression was used to investigate how the fruit and vegetables in the HFI and other home food environment factors were related to the number of days fruit and vegetables were served at dinner during the observation period. Results Availability and accessibility of fruit and vegetables in the home were each found to be significantly associated with the presence of fruits and vegetables at family dinners. Of the fruit and vegetable categories (i.e., fresh, canned, or frozen), having fresh fruit and vegetables available in the home was found to be most strongly associated with serving fruit and vegetables at dinner, respectively. Higher parent intake of vegetables was associated with the presence of vegetables at dinners, and parent meal planning was associated with the presence of fruit at dinners. Conclusions Increasing the availability and accessibility of fresh fruit and vegetables in the home may be an effective approach to increasing the presence of fruits and vegetables at family dinners, especially among low-income, minority households. It is also essential to understand why families are not using all fruits and vegetables (e.g., canned and frozen) available in the home for family meals. Family meals are a place to promote the increased presence of both fruit and vegetables. PMID:26527254

Home food environment factors associated with the presence of fruit and vegetables at dinner: A direct observational study.

PubMed

Trofholz, Amanda C; Tate, Allan D; Draxten, Michelle L; Neumark-Sztainer, Dianne; Berge, Jerica M

2016-01-01

Little research exists about the factors influencing the foods available at family meals. This study examines the home food environment factors contributing to the presence of fruit and vegetables at family meals. Home food inventory (HFI) and survey data were collected from low-income, minority families (n = 120) with children 6-12 years old. Observations from video-recorded family dinner meals, totaling 800 videos, were used to measure the frequency at which fruit and vegetables were served. Multiple regression was used to investigate how the fruit and vegetables in the HFI and other home food environment factors were related to the number of days fruit and vegetables were served at dinner during the observation period. Availability and accessibility of fruit and vegetables in the home were each found to be significantly associated with the presence of fruits and vegetables at family dinners. Of the fruit and vegetable categories (i.e., fresh, canned, or frozen), having fresh fruit and vegetables available in the home was found to be most strongly associated with serving fruit and vegetables at dinner, respectively. Higher parent intake of vegetables was associated with the presence of vegetables at dinners, and parent meal planning was associated with the presence of fruit at dinners. Increasing the availability and accessibility of fresh fruit and vegetables in the home may be an effective approach to increasing the presence of fruits and vegetables at family dinners, especially among low-income, minority households. It is also essential to understand why families are not using all fruits and vegetables (e.g., canned and frozen) available in the home for family meals. Family meals are a place to promote the increased presence of both fruit and vegetables. Copyright © 2015 Elsevier Ltd. All rights reserved.

The right to food, food donation and microbiological problems of food safety: an experience in the territory of Florence.

PubMed

Bonaccorsi, Guglielmo; Lorini, Chiara; Pieralli, Francesca; Pieri, Luca; Sala, Antonino; Tanini, Tommaso; Nasali, Marco; Dall'Olio, Beatrice; Santomauro, Francesca

2016-01-01

The aim of this study is to understand whether the freezing without a rapid blast chiller represents a storage method for food at the end of shelf life that guarantees microbiological food safety, so to be considered an effective tool for the appropriate management of food in charitable organizations. The study has been performed on 90 food samples, among those that a charitable foodservice trust receives by the large-scale distribution. The products have been frozen using a domestic refrigerator. The indicators used were: total aerobic microbial count, Escherichia coli, Salmonella spp, Staphylococcus aureus, Campylobacter spp, sulphite reducing clostridia. The results show that the preservation of the chosen fresh products at the end of shelf life in refrigerators, frozen without the use of chillers, is a potential management strategy to avoid the loss of edible food, while maintaining the safety standards.

Overview of the TREC 2013 Contextual Suggestion Track

DTIC Science & Technology

2013-11-01

trip into Washington on the metro to see the National Mall. The primary goal of this track is to develop evaluation methodologies for such systems...healthiest and best tasting all-natural nonfat frozen yogurt and fresh fruit smoothies . No wonder Zagat ranked us #1, twice. URL http

Enhanced fertility prediction of cryopreserved boar spermatozoa using novel sperm function assessment

USDA-ARS?s Scientific Manuscript database

Cryopreserved semen is seldom used for commercial porcine artificial insemination (AI) despite many advantages that cryopreservation provides. Compared to fresh semen, the fertility of frozen-thawed boar sperm is more variable but usually less. Predicting the fertility of individual ejaculates for s...

Novel and traditional traits of frozen-thawed porcine sperm related to in vitro fertilization success

USDA-ARS?s Scientific Manuscript database

Cryopreserved semen allows the use of single ejaculates for repeated analyses, potentially improving in vitro fertilization (IVF) consistency by eliminating inter-ejaculate variability observed with fresh semen. However, the freezing and thawing processes result in compromised sperm function and IVF...

Alternative of raw material’s suppliers using TOPSIS method in chicken slaughterhouse industry

NASA Astrophysics Data System (ADS)

Sari, R. M.; Rizkya, I.; Syahputri, K.; Anizar; Siregar, I.

2018-02-01

Chicken slaughterhouse industry is one of the fastest growing industries that depends on the freshness of raw materials. The raw materials quality arrive at the company depends heavily on the suppliers. Fresh chicken and frozen chicken meat are the main raw materials for this industry. Problems occurred by the suppliers are catering the amount of raw material needs that are not appropriate and also delay during delivery process. This condition causes disruption of the production process in the company. Therefore, it is necessary to determine the best suppliers to supply the main raw materials of fresh and frozen chicken meat on the slaughterhouse chicken industry. This study analyze the supplier’s capability by using TOPSIS method. This method use to find out the best supplier. The TOPSIS method is performed using the principle that chosen alternative must have the shortest distance from the positive solution and furthest from the ideal solution of the geometric point by using the Euclidean distance to determine the relative proximity of the optimum solution alternative. TOPSIS method found the rank of best supplier’s order is supplier A followed by supplier D, supplier B, supplier C, supplier E, supplier F, and supplier G. Based on the rank order obtained from each company, it will assist the company in prioritizing the order to the supplier with the best rank. Total supply from All suppliers are 885,994 kg per month. Based on the results of research, the top five suppliers have been sufficient to meet the needs of the company.

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

PubMed

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

2015-08-01

Salvia miltiorrhiza polysaccharides (SMPs) were extracted from S. miltiorrhiza in this study. The aim of the present study was to evaluate the effect of SMP on the motility of boar sperm, including the antioxidant effect of SMP on boar sperm and the effect of SMP on the in vivo fertilizing ability of frozen-thawed boar sperm. Fifty ejaculates from 5 Swagger boars were collected and diluted with an extender, which contained 3% glycerol (v/v) with five concentrations of SMP (0.2, 0.4, 0.6, 0.8, and 1.0mg/mL). The semen was frozen in 0.25mL straws at 1.0×10(9) cells/mL. Sixty gilts were inseminated using fresh semen, frozen semen with 0.4mg/mL of SMP and frozen semen without SMP. The results indicate that the addition of SMP to the extender results in a higher percentage of motile sperm post-thaw (P<0.05). The activities of superoxide dismutase, lactate dehydrogenase, glutamic-oxalacetic transaminease and catalase were all determined to be significantly higher than the control group after adding SMP to the extender (P<0.05). The artificial insemination (AI) results demonstrated that the litter size was significantly higher in the 0.4mg/mL of SMP group than in the control group (P<0.05). In conclusion, during the process of freezing, SMP can protect boar sperm from peroxidative damage and increase sperm motility and litter size during the process of freezing-thawing. The optimal concentration of SMP for the frozen extenders in this study was determined to be 0.4mg/mL. Copyright © 2015. Published by Elsevier B.V.

Evaluation of the antioxidant activity of root extract of pepper fruit (Dennetia tripetala), and it's potential for the inhibition of lipid peroxidation.

PubMed

Okolie, Ngozi Paulinus; Falodun, Abiodun; Davids, Oluseyi

2014-01-01

The antioxidant properties of ethanolic root extract of pepper fruit (Donnetia tripetala), and its effect on lipid peroxidation of some fresh beef tissues during frozen storage were investigated. The antioxidant parameters were assessed using standard methods, while malondialdehyde levels of different fresh beef tissue sections treated with the extract prior to freezing, were estimated in a colorimetric reaction with thiobarbituric acid. The H2O2-scavenging ability of the extract was similar to that of ascorbic acid, with a maximum scavenging power of 55.61 ±4.98%, and an IC50 value of 86µg/ml. The extract exhibited a concentration-dependent ferric ion-reducing power, although this was significantly lower relative to that of the ascorbic acid (p < 0.05). The total phenolic content was 212.5 ± 0.002 mg/g, while the nitric oxide-scavenging ability was 64.33 ± 0.2% after 150 min. The capacity of the extract to inhibit lipid peroxidation in frozen heart muscle slices was significantly higher than that of vitamin C (p < 0 .05), but comparable to vitamins C and E in frozen testes and kidney slices. These results suggest that the root extract of D. tripetala is rich in antioxidants which can be applied to meat preservation during refrigerated storage.

Meat Composition and Quality Assessment of King Scallops (Pecten maximus) and Frozen Atlantic Sea Scallops (Placopecten magellanicus) on a Retail Level

PubMed Central

Manthey-Karl, Monika; Lehmann, Ines; Ostermeyer, Ute; Rehbein, Hartmut; Schröder, Ute

2015-01-01

An enlarged range of scallop products on the market allows the consumer to buy lower priced alternatives, which often raises the question of quality and control. Frozen meat of king scallops (Pecten maximus) and Atlantic sea scallops (Placopecten magellanicus) were purchased on the German market and compared with fresh shell-on king scallops of various origin. The approximate composition, inclusive citric acid and phosphates, minerals, free amino acids (FAA) and fatty acid profiles were examined in the muscle to identify changes as a result of processing. The FAA glycine and taurine as well the fatty acids 20:5n-3 (EPA) and 22:6n-3 (DHA) were the most abundant, but were reduced in processed samples. Di- and triphosphate contents were not detectable (<0.01 g·kg−1) in untreated meats. Most frozen scallop products contained added citrates and polyphosphates and had distinctly higher water contents (up to 89%) and an increased moisture to protein ratio (M/P) (up to 9) compared with the fresh king scallops (78%, M/P < 5). Labelling of species, verified by PCR-based DNA analysis, and ingredients were not correct in each case. Overall results indicated no relevant differences in mineral content, except high sodium contents, resulting from additives. Labelling does not readily allow the consumer to recognize the extent of processing effects. PMID:28231221

Effect of cold storage and packaging material on the major aroma components of sweet cream butter.

PubMed

Lozano, Patricio R; Miracle, Evan R; Krause, Andrea J; Drake, Maryanne; Cadwallader, Keith R

2007-09-19

The major aroma compounds of commercial sweet cream AA butter quarters were analyzed by GC-olfactometry and GC-MS combined with dynamic headspace analysis (DHA) and solvent-assisted flavor evaporation (SAFE). In addition, the effect of long-term storage (0, 6, and 12 months) and type of wrapping material (wax parchment paper vs foil) on the aroma components and sensory properties of these butters kept under refrigerated (4 degrees C) and frozen (-20 degrees C) storage was evaluated. The most intense compounds in the aroma of pasteurized AA butter were butanoic acid, delta-octalactone, delta-decalactone, 1-octen-3-one, 2-acetyl-1-pyrroline, dimethyl trisulfide, and diacetyl. The intensities of lipid oxidation volatiles and methyl ketones increased as a function of storage time. Refrigerated storage caused greater flavor deterioration compared with frozen storage. The intensity and relative abundance of styrene increased as a function of time of storage at refrigeration temperature. Butter kept frozen for 12 months exhibited lower styrene levels and a flavor profile more similar to that of fresh butter compared to butter refrigerated for 12 months. Foil wrapping material performed better than wax parchment paper in preventing styrene migration into butter and in minimizing the formation of lipid oxidation and hydroxyl acid products that contribute to the loss of fresh butter flavor.

Sigma metric analysis for performance of creatinine with fresh frozen serum.

PubMed

Kang, Fengfeng; Zhang, Chuanbao; Wang, Wei; Wang, Zhiguo

2016-01-01

Six sigma provides an objective and quantitative methodology to describe the laboratory testing performance. In this study, we conducted a national trueness verification scheme with fresh frozen serum (FFS) for serum creatinine to evaluate its performance in China. Two different concentration levels of FFS, targeted with reference method, were sent to 98 laboratories in China. Imprecision and bias of the measurement procedure were calculated for each participant to further evaluate the sigma value. Quality goal index (QGI) analysis was used to investigate the reason of unacceptable performance for laboratories with σ < 3. Our study indicated that the sample with high concentration of creatinine had preferable sigma values. For the enzymatic method, 7.0% (5/71) to 45.1% (32/71) of the laboratories need to improve their measurement procedures (σ < 3). And for the Jaffe method, the percentages were from 11.5% (3/26) to 73.1% (19/26). QGI analysis suggested that most of the laboratories (62.5% for the enzymatic method and 68.4% for the Jaffe method) should make an effort to improve the trueness (QGI > 1.2). Only 3.1-5.3% of the laboratories should improve both of the precision and trueness. Sigma metric analysis of the serum creatinine assays is disappointing, which was mainly due to the unacceptable analytical bias according to the QGI analysis. Further effort is needed to enhance the trueness of the creatinine measurement.

Optical properties of cells with melanin

NASA Astrophysics Data System (ADS)

Rohde, Barukh; Coats, Israel; Krueger, James; Gareau, Dan

2014-02-01

The optical properties of pigmented lesions have been studied using diffuse reflectance spectroscopy in a noninvasive configuration on optically thick samples such as skin in vivo. However, it is difficult to un-mix the effects of absorption and scattering with diffuse reflectance spectroscopy techniques due to the complex anatomical distributions of absorbing and scattering biomolecules. We present a device and technique that enables absorption and scattering measurements of tissue volumes much smaller than the optical mean-free path. Because these measurements are taken on fresh-frozen sections, they are direct measurements of the optical properties of tissue, albeit in a different hydration state than in vivo tissue. Our results on lesions from 20 patients including melanomas and nevi show the absorption spectrum of melanin in melanocytes and basal keratinocytes. Our samples consisted of fresh frozen sections that were unstained. Fitting the spectrum as an exponential decay between 500 and 1100 nm [mua = A*exp(-B*(lambda-C)) + D], we report on the fit parameters of and their variation due to biological heterogeneity as A = 4.20e4 +/- 1.57e5 [1/cm], B = 4.57e-3 +/- 1.62e-3 [1/nm], C = 210 +/- 510 [nm] , D = 613 +/- 534 [1/cm]. The variability in these results is likely due to highly heterogeneous distributions of eumelanin and pheomelanin.

Frying stability of rapeseed Kizakinonatane (Brassica napus) oil in comparison with canola oil.

PubMed

Ma, Jin-Kui; Zhang, Han; Tsuchiya, Tomohiro; Akiyama, Yoshinobu; Chen, Jie-Yu

2015-04-01

This study was carried out to investigate the frying performance of Kizakinonatane (Brassica napus) oil during deep-fat frying of frozen French fries with/without replenishment. Commercial regular canola oil was used for comparison. The frying oils were used during intermittent frying of frozen French fries at 180, 200, and 220 ℃ for 7 h daily over four consecutive days. The Kizakinonatane oil exhibited lower levels of total polar compounds, carbonyl value, and viscosity as well as comparable color (optical density) values to that of the canola oil. The monounsaturated fatty acid/polyunsaturated fatty acid ratios were lower than that of canola oil, whereas the polyunsaturated fatty acid/saturated fatty acid ratios are higher than that of canola oil after heating. Results showed that fresh Kizakinonatane oil contains higher levels of acid value, viscosity, optical density values, tocopherols, and total phenolics contents than that of canola oil. Replenishment with fresh oil had significant effects on all chemical and physical parameters, except the acid value of the Kizakinonatane oil during frying processes. Based on the results, the Kizakinonatane oil is inherently suitable for preparing deep-fried foods at high temperatures. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

Overdentures on implants placed in bone augmented with fresh frozen bone.

PubMed

Rigo, L; Viscioni, A; Franco, M; Lucchese, A; Zollino, I; Brunelli, G; Carinci, F

2011-01-01

In the last decade several studies have been performed to evaluate the clinical outcome of one or two stage loaded implants supporting overdentures. Since fresh frozen bone (FFB) has an ever-increasing number of clinical applications and few reports are available on implants inserted into FFB, we performed a retrospective study on fixtures inserted in FFB and bearing overdentures. In the period between December 2003 and December 2006, 17 patients (14 females and 3 males with a median age of about 56 years) were grafted and 60 implants inserted thereafter. A total of 17 overdentures were delivered: 8 in the mandible and 9 in the maxilla. Multiple implant systems were used: 22 Double etched, 7 SLA, 9 Anodic oxidized, and 22 CaPo4 ceramic-blasted. Implant diameter ranged from 3.25 to 4.3 mm and length from 11.5 to 16.0 mm. Implants were inserted to replace 23 incisors, 9 cuspids, 20 premolars and 8 molars. No implants were lost (i.e., survival rate=100%) and no differences were detected among the studied variables. Kaplan Meier algorithm and Cox regression did not reveal any statistical differences among the studied variables also as regards the success rate. Implants inserted FFB and bearing overdentures have a high survival rate and success rates, which are comparable to those of implants inserted in non-grafted bone. FFB bone is a reliable material for alveolar ridge augmentation. No difference was detected among removable prostheses supported by two or more implants.

Effect of feeding pomegranate seed oil as a source of conjugated linolenic acid on Arabian stallion semen quality in cooled and postthawed condition.

PubMed

Nouri, Houshang; Shojaeian, Kamal; Jalilvand, Ghasem; Kohram, Hamid

2018-06-11

The objective was to assess the influence of pomegranate seed oil supplementation on the quality of fresh, cooled and frozen-thawed Arabian breed stallion semen. Eight stallions (n = 4 per group) received their normal diet (control group) or normal diet top dressed with 200 ml of pomegranate seed oil (PSO group). Semen was collected every fifteen days for 90 days. Stallions were reversed across the treatments after a sixty-day interval. In cooled and stored condition (2, 12 and 24 hr), spermatozoa motion characteristics, membrane integrity, viability, morphology and lipid peroxidation were analysed. In frozen-thawed semen, sperm dynamic characteristics were analysed by CASA, acrosome status and mitochondrial activity (evaluated by Flow cytometry) determined. The effects of treatment, time, semen type and their interactions were submitted to PROCMIX (SAS ® ), and means compared by the Tukey test. Also, collected semen samples were artificially inseminated to evaluate fertility and pregnancy rate after day 60 of the experiment. The results from fresh condition showed that semen volume, sperm concentration, abnormality and live sperm were not affected by dietary treatment (p > 0.05). In cooled condition, the higher value for sperm plasma membrane integrity and viability was observed in PSO group compared to control after 24 hr cooled and stored in 5°C. In postthawed condition, the higher value for CASA total motility and acrosome status was observed in PSO group compared to control group (p < 0.05). One hundred and twenty-six mares were artificially inseminated for fertility trial using control and PSO groups' fresh semen. The average pregnancy rates were not significantly different between control and treated group (62.88% and 65.90%, respectively) (p > 0.05). We concluded that under the conditions of this study, dietary supplementation of 200 ml pomegranate seed oil seems to relatively improved Arabian horse sperm quality during storage in cooled and frozen condition via increasing plasma membrane integrity, viability and acrosome status, but did not improve the pregnancy rates. © 2018 Blackwell Verlag GmbH.

15 CFR 2014.2 - Definitions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... TRADE REPRESENTATIVE IMPLEMENTATION OF TARIFF-RATE QUOTA FOR IMPORTS OF LAMB MEAT § 2014.2 Definitions... meanings assigned as follows: (a) Lamb meat means fresh, chilled, or frozen lamb meat, provided for in...-quota lamb meat means lamb meat that is entered under the in-quota rate of duty. (c) Participating...

Fresh and Frozen Seafood: Selecting and Serving It Safely

MedlinePlus

... raw seafood from spreading to ready-to-eat food. Take these steps to avoid cross-contamination: When buying unpackaged cooked seafood, make sure it is physically separated from raw seafood. It should be in its own display case ... any raw food. Wash cutting boards, dishes, utensils, and counter tops ...

77 FR 16768 - Export Sales Reporting Requirements

Federal Register 2010, 2011, 2012, 2013, 2014

2012-03-22

... DEPARTMENT OF AGRICULTURE Office of the Secretary 7 CFR Part 20 RIN 0551-AA70 Export Sales... for pork (fresh, chilled, and frozen box/primal cuts) and distillers dried grain (DDG) to the Export...: Contact Peter W. Burr, Branch Chief, Export Sales Reporting Branch, Import Policies and Export Reporting...

7 CFR 220.8 - Meal requirements for breakfasts.

Code of Federal Regulations, 2013 CFR

2013-01-01

... maximum values). h Discretionary sources of calories (solid fats and added sugars) may be added to the... may be plain or flavored, unsweetened or sweetened. Noncommercial and/or non-standardized yogurt... section. Fruits that are fresh; frozen without added sugar; canned in light syrup, water or fruit juice...

9 CFR 94.7 - Disposal of animals, meats, and other articles ineligible for importation.

Code of Federal Regulations, 2011 CFR

2011-01-01

... ANIMAL PRODUCTS RINDERPEST, FOOT-AND-MOUTH DISEASE, EXOTIC NEWCASTLE DISEASE, AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR DISEASE, AND BOVINE SPONGIFORM ENCEPHALOPATHY: PROHIBITED AND...) Ruminants and swine, and fresh (chilled or frozen) meats, prohibited importation under §§ 94.1, 94.8, 94.9...

9 CFR 94.7 - Disposal of animals, meats, and other articles ineligible for importation.

Code of Federal Regulations, 2012 CFR

2012-01-01

... ANIMAL PRODUCTS RINDERPEST, FOOT-AND-MOUTH DISEASE, EXOTIC NEWCASTLE DISEASE, AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR DISEASE, AND BOVINE SPONGIFORM ENCEPHALOPATHY: PROHIBITED AND...) Ruminants and swine, and fresh (chilled or frozen) meats, prohibited importation under §§ 94.1, 94.8, 94.9...

9 CFR 94.7 - Disposal of animals, meats, and other articles ineligible for importation.

Code of Federal Regulations, 2013 CFR

2013-01-01

... ANIMAL PRODUCTS RINDERPEST, FOOT-AND-MOUTH DISEASE, EXOTIC NEWCASTLE DISEASE, AFRICAN SWINE FEVER, CLASSICAL SWINE FEVER, SWINE VESICULAR DISEASE, AND BOVINE SPONGIFORM ENCEPHALOPATHY: PROHIBITED AND...) Ruminants and swine, and fresh (chilled or frozen) meats, prohibited importation under §§ 94.1, 94.8, 94.9...

21 CFR 640.32 - Collection of source material.

Code of Federal Regulations, 2010 CFR

2010-04-01

...) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.32 Collection of source... blood is intended for Plasma, Fresh Frozen Plasma, and Liquid Plasma, until the plasma is removed, the..., Center for Biologics Evaluations and Research. Whole blood intended for Platelet Rich Plasma must be...

21 CFR 640.32 - Collection of source material.

Code of Federal Regulations, 2012 CFR

2012-04-01

...) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.32 Collection of source... blood is intended for Plasma, Fresh Frozen Plasma, and Liquid Plasma, until the plasma is removed, the..., Center for Biologics Evaluations and Research. Whole blood intended for Platelet Rich Plasma must be...

21 CFR 640.32 - Collection of source material.

Code of Federal Regulations, 2011 CFR

2011-04-01

...) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.32 Collection of source... blood is intended for Plasma, Fresh Frozen Plasma, and Liquid Plasma, until the plasma is removed, the..., Center for Biologics Evaluations and Research. Whole blood intended for Platelet Rich Plasma must be...

21 CFR 640.32 - Collection of source material.

Code of Federal Regulations, 2013 CFR

2013-04-01

...) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.32 Collection of source... blood is intended for Plasma, Fresh Frozen Plasma, and Liquid Plasma, until the plasma is removed, the..., Center for Biologics Evaluations and Research. Whole blood intended for Platelet Rich Plasma must be...

21 CFR 640.32 - Collection of source material.

Code of Federal Regulations, 2014 CFR

2014-04-01

...) BIOLOGICS ADDITIONAL STANDARDS FOR HUMAN BLOOD AND BLOOD PRODUCTS Plasma § 640.32 Collection of source... blood is intended for Plasma, Fresh Frozen Plasma, and Liquid Plasma, until the plasma is removed, the..., Center for Biologics Evaluations and Research. Whole blood intended for Platelet Rich Plasma must be...

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

PubMed

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

2017-05-01

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

Semen of spinal cord injured men freezes reliably.

PubMed

Padron, O F; Brackett, N L; Weizman, M S; Lynne, C M

1994-01-01

The objectives of the present study were to: 1) determine the effect of cryopreservation on the percent and the grade of motility of sperm from spinal cord injured (SCI) men and 2) determine which method of freezing yields the best post-thaw motility in sperm from SCI men. Antegrade semen samples were obtained from 9 SCI subjects and 10 age-matched healthy control subjects. Motility in fresh samples was determined and cryopreservative medium was added to each sample. Aliquots of each sample were frozen according to three methods: 1) liquid nitrogen vapor only (V); 2) vapor for 12 minutes followed by submersion into liquid nitrogen (V+N2); and 3) direct submersion into liquid nitrogen (N2). Samples were frozen for 1 week, then thawed. The post-thaw percent and grade of motility was determined. The mean percent motility of fresh samples for SCI subjects (21.0%) was significantly lower than for control subjects (55.7%). After thawing, the mean percent drop in motility for V, V+N2, and N2 for controls was 65.2%, 73.5%, and 79.4%, respectively, and for SCI subjects, it was 64.7%, 74.5%, and 81.6%, respectively. There was no statistically significant difference between control and SCI subjects by method of freezing. Vapor only as a freezing method was superior to all other methods for retention of sperm motility in both control and SCI subjects. We conclude that the semen of SCI men may be frozen reliably and that their sperm retain motility similar to that of normal men. Vapor only, being the most gentle method used, gives the best recovery of sperm motility in either group.

Identification of apoptotic bodies in equine semen.

PubMed

Caselles, A B; Miro-Moran, A; Morillo Rodriguez, A; Gallardo Bolaños, J M; Ortega-Ferrusola, C; Salido, G M; Peña, F J; Tapia, J A; Aparicio, I M

2014-04-01

Apoptosis in the testis is required to ensure an efficient spermatogenesis. However, sometimes, defective germ cells that are marked for elimination during this process escape elimination in the testes, giving rise to ejaculates with increased percentages of abnormal and apoptotic spermatozoa and a high percentage of apoptotic bodies. Apoptosis markers in the ejaculate have been associated with low fertility, either in animals or humans. Therefore, the goal of this study was to investigate whether fresh equine semen contains apoptotic bodies [initially named Merocyanine 540 (M540) bodies] and to study the relationship between the quantity of these bodies and cell concentration, the volume of ejaculate, viability and motility. Moreover, we also studied whether the presence apoptotic bodies in fresh semen was related to the resistance of the stallion spermatozoa to being incubated at 37 °C or being frozen and thawed. Fresh equine semen was stained with fluorescent dyes such as M540 and Annexin-V. Active Caspase 3 was studied in fresh semen through Western blotting and immunofluorescence with a specific antibody. Sperm kinematics was assessed in fresh, incubated and thawed samples using computer-assisted semen analysis, and viability was evaluated with the LIVE/DEAD Sperm Viability Kit. Overall, our results demonstrate for the first time the presence of apoptotic bodies in equine semen. The quantity of apoptotic bodies was highly variable among stallions and was positively correlated with Caspase 3 activity in fresh samples and negatively correlated with the viability and motility of stallion spermatozoa after the cryopreservation process. © 2014 Blackwell Verlag GmbH.

How do cumulative live birth rates and cumulative multiple live birth rates over complete courses of assisted reproductive technology treatment per woman compare among registries?

PubMed

De Neubourg, D; Bogaerts, K; Blockeel, C; Coetsier, T; Delvigne, A; Devreker, F; Dubois, M; Gillain, N; Gordts, S; Wyns, C

2016-01-01

How do the national cumulative (multiple) live birth rates over complete assisted reproduction technology (ART) courses of treatment per woman in Belgium compare to those in other registries? Cumulative live birth rates (CLBRs) remain high with a low cumulative multiple live birth rate when compared with other registries and publications. In ART, a reduction in the multiple live birth rate could be achieved by reducing the number of embryos transferred. It has been shown that by doing so, live birth rates per cycle were maintained, particularly when the augmentation effect of attached frozen-thawed cycles was considered. A retrospective cohort study included all patients with a Belgian national insurance number who were registered in the national ART registry (Belrap) and who started a first fresh ART cycle between 1 July 2009 until 31 December 2011 with follow up until 31 December 2012. We analysed 12 869 patients and 38 008 cycles (both fresh and attached frozen cycles). CLBRs per patient who started a first ART cycle including fresh and consecutive frozen cycles leading to a live birth. Conservative estimates of cumulative live birth assumed that patients who did not return for treatment had no chance of achieving an ART-related live birth, whereas optimal estimates assumed that women discontinuing treatment would have the same chance of achieving a live birth as those continuing treatment. A maximum of six fresh ART cycles with corresponding frozen cycles was investigated and compared with other registries and publications. The CLBR was age dependent and declined from 62.9% for women <35 years, to 51.4% for women 35-37 years, to 34.1% for women 38-40 years and 17.7% for women 41-42 years in the conservative analysis after six cycles. In the optimal estimate, the CLBR declined from 85.9% for women <35 years, to 72.0% for women 35-37 years, to 50.4% for women 38-40 years and 36.4% for women 41-42 years. The cumulative multiple live birth rates for the whole population were 5.1 and 8.6% for the conservative and optimal estimate, respectively. Conservative and optimal estimates use assumptions for the whole ART population and do not take the individual patient into account. These data reinforce the validity of the Belgian model of coupling reimbursement of ART costs to a restriction in the number of embryos transferred. Our data can improve decision-making in medical ART practice both on the patient level and for society at large and could provide health care takers and insurance companies with a valid model. none. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

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

PubMed

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

2012-10-01

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

Development and validation of a modified comet assay to phenotypically assess nucleotide excision repair.

PubMed

Langie, Sabine A S; Knaapen, Ad M; Brauers, Karen J J; van Berlo, Damien; van Schooten, Frederik-Jan; Godschalk, Roger W L

2006-03-01

There is an increasing need for simple and reliable approaches to phenotypically assess DNA repair capacities. Therefore, a modification of the alkaline comet assay was developed to determine the ability of human lymphocyte extracts to perform the initial steps of the nucleotide excision repair (NER) process, i.e. damage recognition and incision. Gel-embedded nucleoids from A549 cells, pre-exposed to 1 microM benzo[a]pyrene-diol-epoxide, were incubated with cell extracts from frozen or freshly isolated lymphocytes. The rate at which incisions are introduced and the subsequent increase in tail moment is indicative for the repair capacity of the extracts. Freshly prepared extracts from lymphocytes of human volunteers (n = 8) showed significant inter-individual variations in their DNA repair capacity, which correlated with the removal of bulky DNA lesions over a period of 48 h determined by (32)P-post-labelling (R(2) = 0.76, P = 0.005). Repeated measurements revealed a low inter-assay variation (11%). Storage of cell extracts for more than 3 weeks significantly reduced (up to 80%) the capacity to incise the damaged DNA as compared to freshly isolated extracts. This reduction was completely restored by addition of ATP to the extracts before use, as it is required for the incision step of NER. In contrast, extracts freshly prepared from frozen lymphocyte pellets can be used without loss of repair activity. DNA repair deficient XPA-/- and XPC-/- fibroblasts were used to further validate the assay. Although some residual capacity to incise the DNA was observed in these cells, the repair activity was restored to normal wild-type levels when a complementary mixture of both extracts (thereby restoring XPA and XPC deficiency) was used. These results demonstrate that this repair assay can be applied in molecular epidemiological studies to assess inter-individual differences in NER.

Endometrial thickness as a predictor of the reproductive outcomes in fresh and frozen embryo transfer cycles: A retrospective cohort study of 1512 IVF cycles with morphologically good-quality blastocyst.

PubMed

Zhang, Tao; Li, Zhou; Ren, Xinling; Huang, Bo; Zhu, Guijin; Yang, Wei; Jin, Lei

2018-01-01

To evaluate the relationship between endometrial thickness during fresh in vitro fertilization (IVF) cycles and the clinical outcomes of subsequent frozen embryo transfer (FET) cycles.FET cycles using at least one morphological good-quality blastocyst conducted between 2012 and 2013 at a university-based reproductive center were reviewed retrospectively. Endometrial ultrasonographic characteristics were recorded both on the oocyte retrieval day and on the day of progesterone supplementation in FET cycles. Clinical pregnancy rate, spontaneous abortion rate, and live birth rate were analyzed.One thousand five hundred twelve FET cycles was included. The results showed that significant difference in endometrial thickness on day of oocyte retrieval (P = .03) was observed between the live birth group (n = 844) and no live birth group (n = 668), while no significant difference in FET endometrial thickness was found (P = .261) between the live birth group and no live birth group. For endometrial thickness on oocyte retrieval day, clinical pregnancy rate ranged from 50.0% among patients with an endometrial thickness of ≤6 mm to 84.2% among patients with an endometrial thickness of >16 mm, with live birth rate from 33.3% to 63.2%. Multiple logistic regression analysis of factors related to live birth indicated endometrial thickness on oocyte retrieval day was associated with improved live birth rate (OR was 1.069, 95% CI: 1.011-1.130, P = .019), while FET endometrial thickness did not contribute significantly to pregnancy outcomes following FET cycles. The ROC curves revealed the cut-off points of endometrial thickness on oocyte retrieval day was 8.75 mm for live birth.Endometrial thickness during fresh IVF cycles was a better predictor of endometrial receptivity in subsequent FET cycles than FET cycle endometrial thickness. For those females with thin endometrium in fresh cycles, additional estradiol stimulation might be helpful for adequate endometrial development.

Fatty acid and cholesterol profiles, hypocholesterolemic, atherogenic, and thrombogenic indices of broiler meat in the retail market.

PubMed

Attia, Youssef A; Al-Harthi, Mohammed A; Korish, Mohamed A; Shiboob, Mohamed M

2017-02-16

Broiler meat is an essential source of food due to its favourable effects on human health derived from its protein, fats, minerals, vitamins and its bioactive components. A total of 90 carcasses were collected from the retail market in Jeddah city, Saudi Arabia during April, May and June 2014 to determine the effects of meat type (frozen vs. fresh) and sources within fresh types (A, B, C) vs. frozen types (D, E and F) on their fatty acid profiles, cholesterol, their hypocholesterolemic, atherogenic and thrombogenic indices, and on their antioxidants' status. The sources of meat had a significant effect on the hypocholesterolemic and atherogenic indices, with the D source of fresh meat having the best indices. Total saturated fatty acids (SFA), unsaturated fatty acids (UFA), the UFA/SAF ratio, and the monounsaturated (MUFA), Omega-6 and Omega-7 fatty acids were significantly affected by the source of meat. The results revealed that the D source of fresh meat showed favourable fatty acid profiles with significant health benefits for human. Correlation analyses showed a significant negative relationship between the SFA and hypocholesterolemic indices, and significant positive relationships with the atherogenic index, the thrombotgenic index and the total antioxidant capacity. In addition, the relationship between UFA and the hypocholesterolemic index was strongly significantly positive, but was highly negative between the atherogenic and thrombotic indices. The correlations between omega-6 and total cholesterol and the atherogenic index was moderately negative, but was moderately positive with the hypocholesterolemic index. Fatty acids profiles and the hypocholesterolemic and atherogenic indices of broiler meat in the retail market in Jeddah city, Saudi Arabia during April-May-June showed significant differences, with the potential for favourable fatty acids to be boosted. Such variability indicates the needs for a feeding strategy to enhance the favourable fatty acids that may positively impact the health of the consumer, lowering the risk of hypercholesterolemia, atherosclerosis, and thrombogenesis although further studies are needed.

Is solvent/detergent plasma better than standard fresh-frozen plasma? A systematic review and an expert consensus document

PubMed Central

Marietta, Marco; Franchini, Massimo; Bindi, M. Lucia; Picardi, Francesco; Ruggeri, Matteo; De Silvestro, Giustina

2016-01-01

Background Only a few studies have compared solvent/detergent plasma (SD-plasma) to standard fresh-frozen plasma (FFP) in terms of efficacy and safety. Materials and methods A systematic review was performed in order to develop a consensus document on the use of SD-plasma. Moreover, a pharmacoeconomic study was performed in order to assess whether the use of SD-plasma can be cost-effective with respect to the use of FFP. A multidisciplinary panel used the systematic review and the GRADE methodology to develop evidence-based recommendations on this topic. Results Based on moderate to very low quality evidence, the panel developed the following consensus statements: (i) the panel suggested that SD-plasma is safer than FFP; (ii) the panel could not express for or against a greater efficacy of SD-plasma as compared to FFP; (iii) the panel suggested that in patients undergoing liver transplantation SD-plasma can be preferred over FFP; (iv) the panel suggested that SD-plasma can be preferred over FFP in patients with thrombotic thrombocytopenic purpura undergoing plasma-exchange procedures; (v) the panel could not recommend for or against preferring SD-plasma over FFP in critical care patients; and (vi) the panel suggested that the use of SD-plasma can be cost-effective with respect to the use of FFP. Discussion Data from additional randomised studies are needed to establish more definitive guidelines on the use of SD-plasma. PMID:27136429

Comparative biochemical studies of fresh frozen plasma and pooled solvent/detergent-treated plasma (octaplasLG® ) with focus on protein S and its impact in different thrombin generation assay set-ups.

PubMed

Heger, A; Janisch, S; Pock, K; Römisch, J

2016-10-01

The solvent/detergent treatment enables effective and robust inactivation of all lipid-enveloped viruses, but also inactivates partly sensitive plasma proteins such as protein S. The aim of this study was to investigate the thrombin generation capacity of octaplasLG ® , in particular focusing on the function of protein S in thrombin generation assay and the impact of assay settings. Sixteen octaplasLG ® batches and 32 units of single donor fresh frozen plasma (FFP) were investigated. For protein S, both functional activity and free antigen levels were measured. Thrombin generation assay was performed using two fluorogenic tests with different triggers. Finally, rotational thromboelastometry was performed. Mean protein S levels were lower in octaplasLG ® , but a wider range of values was found for FFP. Clotting parameters and thrombin generation capacities overlapped between the two plasma groups as demonstrated using both thrombin generation assays and different triggers. Spiking studies with protein S-depleted plasma, human purified protein S or antibodies against protein S confirmed a correlation between protein S and thrombin generation capacity under specific assay conditions, especially in an assay with low tissue factor concentration. Correlation between protein S and thrombin generation capacity was demonstrated in the TGA. Due to higher variability in protein S content in the FFP group, overlapping haemostatic potentials of the two plasma groups were found. © 2016 International Society of Blood Transfusion.

Imaging radar observations of frozen Arctic lakes

NASA Technical Reports Server (NTRS)

Elachi, C.; Bryan, M. L.; Weeks, W. F.

1976-01-01

A synthetic aperture imaging L-band radar flown aboard the NASA CV-990 remotely sensed a number of ice-covered lakes about 48 km northwest of Bethel, Alaska. The image obtained is a high resolution, two-dimensional representation of the surface backscatter cross section, and large differences in backscatter returns are observed: homogeneous low returns, homogeneous high returns and/or low returns near lake borders, and high returns from central areas. It is suggested that a low return indicates that the lake is frozen completely to the bottom, while a high return indicates the presence of fresh water between the ice cover and the lake bed.

Effect of prostatic fluid on the quality of fresh and frozen-thawed canine epididymal spermatozoa.

PubMed

Korochkina, E; Johannisson, A; Goodla, Lavanya; Morrell, J M; Axner, E

2014-12-01

Canine epididymal spermatozoa have a low freeze-tolerance ability compared with ejaculated spermatozoa, which could arise from the absence of prostatic fluid (PF). Therefore, the purpose of this work was to elucidate the influence of PF on the quality of canine epididymal sperm before and after freezing. Caudae epididymides were retrieved from eight dogs after routine castration. Spermatozoa were released by slicing the tissue and were extended in either Tris solution or PF before freezing. Frozen sperm samples were thawed at 70 °C for 8 seconds in a waterbath. Sperm concentration, motility using computer-assisted sperm analysis, morphology, plasma membrane, acrosome and chromatin integrity were assessed in the fresh sperm samples (after 20 minutes incubation) and at 0 and 4 hours after thawing. Progressive motility, distance straight line, distance average path, average path velocity, curvilinear velocity, straight line velocity, straightness, linearity, wobble, and beat cross frequency were significantly increased after extraction into PF. There was a higher proportion of spermatozoa with DNA damage in the PF treatment group at 4 hours after thawing than in the Tris treatment group (15.8% vs. 6.7%, P < 0.05). These results suggest that the addition of PF to canine spermatozoa activates sperm motility in fresh spermatozoa but has a negative effect on chromatin integrity after freezing-thawing. Copyright © 2014 Elsevier Inc. All rights reserved.

Breeding of Freeze-tolerant Yeast and the Mechanisms of Stress-tolerance

NASA Astrophysics Data System (ADS)

Hino, Akihiro

Frozen dough method have been adopted in the baking industry to reduce labor and to produce fresh breads in stores. New freeze-tolerant yeasts for frozen dough preparations were isolated from banana peel and identified. To obtain strains that have fermentative ability even after several months of frozen storage in fermented dough, we attempted to breed new freeze-tolerantstrain. The hybrid between S.cerevisiae, which is a isolated freeze-tolerant strain, and a strain isolated from bakers' yeast with sexual conjugation gave a good quality bread made from frozen dough method. Freeze-tolerant strains showed higher surviving and trehalose accumulating abilities than freeze-sensitive strains. The freeze tolerance of the yeasts was associated with the basal amount of intracellular trehalose after rapid degradation at the onset of the prefermentation period. The complicated metabolic pathway and the regulation system of trehalose in yeast cells are introduced. The trehalose synthesis may act as a metabolic buffer system which contribute to maintain the intracellular inorganic phosphate and as a feedback regulation system in the glycolysis. However, it is not known enough how the trehalose protects yeast cells from stress.

Environmental Load during the Distribution of Cultured Tuna

NASA Astrophysics Data System (ADS)

Sakai, Rirei; Watanabe, Manabu; Suzuki, Toru

Tuna is the most favorite marine products for Japanese people. Most of them are consumed as Sashimi in Japan, and a half of them are imported. Cold transportation technology is essential to keep freshness of tuna during the oversea transportation. In the case of transporting tuna, ship transportation in ultra low temperature such as -60 °C is applied and also high speed transportation even by using airplane is practically used. On the other hand, it is a fact that such transportation processes of tuna are giving huge environmental load, though it has not been so much focused. Evaluation of cold transportation technologies from this environmental viewpoint must be important in the future. In this article, we compared CO2 emission during cold transportations of frozen tuna (marine transportation) and non-frozen tuna (air transportation) by using LCI analysis. As a result, CO2 emission of non-frozen tuna is found to be about four times greater than that of frozen tuna. This is due to the difference in the amount of freight per transportation of the airplane and the ship.

The influence of freezing on the tensile strength of tendon grafts : a biomechanical study.

PubMed

Arnout, Nele; Myncke, Jan; Vanlauwe, Johan; Labey, Luc; Lismont, Daniel; Bellemans, Johan

2013-08-01

We investigated the influence of freezing on the tensile strength of fresh frozen tendon grafts. The biomechanical characteristics of tendons that are less commonly used in knee surgery (tibialis anterior, tibialis posterior, peroneus longus and medial and lateral half of Achilles tendons) were compared to those of a semitendinosus and gracilis graft harvested from the same 10 multi-organ donors. All right side tendons constituted the study group and were frozen at -80 degrees C and thawed at room temperature 5 times. All left side tendons were frozen at -80 degrees C and thawed at room temperature once. There were 59 tendons in the control group and 56 in the study group. The looped grafts were clamped at one side using a custom-made freeze clamp and loaded until failure on an Instron 4505 testing machine. The average ultimate failure load was not significantly different between the control and the study group (p > 0.05). The failure load of the medial tendon Achilles was the lowest in both study and control group (p < 0.001). There was no significant difference in maximum stress, maximum displacement, maximum strain and stiffness between the control and study group (p > 0.05). From our study, we conclude that freezing tendons at -80 degrees C and thawing several times does not influence the maximum load, maximum stress, maximum displacement, maximum strain and stiffness. The medial half of the Achilles tendon is clearly the weakest tendon (p < 0.001). These findings show that tendon grafts can be frozen at -80 degrees C and thawed at room temperature several times without altering their biomechanical properties.

77 FR 11481 - Notice of Request for Approval of an Information Collection; Importation of Fresh (Frozen or...

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-27

... DEPARTMENT OF AGRICULTURE Animal and Plant Health Inspection Service [Docket No. APHIS-2011-0127...) Pork or Pork Products Into the United States AGENCY: Animal and Plant Health Inspection Service, USDA... Reduction Act of 1995, this notice announces the Animal and Plant Health Inspection Service's intention to...

9 CFR 95.23 - Previously used meat covers; importations permitted subject to restrictions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... permitted subject to restrictions. Cloth or burlap which has been used to cover fresh or frozen meats...-mouth disease exists, shall not be imported except under the following conditions: (a) The cloth or... approved for the purpose by the Deputy Administrator, Veterinary Services. (b) The cloth or burlap shall be...

9 CFR 95.23 - Previously used meat covers; importations permitted subject to restrictions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... permitted subject to restrictions. Cloth or burlap which has been used to cover fresh or frozen meats...-mouth disease exists, shall not be imported except under the following conditions: (a) The cloth or... approved for the purpose by the Deputy Administrator, Veterinary Services. (b) The cloth or burlap shall be...

9 CFR 95.23 - Previously used meat covers; importations permitted subject to restrictions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... permitted subject to restrictions. Cloth or burlap which has been used to cover fresh or frozen meats...-mouth disease exists, shall not be imported except under the following conditions: (a) The cloth or... approved for the purpose by the Deputy Administrator, Veterinary Services. (b) The cloth or burlap shall be...

75 FR 48968 - Agency Information Collection Request. 30-Day Public Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-12

... the Advisory Committee on Blood Safety and Availability. Abstract: The NBCUS is a biennial survey of... and regional collections, utilization and safety of all blood products. The objective of the NBCUS is... safety of all blood products--red blood cells, fresh frozen plasma, and platelets, as well as related...

Composition of the edible portion of raw (fresh or frozen) crustaceans, finfish, and mollusks. III. Microelements. [Review

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

Sidwell, V.D.; Loomis, A.L.; Loomis, K.J.

1978-09-01

This report summarizes data from 224 publications referring to the microelements found in the flesh of commonly eaten seafoods. These microelements are: copper, iron, zinc, iodine, manganese, mercury, organic mercury, lead arsenic, fluorine, silver, cadmium, cobalt, selenium, chromium, vanadium, tin, aluminum, nickel, barium, and molybdenum.

76 FR 10266 - Importation of Ovine Meat From Uruguay

Federal Register 2010, 2011, 2012, 2013, 2014

2011-02-24

... Inspection Service 9 CFR Part 94 [Docket No. APHIS-2008-0085] RIN 0579-AD17 Importation of Ovine Meat From... proposing to amend the regulations governing the importation of certain animals, meat, and other animal products to allow, under certain conditions, the importation of fresh (chilled or frozen) ovine meat from...

Chemical and Nutritional Characteristics of long nose skate (Raja rhina) byproducts from Alaska

USDA-ARS?s Scientific Manuscript database

Skates have recently become a small commercial fishery in Alaska and along the western United States coast, but have long been associated with bycatch. The fins are marketed as "skate wings" and mainly sold fresh, frozen, and dried or salted and dehydrated for Asian markets. Byproducts generated inc...

9 CFR 319.145 - Italian sausage products.

Code of Federal Regulations, 2013 CFR

2013-01-01

... Section 319.145 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... accordance with § 319.6. (2) “Italian Sausage with Beef,” “Italian Sausage with Veal,” or “Italian Sausage with Beef and Veal,” shall be prepared so that fresh or frozen pork constitutes the major portion of...

9 CFR 319.145 - Italian sausage products.

Code of Federal Regulations, 2012 CFR

2012-01-01

... Section 319.145 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... accordance with § 319.6. (2) “Italian Sausage with Beef,” “Italian Sausage with Veal,” or “Italian Sausage with Beef and Veal,” shall be prepared so that fresh or frozen pork constitutes the major portion of...

9 CFR 319.145 - Italian sausage products.

Code of Federal Regulations, 2014 CFR

2014-01-01

... Section 319.145 Animals and Animal Products FOOD SAFETY AND INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... accordance with § 319.6. (2) “Italian Sausage with Beef,” “Italian Sausage with Veal,” or “Italian Sausage with Beef and Veal,” shall be prepared so that fresh or frozen pork constitutes the major portion of...

In situ hybridisation of EBV DNA-DNA hybrids using wet heat in polypropylene containers.

PubMed Central

Labrecque, L G

1992-01-01

AIMS: To explore procedures designed to optimise DNA-DNA in situ hybridisation, using cells infected with Epstein-Barr virus (EBV) and tissues and subfragments of the EBV DNA as probes. METHODS: The denaturation step occurred in a polypropylene container, using wet heat generated by a hot water container, the pressure cooker, or the microwave oven, without coverslips, reaching a temperature of 121 degrees C or more in these two last systems. Two different visualisation systems were used. RESULTS: Fixed cells and tumours harbouring a high and medium to low copy number (a few hundreds to 33 copies per cell), were clearly labelled, using a simple reiterated subfragment (BamW) of the EBV DNA, and fresh frozen cells, harbouring a very low copy number (one to two on average) labelled using BamW as well as BamH (single non-reiterated 6 kilobase subfragment). CONCLUSION: This is a valuable alternative technique for DNA-DNA ISH that can be used in fresh frozen samples as well as fixed samples. Images PMID:1336018

Electron spin resonance spectral analysis of irradiated royal jelly.

PubMed

Yamaoki, Rumi; Kimura, Shojiro; Ohta, Masatoshi

2014-01-15

The analysis of unpaired electron components in royal jelly was carried out using electron spin resonance (ESR) with the aim to develop a detection method for irradiated royal jelly. The ESR spectrum of royal jelly had natural signals derived from transition metals, including Fe(3+) and Cu(2+), and a signal line near g=2.00. After irradiation, a new splitting asymmetric spectrum with overall spectrum width ca. 10mT at g=2.004 was observed. The intensities of the signals at g=2.004 increased in proportion to the absorbed dose in samples under different storage conditions: fresh frozen royal jelly and dried royal jelly powder at room temperature. The signal intensity of the fresh frozen sample was stable after irradiation. One year after 10kGy irradiation of dried powder, the signal intensity was sevenfold greater than before irradiation, although the intensity continued to steadily decrease with time. This stable radiation-induced radical component was derived from the poorly soluble constituent of royal jelly. Copyright © 2013 Elsevier Ltd. All rights reserved.

A simple method for collecting eggs of taeniid cestodes from fresh, frozen or ethanol-fixed segments.

PubMed

Takemoto, Y; Negita, T; Ohnishi, K; Suzuki, M; Ito, A

1995-04-01

A simple method was devised for collecting eggs of Taenia taeniaeformis and T. saginata. All gravid segments, either fresh or frozen or 70% ethanol-fixed, were gently scraped using a pestle on a 150 mesh stainless steel sieve. Eggs and tissue debris were washed out all together with mouse tonicity phosphate buffered saline (MTPBS) through the 150 mesh sieve into a glass beaker. Egg suspension with a huge amount of tissue debris in MTPBS was centrifuged 5 min at 3000 r.p.m. (x 1600 g) and the pellet of eggs and tissue debris was resuspended with 1 vol. of MTPBS and 2 vol. of Percoll (Pharmacia) and centrifuged 60 min at 3000 r.p.m. More than 90% of eggs sedimented in the pellet. The supernatant covered with tissue debris was decanted, and the egg pellet was resuspended and centrifuged several times with MTPBS to remove Percoll. It is suggested that this simple method may prove useful for preparation of eggs of biohazardous taeniid cestodes, such as Taenia solium and Echinococcus spp.

Direct Injection of Blood Products Versus Gelatin Sponge as a Technique for Local Hemostasis

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

Haaga, John; Rahim, Shiraz, E-mail: Shiraz.rahim@uhhospitals.org

PurposeTo provide a method of reducing risk of minimally invasive procedures on patients with abnormal hemostasis and evaluate efficacy of direct fresh frozen plasma injection through a procedure needle tract compared to Gelfoam (gelatin sponge) administration.Materials and MethodsEighty patients with elevated international standardized ratio (INR) undergoing minimally invasive procedures using imaging guidance were selected retrospectively. Forty patients had received Gelfoam as a means of tract embolization during the procedure. The other 40 received local fresh frozen plasma (FFP) through the needle tract. The number of complications and clinically significant bleeding events were recorded. A threshold of 30 cc of blood lossmore » after a procedure was used to identify excess bleeding.ResultsNo patients experienced clinically significant bleeding after administration of FFP. Five patients experienced postoperative drops in hemoglobin or hematomas after administration of Gelfoam.ConclusionLocal injection of blood products can reduce postprocedure bleeding in patients undergoing minimally invasive procedures and provides a safe alternative to the use of synthetic fibrin plugs.« less

Transformation of ATLA-negative leukocytes by blood components from anti-ATLA-positive donors in vitro.

PubMed

Miyamoto, K; Tomita, N; Ishii, A; Nishizaki, T; Kitajima, K; Tanaka, T; Nakamura, T; Watanabe, S; Oda, T

1984-06-15

Anti-ATLA-positive blood components transformed healthy human leukocytes in vitro. Blood components examined were packed red cells, whole blood, platelet concentrate and fresh frozen plasma. Leukocytes present in anti-ATLA-positive blood components such as packed red cells, whole blood and platelet concentrate easily transformed anti-ATLA-negative leukocytes. Co-culture in fresh frozen plasma, however, did not transform recipient leukocytes, and leukocytes of anti-ATLA-positive recipients proved refractory to transformation. The transformed cells were morphologically lymphoid, grew in suspension, and possessed normal recipient karyotypes except in the case of three platelet concentrates. A high proportion of all the transformed populations formed E-rosettes with neuraminidase-treated sheep erythrocytes. The cytoplasm of over 90% of each recipient was stained brilliantly with antibodies against ATLV-determined antigens. Electron microscopy of these transformed cells revealed many C-type virus particles in the extracellular space. Blood components, such as packed red cells, whole blood and platelet concentrate, containing leukocytes from anti-ATLA-positive donors, should be used cautiously to prevent the transmission on ATLV to anti-ATLA-negative recipients.

Single-layer centrifugation through PureSperm® 80 selects improved quality spermatozoa from frozen-thawed dog semen.

PubMed

Dorado, J; Alcaraz, L; Gálvez, M J; Acha, D; Ortiz, I; Urbano, M; Hidalgo, M

2013-08-01

The aim of this study was to investigate whether single-layer centrifugation (SLC) with PureSperm® 80 could select good quality spermatozoa, including those with specific motility patterns, from doses of frozen dog semen. Semen from 5 dogs was collected and cryopreserved following a standard protocol. After thawing, semen samples were divided into two aliquots: one of them was used as control and the other one processed by SLC. Assessment of sperm motility (assessed by computer-assisted semen analysis), morphology (Diff-Quick staining) and viability (triple fluorescent stain of propidium iodine/isothiocyanate-labeled peanut (Arachis hypogaea) agglutinin/Rhodamine 123), were performed on aliquots of fresh semen, frozen-thawed control and frozen-thawed SLC treated samples. A multivariate clustering procedure separated 26,051 motile spermatozoa into three subpopulations (sP): sP1 consisting of highly active but non-progressive spermatozoa (40.3%), sP2 consisting of spermatozoa with high velocity and progressive motility (30.0%), and sP3 consisting of poorly active and non-progressive spermatozoa (29.7%). SLC with PureSperm® 80 yielded sperm suspensions with improved motility, morphology, viability and acrosome integrity (P<0.001). The frozen-thawed SLC treated samples were enriched in sP2, reaching a proportion of 44.1% of the present spermatozoa. From these results, we concluded that SLC with PureSperm® 80 may be an alternative and successful method for improving the quality of frozen-thawed dog spermatozoa. Moreover, sP2 (high-speed and progressive spermatozoa) was more frequently observed after SLC. Finally, this study also demonstrated that the general motile sperm structure present in dogs remained constant despite the effect caused by either cryopreservation or separation by SLC through PureSperm® 80. Copyright © 2013 Elsevier B.V. All rights reserved.

Measuring zinc in biological nanovesicles by multiple analytical approaches.

PubMed

Piacenza, Francesco; Biesemeier, Antje; Farina, Marco; Piva, Francesco; Jin, Xin; Pavoni, Eleonora; Nisi, Lorenzo; Cardelli, Maurizio; Costarelli, Laura; Giacconi, Robertina; Basso, Andrea; Pierpaoli, Elisa; Provinciali, Mauro; Hwang, James C M; Morini, Antonio; di Donato, Andrea; Malavolta, Marco

2018-07-01

Exosomes are nanovesicles known to mediate intercellular communication. Although it is established that zinc ions can act as intracellular signaling factors, the measurement of zinc in circulating nanovesicles has not yet been attempted. Providing evidence of the existence of this zinc fraction and methods for its measurement might be important to advance our knowledge of zinc status and its relevance in diseases. Exosomes from 0.5 ml of either fresh or frozen human plasma were isolated by differential centrifugation. A morphological and dimensional evaluation at the nanoscale level was performed by atomic force microscopy (AFM) and Transmission Electron Microscopy (TEM). Energy Dispersive X-Ray Microanalysis (EDX) revealed the elemental composition of exosomes and their respective total Zinc content on a quantitative basis. The zinc mole fraction (in at%) was correlated to the phosphorous mole fraction, which is indicative for exosomal membrane material. Both fresh (Zn/P 0.09 ± 0.01) and frozen exosomes (Zn/P 0.08 ± 0.02) had a significant zinc content, which increased up to 1.09 ± 0.12 for frozen exosomes when treated with increasing amounts of zinc (100-500 μM; each p < 0.05). Interestingly, after zinc addition, the Calcium mole fractions decreased accordingly suggesting a possible exchange by zinc. In order to estimate the intra-exosomal labile zinc content, an Imaging Flow Cytometry approach was developed by using the specific membrane permeable zinc-probe Fluozin-3AM. A labile zinc content of 0.59 ± 0.27 nM was calculated but it is likely that the measurement may be affected by purification and isolation conditions. This study suggests that circulating nano-vesicular-zinc can represent a newly discovered zinc fraction in the blood plasma whose functional and biological properties will have to be further investigated in future studies. Copyright © 2018 Elsevier GmbH. All rights reserved.

Effects of freezing in and out of water on length and weight of Lake Michigan bloaters

USGS Publications Warehouse

Sayers, Richard E.

1987-01-01

The purpose of this study was to determine if freezing significantly alters the length or weight of bloaters Coregonus hoyi. Bloaters were collected from southern Lake Michigan and were frozen for periods of 2-200 d. Freezing in water caused a significant decrease in length and a significant increase in weight. These changes did not vary predictably with time. The mean change in weight was greater for adults than for juveniles, but the mean change in length was not significantly different between juveniles and adults. Regressions for weight or length after freezing versus weight or length before freezing were highly significant and can be used as correction equations for estimating the original lengths and weights of fresh specimens after fish have been frozen. Test fish that were subsequently refrozen in air shrank more than those refrozen in water.

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.

Fruit and vegetable availability and selection: federal food package revisions, 2009.

PubMed

Zenk, Shannon N; Odoms-Young, Angela; Powell, Lisa M; Campbell, Richard T; Block, Daniel; Chavez, Noel; Krauss, Ramona C; Strode, Steven; Armbruster, James

2012-10-01

With nearly 49,000 authorized retailers nationwide, a policy change that added fruits and vegetables (FV) to the U.S. Department of Agriculture's Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) food packages in 2009 had the potential to expand neighborhood FV availability. This study examined changes in availability and selection of commonly consumed and culturally specific FV at authorized retailers (WIC vendors) before and after implementation of the revised WIC food packages. Quasi-experimental, one-group design with two pre-policy observations and one post-policy observation. Trained observers assessed a list of fresh, frozen, and canned FV at each vendor in seven northern Illinois counties. Eight indices of FV availability and selection were derived. Multiple regression estimated relationships. Data were collected in 2008-2010 and analyzed in 2011. Overall, availability and selection of commonly consumed fresh FV and availability of African-American culturally specific fresh FV improved after implementation of the new policy. Modest improvements in the overall availability of canned low-sodium vegetables and frozen FV were observed. Changes differed by vendor type (large vendor, small vendor, and pharmacy). Changes in availability or selection did not differ by neighborhood characteristics (population density, median household income, racial/ethnic composition). Expansion of WIC foods was associated with small positive externalities on the food environment. Larger subsidies to create more demand and more-substantial stocking requirements for retailers may yield significantly larger improvements and thus warrant further investigation. Approaches targeting rural, low-income, and racial/ethnic minority neighborhoods also may be needed. Copyright © 2012 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.

Mitochondrial membrane potential and reactive oxygen species in liquid stored and cryopreserved turkey (Meleagris gallopavo) spermatozoa.

PubMed

Slowinska, M; Liszewska, E; Judycka, S; Konopka, M; Ciereszko, A

2018-06-01

The extensive use of artificial insemination in turkeys has led to the development of in vitro semen storage. However, fertility rates from liquid stored and frozen/thawed turkey semen are still unsatisfactory. The aim of the study was to assess spermatozoa viability, mitochondrial membrane potential (MMP), and reactive oxygen species production (ROS) in liquid stored and cryopreserved turkey semen with the use of flow cytometry. Moreover, motility and adenosine triphosphate (ATP) content in sperm were monitored at the same time to link flow cytometry data with sperm movement and energetics. Liquid storage led to a decrease in sperm motility (80.6 vs. 55.6%, for fresh and stored for 48 h), live sperm with an intact MMP (59.9 vs. 30.5% for fresh and stored for 48 h), and a 20-fold decrease in ATP content after 24 h of storage. A 3-fold increase in ROS+ sperm was observed after 48 h of storage (9.3 vs. 26.8% for fresh and stored for 48 h). Semen equilibration before cryopreservation affected only ATP content. However, freezing/thawing led to a dramatic decrease in all of the studied semen quality parameters. A 5-fold decrease in live sperm with intact MMP (59.8 vs. 11.9%) and a 7-fold increase in sperm ROS+ (10.8 vs. 74.4%) were recorded between fresh and frozen/thawed semen. The results strongly suggested that a significant loss of MMP and a disturbance in sperm ATP production during semen storage can be the main reason for the decline in sperm motility. The disturbance of mitochondria activity during storage seems to be associated with the increase in oxidative stress in turkey semen. Turkey sperm mitochondria also appear to be very sensitive to cryodamage. Diminished energy production in turkey spermatozoa, visible as the low percentage of sperm with an intact MMP and low level of ATP after freezing/thawing, which is associated with high ROS generation, could be responsible for the low fertilizing ability of cryopreserved turkey semen.

Do processed vegetables reduce the socio-economic differences in vegetable purchases? A study in France.

PubMed

Plessz, Marie; Gojard, Séverine

2013-10-01

Vegetable consumption varies highly across households, based on household structure and socio-economic status, but little is known about the share of fresh vs. processed (e.g. frozen or canned) vegetables. Our aim was to compare the social and economic determinants of fresh and processed vegetable consumption. We reviewed detailed data on vegetable purchases for at-home consumption of 2600 French households during 2007. We took into account a wide range of processed vegetables (excluding potatoes) and made a distinction between fresh vegetables, processed vegetables and baby food containing vegetables. We conducted regression analyses to predict consumption of fresh and processed vegetables in kilograms per year and unit values in euros per kilogram. About 60% of the vegetables bought by the sample households were fresh. Fresh vegetable consumption increased with the respondent's income, age and educational level, and with the number of adults but not with the presence of children aged <6 years. The quantity of processed vegetables purchased increased with the household size but was not dependent on age, education or household income, although the richest households spent more per kilogram on processed vegetables. Households with a child aged <6 years also purchased 10 kg of baby foods containing vegetables. We found socio-economic inequalities in the quantities of fresh vegetables, in the spending on fresh and processed vegetables but not in the quantities of processed vegetables. This suggests that monitoring the price and nutritional quality of processed vegetables and providing this information to consumers could help them identify nutritious, affordable and convenient foods.

Use of lactic acid with electron beam irradiation for control of Escherichia coli O157:H7, non-O157 VTEC E. coli, and Salmonella serovars on fresh and frozen beef.

PubMed

Li, Shuliu; Kundu, Devapriya; Holley, Richard A

2015-04-01

Lactic acid pre-treatment was examined to enhance the antimicrobial action of electron (e-) beam irradiation of beef trim. Meat samples were inoculated with Escherichia coli O157:H7, non-O157 VTEC E. coli or Salmonella cocktails and treated with 5% lactic acid at 55 °C. Samples were packaged aerobically or vacuum-packed, kept at 4 °C and treated with 1 kGy e-beam energy. Frozen samples were treated with 1, 3 or 7 kGy and stored at -20 °C for ≤ 5 d. Lactic acid enhanced the antimicrobial action of 1 kGy e-beam treatment against Salmonella by causing an additional <1.8 log CFU/g reduction. One kGy treatment of refrigerated samples reduced VTEC E. coli viability by 4.5 log CFU/g, and while lactic acid did not improve the reduction, after freezing additive effects were found. After 3 kGy irradiation, Salmonella was reduced by 2 and 4 log CFU/g in the irradiated and lactic acid plus irradiated samples, respectively. Lactic acid pre-treatment was of limited value with 1 kGy treatment for improving control of toxigenic E. coli in fresh beef trim, however, it would be useful with low dose irradiation for controlling both VTEC E. coli and Salmonella in frozen product. Copyright © 2014 Elsevier Ltd. All rights reserved.

Autism and mental retardation among offspring born after in vitro fertilization.

PubMed

Sandin, Sven; Nygren, Karl-Gösta; Iliadou, Anastasia; Hultman, Christina M; Reichenberg, Abraham

2013-07-03

Between 1978 and 2010, approximately 5 million infants were born after in vitro fertilization (IVF) treatments. Yet limited information on neurodevelopment after IVF exists, especially after the first year of life. To examine the association between use of any IVF and different IVF procedures and the risk of autistic disorder and mental retardation in the offspring. A population-based, prospective cohort study using Swedish national health registers. Offspring born between 1982 and 2007 were followed up for a clinical diagnosis of autistic disorder or mental retardation until December 31, 2009. The exposure of interest was IVF, categorized according to whether intracytoplasmic sperm injection (ICSI) for male infertility was used and whether embryos were fresh or frozen. For ICSI, whether sperm were ejaculated or surgically extracted was also considered. Relative risks (RRs) for autistic disorder and mental retardation and rates per 100,000 person-years, comparing spontaneously conceived offspring with those born after an IVF procedure and comparing 5 IVF procedures used in Sweden vs IVF without ICSI with fresh embryo transfer, the most common treatment. We also analyzed the subgroup restricted to singletons. Of the more than 2.5 million infants born, 30,959 (1.2%) were conceived by IVF and were followed up for a mean 10 (SD, 6) years. Overall, 103 of 6959 children (1.5%) with autistic disorder and 180 of 15,830 (1.1%) with mental retardation were conceived by IVF. The RR for autistic disorder after any procedure compared with spontaneous conception was 1.14 (95% CI, 0.94-1.39; 19.0 vs 15.6 per 100,000 person-years). The RR for mental retardation was 1.18 (95% CI, 1.01-1.36; 46.3 vs 39.8 per 100,000 person-years). For both outcomes, there was no statistically significant association when restricting analysis to singletons. Compared with IVF without ICSI with fresh embryo transfer, there were statistically significantly increased risks of autistic disorder following ICSI using surgically extracted sperm and fresh embryos (RR, 4.60 [95% CI, 2.14-9.88]; 135.7 vs 29.3 per 100,000 person-years); for mental retardation following ICSI using surgically extracted sperm and fresh embryos (RR, 2.35 [95% CI, 1.01-5.45]; 144.1 vs 60.8 per 100,000 person-years); and following ICSI using ejaculated sperm and fresh embryos (RR, 1.47 [95% CI, 1.03-2.09]; 90.6 vs 60.8 per 100,000 person-years). When restricting the analysis to singletons, the risks of autistic disorder associated with ICSI using surgically extracted sperm were not statistically significant, but the risks associated with ICSI using frozen embryos were significant for mental retardation (with frozen embryos, RR, 2.36 [95% CI, 1.04-5.36], 118.4 vs 50.6 per 100,000 person-years]; with fresh embryos, RR, 1.60 [95% CI, 1.00-2.57], 80.0 vs 50.6 per 100,000 person-years). Compared with spontaneous conception, IVF treatment overall was not associated with autistic disorder but was associated with a small but statistically significantly increased risk of mental retardation. For specific procedures, IVF with ICSI for paternal infertility was associated with a small increase in the RR for autistic disorder and mental retardation compared with IVF without ICSI. The prevalence of these disorders was low, and the increase in absolute risk associated with IVF was small.

A fresh liver agar substrate for rearing small numbers of forensically important blow flies (Diptera: Calliphoridae).

PubMed

Gruner, Susan V; Slone, Daniel H

2014-05-01

Forensically important calliphorids can be reared on a mixture of beef liver and agar. Small pieces of meat, especially fresh or frozen beef liver, will desiccate in 2-6 h, but this simple-to-make feeding substrate remains moist for at least 12 h at 25 and 30 degrees C without desiccation, even in small (5 g) amounts. We determined the survivorship of small numbers of Chrysomya megacephala (F.) (first-instar larvae to adult eclosion) raised on 5 g of liver agar and fresh beef liver. We found that all larvae raised on 5 g of liver died due to desiccation, but survivorship on 5 g of liver agar was equivalent to that on larger (50 g) pieces of either liver agar or beef liver.

Hand-portable gas chromatography-ion mobility spectrometer for the determination of the freshness of fish

NASA Technical Reports Server (NTRS)

Snyder, A. Peter; Harden, Charles S.; Davis, Dennis M.; Shoff, Donald B.; Maswadeh, Waleed M.

1995-01-01

A hand-held, portable gas chromatography-ion mobility spectrometer (GC-IMS) device was used to detect the presence of volatile amine compounds in the headspace of decomposing fish. The Food and Drug Administration (FDA) largely relies on olfactory discrimination with respect to fresh and spoiled, frozen and unfrozen fish. The fish are delivered at ship docks on pallets, and each pallet of fish can range from 30-40 thousand dollars in value. Fresh fish were placed in a teflon bag and the direct headspace was interrogated. In the first three days, only low molecular weight volatile amines were detected. On the fourth day, a number of spectral signatures were observed which indicated the presence of 1,5-diaminopentane, cadaverine. Analyses typically took from 0.5-1 minute.

A methodological study of genome-wide DNA methylation analyses using matched archival formalin-fixed paraffin embedded and fresh frozen breast tumors.

PubMed

Espinal, Allyson C; Wang, Dan; Yan, Li; Liu, Song; Tang, Li; Hu, Qiang; Morrison, Carl D; Ambrosone, Christine B; Higgins, Michael J; Sucheston-Campbell, Lara E

2017-02-28

DNA from archival formalin-fixed and paraffin embedded (FFPE) tissue is an invaluable resource for genome-wide methylation studies although concerns about poor quality may limit its use. In this study, we compared DNA methylation profiles of breast tumors using DNA from fresh-frozen (FF) tissues and three types of matched FFPE samples. For 9/10 patients, correlation and unsupervised clustering analysis revealed that the FF and FFPE samples were consistently correlated with each other and clustered into distinct subgroups. Greater than 84% of the top 100 loci previously shown to differentiate ER+ and ER- tumors in FF tissues were also FFPE DML. Weighted Correlation Gene Network Analyses (WCGNA) grouped the DML loci into 16 modules in FF tissue, with ~85% of the module membership preserved across tissue types. Restored FFPE and matched FF samples were profiled using the Illumina Infinium HumanMethylation450K platform. Methylation levels (β-values) across all loci and the top 100 loci previously shown to differentiate tumors by estrogen receptor status (ER+ or ER-) in a larger FF study, were compared between matched FF and FFPE samples using Pearson's correlation, hierarchical clustering and WCGNA. Positive predictive values and sensitivity levels for detecting differentially methylated loci (DML) in FF samples were calculated in an independent FFPE cohort. FFPE breast tumors samples show lower overall detection of DMLs versus FF, however FFPE and FF DMLs compare favorably. These results support the emerging consensus that the 450K platform can be employed to investigate epigenetics in large sets of archival FFPE tissues.

Robust transcriptional tumor signatures applicable to both formalin-fixed paraffin-embedded and fresh-frozen samples

PubMed Central

Cheng, Jun; He, Jun; Liu, Huaping; Cai, Hao; Hong, Guini; Zhang, Jiahui; Li, Na; Ao, Lu; Guo, Zheng

2017-01-01

Formalin-fixed paraffin-embedded (FFPE) samples represent a valuable resource for clinical researches. However, FFPE samples are usually considered an unreliable source for gene expression analysis due to the partial RNA degradation. In this study, through comparing gene expression profiles between FFPE samples and paired fresh-frozen (FF) samples for three cancer types, we firstly showed that expression measurements of thousands of genes had at least two-fold change in FFPE samples compared with paired FF samples. Therefore, for a transcriptional signature based on risk scores summarized from the expression levels of the signature genes, the risk score thresholds trained from FFPE (or FF) samples could not be applied to FF (or FFPE) samples. On the other hand, we found that more than 90% of the relative expression orderings (REOs) of gene pairs in the FF samples were maintained in their paired FFPE samples and largely unaffected by the storage time. The result suggested that the REOs of gene pairs were highly robust against partial RNA degradation in FFPE samples. Finally, as a case study, we developed a REOs-based signature to distinguish liver cirrhosis from hepatocellular carcinoma (HCC) using FFPE samples. The signature was validated in four datasets of FFPE samples and eight datasets of FF samples. In conclusion, the valuable FFPE samples can be fully exploited to identify REOs-based diagnostic and prognostic signatures which could be robustly applicable to both FF samples and FFPE samples with degraded RNA. PMID:28036264

Reduction of potassium content of green bean pods and chard by culinary processing. Tools for chronic kidney disease.

PubMed

Martínez-Pineda, Montserrat; Yagüe-Ruiz, Cristina; Caverni-Muñoz, Alberto; Vercet-Tormo, Antonio

2016-01-01

In order to prevent a possible hyperkalemia, chronic renal patients, especially in advanced stages, must follow a low potassium diet. So dietary guidelines for chronic kidney disease recommend limiting the consumption of many vegetables, as well as to apply laborious culinary techniques to maximize the reduction of potassium. The aim of this work is to analyze potassium content from several vegetable, fresh products, frozen and preserved, as well as check and compare the effectiveness in potassium reduction of different culinary processes, some of them recommended in dietary guidelines such as soaking or double cooking. Sample potassium content was analyzed by triplicate using flamephotometry. The results showed significant reductions in potassium content in all culinary processes studied. The degree of loss varied depending on the type of vegetable and processing applied. Frozen products achieved greater reductions than the fresh ones, obtaining in some cases losses greater than 90%. In addition, it was observed how in many cases the single application of a normal cooking reached potassium reductions to acceptable levels for its inclusion in renal patient diet. The results shown in this study are very positive because they provide tools for professionals who deal with this kind of patients. They allow them to adapt more easily to the needs and preferences of their patients and increase dietary variety. Copyright © 2016 Sociedad Española de Nefrología. Published by Elsevier España, S.L.U. All rights reserved.

Immunohistochemical diagnosis of Alport's syndrome in paraffin-embedded renal sections: antigen retrieval with autoclave heating.

PubMed

Naito, Ichiro; Ninomiya, Yoshifumi; Nomura, Shinsuke

2003-03-01

Alport's syndrome (AS) is a hereditary renal disease caused by mutations in the genes encoding collagen type IV. Immunohistochemical analysis of the alpha chains of collagen type IV has been found to be useful for the diagnosis of this disease. The monoclonal antibodies (mAbs) generated by us recognize alpha 1(IV) through alpha 6(IV) chains of collagen type IV on fresh-frozen sections but not on paraffin-embedded sections. Antigen retrieval by autoclave heating has been found to restore the epitopes recognized by the mAbs; however the heating conditions had not been well established. In this study, the heating conditions were carefully examined using renal sections obtained from AS and non-AS patients. The heating was performed in an autoclave, at 105 degrees -127 degrees C for 6-8 min. During the heating, the sections were immersed in 0.2 N HCl solution (pH 0.9). Then, the mAbs were applied for 30 min, and the bound mAbs were detected using the LSAB kit. The optimal temperature for the antigen retrieval varied among specimens, and was dependent on the type of basement membrane examined. Thus, it was considered that heating at two or three different temperatures could be helpful for the precise diagnosis of AS. Adopting the antigen retrieval method could extend the possibility of immunohistochemical diagnosis of AS to cases without using fresh-frozen sections.

Development of a compact terahertz time-domain spectrometer for the measurement of the optical properties of biological tissues.

PubMed

Wilmink, Gerald J; Ibey, Bennett L; Tongue, Thomas; Schulkin, Brian; Laman, Norman; Peralta, Xomalin G; Roth, Caleb C; Cerna, Cesario Z; Rivest, Benjamin D; Grundt, Jessica E; Roach, William P

2011-04-01

Terahertz spectrometers and imaging systems are currently being evaluated as biomedical tools for skin burn assessment. These systems show promise, but due to their size and weight, they have restricted portability, and are impractical for military and battlefield settings where space is limited. In this study, we developed and tested the performance of a compact, light, and portable THz time-domain spectroscopy (THz-TDS) device. Optical properties were collected with this system from 0.1 to 1.6 THz for water, ethanol, and several ex vivo porcine tissues (muscle, adipose, skin). For all samples tested, we found that the index of refraction (n) decreases with frequency, while the absorption coefficient (μ(a)) increases with frequency. Muscle, adipose, and frozen/thawed skin samples exhibited comparable n values ranging between 2.5 and 2.0, whereas the n values for freshly harvested skin were roughly 40% lower. Additionally, we found that the freshly harvested samples exhibited higher μ(a) values than the frozen/thawed skin samples. Overall, for all liquids and tissues tested, we found that our system measured optical property values that were consistent with those reported in the literature. These results suggest that our compact THz spectrometer performed comparable to its larger counterparts, and therefore may be a useful and practical tool for skin health assessment.

Changes in microbial communities and quality attributes of white muscle and dark muscle from common carp (Cyprinus carpio) during chilled and freeze-chilled storage.

PubMed

Li, Qian; Zhang, Longteng; Luo, Yongkang

2018-08-01

This study investigated sensory scores, quality attributes and microbial communities in white muscle and dark muscle of common carp (Cyprinus carpio) during chilled (4 °C) and freeze-chilled (-20 °C for 4 weeks prior to 4 °C) storage. Compared to the samples at the end of storage, fresh samples and frozen-thawed samples on day 0 showed greater bacterial diversity and more differences in microbiota. Initially, Aeromonas was the prevalent genus in fresh white muscle and dark muscle. As time progressed, Aeromonas followed by Pseudomonas predominated in white muscle, while Aeromonas, Pseudomonas, and Lactococcus dominated in dark muscle. Paenibacillus was identified as the largest population in frozen-thawed samples of both muscle types, but Pseudomonas increased dramatically to become dominant in the two spoiled samples. Volatile organic compounds (VOCs) of carp muscle consisted mainly of aldehydes and alcohols, and the percentage of ketones in both muscle types increased considerably after storage. Moreover, dark muscle showed more kinds of VOCs, and a slower rate of quality deterioration than white muscle. Based on sensory assessment, the shelf-life of white muscle and dark muscle of common carp for chilled storage was 8 days and 10 days, respectively, as well as 8 days together for freeze-chilled storage. Copyright © 2018 Elsevier Ltd. All rights reserved.

Tumor Touch Imprints as Source for Whole Genome Analysis of Neuroblastoma Tumors

PubMed Central

Brunner, Clemens; Brunner-Herglotz, Bettina; Ziegler, Andrea; Frech, Christian; Amann, Gabriele; Ladenstein, Ruth; Ambros, Inge M.; Ambros, Peter F.

2016-01-01

Introduction Tumor touch imprints (TTIs) are routinely used for the molecular diagnosis of neuroblastomas by interphase fluorescence in-situ hybridization (I-FISH). However, in order to facilitate a comprehensive, up-to-date molecular diagnosis of neuroblastomas and to identify new markers to refine risk and therapy stratification methods, whole genome approaches are needed. We examined the applicability of an ultra-high density SNP array platform that identifies copy number changes of varying sizes down to a few exons for the detection of genomic changes in tumor DNA extracted from TTIs. Material and Methods DNAs were extracted from TTIs of 46 neuroblastoma and 4 other pediatric tumors. The DNAs were analyzed on the Cytoscan HD SNP array platform to evaluate numerical and structural genomic aberrations. The quality of the data obtained from TTIs was compared to that from randomly chosen fresh or fresh frozen solid tumors (n = 212) and I-FISH validation was performed. Results SNP array profiles were obtained from 48 (out of 50) TTI DNAs of which 47 showed genomic aberrations. The high marker density allowed for single gene analysis, e.g. loss of nine exons in the ATRX gene and the visualization of chromothripsis. Data quality was comparable to fresh or fresh frozen tumor SNP profiles. SNP array results were confirmed by I-FISH. Conclusion TTIs are an excellent source for SNP array processing with the advantage of simple handling, distribution and storage of tumor tissue on glass slides. The minimal amount of tumor tissue needed to analyze whole genomes makes TTIs an economic surrogate source in the molecular diagnostic work up of tumor samples. PMID:27560999

Traumatic Hemothorax Blood Contains Elevated Levels of Microparticles that are Prothrombotic but Inhibit Platelet Aggregation.

PubMed

Mitchell, Thomas A; Herzig, Maryanne C; Fedyk, Chriselda G; Salhanick, Marc A; Henderson, Aaron T; Parida, Bijaya K; Prat, Nicolas J; Dent, Daniel L; Schwacha, Martin G; Cap, Andrew P

2017-06-01

Autotransfusion of shed blood from traumatic hemothorax is an attractive option for resuscitation of trauma patients in austere environments. However, previous analyses revealed that shed hemothorax (HX) blood is defibrinated, thrombocytopenic, and contains elevated levels of D-dimer. Mixing studies with normal pooled plasma demonstrated hypercoagulability, evoking concern for potentiation of acute traumatic coagulopathy. We hypothesized that induction of coagulopathic changes by shed HX blood may be due to increases in cellular microparticles (MP) and that these may also affect recipient platelet function. Shed HX blood was obtained from 17 adult trauma patients under an Institutional Review Board approved prospective observational protocol. Blood samples were collected every hour up to 4 h after thoracostomy tube placement. The corresponding plasma was isolated and frozen for analysis. The effects of shed HX frozen plasma (HFP) and isolated HX microparticles (HMP) on coagulation and platelet function were assessed through mixing studies with platelet-rich plasma at various dilutions followed by analysis with thromboelastometry (ROTEM), platelet aggregometry (Multiplate), enzyme-linked immunosorbent assays, and flow cytometry. Furthermore, HFP was assessed for von Willebrand factor antigen levels and multimer content, and plasma-free hemoglobin. ROTEM analysis demonstrated that diluted HFP and isolated HMP samples decreased clotting time, clotting formation time, and increased α angle, irrespective of sample concentrations, when compared with diluted control plasma. Isolated HMP inhibited platelet aggregation in response to adenosine diphosphate, arachidonic acid, and collagen. HFP contained elevated levels of fibrin-degradation products and tissue factor compared with control fresh frozen plasma samples. MP concentrations in HFP were significantly increased and enriched in events positive for phosphatidylserine, tissue factor, CD235, CD45, CD41a, and CD14. von Willebrand factor (vWF) multimer analysis revealed significant loss of high molecular weight multimers in HFP samples. Plasma-free hemoglobin levels were 8-fold higher in HFP compared with fresh frozen plasma. HFP induces plasma hypercoagulability that is likely related to increased tissue factor and phosphatidylserine expression originating from cell-derived MP. In contrast, platelet dysfunction is induced by HMP, potentially aggravated by depletion of high molecular weight multimers of vWF. Thus, autologous transfusion of shed traumatic hemothorax blood may induce a range of undesirable effects in patients with acute traumatic coagulopathy.

Cost-Effectiveness of the Freeze-All Policy.

PubMed

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

2015-08-01

To evaluate the cost-effectiveness of freeze-all cycles when compared to fresh embryo transfer. This was an observational study with a cost-effectiveness analysis. The analysis consisted of 530 intracytoplasmic sperm injection (ICSI) cycles in a private center in Brazil between January 2012 and December 2013. A total of 530 intracytoplasmic sperm injection (ICSI) cycles - 351 fresh embryo transfers and 179 freeze-all cycles - with a gonadotropin-releasing hormone (GnRH) antagonist protocol and day 3 embryo transfers. The pregnancy rate was 31.1% in the fresh group and 39.7% in the freeze-all group. We performed two scenario analyses for costs. In scenario 1, we included those costs associated with the ICSI cycle (monitoring during controlled ovarian stimulation [COS], oocyte retrieval, embryo transfer, IVF laboratory, and medical costs), embryo cryopreservation of supernumerary embryos, hormone measurements during COS and endometrial priming, medication use (during COS, endometrial priming, and luteal phase support), ultrasound scan for frozen- thawed embryo transfer (FET), obstetric ultrasounds, and miscarriage. The total cost (in USD) per pregnancy was statistically lower in the freeze-all cycles (19,156.73 ± 1,732.99) when compared to the fresh cycles (23,059.72 ± 2,347.02). Even in Scenario 2, when charging all of the patients in the freeze-all group for cryopreservation (regardless of supernumerary embryos) and for FET, the fresh cycles had a statistically significant increase in treatment costs per ongoing pregnancy. The results presented in this study suggest that the freeze-all policy is a cost-effective strategy when compared to fresh embryo transfer.

Every Day Lots of Ways. An Interdisciplinary Nutrition Curriculum for Kindergarten-Sixth Grade.

ERIC Educational Resources Information Center

Bagby, Robin; Campbell, V. Star; Achterberg, Cheryl; Probart, Claudia; Ebel, Kimberlyn

This K-6 interdisciplinary nutrition curriculum focuses on vegetables and fruits. The lessons use core subjects to promote consumption of fresh, frozen, and canned vegetables and fruits; vegetable and fruit juices; and dried fruits. The lessons also investigate the Food Guide Pyramid. Each lesson plan has a section entitled Extensions. The…

Attack and success of native and exotic parasitoids on eggs of Halyomorpha halys in three Maryland habitats

USDA-ARS?s Scientific Manuscript database

Egg parasitoids of the exotic invasive brown marmorated stink bug, Halyomorpha halys (Stål), were investigated using lab-reared fresh (live) and frozen (killed) lab-reared sentinel egg masses deployed for 72h on foliage in three habitats -- woods, orchard, and soybean field -- in Maryland, USA, in s...

9 CFR 94.26 - Restrictions on importation of live poultry, poultry meat, and other poultry products from...

Code of Federal Regulations, 2010 CFR

2010-01-01

... poultry, poultry meat, and other poultry products from specified regions. 94.26 Section 94.26 Animals and... poultry, poultry meat, and other poultry products from specified regions. Argentina and the Mexican States... Newcastle disease (END), supplement their meat supply by the importation of fresh (chilled or frozen...

The safety of early fresh, whole blood transfusion among severely battle injured at US Marine Corps forward surgical care facilities in Afghanistan.

PubMed

Auten, Jonathan D; Lunceford, Nicole L; Horton, Jaime L; Galarneau, Mike R; Galindo, Roger M; Shepps, Craig D; Zieber, Tara J; Dewing, Chris B

2015-11-01

In Afghanistan, care of the acutely injured trauma patient commonly occurred in facilities with limited blood banking capabilities. Apheresis platelets were often not available. Component therapy consisted of 1:1 packed red blood cells and fresh frozen plasma. Fresh, whole blood transfusion often augmented therapy in the severely injured patient. This study analyzed the safety of fresh, whole blood use in a resource-limited setting. A retrospective analysis was performed on a prospectively collected data set of US battle injuries presenting to three US Marine Corps (USMC) expeditionary surgical care facilities in Helmand Province, Afghanistan, between January 2010 and July 2012. Included in the review were patients with Injury Severity Scores (ISSs) of 15 or higher receiving blood transfusions. Univariate analyses were performed, followed by multivariable logistic regression to describe the relationship between the treatment group and posttreatment complications such as trauma-induced coagulopathy, infection, mortality, venous thromboembolism, and transfusion reaction. Propensity scores were calculated and included in multivariable models to adjust for potential bias in treatment selection. A total of 61 patients were identified; all were male marines with a mean (SD) age of 23.5 (3.6) years. The group receiving fresh, whole blood was noted to have higher ISSs and lower blood pressure, pH, and base deficits on arrival. Traumatic coagulopathy was significantly less common in the group receiving fresh, whole blood (odds ratio, 0.01; 95% confidence interval, 0.00-0.18). Multivariable models found no other significant differences between the treatment groups. The early use of fresh, whole blood in a resource-limited setting seems to confer a benefit in reducing traumatic coagulopathy. This study's small sample size precludes further statement on the overall safety of fresh, whole blood use. Therapy study, level IV.

Validating a refractometer to evaluate immunoglobulin G concentration in Jersey colostrum and the effect of multiple freeze-thaw cycles on evaluating colostrum quality.

PubMed

Morrill, K M; Robertson, K E; Spring, M M; Robinson, A L; Tyler, H D

2015-01-01

The objectives of this study were to (1) validate a method using refractometry to rapidly and accurately determine immunoglobulin (IgG) concentration in Jersey colostrum, (2) determine whether there should be different refractive index (nD) and %Brix cut points for Jersey colostrum, and (3) evaluate the effect of multiple freeze-thaw (FT) cycles on radial immunodiffusion (RID) and a digital refractometer to determine IgG concentration in Jersey colostrum. Samples (n=58; 3L) of colostrum were collected from a dairy in northwestern Iowa. Samples were analyzed within 2h of collection for IgG concentration by RID, %Brix, and nD by refractometer and an estimate of IgG by colostrometer. Samples were frozen, placed on dry ice, and transported to the laboratory at Iowa State University (Ames). Samples arrived frozen and were placed in a -20°C manual-defrost freezer until further analysis. On d 7 (1FT), d 14 (2FT), and 1yr (3FT) all samples were thawed, analyzed for IgG by RID, %Brix, nD by refractometer, and IgG estimate by colostrometer, and frozen until reanalysis at the next time point. Fresh colostrum had a mean (±SD) IgG concentration of 72.91 (±33.53) mg/mL, 21.24% (±4.43) Brix, and nD 1.3669 (±0.0074). Multiple FT cycles did affect IgG as determined by RID and colostrometer reading. The IgG concentrations were greater in fresh and 1FT samples as compared with 2FT and 3FT samples (72.91, 75.38, 67.20, and 67.31mg of IgG/mL, respectively). The colostrometer reading was lower in 1FT samples compared with fresh and 2FT samples. Multiple FT cycles had no effect on nD or %Brix reading. In fresh samples, IgG concentration was moderately correlated with nD (r=0.79), %Brix (r=0.79), and colostrometer reading (r=0.79). Diagnostic test characteristics using the recommended cut point of 1.35966 nD resulted in similar sensitivities for 1FT and 2 FT samples (94.87 and 94.74%, respectively). Cut points of 18 and 19% Brix resulted in the greatest sensitivities (92.31 and 84.62%) and specificity (94.74 and 94.74%, respectively). The 18% Brix cut point resulted in 94.83% of the samples being correctly classified based on IgG concentration. These data support the use of digital refractometer to accurately and rapidly determine IgG concentration in fresh Jersey colostrum. Additionally, these data suggest that IgG concentration determined by RID is affected by multiple FT cycles, whereas estimates obtained by refractometer are not affected by multiple FT cycles. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

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

PubMed

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

2014-05-01

Long-term storage of human milk (HM) requires freezing at low temperatures, the consequences of which upon macronutrients are unclear. To test the null hypothesis that HM freezing and storage for a range of 1 to 10 weeks at -80 °C does not affect HM fat, protein, lactose and energy contents. Samples of HM were obtained from 20 mothers (60 samples) of preterm infants (25 to 35 weeks gestation), who routinely expressed their milk, every 3 h, using an electric pump, from the second to the seventh week after delivery. All samples were frozen at -80 °C for 8 to 83 days (43.8 days average). After thawing and homogenization, energy and macronutrient contents were measured using an HM analyzer. Fat, carbohydrates and energy contents were significantly lower in thawed HM than in fresh HM (fat, fresh vs thawed: 3.72±1.17 vs 3.36±1.19 g/100 ml, P<0.001; carbohydrates, fresh vs thawed: 5.86±0.71 vs 4.09±0.96 g/100 ml, P<0.001; energy, fresh vs thawed: 64.93±12.97 vs 56.63±16.82 kcal/100 ml, P<0.0001), whereas protein content remained unchanged (protein, fresh vs thawed: 1.14±0.36 vs 1.15±0.37 g/100 ml, P=0.7). The decline in carbohydrates content but not in fat and energy correlated significantly with freezing duration. Freezing at -80 °C significantly decreases the energy content of HM, both from fat and carbohydrates. Since quantitatively the decrease in macronutrients was much higher than that published for HM storage at -20 °C, our results do not support freezing HM at -80 °C as the gold standard for long-term storage. We suggest that caloric intake calculations in preterm infants cannot be established based upon fresh HM data.

Ultrastructure in frozen/etched saline solutions: on the internal cleansing of ice.

PubMed

Menger, Fredric M; Galloway, Ashley L; Chlebowski, Mary E; Apkarian, Robert P

2004-05-19

Seawater, with its 3.5% salt content, freezes into hexagonal ice (Ih) that encloses concentrated brine within its matrix. When unsubmerged sea ice reaches a certain height and temperature, the brine drains downward through narrow channels. This mechanism was now modeled by frozen 2-3.5% saline as investigated by cryo-etch high-resolution secondary electron microscopy. Thus, saline was either plunge-frozen in liquid ethane at -183 degrees C or else high-pressure frozen to -105 degrees C in 5-6 ms. Ice from a freshly exposed surface was then subjected to a high-vacuum sublimation ("etching"), a procedure that removes pure bulk ice in preference to ice from frozen hydrated salt. After chromium-coating the etched surface with a 2-nm film, the sample was examined by cryo-HRSEM. Granular icy "fences" were seen surrounding empty areas where amorphous ice had originally resided. Since the fences, about 1-2 mum high, survived the etching, it is likely that they consist of frozen brine. The presence of such fences suggests that, during freezing, saline can purge itself of salt with remarkable speed (5-6 ms). Alternatively, channels (perhaps routed around submicroscopic crystallites of cubic ice (Ic) embedded in the amorphous ice at -105 degrees C) can guide the migration of salt to the periphery of ice patches. Macromolecules fail to form fences because they diffuse too slowly or because they are too large to pass through the channels.

An audit of fresh frozen plasma usage in a tertiary referral centre in a developing country.

PubMed

Prathiba, R; Jayaranee, S; Ramesh, J C; Lopez, C G; Vasanthi, N

2001-06-01

This paper evaluates the practice of fresh frozen plasma (FFP) transfusion at the University Hospital, Kuala Lumpur, and analyses its usage by the various clinical departments. The aim of this study is to identify where it is inappropriately used and the clinical indications in which such misuse is common. A retrospective analysis of the blood bank request forms and work sheets during a 6-month period between January 1998 and June 1998 formed the basis of this study. Overall, 40% of 2665 units transfused were considered appropriate. However, out of the 931 episodes of FFP transfusions only 31% were for appropriate indications. The average FFP requirement when used for appropriate indication was about 4 units per episode, whereas for inappropriate indication it was 2.5 units per episode. Inappropriate use in terms of the number of units was highest by the surgical services (68%) and Orthopaedics (64%), while the Department of Paediatrics had the lowest incidence of inappropriate use (40%). When Paediatrics was used as the benchmark, the incidence of inappropriate use by other departments was significantly higher (p < 0.01). As for FFP usage in common clinical indications, there was a high incidence of inappropriate use in burns (82%), perioperative period (73%), cardiac surgery (68%), massive bleeding (62%) and trauma (60%). The findings in this study, specifically the use of FFP for volume support in trauma, massive bleeding and burns, routine requests without identified indication in cardiac bypass surgery, and prophylactic use in the perioperative period can be the basis for recommendations to minimize the inappropriate use of FFP in the future.

Early maternal serum ß-human chorionic gonadotropin (ß-hCG) levels and sex-related growth difference of IVF embryos.

PubMed

Esh-Broder, Efrat; Oron, Galia; Son, Weon-Young; Holzer, Hananel; Tulandi, Togas

2015-10-01

Maternal serum ß-human chorionic gonadotropin (ß-hCG) represents the trophoblastic cell mass and is an indirect measurement of embryo development at early implantation stage. Studies in animals and human embryos detected sex-related growth differences (SRGD) in favour of male embryos during the pre-implantation period. The purpose of our study was to correlate SRGD and maternal serum ß-hCG at 16 days after embryo transfer. We retrospectively analysed all (fresh and frozen) non-donor, single embryo transfers (SET), elective and not elective, that were performed between December 2008 and December 2013. We included ß-hCG values from day 16 after oocyte collection of pregnancies resulting in live birth. Neonatal gender was retrieved from patient files. Male and female embryos were further grouped to cleavage and blastocyst stage transfers. Regression analysis for confounding variables included maternal age, maternal body mass index (BMI), use of micromanipulation (ICSI), embryo quality (grade), assisted hatching, day of transfer and fresh or frozen embryo transfer. Seven hundred eighty-six non-donor SETs resulted in live birth. After including only day 16 serum ß-hCG results, 525 SETs were analysed. Neonatal gender was available for 522 cases. Mean maternal serum ß-hCG levels were similar, 347 ± 191 IU/L in the male newborn group and 371 ± 200 IU/L in the female group. The difference between ß-hCG levels remained insignificant after adjusting for confounding variables. Early maternal ß-hCG levels after embryo transfers did not represent SRGD in our study.

Comparative analysis of boar seminal plasma proteome from different freezability ejaculates and identification of Fibronectin 1 as sperm freezability marker.

PubMed

Vilagran, I; Yeste, M; Sancho, S; Castillo, J; Oliva, R; Bonet, S

2015-03-01

Variation in boar sperm freezability (i.e. capacity to withstand cryopreservation) between ejaculates is a limitation largely reported in the literature. Prediction of sperm freezability and classification of boar ejaculates into good (GFEs) and poor freezability ejaculates (PFEs) before cryopreservation takes place may increase the use of frozen-thawed spermatozoa. While markers of boar sperm freezability have been found from sperm cell extracts, little attention has been paid to seminal plasma. On this basis, the present study compared the fresh seminal plasma proteome of 9 GFEs and 9 PFEs through two-dimensional difference gel electrophoresis (2D-DIGE) and liquid chromatography mass spectrometry (LC-MS/MS). The ejaculates were previously classified as GFE or PFE upon their sperm viability and progressive motility assessments at 30 and 240 min post thawing. From a total of 51 spots, four were found to significantly (p < 0.05) differ between GFEs and PFEs, and two were identified as fibronectin-1 (FN1) and glutathione peroxidase 5 (GPX5). These two potential markers were further studied by western blot and correlation analysis between protein relative abundances in fresh seminal plasma and regression factors from principal component analyses (PCA) run using post-thawing sperm quality parameters. Results confirmed that FN1 is a reliable marker of boar sperm freezability, because GFEs presented significantly (p < 0.05) higher FN1-amounts than PFEs and FN1 was found to be correlated with the first PCA component at 240 min post thawing. In contrast, GPX5 was not validated as a boar sperm freezability marker. We can thus conclude that levels of FN1 in fresh seminal plasma from boar semen may be used as a sperm freezability marker, thereby facilitating the use of frozen-thawed boar spermatozoa. © 2015 American Society of Andrology and European Academy of Andrology.

Pregnancy rate in women with adenomyosis undergoing fresh or frozen embryo transfer cycles following gonadotropin-releasing hormone agonist treatment.

PubMed

Park, Chan Woo; Choi, Min Hye; Yang, Kwang Moon; Song, In Ok

2016-09-01

To determine the preferred regimen for women with adenomyosis undergoing in vitro fertilization (IVF), we compared the IVF outcomes of fresh embryo transfer (ET) cycles with or without gonadotropin-releasing hormone (GnRH) agonist pretreatment and of frozen-thawed embryo transfer (FET) cycles following GnRH agonist treatment. This retrospective study included 241 IVF cycles of women with adenomyosis from January 2006 to January 2012. Fresh ET cycles without (147 cycles, group A) or with (105 cycles, group B) GnRH agonist pretreatment, and FET cycles following GnRH agonist treatment (43 cycles, group C) were compared. Adenomyosis was identified by using transvaginal ultrasound at the initial workup and classified into focal and diffuse types. The IVF outcomes were also subanalyzed according to the adenomyotic region. GnRH agonist pretreatment increased the stimulation duration (11.5±2.1 days vs. 9.9±2.0 days) and total dose of gonadotropin (3,421±1,141 IU vs. 2,588±1,192 IU), which resulted in a significantly higher number of retrieved oocytes (10.0±8.2 vs. 7.9±6.8, p=0.013) in group B than in group A. Controlled ovarian stimulation for freezing resulted in a significantly higher number of retrieved oocytes (14.3±9.2 vs. 10.0±8.2, p=0.022) with a lower dose of gonadotropin (2,974±1,112 IU vs. 3,421±1,141 IU, p=0.037) in group C than in group B. The clinical pregnancy rate in group C (39.5%) tended to be higher than those in groups B (30.5%) and A (25.2%) but without a significant difference. FET following GnRH agonist pretreatment tended to increase the pregnancy rate in patients with adenomyosis. Further large-scale prospective studies are required to confirm this result.

The extent of surface contamination of retailed chickens with Campylobacter jejuni serogroups.

PubMed Central

Hood, A. M.; Pearson, A. D.; Shahamat, M.

1988-01-01

Eighty-two chickens purchased at 11 retailers (supplied by 12 wholesalers) in the south of England were cultured for Campylobacter jejuni by a method involving total immersion. The organism was isolated from 22 (48%) of 46 fresh birds, 12 of 12 uneviscerated (New York dressed) birds, but only 1 of 24 frozen birds. Viable counts of up to 1.5 x 10(6)/chicken were obtained from fresh birds and 2.4 x 10(7)/chicken from uneviscerated birds. Surface swabbing of breasts, thighs, wings and vents of fresh chickens showed that contamination was generally distributed over the carcasses. Salmonellas were found in only 2 of 69 of the fresh chickens. The prevalence of several Lior and Penner C. jejuni serogroups was similar in chickens and sporadic human cases of enteritis. Penner serogroup 4 (mostly of Lior serogroup 1) was found in 26% of human isolates and 14% of chicken isolates. The rising incidence of campylobacter enteritis during the last 6 years could well be a reflection of the increasing proportion of fresh chickens consumed over that period (32% higher in 1986 than in 1981). PMID:3338503

The impact of processing meat and fish products on phosphorus intake in chronic kidney disease patients.

PubMed

Lou-Arnal, Luis M; Caverni-Muñoz, Alberto; Arnaudas-Casanova, Laura; Vercet-Tormo, Antonio; Gimeno-Orna, José A; Sanz-París, Alejandro; Caramelo-Gutiérrez, Rocío; Alvarez-Lipe, Rafael; Sahdalá-Santana, Laura; Gracia-García, Olga; Luzón-Alonso, Marta

2013-11-13

The use of phosphate additives in meat and fish processing leads to a phosphorus overload that we cannot quantify through labelling or food composition tables. We analysed this increase by measuring phosphorus content in these products by spectrophotometry. We determined the phosphorus/protein ratio in fresh meat and fish products with varying degrees of processing by spectrophotometry (phosphorus) and the Kjeldahl method (proteins). We contrasted these results with those reflected in the food composition tables. The phosphorus/protein ratio was higher in processed meat products (15.83 mg/g) than in battered (11.04 mg/g) and frozen meat products (10.5mg/g), and was lower in fresh (8.41 mg/g) and refrigerated meat products (8.78 mg/g). Fresh white fish had a phosphorus/protein ratio of 8.58mg/g, while it increased by 22% (10.3mg/g) in frozen white fish and by 46% (12.54 mg/g) in battered fish. The information in the tables was poor and confusing, and no reference is made to the brands tested. Processing meat and fish products poses a serious obstacle to the reduction of phosphorus intake. The current regulatory framework does not assist us in the objective of reducing phosphorus additives, since it considers them safe for public consumption. Overcoming these barriers requires a coordinated effort to demonstrate that a high intake of these additives may be harmful to the general population and it should be more closely examined by regulators.

Freeze-all cycle in reproductive medicine: current perspectives.

PubMed

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

2017-02-01

The freeze-all strategy has emerged as an alternative to fresh embryo transfer (ET) during in vitro fertilization (IVF) cycles. Although fresh ET is the norm during assisted reproductive therapies (ART), there are many concerns about the possible adverse effects of controlled ovarian stimulation (COS) over the endometrium. The supra-physiologic hormonal levels that occur during a conventional COS are associated with modifications in the peri-implantation endometrium, which may be related to a decrease in pregnancy rates and poorer obstetric and perinatal outcomes when comparing fresh to frozen-thawed embryo transfers. The main objective of this study was to assess the available literature regarding the freeze-all strategy in IVF cycles, in regards to effectiveness and safety. Although there are many potential advantages in performing a freeze-all cycle over a fresh ET, it seems that the freeze-all strategy is not designed for all IVF patients. There is a need to develop a non-invasive clinical tool to evaluate the endometrial receptivity during a fresh cycle, which enables the selection of patients that would benefit from this strategy. Today, it is reasonable to perform elective cryopreservation of all oocytes/embryos in cases with a risk of OHSS development, and in patients with supra-physiologic hormonal levels during the follicular phase of COS. It is not clear if all normal responders and poor responders may benefit from this strategy.

Freeze-all cycle in reproductive medicine: current perspectives

PubMed Central

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

2017-01-01

The freeze-all strategy has emerged as an alternative to fresh embryo transfer (ET) during in vitro fertilization (IVF) cycles. Although fresh ET is the norm during assisted reproductive therapies (ART), there are many concerns about the possible adverse effects of controlled ovarian stimulation (COS) over the endometrium. The supra-physiologic hormonal levels that occur during a conventional COS are associated with modifications in the peri-implantation endometrium, which may be related to a decrease in pregnancy rates and poorer obstetric and perinatal outcomes when comparing fresh to frozen-thawed embryo transfers. The main objective of this study was to assess the available literature regarding the freeze-all strategy in IVF cycles, in regards to effectiveness and safety. Although there are many potential advantages in performing a freeze-all cycle over a fresh ET, it seems that the freeze-all strategy is not designed for all IVF patients. There is a need to develop a non-invasive clinical tool to evaluate the endometrial receptivity during a fresh cycle, which enables the selection of patients that would benefit from this strategy. Today, it is reasonable to perform elective cryopreservation of all oocytes/embryos in cases with a risk of OHSS development, and in patients with supra-physiologic hormonal levels during the follicular phase of COS. It is not clear if all normal responders and poor responders may benefit from this strategy. PMID:28333033

Analysis of different thermal processing methods of foodstuffs to optimize protein, calcium, and phosphorus content for dialysis patients.

PubMed

Vrdoljak, Ivica; Panjkota Krbavčić, Ines; Bituh, Martina; Vrdoljak, Tea; Dujmić, Zoran

2015-05-01

To analyze how different thermal processing methods affect the protein, calcium, and phosphorus content of hospital food served to dialysis patients and to generate recommendations for preparing menus that optimize nutritional content while minimizing the risk of hyperphosphatemia. Standard Official Methods of Analysis (AOAC) methods were used to determine dry matter, protein, calcium, and phosphorus content in potatoes, fresh and frozen carrots, frozen green beans, chicken, beef and pork, frozen hake, pasta, and rice. These levels were determined both before and after boiling in water, steaming, stewing in oil or water, or roasting. Most of the thermal processing methods did not significantly reduce protein content. Boiling increased calcium content in all foodstuffs because of calcium absorption from the hard water. In contrast, stewing in oil containing a small amount of water decreased the calcium content of vegetables by 8% to 35% and of chicken meat by 12% to 40% on a dry weight basis. Some types of thermal processing significantly reduced the phosphorus content of the various foodstuffs, with levels decreasing by 27% to 43% for fresh and frozen vegetables, 10% to 49% for meat, 7% for pasta, and 22.8% for rice on a dry weight basis. On the basis of these results, we modified the thermal processing methods used to prepare a standard hospital menu for dialysis patients. Foodstuffs prepared according to the optimized menu were similar in protein content, higher in calcium, and significantly lower in phosphorus than foodstuffs prepared according to the standard menu. Boiling in water and stewing in oil containing some water significantly reduced phosphorus content without affecting protein content. Soaking meat in cold water for 1 h before thermal processing reduced phosphorus content even more. These results may help optimize the design of menus for dialysis patients. Copyright © 2015 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

Effects of Different Pretreatments to Fresh Fruit on Chemical and Thermal Characteristics of Crude Palm Oil.

PubMed

Tang, Minmin; Xia, Qiuyu; Holland, Brendan J; Wang, Hui; Zhang, Yufeng; Li, Rui; Cao, Hongxing

2017-12-01

This study selected 5 methods, including boiling, hot air drying, high-pressurized steam, freezing, and microwave radiation to pretreat fresh oil palm fruit before solvent extraction of the oil. Using fresh fruit as a control, the pretreatment methods were compared for the effects on the activity of the 2 main enzymes in the fruit and some physicochemical properties of the crude palm oil. The results indicated, although all the 5 pretreatments could inactivate lipase and peroxidase in the treated flesh significantly (P < 0.05), the high-pressurized steam was the most effective. There were also differences in the unsaturated fatty acid contents of the 6 oils. The crude oil from frozen fruit contained significantly more vitamin E (37829.33 ppm) than previously reported. Microwave radiation was shown to significantly decrease the free fatty acid content and the peroxide value, while increasing the oxidative stability index. Thermal behaviors of the oils were significantly different to each other with the exception a few parameters (P < 0.05). © 2017 Institute of Food Technologists®.

Simultaneous accumulation of proline and trehalose in industrial baker's yeast enhances fermentation ability in frozen dough.

PubMed

Sasano, Yu; Haitani, Yutaka; Hashida, Keisuke; Ohtsu, Iwao; Shima, Jun; Takagi, Hiroshi

2012-05-01

Freeze tolerance is a necessary characteristic for industrial baker's yeast because frozen-dough baking is one of the key technologies for supplying oven-fresh bakery products to consumers. Both proline and trehalose are known to function as cryoprotectants in yeast cells. In order to enhance the freeze tolerance of yeast cells, we constructed a self-cloning diploid baker's yeast strain with simultaneous accumulation of proline, by expressing the PRO1-I150T allele, encoding the proline-feedback inhibition-less sensitive γ-glutamyl kinase, and trehalose, by disrupting the NTH1 gene, encoding neutral trehalase. The resultant strain retained higher tolerance to oxidative and freezing stresses than did the single proline- or trehalose-accumulating strain. Interestingly, our results suggest that proline and trehalose protect yeast cells from short-term and long-term freezing, respectively. Simultaneous accumulation of proline and trehalose in industrial baker's yeast also enhanced the fermentation ability in the frozen dough compared with the single accumulation of proline or trehalose. These results indicate that baker's yeast that accumulates both proline and trehalose is applicable for frozen-dough baking. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Specimen preparation for cryogenic coherent X-ray diffraction imaging of biological cells and cellular organelles by using the X-ray free-electron laser at SACLA

PubMed Central

Kobayashi, Amane; Sekiguchi, Yuki; Oroguchi, Tomotaka; Okajima, Koji; Fukuda, Asahi; Oide, Mao; Yamamoto, Masaki; Nakasako, Masayoshi

2016-01-01

Coherent X-ray diffraction imaging (CXDI) allows internal structures of biological cells and cellular organelles to be analyzed. CXDI experiments have been conducted at 66 K for frozen-hydrated biological specimens at the SPring-8 Angstrom Compact Free-Electron Laser facility (SACLA). In these cryogenic CXDI experiments using X-ray free-electron laser (XFEL) pulses, specimen particles dispersed on thin membranes of specimen disks are transferred into the vacuum chamber of a diffraction apparatus. Because focused single XFEL pulses destroy specimen particles at the atomic level, diffraction patterns are collected through raster scanning the specimen disks to provide fresh specimen particles in the irradiation area. The efficiency of diffraction data collection in cryogenic experiments depends on the quality of the prepared specimens. Here, detailed procedures for preparing frozen-hydrated biological specimens, particularly thin membranes and devices developed in our laboratory, are reported. In addition, the quality of the frozen-hydrated specimens are evaluated by analyzing the characteristics of the collected diffraction patterns. Based on the experimental results, the internal structures of the frozen-hydrated specimens and the future development for efficient diffraction data collection are discussed. PMID:27359147

Specimen preparation for cryogenic coherent X-ray diffraction imaging of biological cells and cellular organelles by using the X-ray free-electron laser at SACLA.

PubMed

Kobayashi, Amane; Sekiguchi, Yuki; Oroguchi, Tomotaka; Okajima, Koji; Fukuda, Asahi; Oide, Mao; Yamamoto, Masaki; Nakasako, Masayoshi

2016-07-01

Coherent X-ray diffraction imaging (CXDI) allows internal structures of biological cells and cellular organelles to be analyzed. CXDI experiments have been conducted at 66 K for frozen-hydrated biological specimens at the SPring-8 Angstrom Compact Free-Electron Laser facility (SACLA). In these cryogenic CXDI experiments using X-ray free-electron laser (XFEL) pulses, specimen particles dispersed on thin membranes of specimen disks are transferred into the vacuum chamber of a diffraction apparatus. Because focused single XFEL pulses destroy specimen particles at the atomic level, diffraction patterns are collected through raster scanning the specimen disks to provide fresh specimen particles in the irradiation area. The efficiency of diffraction data collection in cryogenic experiments depends on the quality of the prepared specimens. Here, detailed procedures for preparing frozen-hydrated biological specimens, particularly thin membranes and devices developed in our laboratory, are reported. In addition, the quality of the frozen-hydrated specimens are evaluated by analyzing the characteristics of the collected diffraction patterns. Based on the experimental results, the internal structures of the frozen-hydrated specimens and the future development for efficient diffraction data collection are discussed.

A fresh liver agar substrate for rearing small numbers of forensically important blow flies (Diptera: Calliphoridae)

USGS Publications Warehouse

Gruner, Susan V.; Slone, Daniel H.

2014-01-01

Forensically important calliphorids can be reared on a mixture of beef liver and agar. Small pieces of meat, especially fresh or frozen beef liver, will desiccate in 2–6 h, but this simple-to-make feeding substrate remains moist for at least 12 h at 25 and 30°C without desiccation, even in small (5 g) amounts. We determined the survivorship of small numbers of Chrysomya megacephala (F.) (first-instar larvae to adult eclosion) raised on 5 g of liver agar and fresh beef liver. We found that all larvae raised on 5 g of liver died due to desiccation, but survivorship on 5 g of liver agar was equivalent to that on larger (50 g) pieces of either liver agar or beef liver.

The definition of massive transfusion in trauma: a critical variable in examining evidence for resuscitation.

PubMed

Mitra, Biswadev; Cameron, Peter A; Gruen, Russell L; Mori, Alfredo; Fitzgerald, Mark; Street, Alison

2011-06-01

'Massive' transfusion is a poorly defined inclusion criteria for studies examining the blood and blood product that are used during trauma resuscitation. We aimed to compare the traditional definition of massive transfusion (≥10 units in 24 h) to a more acute definition of at least 5 units in 4 h. Multitrauma patients were subgrouped according to the traditional definition and compared with the acute definition. Demographics, presenting vital signs and blood results, management including transfusion practice and outcomes were retrospectively studied. Associations of transfused fresh frozen plasma:packed red blood cells (PRBC) ratios with mortality were studied. There were 927 patients who received PRBCs in the first 24 h, with 314 patients identified using the traditional definition and 303 patients using the acute definition. The patients identified using the traditional definition received 18 (12-29) units of PRBC in 24 h, significantly higher than those identified using the acute definition [15 (9-29) units, P<0.001]. The traditional definition excluded a significant proportion of patients who died in the emergency department. By using the acute definition to select a study sample, there seems to be an increase in mortality with fresh frozen plasma:PRBC ratio of 1 : 1 ratio compared with a 1 : 2 ratio. The traditional 'massive' transfusion definition not only 'dilutes' the potential study samples with a less acute group of patients, but also further excludes patients who die early. This latter group is most likely to be benefitted from any change to resuscitation practice. An acute definition of massive transfusion should be adopted when examining clinical practice during initial trauma resuscitation.

[Study of the reineta protein modifications (Brama australis), put under freezing and storage to -18 degrees C and -30 degrees C].

PubMed

Abugoch, Lilian; Quitral, Vilma; Larraín, M Angélica; Vinagre, Julia; Kriukov, Andrei; Chávez, Gloria

2006-12-01

The objective of the present work was to study functional and thermal properties of reineta (Brama australis) frozen meat, analysed by water retention capacity (WRC), gel forming capacity (GFC), texture, emulsifying capacity and differential scanning calorimetry (DSC). For this study, reineta fillets were obtained and extracted by the same conditions, and cutted, packaged, frozen and stored at -18 degrees C and -30 degrees C for 7 months. The results obtained, showed that there were no signifficant differences in the responses to thermal treatment for all the specimens. For samples frozen at -18 degrees C and -30 degrees C, the protein contents were 23.5 + 0.0 and 25.4 + 1.0%, respectively. The WRC values were 0.45 + 0.1 and 1.59 +/- 0.0 g water/g protein, respectively. The gel forming capacity was only present in the fresh samples, whereas the frozen stored ones only form protein aggregates. The emulsifying capacity was between 960 and 1400 g oil / g protein, and the storage time increased this value. The miosin denaturation temperature (Td) and denaturation enthalpy (?H), obtained by DSC, fluctuated between 39.2 +/- 0.5 to 44.8 +/- 0.8 degrees C and 1.12 +/- 0.3 to 0.52 +/- 0.2 J/g, respectively. The actina values were between 71.0 +/- 0.6 to 75.3 +/- 0.5 degrees C and between 0.5 +/- 0.1 to 0.7 +/- 0.1 J/g. Cooperativity decreased as the storage time increased. This is showing a certain degree of protein displacement. The values found by thermal analyses showed a direct relationship with the functional properties, both decreasing with storage time.

A reduction in public funding for fertility treatment--an econometric analysis of access to treatment and savings to government.

PubMed

Chambers, Georgina M; Hoang, Van Phuong; Zhu, Rong; Illingworth, Peter J

2012-06-08

Almost all assisted reproductive technology (ART) and intrauterine insemination (IUI) treatments performed in Australia are subsidized through the Australian Government's universal insurance scheme, Medicare. In 2010 restrictions on the amount Medicare paid in benefits for these treatments were introduced, increasing patient out-of-pocket payments for fresh and frozen embryo ART cycles and IUI. The aim of this study was to evaluate the impact of the policy on access to treatment, savings in Medicare benefits and the number of ART conceived children not born. Pooled quarterly cross-sectional Medicare data from 2007 and 2011 where used to construct a series of Ordinary Least Squares (OLS) regression models to evaluate the impact of the policy on access to treatment by women of different ages. Government savings in the 12 months after the policy was calculated as the difference between the predicted and observed Medicare benefits paid. After controlling for underlying time trends and unobserved factors the policy change reduced the number of fresh embryo cycles by almost 8600 cycles over 12 months (a 16% reduction in cycles, p < 0.001). The policy effect was greatest on women aged 40 years and older (38% reduction in cycles, p < 0.001). Younger women engaged in relatively more anticipatory behaviour by bringing forward their fresh cycles to 2009. Frozen embryo cycles, which are approximately one quarter of the cost of a fresh cycle, were only marginally impacted by the policy. Utilisation of IUI cycles were not impacted by the policy. After adjusting for anticipatory behaviour, $76 million in Medicare benefits was saved in the 12 months after the policy change (0.47% of annual Medicare benefits). Between 1200 and 1500 ART conceived children were not born in 2010 as a consequence of the policy. The introduction of the policy resulted in a significant reduction in fresh ART cycles in the first 15 months after its introduction. Further evaluation on the long-term impact of the policy with regard access to treatment and on clinical practice, particularly the number of embryos transferred, is crucial to ensuring equitable access to fertility treatment and the health and welfare of ART children.

A reduction in public funding for fertility treatment - an econometric analysis of access to treatment and savings to government

PubMed Central

2012-01-01

Background Almost all assisted reproductive technology (ART) and intrauterine insemination (IUI) treatments performed in Australia are subsidized through the Australian Government’s universal insurance scheme, Medicare. In 2010 restrictions on the amount Medicare paid in benefits for these treatments were introduced, increasing patient out-of-pocket payments for fresh and frozen embryo ART cycles and IUI. The aim of this study was to evaluate the impact of the policy on access to treatment, savings in Medicare benefits and the number of ART conceived children not born. Methods Pooled quarterly cross-sectional Medicare data from 2007 and 2011 where used to construct a series of Ordinary Least Squares (OLS) regression models to evaluate the impact of the policy on access to treatment by women of different ages. Government savings in the 12 months after the policy was calculated as the difference between the predicted and observed Medicare benefits paid. Results After controlling for underlying time trends and unobserved factors the policy change reduced the number of fresh embryo cycles by almost 8600 cycles over 12 months (a 16% reduction in cycles, p < 0.001). The policy effect was greatest on women aged 40 years and older (38% reduction in cycles, p < 0.001). Younger women engaged in relatively more anticipatory behaviour by bringing forward their fresh cycles to 2009. Frozen embryo cycles, which are approximately one quarter of the cost of a fresh cycle, were only marginally impacted by the policy. Utilisation of IUI cycles were not impacted by the policy. After adjusting for anticipatory behaviour, $76 million in Medicare benefits was saved in the 12 months after the policy change (0.47% of annual Medicare benefits). Between 1200 and 1500 ART conceived children were not born in 2010 as a consequence of the policy. Conclusions The introduction of the policy resulted in a significant reduction in fresh ART cycles in the first 15 months after its introduction. Further evaluation on the long term impact of the policy with regard access to treatment and on clinical practice, particularly the number of embryos transferred, is crucial to ensuring equitable access to fertility treatment and the health and welfare of ART children. PMID:22682009

Relative influence upon microwave emissivity of fine-scale stratigraphy, internal scattering, and dielectric properties

USGS Publications Warehouse

England, A.W.

1976-01-01

The microwave emissivity of relatively low-loss media such as snow, ice, frozen ground, and lunar soil is strongly influenced by fine-scale layering and by internal scattering. Radiometric data, however, are commonly interpreted using a model of emission from a homogeneous, dielectric halfspace whose emissivity derives exclusively from dielectric properties. Conclusions based upon these simple interpretations can be erroneous. Examples are presented showing that the emission from fresh or hardpacked snow over either frozen or moist soil is governed dominantly by the size distribution of ice grains in the snowpack. Similarly, the thickness of seasonally frozen soil and the concentration of rock clasts in lunar soil noticeably affect, respectively, the emissivities of northern latitude soils in winter and of the lunar regolith. Petrophysical data accumulated in support of the geophysical interpretation of microwave data must include measurements of not only dielectric properties, but also of geometric factors such as finescale layering and size distributions of grains, inclusions, and voids. ?? 1976 Birkha??user Verlag.

A SIMPLE FREEZE-FRACTURE REPLICATION METHOD FOR ELECTRON MICROSCOPY

PubMed Central

Bullivant, Stanley; Ames, Adelbert

1966-01-01

A simple method to achieve results similar to the freeze-etching technique of Moor et al. (1961) is described. The frozen tissue is cut under liquid nitrogen with a razor blade outside the evaporator rather than inside with a cooled microtome. The conditions of the experiment do not favor sublimation, and it is proposed that the structure of the replica be explained by local faults in the cleavage plane which leaves structures, such as membranes, standing above the ice. Micrographs of replicas of glycerol-protected frozen small intestine of mouse prepared by the method are presented and the structural details they show are discussed. The problem of vapor-deposited contamination is discussed. It is concluded that this is a practical method for obtaining electron micrographs that are relatively free of artifact, and that further improvements may be expected from the use of rapidly frozen fresh tissue and a clean vacuum system, possibly of the ion-pumped type. PMID:5962938

Initial experience with a donor egg bank.

PubMed

Akin, James W; Bell, Katrina A; Thomas, Diana; Boldt, Jeffrey

2007-08-01

To report on the establishment of a commercial donor egg bank (CryoEggs International, LP) and to present our initial experience from the first four patients to receive eggs. Case report. Private fertility clinic. The four recipient women were aged 43, 43, 40, and 33 years. All had cycle day FSH levels greater than 25 mIU/mL. All were given the option of fresh donor egg IVF but opted to use frozen donor oocytes. Purchased and quarantined frozen donor eggs were thawed and inseminated using intracytoplasmic sperm injection (ICSI). Subsequent embryos were transferred on day 3. Clinical pregnancy as defined by presence of cardiac activity. There was a thawed egg survival rate of 76%, a fertilization rate of 74%, a pregnancy rate (PR) of 50%, with an average of 2.75 embryos per transfer and an implantation rate of 27%. Although very preliminary, these results indicate that more widespread use of frozen donor eggs obtained from a commercial egg bank may be feasible in the future, changing the landscape of donor egg IVF.

9 CFR 94.7 - Disposal of animals, meats, and other articles ineligible for importation.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Disposal of animals, meats, and other... RESTRICTED IMPORTATIONS § 94.7 Disposal of animals, meats, and other articles ineligible for importation. (a) Ruminants and swine, and fresh (chilled or frozen) meats, prohibited importation under §§ 94.1, 94.8, 94.9...

Seafood Packaging

NASA Technical Reports Server (NTRS)

1996-01-01

NASA's Technology Transfer Office at Stennis Space Center worked with a New Orleans seafood packaging company to develop a container to improve the shipping longevity of seafood, primarily frozen and fresh fish, while preserving the taste. A NASA engineer developed metalized heat resistant polybags with thermal foam liners using an enhanced version of the metalized mylar commonly known as 'space blanket material,' which was produced during the Apollo era.

Immunological responsiveness of frozen-thawed human lymphocytes.

PubMed Central

Strong, D M; Woody, J N; Factor, M A; Ahmed, A; Sell, K W

1975-01-01

Mononuclear cells (10--20 X 10(6)) obtained from human peripheral blood by a standard Ficoll-Hypaque technique were suspended in RPMI 1640 media at 4 degrees C containing 10% foetal calf serum and 7-5% dimethyl sulphoxide (DMSO). Two-millilitre aliquots were cooled at -1 degree C/min in a Cryoson BV-4 programmed freezing system to -30 degrees C, then -5 degrees C/min to -80 degrees C and stored in liquid nitrogen vapor. On the day of testing, cell suspensions were thawed rapidly in a 37 degree C water bath. DMSO was diluted slowly out of the sample and cells resuspended in fresh RPMI 1640. It was found that frozen stored human lymphocytes (FSHL) demonstrated all the characteristics of fresh unfrozen cells. These included their ability to form spontaneous rosettes with sheep erythrocytes ('E' rosettes) and sheep erythrocyte--antibody--complement rosettes ('EAC' rosettes). The presence of surface immunoglobulins and Fc receptors were shown by membrane immunofluorescence to be comparable. In addition, the results show that FSHL respond to mitogens, specific antigens; act as both stimulators and responders in the mixed lymphocyte culture reaction; and exhibit cell-mediated lymphocytotoxicity following in vitro sensitization, or against antibody-coated target cells. PMID:128429

Cost analysis of fresh-frozen femoral head allografts: is it worthwhile to run a bone bank?

PubMed

Benninger, E; Zingg, P O; Kamath, A F; Dora, C

2014-10-01

To assess the sustainability of our institutional bone bank, we calculated the final product cost of fresh-frozen femoral head allografts and compared these costs with the use of commercial alternatives. Between 2007 and 2010 all quantifiable costs associated with allograft donor screening, harvesting, storage, and administration of femoral head allografts retrieved from patients undergoing elective hip replacement were analysed. From 290 femoral head allografts harvested and stored as full (complete) head specimens or as two halves, 101 had to be withdrawn. In total, 104 full and 75 half heads were implanted in 152 recipients. The calculated final product costs were €1367 per full head. Compared with the use of commercially available processed allografts, a saving of at least €43 119 was realised over four-years (€10 780 per year) resulting in a cost-effective intervention at our institution. Assuming a price of between €1672 and €2149 per commercially purchased allograft, breakeven analysis revealed that implanting between 34 and 63 allografts per year equated to the total cost of bone banking. ©2014 The British Editorial Society of Bone & Joint Surgery.

Lipid class and fatty acid composition of a little-known and rarely collected alga Exophyllum wentii Weber-van Bosse from Bali Island, Indonesia.

PubMed

Illijas, Muhammad I; Indy, Jeane R; Yasui, Hajime; Itabashi, Yutaka

2009-01-01

The lipid class and fatty acid composition of a little-known and rarely collected alga Exophyllum wentii from Bali Island, Indonesia were determined for fresh and frozen-thawed samples using thin-layer chromatography, gas-liquid chromatography, and high-performance liquid chromatography. Glycoglycerolipids, which mainly consisted of mongalactosyldiacylglycerols (MGDG) and digalactosyldiacylglycerols (DGDG), were the predominant lipid components, accounting for 67% and 56% of the total polar lipid content in the fresh and frozen-thawed samples, respectively. Phospholipids, including phosphatidylcholines (PC) and phosphatidylglycerols (PG), were detected with lesser amounts in both samples (16-17% of the total polar lipid content). Free fatty acids (FFA), sterols and triacylglycerols (TAG) were also detected in minor quantities; however, the FFA content in the frozen-thawed sample increased to up to 20% of the total lipid content, suggesting that hydrolysis of the membrane lipids had occurred. A crude enzyme preparation from the alga showed activities for hydrolyzing the acyl groups of the phospholipids and glycoglycerolipids. Palmitic acid (16:0) and arachidonic acid (20:4n-6) were the major fatty acids in both the total lipid and in individual polar lipid classes as well as the dominant fatty acids released from the membrane lipids by enzymatic hydrolysis. The high level of 20:4n-6 (29%) in the total lipid and the presence of considerable amounts of PC (11% of the total polar lipid) and PG (6.2%) support classification of E. wentii into the Division Rhodophyta.

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

PubMed

van Loendersloot, Laura L; Moolenaar, Lobke M; van Wely, Madelon; Repping, Sjoerd; Bossuyt, Patrick M; Hompes, Peter G A; van der Veen, Fulco; Mol, Ben Willem J

2017-07-01

To evaluate the cost-effectiveness of single embryo transfer followed by an additional frozen-thawed single embryo transfer, if more embryos are available, as compared to double embryo transfer in relation to female age. We used a decision tree model to evaluate the costs from a healthcare provider perspective and the pregnancy rates of two embryo transfer policies: one fresh single embryo transfer followed by an additional frozen-thawed single embryo transfer, if more embryos are available (strategy I), and double embryo transfer (strategy II). The analysis was performed on an intention-to-treat basis. Sensitivity analyses were carried out to evaluate the robustness of our model and to identify which model parameters had the strongest impact on the results. SET followed by an additional frozen-thawed single embryo transfer if available was dominant, less costly and more effective, over DET in women under 32 years. In women aged 32 or older DET was more effective than SET followed by an additional frozen-thawed single embryo transfer if available but also more costly. SET followed by an additional frozen-thawed single embryo transfer should be the preferred strategy in women under 32 undergoing IVF. The choice for SET followed by an additional frozen-thawed single embryo transfer or DET in women aged 32 or older depends on individual patient preferences and on how much society is willing to pay for an extra child. There is a strong need for a randomized clinical trial comparing the cost and effects of SET followed by an additional frozen-thawed single embryo transfer and DET in the latter category of women. Copyright © 2017 Elsevier B.V. All rights reserved.

Description of the EuroTARGET cohort: A European collaborative project on TArgeted therapy in renal cell cancer-GEnetic- and tumor-related biomarkers for response and toxicity.

PubMed

van der Zanden, Loes F M; Vermeulen, Sita H; Oskarsdottir, Arna; Maurits, Jake S F; Diekstra, Meta H M; Ambert, Valentin; Cambon-Thomsen, Anne; Castellano, Daniel; Fritsch, Achim; Garcia Donas, Jesus; Guarch Troyas, Rosa; Guchelaar, Henk-Jan; Hartmann, Arndt; Hulsbergen-van de Kaa, Christina; Jaehde, Ulrich; Junker, Kerstin; Martinez-Cardus, Anna; Masson, Gisli; Oosterwijk-Wakka, Jeannette; Radu, Marius T; Rafnar, Thorunn; Rodriguez-Antona, Cristina; Roessler, Max; Ruijtenbeek, Rob; Stefansson, Kari; Warren, Anne; Wessels, Lodewyk; Eisen, Tim; Kiemeney, Lambertus A L M; Oosterwijk, Egbert

2017-08-01

For patients with metastatic renal cell cancer (mRCC), treatment choice is mainly based on clinical parameters. With many treatments available and the limited response to treatment and associated toxicities, there is much interest in identifying better biomarkers for personalized treatment. EuroTARGET aims to identify and characterize host- and tumor-related biomarkers for prediction of response to tyrosine kinase inhibitor therapy in mRCC. Here, we describe the EuroTARGET mRCC patient cohort. EuroTARGET is a European collaborative project designed as an observational study for which patients with mRCC were recruited prospectively in 62 centers. In addition, 462 patients with mRCC from previous studies were included. Detailed clinical information (baseline and follow-up) from all patients was entered in web-based case record forms. Blood was collected for germline DNA and pharmacokinetic/pharmacodynamic analyses and, where available, fresh-frozen tumor material was collected to perform tumor DNA, RNA, kinome, and methylome analyses. In total, 1,210 patients with mRCC were included. Of these, 920 received a tyrosine kinase inhibitor as first-line targeted treatment (sunitinib [N = 713, 78%], sorafenib [N = 41, 4%], or pazopanib [N = 166, 18%]) and had at least 6 months of outcome assessment (median follow-up 15.3 months [interquartile range: 8.5-30.2 months]). Germline DNA samples were available from 824 of these patients, fresh-frozen tumor material from 142 patients, fresh-frozen normal kidney tissue from 95 patients, and tissue microarrays created from formalin-fixed paraffin-embedded tumor material from 247 patients. Of the 920 patients, germline DNA variant chip data were successfully generated for 811 patients (Illumina HumanOmniExpress BeadChip). For 80 patients, next-generation exome sequencing of germline and tumor DNA was performed, tumor RNA sequencing was performed for 124 patients, kinome activity measured and processed for 121 patients (PamChip), and methylome data (Illumina Infinium HumanMethylation450 BeadChip) were created for 116 RCC tissues (and 23 normal kidney tissues). For 73 out of the 920 patients, all platform data types were generated. In addition, 40 patients were included in a pharmacokinetic/pharmacodynamic phase IV substudy. Analysis of EuroTARGET cohort data will contribute to personalization of therapy for patients with mRCC. The extensive clinical data and multiplatform EuroTARGET data will be freely available. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Evaluation of breast tissue with confocal strip-mosaicking microscopy: a test approach emulating pathology-like examination

PubMed Central

Abeytunge, Sanjee; Larson, Bjorg; Peterson, Gary; Morrow, Monica; Rajadhyaksha, Milind

2017-01-01

Abstract. Confocal microscopy is an emerging technology for rapid imaging of freshly excised tissue without the need for frozen- or fixed-section processing. Initial studies have described imaging of breast tissue using fluorescence confocal microscopy with small regions of interest, typically 750×750  μm2. We present exploration with a microscope, termed confocal strip-mosaicking microscope (CSM microscope), which images an area of 2×2  cm2 of tissue with cellular-level resolution in 10 min of excision. Using the CSM microscope, we imaged 34 fresh, human, large breast tissue specimens from 18 patients, blindly analyzed by a board-certified pathologist and subsequently correlated with the corresponding standard fixed histopathology. Invasive tumors and benign tissue were clearly identified in CSM strip-mosaic images. Thirty specimens were concordant for image-to-histopathology correlation while four were discordant. PMID:28327961

The best source of isolated stromal cells for the artificial ovary: medulla or cortex, cryopreserved or fresh?

PubMed

Soares, M; Sahrari, K; Chiti, M C; Amorim, C A; Ambroise, J; Donnez, J; Dolmans, M-M

2015-07-01

What is the best source of ovarian cells for the artificial ovary: medulla or cortex, cryopreserved or fresh? Ovarian cells from fresh medullary tissue, which can be isolated in larger numbers, show higher viability and are able to improve graft vascularization. In a previous study, addition of endothelial cells along with ovarian cells was found to be crucial for formation of a well-vascularized ovary-like structure. This study is the first to evaluate both the effect of cryopreservation and the source of ovarian tissue on isolated ovarian cells. Prospective experimental study in an academic research unit using ovarian tissue from seven patients undergoing surgery for benign gynecologic disease. Ovarian tissue was retrieved from seven patients, with one half processed as fresh (fresh group) and the other half frozen and thawed before processing (frozen group). In each group, ovarian cells from the cortex and medulla were isolated separately, and their viability was tested using a calcein AM/ethidium homodimer viability assay. Fifty thousand cells were then encapsulated in fibrin and grafted to peritoneal pockets in nude mice (14 in all). Grafts recovered after 7 days were analyzed by immunohistochemistry for the presence of ovarian cells (vimentin), proliferation (Ki67) and graft vascularization (double CD34). Cell apoptosis was analyzed by TUNEL assay. Cryopreservation decreased ovarian cell yield (-2804 cells/mg, P = 0.015) and viability (-9.72%, P = 0.052) before grafting and had a considerable (5-fold, P = 0.2) but non-significant negative impact on ovarian cell presence in grafts. The medulla yielded many more cells (+3841 cells/mg, P < 0.001) with higher viability (+18.23%, P < 0.001) than did the cortex. Moreover, grafts with cells from the medulla exhibited a statistically significant 6.44- and 2.47-fold increase in human and total vascular surface area, respectively. P-values were adjusted for multiple testing using the Benjamini-Hochberg method to achieve a 10% false discovery rate and adjusted P-values < 0.1 were therefore considered significant. Pilot study involving a limited number of experiments. Knowing that fresh medullary tissue is the best source of stromal cells is important for construction of the artificial ovary, as isolated follicles require structural support and a rich vascular network for their survival and development. This work was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (5/4/150/5 and 7.4518.12F), Fonds Spéciaux de Recherche, Fondation Saint Luc and Foundation Against Cancer, and donations from Mr Pietro Ferrero, Baron Frère and Viscount Philippe de Spoelberch. None of the authors have any conflicting interests to declare. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Prediction of fracture load and stiffness of the proximal femur by CT-based specimen specific finite element analysis: cadaveric validation study.

PubMed

Miura, Michiaki; Nakamura, Junichi; Matsuura, Yusuke; Wako, Yasushi; Suzuki, Takane; Hagiwara, Shigeo; Orita, Sumihisa; Inage, Kazuhide; Kawarai, Yuya; Sugano, Masahiko; Nawata, Kento; Ohtori, Seiji

2017-12-16

Finite element analysis (FEA) of the proximal femur has been previously validated with large mesh size, but these were insufficient to simulate the model with small implants in recent studies. This study aimed to validate the proximal femoral computed tomography (CT)-based specimen-specific FEA model with smaller mesh size using fresh frozen cadavers. Twenty proximal femora from 10 cadavers (mean age, 87.1 years) were examined. CT was performed on all specimens with a calibration phantom. Nonlinear FEA prediction with stance configuration was performed using Mechanical Finder (mesh,1.5 mm tetrahedral elements; shell thickness, 0.2 mm; Poisson's coefficient, 0.3), in comparison with mechanical testing. Force was applied at a fixed vertical displacement rate, and the magnitude of the applied load and displacement were continuously recorded. The fracture load and stiffness were calculated from force-displacement curve, and the correlation between mechanical testing and FEA prediction was examined. A pilot study with one femur revealed that the equations proposed by Keller for vertebra were the most reproducible for calculating Young's modulus and the yield stress of elements of the proximal femur. There was a good linear correlation between fracture loads of mechanical testing and FEA prediction (R 2 = 0.6187) and between the stiffness of mechanical testing and FEA prediction (R 2 = 0.5499). There was a good linear correlation between fracture load and stiffness (R 2 = 0.6345) in mechanical testing and an excellent correlation between these (R 2 = 0.9240) in FEA prediction. CT-based specimen-specific FEA model of the proximal femur with small element size was validated using fresh frozen cadavers. The equations proposed by Keller for vertebra were found to be the most reproducible for the proximal femur in elderly people.

A methodological study of genome-wide DNA methylation analyses using matched archival formalin-fixed paraffin embedded and fresh frozen breast tumors

PubMed Central

Yan, Li; Liu, Song; Tang, Li; Hu, Qiang; Morrison, Carl D.; Ambrosone, Christine B.; Higgins, Michael J.; Sucheston-Campbell, Lara E.

2017-01-01

Background DNA from archival formalin-fixed and paraffin embedded (FFPE) tissue is an invaluable resource for genome-wide methylation studies although concerns about poor quality may limit its use. In this study, we compared DNA methylation profiles of breast tumors using DNA from fresh-frozen (FF) tissues and three types of matched FFPE samples. Results For 9/10 patients, correlation and unsupervised clustering analysis revealed that the FF and FFPE samples were consistently correlated with each other and clustered into distinct subgroups. Greater than 84% of the top 100 loci previously shown to differentiate ER+ and ER– tumors in FF tissues were also FFPE DML. Weighted Correlation Gene Network Analyses (WCGNA) grouped the DML loci into 16 modules in FF tissue, with ~85% of the module membership preserved across tissue types. Materials and Methods Restored FFPE and matched FF samples were profiled using the Illumina Infinium HumanMethylation450K platform. Methylation levels (β-values) across all loci and the top 100 loci previously shown to differentiate tumors by estrogen receptor status (ER+ or ER−) in a larger FF study, were compared between matched FF and FFPE samples using Pearson's correlation, hierarchical clustering and WCGNA. Positive predictive values and sensitivity levels for detecting differentially methylated loci (DML) in FF samples were calculated in an independent FFPE cohort. Conclusions FFPE breast tumors samples show lower overall detection of DMLs versus FF, however FFPE and FF DMLs compare favorably. These results support the emerging consensus that the 450K platform can be employed to investigate epigenetics in large sets of archival FFPE tissues. PMID:28118602

Inhibition of lipid oxidation in frozen farmed ovate pompano (Trachinotus ovatus L.) fillets stored at -18 °C by chitosan coating incorporated with citric acid or licorice extract.

PubMed

Qiu, Xujian; Chen, Shengjun; Liu, Guangming; Lin, Hong

2016-08-01

Lipid oxidation can occur in fish fillets during long-term frozen storage and cause quality and nutrition loss, which is a major concern in the seafood industry. Our previous study showed that chitosan combined with citric acid or licorice extract can have a preserving effect on fresh fish fillets stored at 4 °C. It is of interest to further study their antioxidant effects on fish fillets during frozen storage. Chitosan, chitosan and citric acid, chitosan and licorice extract can inhibit primary and secondary lipid oxidation, as indicated by lower peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) values compared to the control samples. In addition, drip loss was decreased in the treatment samples. Both citric acid and licorice extract enhanced the antioxidant effects of chitosan. Among all the three treatments, chitosan and licorice extract showed the best antioxidant effects, reducing both PV and TBARS significantly at the end of storage. The combination of chitosan and citric acid or licorice extract showed significant antioxidant effects on ovate pompano fillets at -18 °C during 6 months of storage. They could be applied as natural antioxidant preservatives for use in seafood products or other meat products. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

Anatomy of the interosseous region of the sacroiliac joint.

PubMed

Rosatelli, Alessandro L; Agur, Anne M; Chhaya, Sam

2006-04-01

Anatomical study of the interosseous region of the sacroiliac joint (SIJ) complex. To document and quantify the surface topography of the interosseous region of the SIJ. A review of the literature reveals that little consideration has been given to the interosseous region of the SIJ anatomically, biomechanically, and clinically. The interosseous region of 11 cadaveric specimens (9 formalin embalmed and 2 fresh frozen) were studied. Ten specimens were 55 years of age or older and 1 was 20 years old. To view the interosseous surfaces of the sacrum and ilium the specimens were either axially sectioned (1-cm slices) or disarticulated. One fresh-frozen and 6 embalmed specimens were disarticulated and the remainder axially sectioned. The topography (surface ridging and areas of ossification) of the interosseous region was documented in all specimens and in 2 specimens the surfaces were 3-dimensionally reconstructed using modeling and animation software (MAYA; Autodesk, Inc, San Rafael, CA). Surface characteristics of the SIJ complex observed in specimens 55 years of age or older included moderate to extensive ridging of the interosseous region of the sacrum and ilium in 100% of specimens and ossification of the central interosseous region of the sacroiliac (SI) ligament in 60% of specimens. Central region ossification of the interosseous SI ligament and the presence of ridges and depressions over the opposing interosseous surfaces of the sacrum and ilium are features common to specimens that are in or beyond their sixth decade. These findings further support the contention that there is little to no movement available at this joint in older individuals.

Health Technology Assessment of pathogen reduction technologies applied to plasma for clinical use

PubMed Central

Cicchetti, Americo; Berrino, Alexandra; Casini, Marina; Codella, Paola; Facco, Giuseppina; Fiore, Alessandra; Marano, Giuseppe; Marchetti, Marco; Midolo, Emanuela; Minacori, Roberta; Refolo, Pietro; Romano, Federica; Ruggeri, Matteo; Sacchini, Dario; Spagnolo, Antonio G.; Urbina, Irene; Vaglio, Stefania; Grazzini, Giuliano; Liumbruno, Giancarlo M.

2016-01-01

Although existing clinical evidence shows that the transfusion of blood components is becoming increasingly safe, the risk of transmission of known and unknown pathogens, new pathogens or re-emerging pathogens still persists. Pathogen reduction technologies may offer a new approach to increase blood safety. The study is the output of collaboration between the Italian National Blood Centre and the Post-Graduate School of Health Economics and Management, Catholic University of the Sacred Heart, Rome, Italy. A large, multidisciplinary team was created and divided into six groups, each of which addressed one or more HTA domains. Plasma treated with amotosalen + UV light, riboflavin + UV light, methylene blue or a solvent/detergent process was compared to fresh-frozen plasma with regards to current use, technical features, effectiveness, safety, economic and organisational impact, and ethical, social and legal implications. The available evidence is not sufficient to state which of the techniques compared is superior in terms of efficacy, safety and cost-effectiveness. Evidence on efficacy is only available for the solvent/detergent method, which proved to be non-inferior to untreated fresh-frozen plasma in the treatment of a wide range of congenital and acquired bleeding disorders. With regards to safety, the solvent/detergent technique apparently has the most favourable risk-benefit profile. Further research is needed to provide a comprehensive overview of the cost-effectiveness profile of the different pathogen-reduction techniques. The wide heterogeneity of results and the lack of comparative evidence are reasons why more comparative studies need to be performed. PMID:27403740

Preliminarily comparison of nutritional composition of some fresh and processed seafood.

PubMed

Aberoumand, Ali

2012-10-01

Processing made fish less susceptible to spoilage. Fish are rich in protein content but the protein content is reduced with processing gave a better result when long-time preservation was carried out. Aim of this study was comparison of proximate analysis of some fresh and processed seafoods. Raw materials and processed seafoods (canned mackerel tuna, frozen Sea-Bream and Pressed caviar) were obtained from different firms and analyzed. Analysis carried out according AOAC methods. Moisture, protein and fat values of tuna fish were estimated to be 51, 23.9 and 21.4%, respectively. In this study, moisture content of pressed caviar was 36%, protein content was 34.4% and fat content was 16.7%, carbohydrate and energy values were 4.9% and 316 kcal/100 g, respectively. Pressed and smoked seafoods contained lower amount of moisture but higher amounts of the other components than raw materials (p < 0.05). Canned mackerel tuna, frozen sea bream and pressed caviar also contained higher amounts of fat, carbohydrate and energy, respectively (p < 0.05) than raw material. Except canning with water, all processing technologies decreased the moisture content but increased energy values (p < 0.05) of the fish. It is concluded that processed seafoods are rich in chemical components and very nutritive. Canned tuna with salted water may be advised for low-calorie diets. Caviar pressed was one the best sea foods that was produced in Iran. Since fishes are consumed as a major protein source in food, it is very important that the protein content should not be compromised during table preparation.

Ejaculate and type of freezing extender affect rates of fertilization of horse oocytes in vitro.

PubMed

Roasa, L M; Choi, Y H; Love, C C; Romo, S; Varner, D D; Hinrichs, K

2007-09-01

In vitro fertilization (IVF) was performed on in vitro-matured equine oocytes in three experiments. Frozen-thawed sperm were prepared using swim-up separation and heparin treatment. In Experiment 1, fertilization was achieved with sperm from only one frozen ejaculate of four obtained from the same stallion. Within this ejaculate, fertilization rates were higher with fresh media, as compared to media held for 6-8 days before use (39.6% versus 7.3%, respectively; P<0.001). The type of bovine serum albumin used affected fertilization rates (4% versus 39.6%; P<0.001). To determine if IVF rates were influenced by factors associated with the freezing process (Experiment 2), a single ejaculate from a second stallion was frozen using eight variations in timing of steps in the freezing protocol. There were no differences among treatments in fertilization rates (range, 0-3%). In Experiment 3, fertilization rates of semen frozen in an extender containing 21.5% egg yolk were lower than fertilization rates of semen from the same ejaculate but frozen with a 3% egg-yolk extender (0% versus 15%, respectively; P<0.01). We inferred that rates of equine IVF with frozen-thawed sperm were influenced by ejaculate, the composition and age of the media used, and freezing extender. To our knowledge, this is the first report of ejaculate or extender differences affecting in vitro fertilization in this species. These factors may help to explain the great variability in fertilization rates reported with equine IVF, both among and within laboratories.

Estimate of colostral immunoglobulin G concentration using refractometry without or with caprylic acid fractionation.

PubMed

Morrill, K M; Conrad, E; Polo, J; Lago, A; Campbell, J; Quigley, J; Tyler, H

2012-07-01

Our objectives were to evaluate the use of refractometry as a means of estimating immunoglobulin G (IgG) concentration of bovine maternal colostrum (MC) and determine if fractionation of MC using caprylic acid (CA) improved estimates of IgG. Samples (n=85) of MC were collected from a single dairy in California and used to determine the method of CA fraction that produced the best prediction of IgG based on CA fractionation followed by refractometry. Subsequently, samples of MC (n=827) were collected from 67 farms in 12 states to compare refractometry with or without CA fractionation as methods to estimate IgG concentration. Samples were collected from the feeding pool and consisted of fresh (n=196), previously frozen (n=479), or refrigerated (n=152) MC. Samples were further classified by the number freeze-thaw cycles before analysis. Fractionation with CA was conducted by adding 1 mL of MC to a tube containing 75 μL of CA and 1 mL of 0.06 M acetic acid. The tube was shaken and allowed to react for 1 min. Refractive index of the IgG-rich supernatant (nDf) was determined using a digital refractometer. Whole, nonfractionated MC was analyzed for IgG by radial immunodiffusion (RID) and refractive index (nDw). The relationship between nDf and IgG (r=0.53; n=805) was weak, whereas that between nDw and IgG was stronger (r=0.73; n=823). Fresh samples analyzed by refractometry that subsequently went through 1 freeze-thaw cycle before RID analysis resulted in the strongest relationship between IgG and nDf or nDw (r=0.93 and 0.90, respectively). The MC samples collected fresh on the farm but frozen 2 or more times before analysis by refractometry or RID had low correlations between IgG and nDf and nDw (r=0.09 and 0.01). Samples refrigerated or frozen on the farm before analysis had weaker relationships between RID and nDf or nDw (r=0.38 to 0.80), regardless of the number of freeze-thaw cycles. Breed and lactation number did not affect the accuracy of either test. These results indicated that refractometry, without or with CA fractionation, was an accurate and rapid method to determine IgG concentration when samples of MC were not previously stored before refractometry and frozen only once before RID analysis. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Serum hCG-β levels of postovulatory day 12 and 14 with the sequential application of hCG-β fold change significantly increased predictability of pregnancy outcome after IVF-ET cycle.

PubMed

Sung, Nayoung; Kwak-Kim, Joanne; Koo, H S; Yang, K M

2016-09-01

To investigate hCG-β level on postovulatory day (POD) 12 and its fold increase as predictors for pregnancy outcome after in vitro fertilization (IVF) cycles. A retrospective cohort study was performed in total 1408 fresh and 598 frozen cycles between November 2008 and October 2011, which resulted in biochemical pregnancy, early pregnancy loss, or live birth of singleton pregnancy. The serum hCG-β levels of POD 12 and 14 were compared among biochemical pregnancy, early pregnancy loss, and live birth groups. The cutoff values of POD 12 and 14 hCG-β levels and the degree of hCG-β increase from POD 12 to 14 were determined for each pregnancy outcome. POD 12 and 14 hCG-β levels stratified based on pregnancy outcomes were significantly different among the biochemical pregnancy, early pregnancy loss, and live birth in both fresh and frozen cycles. Serum hCG-β levels of POD 12 and 14 and the fold increase of hCG-β levels from POD 12 to 14 significantly predict pregnancy outcomes after fresh and frozen cycles. Among these, the cutoff value of POD 14 hCG-β had the highest sensitivity and positive predictive value (PPV). In fresh cycles, the cutoff values of POD 12 and 14 serum hCG-β levels for clinical pregnancies were 30.2 mIU/mL (sensitivity 81.3 %, specificity 79.6 %, and PPV 92.3 %) and 70.5 mIU/mL (sensitivity 88.4 %, specificity 85.2 %, and PPV 94.7 %). In pregnancies with POD 12 serum hCG-β levels ≥30.2 mIU/mL, the cutoff level of increase of hCG-β for clinical pregnancy was 2.56 (sensitivity 73.6 %, specificity 72.4 %, and PPV 97.8 %). Sequential application of cutoff values such as POD 12 hCG-β and fold increase of hCG-β improved predictability of pregnancy outcome as compared with that of POD 12 hCG-β alone. The cutoff values of POD 12 and 14 serum hCG-β levels for live birth were 40.5 mIU/mL (sensitivity 75.2 %, specificity 72.6 %, PPV 78.9 %) and 104.5 mIU/mL (sensitivity 80.3 %, specificity 74.1 %, PPV 80.8 %). In the frozen cycles, the cutoff values of POD 12 and 14 serum hCG-β level for clinical pregnancy were 31.5 IU/L (sensitivity 80.4 %, specificity 71.1 % and PPV 90 %) and 43.5 mIU/mL (sensitivity 72.6 %, specificity 71.7 %, PPV 77.2 %). In pregnancies with POD 12 serum hCG-β level ≥31.5 mIU/mL, the cutoff value for fold increase of hCG-β was 2.38 for clinical pregnancy (sensitivity 81.6 %, specificity 71.4 % and PPV 87.9 %). The cutoff values of POD 12 and 14 for live birth were 43.5 mIU/mL (sensitivity 72.6 %, specificity 71.7 %, PPV 77.2 %) and 101.6 mIU/mL (sensitivity 79.6 %, specificity 71.1 %, PPV 78.4 %). Sequential application of cutoff values for POD 12 hCG-β level and fold increase of hCG-β significantly increased PPV for live birth but not clinical pregnancy in frozen cycles. Early prediction of pregnancy outcome by using POD 12 and 14 cutoff levels and sequential application of cutoff value of fold increase could provide appropriate reference to health care providers to initiate earlier management of high-risk pregnancies and precise follow-up of abnormal pregnancies.

Development of a One-Step Duplex RT-PCR Method for the Simultaneous Detection of VP3/VP1 and VP1/P2B Regions of the Hepatitis A Virus.

PubMed

Kim, Mi-Ju; Lee, Shin-Young; Kim, Hyun-Joong; Lee, Jeong Su; Joo, In Sun; Kwak, Hyo Sun; Kim, Hae-Yeong

2016-08-28

The simultaneous detection and accurate identification of hepatitis A virus (HAV) is critical in food safety and epidemiological studies to prevent the spread of HAV outbreaks. Towards this goal, a one-step duplex reverse-transcription (RT)-PCR method was developed targeting the VP1/P2B and VP3/VP1 regions of the HAV genome for the qualitative detection of HAV. An HAV RT-qPCR standard curve was produced for the quantification of HAV RNA. The detection limit of the duplex RT-PCR method was 2.8 × 10(1) copies of HAV. The PCR products enabled HAV genotyping analysis through DNA sequencing, which can be applied for epidemiological investigations. The ability of this duplex RT-PCR method to detect HAV was evaluated with HAV-spiked samples of fresh lettuce, frozen strawberries, and oysters. The limit of detection of the one-step duplex RT-PCR for each food model was 9.4 × 10(2) copies/20 g fresh lettuce, 9.7 × 10(3) copies/20 g frozen strawberries, and 4.1 × 10(3) copies/1.5 g oysters. Use of a one-step duplex RT-PCR method has advantages such as shorter time, decreased cost, and decreased labor owing to the single amplification reaction instead of four amplifications necessary for nested RT-PCR.

Growing and processing conditions lead to changes in the carotenoid profile of spinach.

PubMed

Heymann, Thomas; Westphal, Lore; Wessjohann, Ludger; Glomb, Marcus A

2014-05-28

This study aimed to evaluate the influence of different light regimens during spinach cultivation on the isomeric composition of β-carotene. Irradiation with a halogen lamp, which has a wavelength spectrum close to that of daylight, was used to mimic field-grown conditions. The additional use of optical filters was established as a model system for greenhouse cultivation. Field-grown model systems led to a preferential increase of 9-cis-β-carotene, whereas 13-cis-β-carotene was just formed at the beginning of irradiation. Additionally 9,13-di-cis-β-carotene decreased significantly in the presence of energy-rich light. Isomerization of β-carotene was strongly suppressed during irradiation in greenhouse-grown model systems and led to significant differences. These results were verified in biological samples. Authentic field-grown spinach (Spinacia oleracea L.) showed among changes of other isomers a significantly higher level of 9-cis-isomers (7.52 ± 0.14%) and a significantly lower level of 9,13-di-cis-isomers (0.25 ± 0.03%) compared to authentic greenhouse-grown spinach (6.49 ± 0.11 and 0.76 ± 0.05%). Almost all analyzed commercial spinach samples (fresh and frozen) were identified as common field-grown cultivation. Further investigations resulted in a clear differentiation of frozen commercial samples from fresh spinach, caused by significantly higher levels of 13-cis- and 15-cis-β-carotene as a result of industrial blanching processes.

Vegetable cost metrics show that potatoes and beans provide most nutrients per penny.

PubMed

Drewnowski, Adam; Rehm, Colin D

2013-01-01

Vegetables are important sources of dietary fiber, vitamins and minerals in the diets of children. The United States Department of Agriculture (USDA) National School Lunch Program has new requirements for weekly servings of vegetable subgroups as well as beans and peas. This study estimated the cost impact of meeting the USDA requirements using 2008 national prices for 98 vegetables, fresh, frozen, and canned. Food costs were calculated per 100 grams, per 100 calories, and per edible cup. Rank 6 score, a nutrient density measure was based on six nutrients: dietary fiber; potassium; magnesium; and vitamins A, C, and K. Individual nutrient costs were measured as the monetary cost of 10% daily value of each nutrient per cup equivalent. ANOVAs with post hoc tests showed that beans and starchy vegetables, including white potatoes, were cheaper per 100 calories than were dark-green and deep-yellow vegetables. Fresh, frozen, and canned vegetables had similar nutrient profiles and provided comparable nutritional value. However, less than half (n = 46) of the 98 vegetables listed by the USDA were were consumed >5 times by children and adolescents in the 2003-4 National Health and Nutrition Examination Survey database. For the more frequently consumed vegetables, potatoes and beans were the lowest-cost sources of potassium and fiber. These new metrics of affordable nutrition can help food service and health professionals identify those vegetable subgroups in the school lunch that provide the best nutritional value per penny.

Use of Occult Blood Detection Cards for Real-Time PCR-Based Diagnosis of Schistosoma Mansoni Infection

PubMed Central

Schunk, Mirjam; Kebede Mekonnen, Seleshi; Wondafrash, Beyene; Mengele, Carolin; Fleischmann, Erna; Herbinger, Karl-Heinz; Verweij, Jaco J.; Geldmacher, Christof; Bretzel, Gisela; Löscher, Thomas; Zeynudin, Ahmed

2015-01-01

Background In Schistosoma mansoni infection, diagnosis and control after treatment mainly rely on parasitological stool investigations which are laborious and have limited sensitivity. PCR methods have shown equal or superior sensitivity but preservation and storage methods limit their use in the field. Therefore, the use of occult blood detection cards (fecal cards) for easy sampling and storage of fecal samples for further PCR testing was evaluated in a pilot study. Methodology Stool specimens were collected in a highly endemic area for S. mansoni in Ethiopia and submitted in an investigator-blinded fashion to microscopic examination by Kato-Katz thick smear as well as to real-time PCR using either fresh frozen stool samples or stool smears on fecal cards which have been stored at ambient temperature for up to ten months. Principal Findings Out of 55 stool samples, 35 were positive by microscopy, 33 and 32 were positive by PCR of frozen samples and of fecal card samples, respectively. When microscopy was used as diagnostic “gold standard”, the sensitivity of PCR on fresh stool was 94.3% (95%-CI: 86.6; 100) and on fecal cards 91.4% (95%-CI: 82.2; 100). Conclusions The use of fecal cards proved to be a simple and useful method for stool collection and prolonged storage prior to PCR based diagnosis of S. mansoni infection. This technique may be a valuable approach for large scale surveillance and post treatment assessments PMID:26360049

Use of Occult Blood Detection Cards for Real-Time PCR-Based Diagnosis of Schistosoma Mansoni Infection.

PubMed

Schunk, Mirjam; Kebede Mekonnen, Seleshi; Wondafrash, Beyene; Mengele, Carolin; Fleischmann, Erna; Herbinger, Karl-Heinz; Verweij, Jaco J; Geldmacher, Christof; Bretzel, Gisela; Löscher, Thomas; Zeynudin, Ahmed

2015-01-01

In Schistosoma mansoni infection, diagnosis and control after treatment mainly rely on parasitological stool investigations which are laborious and have limited sensitivity. PCR methods have shown equal or superior sensitivity but preservation and storage methods limit their use in the field. Therefore, the use of occult blood detection cards (fecal cards) for easy sampling and storage of fecal samples for further PCR testing was evaluated in a pilot study. Stool specimens were collected in a highly endemic area for S. mansoni in Ethiopia and submitted in an investigator-blinded fashion to microscopic examination by Kato-Katz thick smear as well as to real-time PCR using either fresh frozen stool samples or stool smears on fecal cards which have been stored at ambient temperature for up to ten months. Out of 55 stool samples, 35 were positive by microscopy, 33 and 32 were positive by PCR of frozen samples and of fecal card samples, respectively. When microscopy was used as diagnostic "gold standard", the sensitivity of PCR on fresh stool was 94.3% (95%-CI: 86.6; 100) and on fecal cards 91.4% (95%-CI: 82.2; 100). The use of fecal cards proved to be a simple and useful method for stool collection and prolonged storage prior to PCR based diagnosis of S. mansoni infection. This technique may be a valuable approach for large scale surveillance and post treatment assessments.

Clinical experience with three-factor prothrombin complex concentrate to reverse warfarin anticoagulation in intracranial hemorrhage.

PubMed

Switzer, Jeffrey A; Rocker, Jody; Mohorn, Phillip; Waller, Jennifer L; Hughes, Douglas; Bruno, Askiel; Nichols, Fenwick T; Hess, David C; Natarajan, Kavita; Fagan, Susan C

2012-09-01

The effectiveness of prothrombin complex concentrate (PCC) products available in the United States that contain low levels of factor VII (3-factor PCC) has not been tested. The purpose of this study was to review our experience with 3-factor PCC (Profilnine) in the setting of warfarin-associated intracranial hemorrhage (wICH). In November 2007, we implemented a protocol for reversal of anticoagulation in wICH using Profilnine. Additional treatment with fresh-frozen plasma was at the discretion of the treating physician. Medical records of all patients receiving PCC for wICH between November 1, 2007, and December 7, 2011 were reviewed. Correction of the international normalized rate (INR) was defined as an INR <1.4. Seventy wICH patients were treated with Profilnine, including 46 (66%) with intraparenchymal hemorrhage, 22 (31%) with subdural hemorrhage, and 2 (3%) with subarachnoid hemorrhage. Mean INR was reduced from 3.36 to 1.96, and in 44 (62.9%) patients the INR corrected to <1.4. Baseline INR ≥3.0 decreased the likelihood of INR correction. Concomitant administration of fresh-frozen plasma (mean, 2.6 U) did not increase the likelihood of INR correction. Seven (10%) patients had serious adverse events during their hospital course, including 2 sudden deaths from suspected pulmonary embolism. Reversal of coagulopathy in wICH with Profilnine was incomplete and associated with serious adverse events. In the absence of available 4-factor PCC, options for urgent reversal of anticoagulation in wICH remain limited.

Freezing of meat raw materials affects tyramine and diamine accumulation in spontaneously fermented sausages.

PubMed

Bover-Cid, Sara; Miguelez-Arrizado, M Jesús; Luz Latorre Moratalla, L; Vidal Carou, M Carmen

2006-01-01

Biogenic amine accumulation was studied in spontaneously fermented sausages (Fuet) manufactured from unfrozen-fresh meat (U-sausages) and frozen-thawed meat (F-sausages). The aim was to investigate whether the frozen storage of raw materials affects the microbial composition and its aminogenic activity during sausage fermentation. Tyramine was the major amine in all sausages. Although the final levels were similar, tyramine accumulated more rapidly in F-sausages, which contained putrescine as the second amine. By contrast, U-sausages accumulated much more cadaverine than putrescine. F-sausages showed a slightly lower pH and free amino acid content as well as higher counts of technological flora (lactic acid and gram positive catalase positive bacteria) and lower counts of enterobacteria. Therefore, to freeze the meat raw materials for few days before sausage manufacture could be a useful practice, especially for the artisan fermented sausages (without starter), because it helps to reduce enterobacteria development and cadaverine production.

Quality assessment of baby food made of different pre-processed organic raw materials under industrial processing conditions.

PubMed

Seidel, Kathrin; Kahl, Johannes; Paoletti, Flavio; Birlouez, Ines; Busscher, Nicolaas; Kretzschmar, Ursula; Särkkä-Tirkkonen, Marjo; Seljåsen, Randi; Sinesio, Fiorella; Torp, Torfinn; Baiamonte, Irene

2015-02-01

The market for processed food is rapidly growing. The industry needs methods for "processing with care" leading to high quality products in order to meet consumers' expectations. Processing influences the quality of the finished product through various factors. In carrot baby food, these are the raw material, the pre-processing and storage treatments as well as the processing conditions. In this study, a quality assessment was performed on baby food made from different pre-processed raw materials. The experiments were carried out under industrial conditions using fresh, frozen and stored organic carrots as raw material. Statistically significant differences were found for sensory attributes among the three autoclaved puree samples (e.g. overall odour F = 90.72, p < 0.001). Samples processed from frozen carrots show increased moisture content and decrease of several chemical constituents. Biocrystallization identified changes between replications of the cooking. Pre-treatment of raw material has a significant influence on the final quality of the baby food.

An exploration and comparison of food and drink availability in homes in a sample of families of White and Pakistani origin within the UK.

PubMed

Bryant, Maria; Sahota, Pinki; Santorelli, Gillian; Hill, Andrew

2015-05-01

Knowledge of the types and quantities of foods and drinks available in family homes supports the development of targeted intervention programmes for obesity prevention or management, or for overall diet improvement. In the UK, contemporary data on foods that are available within family homes are lacking. The present study aimed to explore home food and drink availability in UK homes. An exploratory study using researcher-conducted home food availability inventories, measuring all foods and drinks within the categories of fruits, vegetables, snack foods and beverages. Bradford, a town in the north of the UK. Opportunistic sample of mixed ethnicity families with infants approximately 18 months old from the Born in Bradford birth cohort. All homes had at least one type of fruit, vegetable and snack available. Fresh fruits commonly available were oranges, bananas, apples, satsumas and grapes. Commonly available fresh vegetables included potatoes, cucumber, tomatoes and carrots. The single greatest non-fresh fruit available in homes was raisins. Non-fresh vegetables contributing the most were frozen mixed vegetables, tinned tomatoes and tinned peas. Ethnic differences were found for the availability of fresh fruits and sugar-sweetened beverages, which were both found in higher amounts in Pakistani homes compared with White homes. These data contribute to international data on availability and provide an insight into food availability within family homes in the UK. They have also supported a needs assessment of the development of a culturally specific obesity prevention intervention in which fruits and vegetables and sugar-sweetened beverages are targeted.

Dietary Supplementation with Fresh Pineapple Juice Decreases Inflammation and Colonic Neoplasia in IL-10-deficient Mice with Colitis

PubMed Central

Hale, Laura P.; Chichlowski, Maciej; Trinh, Chau T.; Greer, Paula K.

2010-01-01

Background Bromelain, a mixture of proteolytic enzymes typically derived from pineapple stem, decreases production of pro-inflammatory cytokines and leukocyte homing to sites of inflammation. We previously showed that short-term oral treatment with bromelain purified from pineapple stem decreased the severity of colonic inflammation in C57BL/6 Il10−/− mice with chronic colitis. Since fresh pineapple fruit contains similar bromelain enzymes but at different proportions, this study aimed to determine whether long-term dietary supplementation with pineapple (supplied as juice) could decrease colon inflammation and neoplasia in Il10−/− mice with chronic colitis as compared with bromelain derived from stem. Results Experimental mice readily consumed fresh pineapple juice at a level that generated mean stool proteolytic activities equivalent to 16 mg bromelain purified from stem, while control mice received boiled juice with inactive enzymes. Survival was increased in the group supplemented with fresh rather than boiled juice (p = 0.01). Mice that received fresh juice also had decreased histologic colon inflammation scores and a lower incidence of inflammation-associated colonic neoplasia (35% vs. 66%; p< 0.02), with fewer neoplastic lesions/colon (p = 0.05). Flow cytometric analysis of murine splenocytes exposed to fresh pineapple juice in vitro demonstrated proteolytic removal of cell surface molecules that can affect leukocyte trafficking and activation. Conclusions These results demonstrate that long-term dietary supplementation with fresh or unpasteurized frozen pineapple juice with proteolytically active bromelain enzymes is safe and decreases inflammation severity and the incidence and multiplicity of inflammation-associated colonic neoplasia in this commonly used murine model of inflammatory bowel disease. PMID:20848493

Tactical Damage Control Resuscitation.

PubMed

Fisher, Andrew D; Miles, Ethan A; Cap, Andrew P; Strandenes, Geir; Kane, Shawn F

2015-08-01

Recently the Committee on Tactical Combat Casualty Care changed the guidelines on fluid use in hemorrhagic shock. The current strategy for treating hemorrhagic shock is based on early use of components: Packed Red Blood Cells (PRBCs), Fresh Frozen Plasma (FFP) and platelets in a 1:1:1 ratio. We suggest that lack of components to mimic whole blood functionality favors the use of Fresh Whole Blood in managing hemorrhagic shock on the battlefield. We present a safe and practical approach for its use at the point of injury in the combat environment called Tactical Damage Control Resuscitation. We describe pre-deployment preparation, assessment of hemorrhagic shock, and collection and transfusion of fresh whole blood at the point of injury. By approaching shock with goal-directed therapy, it is possible to extend the period of survivability in combat casualties. Reprint & Copyright © 2015 Association of Military Surgeons of the U.S.

Spermaurin, an La1-like peptide from the venom of the scorpion Scorpio maurus palmatus, improves sperm motility and fertilization in different mammalian species.

PubMed

Martinez, Guillaume; Hograindleur, Jean-Pascal; Voisin, Sébastien; Abi Nahed, Roland; Abd El Aziz, Tarek M; Escoffier, Jessica; Bessonnat, Julien; Fovet, Claire-Maëlle; De Waard, Michel; Hennebicq, Sylviane; Aucagne, Vincent; Ray, Pierre F; Schmitt, Eric; Bulet, Philippe; Arnoult, Christophe

2017-02-10

Is it possible to identify original compounds that are able to enhance sperm motility from the venom of the scorpion Scorpio maurus palmatus? We identified a potent disulfide-rich peptide (DRP) of 73 amino acids that significantly improved the motility of fresh and frozen-thawed sperm in different mammalian species, including human, and improved fertilization outcome in mouse IVF experiments. Any disturbance of sperm motility has a strong impact on fertilization and can lead to subfertility or infertility. Significant efforts have, therefore,  been made to identify pharmacological drugs that might improve sperm motility. Such compounds are particularly useful in azoospermia to improve testicular sperm extraction and in the domain of cryopreservation because the motility of frozen-thawed sperm is reduced. This was a basic science/medical research study aimed at identifying original compounds from a library of venoms able to enhance mammalian sperm motility, including human. We first identified in the venom of a scorpion S. m. palmatus a fraction able to potently activate sperm motility. We next purified and characterized the compound by liquid chromatography, mass spectrometry and peptide synthesis. Finally, the potency and toxicity of both purified and synthetic versions of the identified compound on sperm motility were assessed using different in vitro tests in different mammalian species. For human sperm, biological samples were collected from normozoospermic donors and subfertile patients attending a reproduction department for diagnostic semen analysis. Testicular sperm was collected from cynomolgus monkeys (Macaca fascicularis) euthanized for the needs of specific authorized research projects. The peptide was also tested on bovine and mouse epidydimal sperm. We measured different sperm motility parameters with a computer-assisted sperm analysis system in the presence or absence of the peptide. Size exclusion chromatography enabled us to isolate a fraction of the venom of S. m. palmatus able to increase sperm motility. By liquid chromatography and mass spectrometry, a peptide comprising 73 amino acids with 4 disulfide bridges was identified as responsible for the biological activity and called 'spermaurin'. The identity of spermaurin was confirmed by chemical synthesis. We showed that the peptide increased the motility of fresh and frozen-thawed human sperm. We observed that the potency of the peptide was higher on fresh ejaculated spermatozoa with a low motility, achieving a 100% increase of curvilinear velocity in poorly performing sperm. We also demonstrated that peptide is effective on bovine and mouse fresh epididymal, bovine frozen-thawed ejaculated and fresh non-human primate testicular sperm. Finally, in mouse IVF, the production of 2-cell embryos was increased by 24% when sperm were treated with the peptide. This work is an in vitro evaluation of the ability of spermaurin to improve sperm motility parameters. Another limitation of this study is the small number of human sperm samples tested with the natural (n = 36) and synthetic (n = 12) peptides. Moreover, the effect of the peptide on IVF outcome was only tested in mouse and further tests with human and bovine gametes are required to confirm and extend this result in other mammalian species. This work confirms our initial study showing that venoms represent an interesting source of molecules that are able to modify sperm physiology. Moreover, this work presents the first demonstrated biological action of a venom peptide from the scorpion S. m. palmatus with sequence similarities to La1 peptide from Liocheles australasiae (Wood scorpion), a widespread family of DRPs. Not applicable. This work is part of the project 'LAB COM-14 LAB7 0004 01-LIPAV', funded by the program LabCom 2014 from the French Research Agency (ANR). Dr Arnoult reports grants from IMV Technologies during the conduct of the study. In addition, Drs Arnoult, Martinez, Ray and Schmitt have a patent EP16305642.7 pending containing some of the information presented in this manuscript. © 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

Effects of preservation conditions of canine feces on in vitro gas production kinetics and fermentation end products.

PubMed

Bosch, G; Wrigglesworth, D J; Cone, J W; Pellikaan, W F; Hendriks, W H

2013-01-01

This study investigated the effect of chilling and freezing (for 24 h) canine feces on in vitro gas production kinetics and fermentation end product profiles from carbohydrate-rich (in vitro run 1) and protein-rich (in vitro run 2) substrates. Feces were collected from 3 adult retriever-type dogs fed a canned diet for at least 2 wk. Each fecal sample was divided into 3 portions: 1 portion was used immediately as an inoculum (fresh) and the other 2 portions were used after either chilling to 5°C for 30 min and storage in crushed ice for 23.5 h (chilling) or freezing to -20°C for 30 min and storage in a prefrozen (-20°C) container for 23.5 h (freezing). The medium solution for run 1 contained N whereas that for run 2 was N free. Substrates included fructooligosaccharide (FOS), sugar beet pulp, and wheat middlings in run 1 and soybean meal, poultry meat meal, and feather meal in run 2. Gas production kinetics were calculated from cumulative gas production data measured for 72 h. After incubation, fermentation liquids were analyzed for short-chain fatty acids, NH3, and aromatic compounds. For both in vitro runs, chilling feces did not affect gas production kinetics and end product profiles of substrates compared with inocula from fresh feces. Freezing feces decreased the maximum rate of gas production in phase 2 for FOS (P<0.001) and across substrates increased gas produced (P≤0.005) and time of maximum gas production in phase 2 (P<0.001). Furthermore, compared with fresh fecal inocula, inocula from frozen feces resulted in increased overall indole concentrations in run 1 (P=0.006) and indole concentrations from soybean meal and poultry meat meal in run 2 (P<0.001). In run 2, phenol concentrations were greater (P=0.015) for frozen feces than for fresh feces (P=0.015). In conclusion, freezing canine feces for 24 h slightly altered fermentative characteristics of fecal inoculum whereas chilling feces in crushed ice for 24 h maintained fermentative characteristics. Chilling feces in crushed ice is a practical method to preserve feces during transport between laboratories within 24 h for in vitro fermentation studies evaluating dietary ingredients.

A simple method to recover Norovirus from fresh produce with large sample size by using histo-blood group antigens conjugated magnetic beads re-circulating immunomagnetic separation system

USDA-ARS?s Scientific Manuscript database

Noroviruses (NoV) annually cause millions of cases of gastrointestinal disease in the United States. Although NoV outbreaks are generally associated with raw shellfish, particularly oysters, outbreaks have also been known to occur from other common-source food-borne vehicles such as lettuce, frozen...

21 CFR 600.15 - Temperatures during shipment.

Code of Federal Regulations, 2010 CFR

2010-04-01

... Virus Vaccine Live Do. Measles Virus Vaccine Live Do. Mumps Virus Vaccine Live Do. Fresh Frozen Plasma −18 °C or colder. Liquid Plasma 1 to 10 °C. Plasma −18 °C or colder. Platelet Rich Plasma Between 1... Vaccine (Liquid Product) 0 °C or colder. Source Plasma −5 °C or colder. Source Plasma Liquid 10 °C or...

21 CFR 600.15 - Temperatures during shipment.

Code of Federal Regulations, 2012 CFR

2012-04-01

... Virus Vaccine Live Do. Measles Virus Vaccine Live Do. Mumps Virus Vaccine Live Do. Fresh Frozen Plasma −18 °C or colder. Liquid Plasma 1 to 10 °C. Plasma −18 °C or colder. Platelet Rich Plasma Between 1... Vaccine (Liquid Product) 0 °C or colder. Source Plasma −5 °C or colder. Source Plasma Liquid 10 °C or...

21 CFR 600.15 - Temperatures during shipment.

Code of Federal Regulations, 2011 CFR

2011-04-01

... Virus Vaccine Live Do. Measles Virus Vaccine Live Do. Mumps Virus Vaccine Live Do. Fresh Frozen Plasma −18 °C or colder. Liquid Plasma 1 to 10 °C. Plasma −18 °C or colder. Platelet Rich Plasma Between 1... Vaccine (Liquid Product) 0 °C or colder. Source Plasma −5 °C or colder. Source Plasma Liquid 10 °C or...

21 CFR 600.15 - Temperatures during shipment.

Code of Federal Regulations, 2013 CFR

2013-04-01

... Virus Vaccine Live Do. Measles Virus Vaccine Live Do. Mumps Virus Vaccine Live Do. Fresh Frozen Plasma −18 °C or colder. Liquid Plasma 1 to 10 °C. Plasma −18 °C or colder. Platelet Rich Plasma Between 1... Vaccine (Liquid Product) 0 °C or colder. Source Plasma −5 °C or colder. Source Plasma Liquid 10 °C or...