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Sample records for higher cryopreservation efficiency

  1. Highly efficient vitrification method for cryopreservation of human oocytes.

    Science.gov (United States)

    Kuwayama, Masashige; Vajta, Gábor; Kato, Osamu; Leibo, Stanley P

    2005-09-01

    Two experiments were performed to develop a method to cryopreserve MII human oocytes. In the first experiment, three vitrification methods were compared using bovine MII oocytes with regard to their developmental competence after cryopreservation: (i) vitrification within 0.25-ml plastic straws followed by in-straw dilution after warming (ISD method); (ii) vitrification in open-pulled straws (OPS method); and (iii) vitrification in plastic handle (Cryotop method). In the second experiment, the Cryotop method, which had yielded the best results, was used to vitrify human oocytes. Out of 64 vitrified oocytes, 58 (91%) exhibited normal morphology after warming. After intracytoplasmic sperm injection, 52 became fertilized, and 32 (50%) developed to the blastocyst stage in vitro. Analysis by fluorescence in-situ hybridization of five blastocysts showed that all were normal diploid embryos. Twenty-nine embryo transfers with a mean number of 2.2 embryos per transfer on days 2 and 5 resulted in 12 initial pregnancies, seven healthy babies and three ongoing pregnancies. The results suggest that vitrification using the Cryotop is the most efficient method for human oocyte cryopreservation.

  2. Higher Efficiency HVAC Motors

    Energy Technology Data Exchange (ETDEWEB)

    Flynn, Charles Joseph [QM Power, Inc., Kansas City, MO (United States)

    2018-02-13

    failure prone capacitors from the power stage. Q-Sync’s simpler electronics also result in higher efficiency because it eliminates the power required by the PCB to perform the obviated power conversions and PWM processes after line synchronous operating speed is reached in the first 5 seconds of operation, after which the PWM circuits drop out and a much less energy intensive “pass through” circuit takes over, allowing the grid-supplied AC power to sustain the motor’s ongoing operation.

  3. Efficient cryopreservation of human pluripotent stem cells by surface-based vitrification

    NARCIS (Netherlands)

    Neubauer, Julia C; Beier, Axel F; Geijsen, Niels; Zimmermann, Heiko

    2015-01-01

    Efficient cryopreservation of human stem cells is crucial for guaranteeing a permanent supply of high-quality cell material for drug discovery or regenerative medicine. Conventionally used protocols usually employing slow freezing rates, however, result in low recovery rates for human pluripotent

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

    Science.gov (United States)

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

    2014-06-01

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

  5. Comparison of the interleukin-1β-inducing potency of allergenic spores from higher fungi (Basidiomycetes) in a cryopreserved human whole blood system

    Science.gov (United States)

    Rivera-Mariani, Félix E.; Vysyaraju, Kranthi; Negherbon, Jesse; Levetin, Estelle; Horner, W. Elliot; Hartung, Thomas; Breysse, Patrick N.

    2014-01-01

    Background Spores from basidiomycete fungi (basidiospores) are highly prevalent in the atmosphere of urban and rural settings. Studies have confirmed their potential to affect human health as allergens. Less is known about their potential to serve as stimuli of the innate immune system and induce pro-inflammatory reactions. Methods In this study, we evaluated the pro-inflammatory potential of spores from 11 allergenic gilled (Pleurotus ostreatus, Oudemansiella radicata, Armillaria tabescens, Coprinus micaceus, Pluteus cervinus, Chlorophyllum molybdites) and non-gilled (Pisolithus arhizus, Merulius tremullosus, Calvatia cyathiformis, Lycoperdon pyriforme, Boletus bicolor) basidiomycetes fungi based on their potency to induce the release of the pro-inflammatory cytokine interleukin (IL)-1β in a cryopreserved human whole blood system. In addition, the role of morphological features of the spores (surface area, shape, and pigmentation) were examined for their role in the spores’ interleukin (IL)-1β-including potency. Peripheral blood from healthy volunteers was collected, pooled, and cryopreserved. After stimulating the cryopreserved pooled blood with 106 to 103 basidiospores/ml, the concentration of IL-1β in culture supernatants was determined with ELISA. Results Basidiospores manifested concentration-dependent IL-1β-inducing potency, which was more noteworthy among basidiospores from gilled basidiomycetes. At higher concentrations of basidiospores, the IL-1β-inducing potency was able to be differentiated in the cryopreserved human whole blood system. Morphological features did not correlate with the IL-1β-inducing potency of the basidiospores, suggesting that non-morphological properties modulate the IL-1β-inducing potency. Conclusion Our data provides evidence of the pro-inflammatory potential of basidiospores, and the utility of cryopreserved human whole blood as a human-based in-vitro system to study the immune reactivity of allergenic basidiospores. PMID

  6. Cost Efficiency in Public Higher Education.

    Science.gov (United States)

    Robst, John

    This study used the frontier cost function framework to examine cost efficiency in public higher education. The frontier cost function estimates the minimum predicted cost for producing a given amount of output. Data from the annual Almanac issues of the "Chronicle of Higher Education" were used to calculate state level enrollments at two-year and…

  7. Retrofitting the 5045 Klystron for Higher Efficiency

    International Nuclear Information System (INIS)

    Jensen, Aaron; Fazio, Michael; Haase, Andy; Jongewaard, Erik; Kemp, Mark; Neilson, Jeff

    2015-01-01

    The 5045 klystron has been in production and accelerating particles at SLAC National Accelerator Laboratory for over 25 years. Although the design has undergone some changes there are still significant opportunities for improvement in performance. Retrofitting the 5045 for higher efficiencies and a more mono-energetic spent beam profile is presented.

  8. Cryopreservation of oocytes

    International Nuclear Information System (INIS)

    Jadoon, S.

    2015-01-01

    Various approaches have been utilized in attempting to cryopreserve oocytes, beginning with slow cooling and more recently the advent of technique of vitrification. Now it seems that oocyte cryopreservation is no longer an experimental technique and it is being increasingly utilized in clinics around the world. As successful outcome in oocyte cryopreservation can be assessed by survival through the freeze-thaw process, potential for fertilization, embryo development and dynamics of meiotic spindles. This study aimed to analyse these features in context of vitrification and slow freezing. Methods: In this laboratory based study, mature MII mouse oocytes from F1(C57BL6/J X CBA) mice (n=43) were divided randomly into two groups of equal numbers and were cryopreserved by slow freezing and by vitrification. Upon re-warming these oocytes were assessed for survival and for fertilization potential. Oocytes were fixed and stained to compare the effect of both protocols on spindle reassembly and chromosome configuration 10min, 1h and 3h after warming. Unfrozen oocytes were used as controls. Results: A greater number of vitrified oocytes survived cryopreservation than slow frozen oocytes (70.3% vs. 12.5%, p=0.024). After insemination, fertilization rates were higher for vitrified oocytes as compared to slow frozen oocytes (15.86% vs. 4.6%, p=0.046). Morphology of the meiotic spindle was found to be in a disorganized configuration in slow frozen oocytes at all-time points 10 mins, 1 h and 3h), whereas in vitrified oocytes the spindles were found to be aligned at all-time points. Chromosomes were seen to be displaced from equatorial region in both groups. Conclusion: Cryopreservation of mouse oocytes was conducted with greater success using vitrification, compared to slow freezing, with survival, fertilization, and spindle assembly more favourable to a successful outcome in this model. (author)

  9. Effect of Antioxidants and Apoptosis Inhibitors on Cryopreservation of Murine Germ Cells Enriched for Spermatogonial Stem Cells.

    Directory of Open Access Journals (Sweden)

    Seung-Jung Ha

    Full Text Available Spermatogonial stem cells (SSCs are germline stem cells that serve as the foundation of spermatogenesis to maintain fertility throughout a male's lifetime. To treat male infertility using stem cell banking systems and transplantation, it is important to be able to preserve SSCs for long periods of time. Therefore, this study was conducted to develop an optimal cryopreservation protocol for SSCs using antioxidants and apoptosis inhibitors in freezing medium. No differences were observed compared to controls when SSCs were cryopreserved in the presence of apoptosis inhibitors by themselves. However, mouse germ cells cryopreserved in basal medium containing the antioxidant hypotaurine (14 mM resulted in significantly greater proliferation potential and mitochondrial activity. Furthermore, treatment groups with combinations containing 200 mM trehalose and 14 mM hypotaurine showed higher proliferation rates compared to controls. In addition, several serum free conditions were evaluated for SSC cryopreservation. Treatment media containing 10% or 20% knockout serum replacement resulted in similar cryopreservation results compared to media containing FBS. SSC transplantation was also performed to confirm the functionality of SSCs frozen in 14 mM hypotaurine. Donor SSCs formed normal spermatogenic colonies and sperm in the recipient testis. These data indicate that inclusion of 14 mM hypotaurine in cryopreservation media is an effective way to efficiently cryopreserve germ cells enriched for SSCs and that knockout serum replacement can replace FBS in germ cell cryopreservation media.

  10. Phosphorus Processing—Potentials for Higher Efficiency

    OpenAIRE

    Ludwig Hermann; Fabian Kraus; Ralf Hermann

    2018-01-01

    In the aftermath of the adoption of the Sustainable Development Goals (SDGs) and the Paris Agreement (COP21) by virtually all United Nations, producing more with less is imperative. In this context, phosphorus processing, despite its high efficiency compared to other steps in the value chain, needs to be revisited by science and industry. During processing, phosphorus is lost to phosphogypsum, disposed of in stacks globally piling up to 3–4 billion tons and growing by about 200 million ...

  11. Phosphorus Processing—Potentials for Higher Efficiency

    Directory of Open Access Journals (Sweden)

    Ludwig Hermann

    2018-05-01

    Full Text Available In the aftermath of the adoption of the Sustainable Development Goals (SDGs and the Paris Agreement (COP21 by virtually all United Nations, producing more with less is imperative. In this context, phosphorus processing, despite its high efficiency compared to other steps in the value chain, needs to be revisited by science and industry. During processing, phosphorus is lost to phosphogypsum, disposed of in stacks globally piling up to 3–4 billion tons and growing by about 200 million tons per year, or directly discharged to the sea. Eutrophication, acidification, and long-term pollution are the environmental impacts of both practices. Economic and regulatory framework conditions determine whether the industry continues wasting phosphorus, pursues efficiency improvements or stops operations altogether. While reviewing current industrial practice and potentials for increasing processing efficiency with lower impact, the article addresses potentially conflicting goals of low energy and material use as well as Life Cycle Assessment (LCA as a tool for evaluating the relative impacts of improvement strategies. Finally, options by which corporations could pro-actively and credibly demonstrate phosphorus stewardship as well as options by which policy makers could enforce improvement without impairing business locations are discussed.

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

    Science.gov (United States)

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

    2016-06-01

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

  13. Trehalose preincubation increases mesenchymal (CD271+ stem cells post-cryopreservation viability

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    Indra Kusuma

    2016-10-01

    Full Text Available Background: Dimethyl sulfoxide (Me2SO is a common cryoprotective agent widely used in cell preservation system. Me2SO is currently known to cause epigenetic changes which are  critical in stem cells development and cellular differentiation. Therefore, it is imperative to develop cryopreservation techniques that protect cellular functions and avert Me2SO adverse effect. Trehalose was able to protect organism in extreme condition such as dehydration and cold. This study aimed to verify the protective effect of trehalose preincubation procedure in cryopreservation.Methods: The study was conducted using experimental design. Thawed mesenchymal (CD271+ stem cells from YARSI biorepository were used for the experiment. Trehalose preincubation was performed for 1 hour, internalized trehalose was confirmed by FTIR-ATR measurement. Three groups consisted of (1 cryopreserved without trehalose preincubation, (2 cryopreserved with trehalose preincubation, and (3 did not undergo cryopreservation were evaluated after 24 hours in LN2 for viability in culture. The absorbance from each group was measured at 450 nm. The analysis performed using paired student t test.Results: Viability of thawed mesenchymal (CD271+ stem cells that undergo trehalose preincubation prior cryopreservation was significantly higher (p<0.05 compared to group without trehalose preincubation. Higher viability observed between group with trehalose preincubation compared with controlled group suggests protection to trypsinization. Mesenchymal (CD271+ stem cells incubated for 1 hour in 100 mM trehalose supplemented medium  results in 15%  trehalose loading efficiency.Conclusion: These findings confirm the protective effect of trehalose preincubation in cryopreservation. Future research should be directed to elucidate the trehalose internalization mechanism and eventually the protective mechanism of trehalose in mammalian cell cryopreservation.

  14. Embryo cryopreservation and preeclampsia risk.

    Science.gov (United States)

    Sites, Cynthia K; Wilson, Donna; Barsky, Maya; Bernson, Dana; Bernstein, Ira M; Boulet, Sheree; Zhang, Yujia

    2017-11-01

    To determine whether assisted reproductive technology (ART) cycles involving cryopreserved-warmed embryos are associated with the development of preeclampsia. Retrospective cohort study. IVF clinics and hospitals. A total of 15,937 births from ART: 9,417 singleton and 6,520 twin. We used linked ART surveillance, birth certificate, and maternal hospitalization discharge data, considering resident singleton and twin births from autologous or donor eggs from 2005-2010. We compared the frequency of preeclampsia diagnosis for cryopreserved-warmed versus fresh ET and used multivariable logistic regression to adjust for confounders. Among pregnancies conceived with autologous eggs resulting in singletons, preeclampsia was greater after cryopreserved-warmed versus fresh ET (7.51% vs. 4.29%, adjusted odds ratio = 2.17 [95% CI 1.67-2.82]). Preeclampsia without and with severe features, preeclampsia with preterm delivery, and chronic hypertension with superimposed preeclampsia were more frequent after cryopreserved-warmed versus fresh ET (3.99% vs. 2.55%; 2.95% vs. 1.41%; 2.76 vs. 1.48%; and 0.95% vs. 0.43%, respectively). Among pregnancies from autologous eggs resulting in twins, the frequency of preeclampsia with severe features (9.26% vs. 5.70%) and preeclampsia with preterm delivery (14.81% vs. 11.74%) was higher after cryopreserved versus fresh transfers. Among donor egg pregnancies, rates of preeclampsia did not differ significantly between cryopreserved-warmed and fresh ET (10.78% vs. 12.13% for singletons and 28.0% vs. 25.15% for twins). Among ART pregnancies conceived using autologous eggs resulting in live births, those involving transfer of cryopreserved-warmed embryos, as compared with fresh ETs, had increased risk for preeclampsia with severe features and preeclampsia with preterm delivery. Copyright © 2017 American Society for Reproductive Medicine. All rights reserved.

  15. CRYOPRESERVATION OF REPRODUCTIVE PRODUCTS AS AN EFFECTIVE METHOD FOR PRESERVING THE BIODIVERSITY OF STURGEON FISH SPECIES (REVIEW

    Directory of Open Access Journals (Sweden)

    I. Kononenko

    2016-06-01

    Full Text Available Purpose. In recent years, cryopreservation of reproductive products has widely been used as one of the accessible and in some cases the only ways of preserving and supporting the number of endangered fish species including sturgeon fish species. Currently, the method of cryopreservation is represented by various techniques and ways using individual and species approaches. Numerous publications on the issue of fish sperm cryopreservation mostly contain inconsistent results and ambiguous data. Thus, the analysis of the existing information about principles and methods of fish sperm cryopreservation is an important issue for further studies. Moreover, summarizing the existing information will enable us to plan the experiment more efficiently and reasonably and to get the desired outcomes with higher reliability. Findings. The study presents main principles of widely used methods of fish sperm cryopreservation, the analysis of main factors of influence on outcomes of freezing or unfreezing as well as the analysis of results received when using various ways and methods of cryopreservation. Besides, the paper shows the importance of forming and full functioning of fish sperm cryobanks. Originality. The paper summarizes the existing information on the issue of fish sperm low temperature cryopreservation. The information is given in the form of successive presentation of the research outcomes received at each point of freezing or unfreezing when using different techniques as well as results of different factors influence on it. Moreover, a review of achievements in the field of cryopreservation and main principles of forming fish sperm cryobanks are given. Practical value. The presented review of traditional and modern literature data in the issue of cryopreservation can be used when planning, redesigning and experimenting fish sperm freezing or unfreezing.

  16. Coconut (Cocos nucifera l.) pollen cryopreservation.

    Science.gov (United States)

    Karun, A; Sjini, K K; Niral, V; Amarnth, C H; Remya, P; Rajesh, M K; Samsudeen, K; Jerard, B A; Engelmann, F

    2014-01-01

    Coconut genetic resources are threatened by pests and pathogens, natural hazards and human activities. Cryopreservation is the only method allowing the safe and cost-effective long-term conservation of recalcitrant seed species such as coconut. The objective of this work was to test the effect of cryopreservation and of cryostorage duration on coconut pollen germination and fertility. Pollen of two coconut varieties (West Coast Tall WWCTW and Chowghat Orange Dwarf CODC) was collected in March-May over three successive years, desiccated to 7.5 % moisture content (FW) and cryopreserved by direct immersion in liquid nitrogen. Germination and pollen tube length (PTL) of desiccated and cryopreserved pollen were not significantly different for both WCT and COD over the three harvest months of the three consecutive years of study. Pollen germination ranged from 24 to 32 % in desiccated pollen whereas it was between 26 and 29 % in cryopreserved COD pollen. In the case of WCT, germination ranged from 30 to 31 % in desiccated pollen, while it was between 28 and 32 % in cryopreserved pollen. PTL of cryopreserved pollen ranged between 224-390 nm and 226-396 mm for COD and WCT, respectively. Germination of COD pollen varied between 29.0 and 44.1 % after 4 years and 1.0/1.5 years cryostorage, respectively. Germination of WCT pollen did not change significantly between 0 and 6 years cryostorage, being comprised between 32 (24 h) and 40 % (1.5 years). Germination and vigour of cryopreserved pollen were generally higher compared to that of pollen dried in oven and non-cryopreserved. Normal seed set was observed in COD and WCT palms using pollen cryostored for 6 months and 4 years. Cryopreserved pollen of five Tall and five Dwarf accessions displayed 24-31 % and 25-49 % germination, respectively. These results show that it is now possible to establish pollen cryobanks to contribute to coconut germplasm long-term conservation.

  17. Higher Education and Efficiency in Europe: A Comparative Analysis

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    Sánchez-Pérez, Rosario

    2012-01-01

    This paper analyses the efficiency of higher education in equalizing the feasible wages obtained for men and women in the labour market. To do that, It is estimated two stochastic frontiers. The first one measures the effect of higher education inside the group of men and women for six European countries. The results indicate that in Denmark,…

  18. IMPACT OF ROMANIAN HIGHER EDUCATION FUNDING POLICY ON UNIVERSITY EFFICIENCY

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    CRETAN Georgiana Camelia

    2015-07-01

    Full Text Available The issues of higher education funding policy and university operating efficiency are hot points on the actual public agenda worldwide as the pressures exercised upon the public resources increased, especially in the aftermath of the last economic crisis. Concerned with the improvement of the funding mechanism through which government allocates the public funds in order to meet the national core objectives within the area of higher education, the policy makers adjusted the funding policy by diversifying the criteria used in distributing the funds to public universities. Thus, the aim of this research is to underline both the impact and the consequences the public funding patterns of higher education have on the relative efficiency of public funded higher education institutions, across time. Moreover, the research conducted aims to determine whether the changes occurred within the Romanian public funding methodology of higher education institutions improved the relative efficiency scores of public funded universities, before and after the economic crisis of 2008. Thus, on one hand we have underlined the changes brought to the Romanian public funding mechanism of higher education during the years of 2007, 2009 and 2010 compared to the year of 2006, using the content analysis, and on the other hand we assessed and compared the relative efficiency scores of each selected public funded university using a multiple input - multiple output linear programming model, by employing the Data Envelopment Analysis technique. The findings of the research undertaken emphasized that a more performance oriented funding mechanism improves the efficiency scores of public universities. The results of the research undertaken could be used either by the policy makers within the area of higher education or by the administrative management of public universities in order to correlate the funding with the results obtained and/or the objectives assumed by both the

  19. ALTERNATIVE APPROACHES TO EFFICIENCY EVALUATION OF HIGHER EDUCATION INSTITUTIONS

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    Furková, Andrea

    2013-09-01

    Full Text Available Evaluation of efficiency and ranking of higher education institutions is very popular and important topic of public policy. The assessment of the quality of higher education institutions can stimulate positive changes in higher education. In this study we focus on assessment and ranking of Slovak economic faculties. We try to apply two different quantitative approaches for evaluation Slovak economic faculties - Stochastic Frontier Analysis (SFA as an econometric approach and PROMETHEE II as multicriteria decision making method. Via SFA we examine faculties’ success from scientific point of view, i.e. their success in area of publications and citations. Next part of analysis deals with assessing of Slovak economic sciences faculties from overall point of view through the multicriteria decision making method. In the analysis we employ panel data covering 11 economic faculties observed over the period of 5 years. Our main aim is to point out other quantitative approaches to efficiency estimation of higher education institutions.

  20. Energy Efficiency of Higher Education Buildings: A Case Study

    Science.gov (United States)

    Soares, Nelson; Pereira, Luísa Dias; Ferreira, João; Conceição, Pedro; da Silva, Patrícia Pereira

    2015-01-01

    Purpose: This paper aims to propose an energy efficiency plan (with technical and behavioural improvement measures) for a Portuguese higher education building--the Teaching Building of the Faculty of Economics of the University of Coimbra (FEUC). Design/methodology/approach: The study was developed in the context of both the "Green…

  1. Oocyte cryopreservation for donor egg banking.

    Science.gov (United States)

    Cobo, Ana; Remohí, José; Chang, Ching-Chien; Nagy, Zsolt Peter

    2011-09-01

    Oocyte donation is an efficient alternative to using own oocytes in IVF treatment for different indications. Unfortunately, 'traditional' (fresh) egg donations are challenged with inefficiency, difficulties of synchronization, very long waiting periods and lack of quarantine measures. Given the recent improvements in the efficiency of oocyte cryopreservation, it is reasonable to examine if egg donation through oocyte cryopreservation has merits. The objective of the current manuscript is to review existing literature on this topic and to report on the most recent outcomes from two established donor cryobank centres. Reports on egg donation using slow freezing are scarce and though results are encouraging, outcomes are not yet comparable to a fresh egg donation treatment. Vitrification on the other hand appears to provide high survival rates (90%) of donor oocytes and comparable fertilization, embryo development, implantation and pregnancy rates to traditional (fresh) egg donation. Besides the excellent outcomes, the ease of use for both donors and recipients, higher efficiency, lower cost and avoiding the problem of synchronization are all features associated with the benefit of a donor egg cryobank and makes it likely that this approach becomes the future standard of care. Oocyte donation is one of the last resorts in IVF treatment for couples challenged with infertility problems. However, traditional (fresh) egg donation, as it is performed today, is not very efficient, as typically all eggs from one donor are given to only one recipient, it is arduous as it requires an excellent synchronization between the donor and recipient and there are months or years of waiting time. Because of the development of an efficient oocyte cryopreservation technique, it is now possible to cryo-store donor (as well as non-donor) eggs, maintaining their viability and allowing their use whenever there is demand. Therefore, creating a donor oocyte cryobank would carry many advantages

  2. Stress preconditioning of rooster semen before cryopreservation improves fertility potential of thawed sperm.

    Science.gov (United States)

    Feyzi, S; Sharafi, M; Rahimi, S

    2018-03-22

    Avian semen cryopreservation is not as successful as that seen in mammals. This failure is mostly attributed to unique physiological characteristics of poultry semen that make it susceptible to cryo-damages. Utilization of sublethal oxidative stress for preconditioning of sperm, as an innovative approach, improves the cryo-survival of sperm in certain mammalian species. The purpose of this study was to investigate the effects of preconditioning of rooster semen with sublethal oxidative stress [very low concentrations of nitric oxide (NO)] before cryopreservation on the quality and fertility potential of thawed sperm. Semen samples were collected from 20 roosters, twice a wk, and different concentrations of NO [0 (NO-0), 0.01 (NO-0.01), 0.1 (NO-0.1), 1 (NO-1), 10 (NO-10), and 100 μM (NO-100)] were used to investigate the effects of controlled induction of sublethal stress before semen cryopreservation on the thawed sperm performance. A significantly higher (P 0.05) affected by different concentrations of NO. Sperm exposed to NO-1 produced the highest percentage of viable spermatozoa (Annexin-/PI-), which was significantly different from the other samples. Finally, rate of fertility after artificial insemination was significantly higher (P < 0.05) following treatment with NO-1 compared to NO-0 and NO-0.1. Application of 1 μM NO as a sublethal oxidative stress before cryopreservation of sperm efficiently increased numerous quality indices of thawed sperm as well as its fertility potential.

  3. Detection of tumor-associated cells in cryopreserved peripheral blood mononuclear cell samples for retrospective analysis.

    Science.gov (United States)

    Zhu, Peixuan; Stanton, Melissa L; Castle, Erik P; Joseph, Richard W; Adams, Daniel L; Li, Shuhong; Amstutz, Platte; Tang, Cha-Mei; Ho, Thai H

    2016-07-02

    Cryopreserved peripheral blood mononuclear cells (PBMCs) are commonly collected in biobanks. However, little data exist regarding the preservation of tumor-associated cells in cryopreserved collections. The objective of this study was to determine the feasibility of using the CellSieve™ microfiltration assay for the isolation of circulating tumor cells (CTCs) and circulating cancer-associated macrophage-like cells (CAMLs) from cryopreserved PBMC samples. Blood samples spiked with breast (MCF-7), prostate (PC-3), and renal (786-O) cancer cell lines were used to establish analytical accuracy, efficiency, and reproducibility after cryopreservation. The spiked samples were processed through Ficoll separation, and cryopreservation was followed by thawing and microfiltration. MCF-7 cells were successfully retrieved with recovery efficiencies of 90.5 % without cryopreservation and 87.8 and 89.0 %, respectively, on day 7 and day 66 following cryopreservation. The corresponding recovery efficiencies of PC-3 cells were 83.3 % without cryopreservation and 85.3 and 84.7 %, respectively, after cryopreservation. Recovery efficiencies of 786-O cells were 92.7 % without cryopreservation, and 82.7 and 81.3 %, respectively, after cryopreservation. The recovered cells retained the morphologic characteristics and immunohistochemical markers that had been observed before freezing. The protocols were further validated by quantitation of CAMLs in blood samples from two patients with renal cell carcinoma (RCC). The recovery rates of CTCs and CAMLs from cryopreserved samples were not statistically significant different (P > 0.05) from matched fresh samples. To our knowledge, this is the first report that CAMLs could be cryopreserved and analyzed after thawing with microfiltration technology. The application of microfiltration technology to cryopreserved samples will enable much greater retrospective study of cancer patients in relation to long-term outcomes.

  4. Cryopreservation of crane semen

    Science.gov (United States)

    Gee, G.F.; Harris, James

    1991-01-01

    The method for the cryopreservation of crane semen at Patuxent Wildlife Research Center is described in detail. Cryopreservation is useful for the long-term storage of crane semen and for specialized propagation needs. A 50% fertility rate from most sandhill cranes, Grus canadensis, inseminated with frozen-thawed semen can be expected. Additional research should improve the fertility rate and determine how applicable the technique is to other crane species.

  5. Energy efficiency interventions in UK higher education institutions

    International Nuclear Information System (INIS)

    Altan, Hasim

    2010-01-01

    This paper provides an insight into energy efficiency interventions studies, focusing on issues arising in UK higher education institutions (HEIs) in particular. Based on a review of the context for energy efficiency and carbon reduction programmes in the UK and the trends in higher education sector, existing external and internal policies and initiatives and their relevant issues are extensively discussed. To explore the efficacy of some internal intervention strategies, such as technical, non-technical and management interventions, a survey was conducted among UK higher education institutions between February and April 2008. Consultation responses show that there are a relatively high percentage of institutions (83%) that have embarked on both technical and non-technical initiatives, which is a demonstration to the joined-up approach in such area. Major barriers for intervention studies are also identified, including lack of methodology, non-clarity of energy demand and consumption issues, difficulty in establishing assessment boundaries, problems with regards to indices and their effectiveness and so on. Besides establishing clear targets for carbon reductions within the sector, it is concluded that it is important to develop systems for effectively measuring and evaluating the impact of different policies, regulations and schemes in the future as the first step to explore. - Research Highlights: → The research provides an insight into energy efficiency interventions studies, focusing particularly on issues arising in UK higher education institutions (HEIs). → Based on a review of the context for energy efficiency and carbon reduction programmes in the UK and the trends in higher education sector, existing external and internal policies and initiatives, and their relevant issues are extensively discussed. → To explore the efficacy of some internal intervention strategies, such as technical, non-technical and management interventions, a survey was conducted

  6. Cryopreservation of Human Mucosal Leukocytes.

    Directory of Open Access Journals (Sweden)

    Sean M Hughes

    Full Text Available Understanding how leukocytes in the cervicovaginal and colorectal mucosae respond to pathogens, and how medical interventions affect these responses, is important for developing better tools to prevent HIV and other sexually transmitted infections. An effective cryopreservation protocol for these cells following their isolation will make studying them more feasible.To find an optimal cryopreservation protocol for mucosal mononuclear leukocytes, we compared cryopreservation media and procedures using human vaginal leukocytes and confirmed our results with endocervical and colorectal leukocytes. Specifically, we measured the recovery of viable vaginal T cells and macrophages after cryopreservation with different cryopreservation media and handling procedures. We found several cryopreservation media that led to recoveries above 75%. Limiting the number and volume of washes increased the fraction of cells recovered by 10-15%, possibly due to the small cell numbers in mucosal samples. We confirmed that our cryopreservation protocol also works well for both endocervical and colorectal leukocytes. Cryopreserved leukocytes had slightly increased cytokine responses to antigenic stimulation relative to the same cells tested fresh. Additionally, we tested whether it is better to cryopreserve endocervical cells on the cytobrush or in suspension.Leukocytes from cervicovaginal and colorectal tissues can be cryopreserved with good recovery of functional, viable cells using several different cryopreservation media. The number and volume of washes has an experimentally meaningful effect on the percentage of cells recovered. We provide a detailed, step-by-step protocol with best practices for cryopreservation of mucosal leukocytes.

  7. ICT energy efficiency in higher education. Continuous measurement and monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Ter Hofte, H. [Novay, Enschede (Netherlands)

    2011-11-15

    Power consumption of information and communications technology (ICT) is rising rapidly worldwide. Reducing (the growth in) energy demand helps to achieve sustainability goals in the area of energy resource depletion, energy security, economy, and ecology. Various governments and industry consortia have set out policies and agreements to reduce the (growth in) demand for energy. In the MJA3 agreements in the Netherlands, various organizations, including all 14 universities and 39 universities of applied sciences pledged to achieve 30% increase in energy efficiency in 2020 compared to 2005. In this report, we argue that using the number of kilowatt-hours of final electricity used for ICT per enrolled student per day (kWh/st/d), should be used as the primary metric for ICT energy efficiency in higher education. For other uses of electricity than ICT in higher education, we express electricity use in kilowatthours per person per day (kWh/p/d). Applying continuous monitoring and management of ICT energy is one approach one could take to increase ICT energy efficiency in education. In households, providing direct (i.e. real-time) feedback about energy use typically results in 5-15% energy savings, whereas indirect feedback (provided some time after consumption occurs), results in less energy savings, typically 0-10%. Continuous measurement of ICT electricity use can be done in a variety of ways. In this report, we distinguish and describe four major measurement approaches: (1) In-line meters, which require breaking the electrical circuit to install the meter; (2) clamp-on-meters, which can be wrapped around a wire; (3) add-ons to existing energy meters, which use analog or digital ports of existing energy meters; (4) software-only measurement, which uses existing network interfaces, protocols and APIs. A measurement approach can be used at one or more aggregation levels: at building level (to measure all electrical energy used in a building, e.g. a datacenter); at

  8. Considerations for higher efficiency and productivity in research activities.

    Science.gov (United States)

    Forero, Diego A; Moore, Jason H

    2016-01-01

    There are several factors that are known to affect research productivity; some of them imply the need for large financial investments and others are related to work styles. There are some articles that provide suggestions for early career scientists (PhD students and postdocs) but few publications are oriented to professors about scientific leadership. As academic mentoring might be useful at all levels of experience, in this note we suggest several key considerations for higher efficiency and productivity in academic and research activities. More research is needed into the main work style features that differentiate highly productive scientists and research groups, as some of them could be innate and others could be transferable. As funding agencies, universities and research centers invest large amounts of money in order to have a better scientific productivity, a deeper understanding of these factors will be of high academic and societal impact.

  9. [Cryopreservation of teeth].

    Science.gov (United States)

    Zimmerli, Melanie; Filippi, Andreas

    2010-01-01

    After tooth loss dental implants or fixed prosthetic restorations are not indicated in children and adolescents due to incomplete maxillary and mandibular development. Cryopreservation is a method for long-term storage of healthy teeth which were removed for orthodontic reasons or due to traumatic origin. These preserved teeth can be used as autogenous replants or transplants after tooth loss. During transport to and from the freezing facilities prior to freezing the teeth are stored in a cell culture medium. The tooth is transferred into a freezing tube containing cell culture medium and cryoprotectant DMSO. Teeth autotransplanted after cryopreservation show vitality of the PDL cells. Usually no enamel and/or dentinal cracks can be observed. After tooth loss transplantation of cryopreserved teeth could be an effective and biological therapy for tooth replacement.

  10. Efficiency assessment models of higher education institution staff activity

    Directory of Open Access Journals (Sweden)

    K. A. Dyusekeyev

    2016-01-01

    Full Text Available The paper substantiates the necessity of improvement of university staff incentive system under the conditions of competition in the field of higher education, the necessity to develop a separate model for the evaluation of the effectiveness of the department heads. The authors analysed the methods for assessing production function of units. The advantage of the application of the methods to assess the effectiveness of border economic structures in the field of higher education is shown. The choice of the data envelopment analysis method to solve the problem has proved. The model for evaluating of university departments activity on the basis of the DEAmethodology has developed. On the basis of operating in Russia, Kazakhstan and other countries universities staff pay systems the structure of the criteria system for university staff activity evaluation has been designed. For clarification and specification of the departments activity efficiency criteria a strategic map has been developed that allowed us to determine the input and output parameters of the model. DEA-methodology using takes into account a large number of input and output parameters, increases the assessment objectivity by excluding experts, receives interim data to identify the strengths and weaknesses of the evaluated object.

  11. Sperm cryopreservation in fish and shellfish.

    Science.gov (United States)

    Tiersch, Terrence R; Yang, Huiping; Jenkins, Jill A; Dong, Qiaoxiang

    2007-01-01

    Initial success in sperm cryopreservation came at about the same time for aquatic species and livestock. However, in the 50-plus years since then cryopreserved sperm of livestock has grown into a billion-dollar global industry, while despite work in some 200 species with well over 200 published reports, cryopreservation of aquatic species sperm remains essentially a research activity with little commercial application. Most research has focused on large-bodied culture and sport fishes, such as salmonids, carps, and catfishes, and mollusks such as commercially important oyster and abalone species. However, only a handful of studies have addressed sperm cryopreservation in small fishes, such as zebrafish, and in endangered species. Overall, this work has yielded techniques that are being applied with varying levels of success around the world. Barriers to expanded application include a diverse and widely distributed literature base, technical problems, small sperm volumes, variable results, a general lack of access to the technology, and most importantly, the lack of standardization in practices and reporting. The benefits of cryopreservation include at least five levels of improvements for existing industries and for creation of new industries. First, cryopreservation can be used to improve existing hatchery operations by providing sperm on demand and simplifying the timing of induced spawning. Second, frozen sperm can enhance efficient use of facilities and create new opportunities in the hatchery by eliminating the need to maintain live males, potentially freeing resources for use with females and larvae. Third, valuable genetic lineages such as endangered species, research models, or improved farmed strains can be protected by storage of frozen sperm. Fourth, cryopreservation opens the door for rapid genetic improvement. Frozen sperm can be used in breeding programs to create improved lines and shape the genetic resources available for aquaculture. Finally

  12. Strategies for commercialization of cryopreserved fish semen

    Directory of Open Access Journals (Sweden)

    Terrence R. Tiersch

    2008-07-01

    Full Text Available Initial success in sperm cryopreservation occurred at about the same time for aquatic species and livestock. However, in the 50 plus years since then cryopreserved sperm of livestock has grown into a billion-dollar global industry, while cryopreserved sperm of aquatic species remains a research activity with little commercial application despite work in more than 90 species and more than 200 published reports. Most research work has focused on large-bodied culture and sport fishes, such as salmon, trout, carp, and catfish, and mollusks such as commercially important oyster and abalone species. However, only a few studies have addressed sperm cryopreservation in small fishes such as zebrafish, or in endangered species. Overall, this work has yielded techniques that are being applied with varied levels of success around the world. Barriers to expanded application include a diverse and widely distributed literature base, technical problems, small sperm volumes, variable results, a general lack of access to the technology, and most importantly, a lack of standardization in practices and reporting. The benefits of cryopreservation include at least five levels of improvements for existing industries and for creation of new industries. First, cryopreservation can be used to improve existing hatchery operations by providing sperm on demand and simplifying the timing of induced spawning. Second, frozen sperm can enhance efficient use of facilities and create new opportunities in the hatchery by eliminating the need to maintain live males, potentially freeing resources for use with females and larvae. Third, valuable genetic lineages such as endangered species, research models or improved farmed strains can be protected by storage of frozen sperm. Fourth, cryopreservation opens the door for rapid genetic improvement. Frozen sperm can be used in breeding programs to create improved lines and shape the genetic resources available for aquaculture. Finally

  13. Cryopreservation of microencapsulated canine sperm.

    Science.gov (United States)

    Shah, Shambhu; Otsuki, Tsubasa; Fujimura, Chika; Yamamoto, Naoki; Yamashita, Yasuhisa; Higaki, Shogo; Hishinuma, Mitsugu

    2011-03-01

    The objective was to develop a method for cryopreserving microencapsulated canine sperm. Pooled ejaculates from three beagle dogs were extended in egg yolk tris extender and encapsulated using alginate and poly-L-lysine at room temperature. The microcapsules were cooled at 4 °C, immersed in pre-cooled extender (equivalent in volume to the microcapsules) to reach final concentration of 7% (v/v) glycerol and 0.75% (v/v) Equex STM paste, and equilibrated for 5, 30 and 60 min at 4 °C. Thereafter, microcapsules were loaded into 0.5 mL plastic straws and frozen in liquid nitrogen. In Experiment 1, characteristics of microencapsulated canine sperm were evaluated after glycerol addition at 4 °C. Glycerol exposure for 5, 30 and 60 min did not significantly affect progressive motility, viability, or acrosomal integrity of microencapsulated sperm compared with pre-cooled unencapsulated sperm (control). In Experiment 2, characteristics of frozen-thawed canine microencapsulated sperm were evaluated at 0, 3, 6, and 9 h of culture at 38.5 °C. Pre-freeze glycerol exposure for 5, 30, and 60 min at 4 °C did not influence post-thaw quality in unencapsulated sperm. Post-thaw motility and acrosomal integrity of microencapsulated sperm decreased more than those of unencapsulated sperm (P < 0.05) following glycerol exposure for 5 min. However, motility, viability and acrosomal integrity of microencapsulated sperm after 30 and 60 min glycerol exposure were higher than unencapsulated sperm cultured for 6 or 9 h (P < 0.05). In conclusion, since microencapsulated canine sperm were successfully cryopreserved, this could be a viable alternative to convention sperm cryopreservation in this species. Copyright © 2011 Elsevier Inc. All rights reserved.

  14. Optimization of Artificial Propagation in Piracanjuba Fish Brycon orbignyanus Using Cryopreserved Semen.

    Science.gov (United States)

    Felizardo, V O; Melo, C C V; Murgas, L D S; Andrade, E S; Navarro, R D; Ftreitas, T F

    BACKGROUND: Cryopreserved semen could facilitate procedures during the artificial reproduction in fish. Factors affecting cryopreservation efficiency are important to define efficient protocols. This study investigated the application of cryoprotectants on the quality of piracanjuba fish semen, the sperm concentration required for oocyte fertilization and spermatic activation. We evaluated two intracellular cryoprotectant solutions (DMSO and methanol) and two extracellular cryoprotectant solutions (egg yolk and lactose) to cryopreserved piracanjuba semen. Sperm motility rate, motility duration and spermatic alterations were assessed. The protocol for piracanjuba semen cryopreservation can use solutions including either DMSO or methanol as intracellular cryoprotectant and egg yolk or lactose as extracellular cryoprotectants.

  15. Comparison of conventional freezing and vitrification with dimethylformamide and ethylene glycol for cryopreservation of ovine embryos.

    Science.gov (United States)

    Varago, F C; Moutacas, V S; Carvalho, B C; Serapião, R V; Vieira, F; Chiarini-Garcia, H; Brandão, F Z; Camargo, L S; Henry, M; Lagares, M A

    2014-10-01

    The aim of this work was to evaluate the efficiency of the cryoprotectants dimethylformamide and ethylene glycol for cryopreservation of ovine embryos using vitrification and conventional freezing. The recovered embryos were distributed randomly in three treatment groups: Gr. 1: conventional freezing (n = 44), Gr. 2: vitrification with ethylene glycol (n = 39) and Gr. 3: vitrification with dimethylformamide (n = 38). Quality of fresh embryos in control group as well as of frozen and vitrified embryos was examined by three methodologies: staining with propidium iodide and Hoechst 33258 and evaluation under fluorescent microscopy, evaluation of re-expansion and hatching rates after culture, and determination of apoptotic index with TUNEL technique. It was established that re-expansion rate in all treatment groups was similar. In the same time, hatching rates were higher in Gr. 1 (40.5%) and Gr. 2 (35.3%) in comparison with Gr. 3 (15.5%, p conventional freezing, 10.1 ± 8.5, p conventional freezing) and fresh embryos. In conclusion, the dimethylformamide and ethylene glycol used as cryoprotectant to vitrify ovine embryos, in the concentrations and exposition time tested in this work, were not as efficient as the conventional freezing for cryopreservation of ovine embryos Thus, the conventional freezing with ethylene glycol was the most efficient method to cryopreserve ovine embryos in comparison with vitrification. © 2014 Blackwell Verlag GmbH.

  16. MODEL TESTING OF LOW PRESSURE HYDRAULIC TURBINE WITH HIGHER EFFICIENCY

    Directory of Open Access Journals (Sweden)

    V. K. Nedbalsky

    2007-01-01

    Full Text Available A design of low pressure turbine has been developed and it is covered by an invention patent and a useful model patent. Testing of the hydraulic turbine model has been carried out when it was installed on a vertical shaft. The efficiency was equal to 76–78 % that exceeds efficiency of the known low pressure blade turbines. 

  17. Viability of ectomycorrhizal fungi following cryopreservation.

    Science.gov (United States)

    Crahay, Charlotte; Declerck, Stéphane; Colpaert, Jan V; Pigeon, Mathieu; Munaut, Françoise

    2013-02-01

    The use of ectomycorrhizal (ECM) fungi in biotechnological processes requires their maintenance over long periods under conditions that maintain their genetic, phenotypic, and physiological stability. Cryopreservation is considered as the most reliable method for long-term storage of most filamentous fungi. However, this technique is not widespread for ECM fungi since many do not survive or exhibit poor recovery after freezing. The aim of this study was to develop an efficient cryopreservation protocol for the long-term storage of ECM fungi. Two cryopreservation protocols were compared. The first protocol was the conventional straw protocol (SP). The mycelium of the ECM isolates was grown in Petri dishes on agar and subsequently collected by punching the mycelium into a sterile straw before cryopreservation. In the second protocol, the cryovial protocol (CP), the mycelium of the ECM isolates was grown directly in cryovials filled with agar and subsequently cryopreserved. The same cryoprotectant solution, freezing, and thawing process, and re-growth conditions were used in both protocols. The survival (positive when at least 60 % of the replicates showed re-growth) was evaluated before and immediately after freezing as well as after 1 week, 1 m, and 6 m of storage at -130 °C. Greater survival rate (80 % for the CP as compared to 25 % for the SP) and faster re-growth (within 10 d for the CP compared to the 4 weeks for the SP) were observed for most isolates with the CP suggesting that the preparation of the cultures prior to freezing had a significant impact on the isolates survival. The suitability of the CP for cryopreservation of ECM fungi was further confirmed on a set of 98 ECM isolates and displayed a survival rate of 88 % of the isolates. Only some isolates belonging to Suillus luteus, Hebeloma crustuliniforme, Paxillus involutus and Thelephora terrestris failed to survive. This suggested that the CP is an adequate method for the ultra-low cryopreservation of

  18. Cryopreservation of Mammalian Oocyte for Conservation of Animal Genetics

    Directory of Open Access Journals (Sweden)

    Jennifer R. Prentice

    2011-01-01

    Full Text Available The preservation of the female portion of livestock genetics has become an international priority; however, in situ conservation strategies are extremely expensive. Therefore, efforts are increasingly focusing on the development of a reliable cryopreservation method for oocytes, in order to establish ova banks. Slow freezing, a common method for cryopreservation of oocytes, causes osmotic shock (solution effect and intracellular ice crystallization leading to cell damage. Vitrification is an alternative method for cryopreservation in which cells are exposed to a higher concentration of cryoprotectants and frozen with an ultra rapid freezing velocity, resulting in an ice crystal free, solid glass-like structure. Presently, vitrification is a popular method for cryopreservation of embryos. However, vitrification of oocytes is still challenging due to their complex structure and sensitivity to chilling.

  19. Cryopreservation of Parathyroid Glands

    Directory of Open Access Journals (Sweden)

    Marlon A. Guerrero

    2010-01-01

    Full Text Available The risk of permanent hypoparathyroidism following thyroid and parathyroid surgery is around 1% in the hands of experienced endocrine surgeons. Although this complication is rare, rendering a patient permanently aparathyroid has significant consequences on the health and quality of life of the patient. Immediate autotransplantation of parathyroid glands that are injured or unintentionally removed offers the best possibility of graft viability and functionality. However, since the majority of cases of hypoparathyroidism are transient, immediate autotransplantation can complicate postoperative surveillance in certain patients, especially those with primary hyperparathyroidism. Cryopreservation of parathyroid tissue is an alternate technique that was developed to treat patients with permanent hypoparathyroidism. This method allows for parathyroid tissue to be stored and then autotransplanted in a delayed fashion once permanent hypoparathyroidism is confirmed. This article provides a contemporary review on cryopreservation of parathyroid tissue and its current role in thyroid and parathyroid surgery.

  20. HTR plus modern turbine technology for higher efficiencies

    International Nuclear Information System (INIS)

    Barnert, H.; Kugeler, K.

    1996-01-01

    The recent efficiency race for natural gas fired power plants with gas-plus steam-turbine-cycle, is shortly reviewed. The question 'can the HTR compete with high efficiencies?' is answered: Yes, it can - in principle. The gas-plus steam-turbine cycle, also called combi-cycle, is proposed to be taken into consideration here. A comparative study on the efficiency potential is made; it yields 54.5% at 1,050 deg. C gas turbine-inlet temperature. The mechanisms of release versus temperature in the HTR are summarized from the safety report of the HTR MODUL. A short reference is made to the experiences from the HTR-Helium Turbine Project HHT, which was performed in the Federal Republic of Germany in 1968 to 1981. (author). 8 figs,. 1 tab

  1. HTR plus modern turbine technology for higher efficiencies

    Energy Technology Data Exchange (ETDEWEB)

    Barnert, H; Kugeler, K [Forschungszentrum Juelich GmbH (Germany). Inst. fuer Sicherheitsforschung und Reaktortechnik

    1996-08-01

    The recent efficiency race for natural gas fired power plants with gas-plus steam-turbine-cycle, is shortly reviewed. The question `can the HTR compete with high efficiencies?` is answered: Yes, it can - in principle. The gas-plus steam-turbine cycle, also called combi-cycle, is proposed to be taken into consideration here. A comparative study on the efficiency potential is made; it yields 54.5% at 1,050 deg. C gas turbine-inlet temperature. The mechanisms of release versus temperature in the HTR are summarized from the safety report of the HTR MODUL. A short reference is made to the experiences from the HTR-Helium Turbine Project HHT, which was performed in the Federal Republic of Germany in 1968 to 1981. (author). 8 figs,. 1 tab.

  2. Fertility and flow cytometry study of frozen-thawed sperm in cryopreservation medium supplemented with soybean lecithin.

    Science.gov (United States)

    Masoudi, R; Sharafi, M; Zareh Shahneh, A; Towhidi, A; Kohram, H; Esmaeili, V; Shahverdi, A; Davachi, N Dadashpour

    2016-08-01

    Semen cryopreservation can provide genetic resources for a large number of females from a small number of superior males. Optimization of cryopreservation media to achieve the highest quality of post-thaw semen is crucial. Soybean lecithin has evaluated as a plant-based cryoprotectant for substitution of egg yolk in ram semen extender. Flow cytometric and fertility assessments were applied following cryopreservation procedure in two experimental groups (SL group: extender containing 1% w/v soybean lecithin and EY group: extender containing 20% v/v egg yolk). The higher percentage of live sperm and the lower percentage of dead sperm were obtained in SL (47.66 ± 1.38, 52.33 ± 1.69, respectively) extender compared to EY (41.16 ± 1.38, 58.83 ± 1.69). For motion characteristics, plasma membrane integrity, acrosome integrity and mitochondria activity, no significant difference was observed between SL and EY extenders. In artificial insemination experiment, there was no significant difference in pregnancy rate, lambing rate and twining rate between SL and EY extenders. It can be concluded that SL extender can be an efficient alternative extender to preserve ram sperm during cryopreservation procedure without adverse effects. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. COBRE Research Workshop on Higher Education: Equity and Efficiency.

    Science.gov (United States)

    Chicago Univ., IL.

    This document comprises 8 papers presented at the COBRE Research Workshop on Higher Education. The papers are: (1) "Schooling and Equality from Generation to Generation;" (2) "Time Series Changes in Personal Income Inequality: The United States Experience, 1939 to 1985;" (3) "Education, Income, and Ability;" (4) "Proposals for Financing Higher…

  4. Exploiting Software Tool Towards Easier Use And Higher Efficiency

    Science.gov (United States)

    Lin, G. H.; Su, J. T.; Deng, Y. Y.

    2006-08-01

    In developing countries, using data based on instrument made by themselves in maximum extent is very important. It is not only related to maximizing science returns upon prophase investment -- deep accumulations in every aspects but also science output. Based on the idea, we are exploiting a software (called THDP: Tool of Huairou Data Processing). It is used for processing a series of issues, which is met necessary in processing data. This paper discusses its designed purpose, functions, method and specialities. The primary vehicle for general data interpretation is through various techniques of data visualization, techniques of interactive. In the software, we employed Object Oriented approach. It is appropriate to the vehicle. it is imperative that the approach provide not only function, but do so in as convenient a fashion as possible. As result of the software exploiting, it is not only easier to learn data processing for beginner and more convenienter to need further improvement for senior but also increase greatly efficiency in every phrases include analyse, parameter adjusting, result display. Under frame of virtual observatory, for developing countries, we should study more and newer related technologies, which can advance ability and efficiency in science research, like the software we are developing

  5. An efficient higher order family of root finders

    Science.gov (United States)

    Petkovic, Ljiljana D.; Rancic, Lidija; Petkovic, Miodrag S.

    2008-06-01

    A one parameter family of iterative methods for the simultaneous approximation of simple complex zeros of a polynomial, based on a cubically convergent Hansen-Patrick's family, is studied. We show that the convergence of the basic family of the fourth order can be increased to five and six using Newton's and Halley's corrections, respectively. Since these corrections use the already calculated values, the computational efficiency of the accelerated methods is significantly increased. Further acceleration is achieved by applying the Gauss-Seidel approach (single-step mode). One of the most important problems in solving nonlinear equations, the construction of initial conditions which provide both the guaranteed and fast convergence, is considered for the proposed accelerated family. These conditions are computationally verifiable; they depend only on the polynomial coefficients, its degree and initial approximations, which is of practical importance. Some modifications of the considered family, providing the computation of multiple zeros of polynomials and simple zeros of a wide class of analytic functions, are also studied. Numerical examples demonstrate the convergence properties of the presented family of root-finding methods.

  6. Oocyte cryopreservation beyond cancer: tools for ethical reflection.

    Science.gov (United States)

    Linkeviciute, Alma; Peccatori, Fedro A; Sanchini, Virginia; Boniolo, Giovanni

    2015-08-01

    This article offers physicians a tool for structured ethical reflection on challenging situations surrounding oocyte cryopreservation in young healthy women. A systematic literature review offers a comprehensive overview of the ethical debate surrounding the practice. Ethical Counseling Methodology (ECM) offers a practical approach for addressing ethical uncertainties. ECM consists of seven steps: (i) case presentation; (ii) analysis of possible implications; (iii) presentation of ethical question(s); (iv) explanation of ethical terms; (v) presentation of the ethical arguments in favor of and against the procedure; (vi) examination of the individual patient's beliefs and wishes; and (vii) conclusive summary. The most problematic aspects in the ethical debate include the distinction between medical and non-medical use of oocyte cryopreservation, safety and efficiency of the procedure, and marketing practices aimed at healthy women. Female empowerment and enhanced reproductive choices (granted oocyte cryopreservation is a safe and efficient technique) are presented as ethical arguments supporting the practice, while ethical reservations towards oocyte cryopreservation are based on concerns about maternal and fetal safety and wider societal implications. Oocyte cryopreservation is gaining popularity among healthy reproductive age women. However, despite promised benefits it also involves risks that are not always properly communicated in commercialized settings. ECM offers clinicians a tool for structured ethical analysis taking into consideration a wide range of implications, various ethical standpoints, and patients' perceptions and beliefs.

  7. Cryopreservation of mutton snapper ( Lutjanus analis sperm

    Directory of Open Access Journals (Sweden)

    EDUARDO G. SANCHES

    2013-09-01

    Full Text Available This study aimed to develop a protocol of semen cryopreservation of the mutton snapper Lutjanus analis. The interaction between three extenders ( pH 6.1; 7.8 and 8.2 , two concentrations of dimethyl sulfoxide ( DMSO, 5 and 10% and three cooling rates ( -90; -60 and -30°C.min−1 on the sperm motility rate and motility time were analyzed by a factorial experiment. A sample of 30 fishes ( 1,261 ± 449 g collected in the nature was kept in floating net cages. The semen was frozen by using cryogenic straws, in nitrogen vapour and transferred, later, to liquid nitrogen. Fertilization test was accomplished to evaluate the viability of the cryopreserved sperm. The highest sperm motility rate and motility time ( P < 0.05 was achieved by combining extender C ( pH 8.2 with DMSO ( 10% and cooling rate of -60°C.min−1 ( P < 0.05 . The use of cryopreserved sperm presented fertilization rates higher than 59% validating the present protocol for mutton snapper.

  8. Survival, growth and reproduction of cryopreserved larvae from a marine invertebrate, the Pacific oyster (Crassostrea gigas.

    Directory of Open Access Journals (Sweden)

    Marc Suquet

    Full Text Available This study is the first demonstration of successful post-thawing development to reproduction stage of diploid cryopreserved larvae in an aquatic invertebrate. Survival, growth and reproductive performances were studied in juvenile and adult Pacific oysters grown from cryopreserved embryos. Cryopreservation was performed at three early stages: trochophore (13±2 hours post fertilization: hpf, early D-larvae (24±2 hpf and late D-larvae (43±2 hpf. From the beginning (88 days at the end of the ongrowing phase (195 days, no mortality was recorded and mean body weights did not differ between the thawed oysters and the control. At the end of the growing-out phase (982 days, survival of the oysters cryopreserved at 13±2 hpf and at 43±2 hpf was significantly higher (P<0.001 than those of the control (non cryopreserved larvae. Only the batches cryopreserved at 24±2 hpf showed lower survival than the control. Reproductive integrity of the mature oysters, formely cryopreserved at 13±2 hpf and 24±2 hpf, was estimated by the sperm movement and the larval development of their offspring in 13 crosses gamete pools (five males and five females in each pool. In all but two crosses out of 13 tested (P<0.001, development rates of the offspring were not significantly different between frozen and unfrozen parents. In all, the growth and reproductive performances of oysters formerly cryopreserved at larval stages are close to those of controls. Furthermore, these performances did not differ between the three initial larval stages of cryopreservation. The utility of larvae cryopreservation is discussed and compared with the cryopreservation of gametes as a technique for selection programs and shellfish cryobanking.

  9. Dehydration improves cryopreservation of coconut (Cocos nucifera L.).

    Science.gov (United States)

    Sisunandar; Sopade, Peter A; Samosir, Yohannes M S; Rival, Alain; Adkins, Steve W

    2010-12-01

    Cryopreservation of coconut can be used as a strategy to back up the establishment of living collections which are expensive to maintain and are under constant threat from biotic and abiotic factors. Unfortunately, cryopreservation protocols still need to be developed that are capable of producing a sizeable number of field-grown plants. Therefore, we report on the development of an improved cryopreservation protocol which can be used on a wide range of coconut cultivars. The cryopreservation of zygotic embryos and their recovery to soil-growing plants was achieved through the application of four optimised steps viz.: (i) rapid dehydration; (ii) rapid cooling; (iii) rapid warming and recovery in vitro and (iv) acclimatization and soil-supported growth. The thermal properties of water within the embryos were monitored using differential scanning calorimetry (DSC) in order to ensure that the freezable component was kept to a minimum. The feasibility of the protocol was assessed using the Malayan Yellow Dwarf (MYD) cultivar in Australia and then tested on a range of cultivars which were freshly harvested and studied in Indonesia. The most efficient protocol was one based on an 8-h rapid dehydration step followed by rapid cooling step. Best recovery percentages were obtained when a rapid warming step and an optimised in vitro culture step were used. Following this protocol, 20% (when cryopreserved 12 days after harvesting) and 40% (when cryopreserved at the time of harvest) of all MYD embryos cryopreserved could be returned to normal seedlings growing in soil. DSC showed that this protocol induced a drop in embryo fresh weight to 19% and significantly reduced the amount of water remaining that could produce ice crystals (0.1%). Of the 20 cultivars tested, 16 were found to produce between 10% and 40% normal seedlings while four cultivars generated between 0% and 10% normal seedlings after cryopreservation. This new protocol is applicable to a wide range of coconut

  10. CHARACTERISTICS AND FERTILITY OF SUMATRAN TIGER SPERMATOZOA CRYOPRESERVED WITH DIFFERENT SUGARS.

    Science.gov (United States)

    Wayan Kurniani Karja, Ni; Fahrudin, Mokhamad; Setiadi, Mohamad Agus; Tumbelaka, Ligaya Ita; Sudarwati, Retno; Hastuti, Yohana Tri; Mulia, Bongot Huas; Widianti, Ardyta; Sultan, Keni; Terazono, Tsukasa; Namula, Zhao; Taniguchi, Masayasu; Tanihara, Fuminori; Takemoto, Tatsuya; Kikuchi, Kazuhiro; Sato, Yoko; Otoi, Takeshige

    Cryopreservation of semen is one of the most important methods for the preservation of endangered tigers. This study evaluated the effects of sugar supplementation on the cryosurvival of spermatozoa from Sumatran tigers (Panthera tigris sumatrae). The post-thaw characteristics and fertility of spermatozoa cryopreserved with different sugars (glucose, lactose, and trehalose) were evaluated using heterologous in-vitro fertilisation with cat oocytes. All parameters of post-thaw spermatozoa significantly decreased as compared with those of fresh spermatozoa. The index of sperm motility for semen cryopreserved with lactose was significantly higher than that for semen cryopreserved with trehalose. The percentage of total fertilisation for tiger spermatozoa cryopreserved with trehalose was significantly lower than that for control cat spermatozoa. Our findings indicated that supplementation with lactose or glycerol as the main sugar in the egg yolk extender resulted in a better motility and fertility potential for post-thawed spermatozoa.

  11. Cryopreservation of Sambar deer semen in Thailand.

    Science.gov (United States)

    Vongpralub, Thevin; Chinchiyanond, Wittaya; Hongkuntod, Pornchai; Sanchaisuriya, Pitcharat; Liangpaiboon, Sanan; Thongprayoon, Areeya; Somphol, Noppadon

    2015-01-01

    Little is known of the different freezing and thawing techniques for post-thaw survival of spermatozoa in Sambar deer. So, this study determined the effect of seminal plasma, egg yolk and glycerol extenders and their concentrations, plus cooling, freezing, and thawing protocols on the post-thaw quality of their semen. Semen samples were collected by electro-ejaculation from four Thai Sambar deer stags (Cervus unicolor equinus). As evaluated by post-thaw progressive motility and acrosome integrity removal of seminal plasma was beneficial; Tris-egg yolk was the most efficient extender; a 20% egg yolk concentration was better than the 0%, 10%, or 30%; and a 3% glycerol concentration was better than 5%, 7%, or 9%. Using the optimum dilution techniques, semen was loaded in 0.5 ml plastic straws. Cooling times from ambient temperature to 5°C in 3 hr resulted in higher post-thaw progressive motility and acrosome integrity than 1, 2, or 4 hr. Suspending the straws 4 cm above the surface for 15 min before plunging into liquid nitrogen was better than suspending at 2 or 6 cm. For thawing frozen semen, an intermediate thawing (50°C, 8 sec) protocol was more effective than the slower (37°C, 10 sec) or faster (70°C, 5 sec) thawing rates. Timed insemination following estrus synchronization of 10 hinds resulted in six confirmed pregnancies at 60 days. Five hinds delivered live fawns. This study provides an effective approach for semen cryopreservation and artificial insemination (AI), which should be valuable to scientists for genetics and reproductive management of Sambar deer in developing countries. © 2015 Wiley Periodicals, Inc.

  12. Serum-Free Cryopreservation of Five Mammalian Cell Lines in Either a Pelleted or Suspended State

    Directory of Open Access Journals (Sweden)

    Corsini Joe

    2004-01-01

    Full Text Available Herein we have explored two practical aspects of cryopreserving cultured mammalian cells during routine laboratory maintenance. First, we have examined the possibility of using a serum-free, hence more affordable, cryopreservative. Using five mammalian lines (Crandell Feline Kidney, MCF7, A72, WI 38 and NB324K, we found that the serum-free alternative preserves nearly as efficiently as the serum-containing preservatives. Second, we compared cryostorage of those cells in suspended versus a pellet form using both aforementioned cryopreservatives. Under our conditions, cells were in general recovered equally well in a suspended versus a pellet form.

  13. Impact of starting material (fresh versus cryopreserved marrow) on mesenchymal stem cell culture.

    Science.gov (United States)

    Kaplan, Alesia; Sackett, Katie; Sumstad, Darin; Kadidlo, Dianne; McKenna, David H

    2017-09-01

    Mesenchymal stem cells (MSCs) continue to be investigated in multiple clinical trials as potential therapy for different disorders. There is ongoing controversy surrounding the clinical use of cryopreserved versus fresh MSCs. However, little is known about how cryopreservation affects marrow as starting material. The growth kinetics of MSC cultures derived from fresh versus cryopreserved marrow were compared. Data were reviewed on the growth kinetics of MSCs derived from fresh versus cryopreserved marrow of nine donors. Marrow harvested from each donor was separated into four aliquots (one fresh and three cryopreserved for culture). Data on the date of mononuclear cell cryopreservation/thaw, MSC counts at Passages 1 and 2, MSC doubling, MSC fold expansion, viability (of mononuclear cells and final MSCs), and on flow cytometry markers of mononuclear cells and final MSCs were analyzed for the fresh and cryopreserved marrow groups. In total, 21 MSC lots (seven fresh and 14 cryopreserved) were obtained. The average age of cryopreserved mononuclear cell product was 295 days (range, 18-1241 days). There were no significant differences between MSC numbers at Passage 1 (p = 0.1), final MSC numbers (p = 0.5), MSC doubling (p = 0.7), or MSC fold expansion (p = 0.7). A significant difference was observed in viability by flow cytometry for both mononuclear cells (p = 0.002) and final MSCs (p = 0.009), with higher viability in the fresh marrow group. This study demonstrates that MSCs derived from cryopreserved marrow have the same growth characteristics as fresh marrow-derived MSCs. Further studies are needed to explore potential differences in clinical efficacy. © 2017 AABB.

  14. Cryopreservation of mammalian semen.

    Science.gov (United States)

    Curry, Mark R

    2007-01-01

    Mammalian spermatozoa were among the very first cells to be successfully cryopreserved and over the last five decades the use of frozen-thawed semen for artificial insemination has come to play an important role in domestic livestock production. More recently, semen freezing has increasingly been utilized in the establishment of genetic resource banks for endangered species. Semen is collected, most commonly either by use of an artificial vagina or by electroejaculation of an anaesthetized animal, and basic sperm parameters assessed. Semen is extended using a TEST-egg yolk-glycerol diluent, packaged in 0.25-mL plastic straws and slowly cooled to 5 degrees C over a period of 1-2 h. Cooled straws are frozen by suspending within liquid nitrogen vapor above the liquid nitrogen surface before plunging into the liquid phase. Straws are thawed briefly in air before immersing in a 35 degrees C water bath for 15 s, and often are used directly for insemination without any further processing.

  15. Efficiency and Effectiveness in Higher Education: A Report by the Universities UK Efficiency and Modernisation Task Group

    Science.gov (United States)

    Universities UK, 2011

    2011-01-01

    Effectiveness, efficiency and value for money are central concerns for the higher education sector. In England, decisions made by the current Government will effect a radical change in the funding for teaching. Institutions will be managing a reduction in public funding for teaching and the transition to the new system of graduate contributions,…

  16. In vitro propagation and cryopreservation of Aerides odorata Lour. (Orchidaceae).

    Science.gov (United States)

    Hongthongkham, J; Bunnag, S

    2014-05-01

    An efficient method for in vitro propagation and cryopreservation of Aerides odorata was established. Leaf segments were cultured on New Dogashima (ND) mediums supplemented with various concentrations of Benzyladenine (BA) (0-5 mg L(-1)) combined with Naphthaleneacetic Acid (NAA) (0-2 mg L(-1)). The optimal treatment for inducing Protocorm-like Bodies (PLBs) from leaf segments was obtained from the combination of 1 or 3 mg L(-1) BA and 0.5 or 1 mg L(-1) NAA; whereas, the addition of BA or NAA alone induced shoot and/or root initiation rather than PLB or callus formation. Shoots rapidly developed on ND mediums containing 5 mg L(-1) BA. Cryopreservation of leaf segment-derived PLBs was successful using the encapsulation-dehydration method. The maximum survival percentage of Cryopreserved (Cryp) PLBs was achieved by encapsulating PLBs with 2% Na-alginate combined with 2 M glycerol and 0.4 M sucrose. The encapsulated PLBs were then precultured in 0.75 M sucrose for 24 h and dehydrated for 6 h before plunging into liquid nitrogen. Genetic stability of Cryp PLBs after regrowth was assessed by flow cytometry. The findings showed no different patterns of ploidy levels and morphology between Cryp and non-cryopreserved (Ncryp) control plantlets.

  17. Semen cryopreservation in fish: effects on sperm motility and fertility

    International Nuclear Information System (INIS)

    Martinez, Jose Gregorio; Pardo Carrasco, Sandra

    2010-01-01

    The cryopreservation of semen in fish, as in many species even shows effects that decrease sperm quality and directly engage cell ability to successfully participate in the processes of fertilization and embryonic development. the characteristics such as mobility and fertilizing capacity of fertilization of sperm are considered to be quality criteria that allow to measure the success or failure of the process, since they are considered integrative variables, being indicators that depend not on a single factor, but on the stability and welfare of all structures, enzymes and subcellular functional compounds that give place to these spermatic characteristics. membrane damage (Adenylate cyclase, ion channels, grouping of other proteins, among others) and their implication in the route of signaling pathway leading to spermatic activation, ATP degradation and fragmentation of nuclear and mitochondrial DNA (genome), degradation of kinase enzymes and other cytosolic proteins (proteome) are considered today, as some of the molecular factors that most affect during cryopreservation and markedly decreasing the fertilizing capacity and mobility of sperm in fish. Proposals on the molecular mechanisms, by which these subcellular factors interact and act as consequence of cryopreservation, are some of the topics covered in this review. Understanding the principles and factors that are involved in the origin of such damages, will allow to improved cryopreservation processes, making them less harmful and more efficient.

  18. Effectiveness of human spermatozoa biomarkers as indicators of structural damage during cryopreservation.

    Science.gov (United States)

    Gómez-Torres, María José; Medrano, Llanos; Romero, Alejandro; Fernández-Colom, Pedro José; Aizpurúa, Jon

    2017-10-01

    Human spermatozoa cryopreservation techniques are used to maintain and protect male fertility in cases such as infertility and malignancy treatments. However, during cryopreservation, the spermatozoa's metabolic rate is reduced and they undergo dramatic functional and structural changes owing to exposure to cryoprotectants and freezing-thawing procedures. While the effects of cryopreservation on cells are documented, to date the induced cryodamage on structural and/or functional sperm biomarkers is not well established at multivariate scale. To address this question, we performed basic sperm analysis, sperm DNA fragmentation assessment, spontaneous acrosome reaction measurement, and cytoskeleton evaluation after thawing samples from subjects with normal and low-quality semen. A cryodamage rate was used to determine the effects of the freeze-thaw process on spermatozoa. In addition, a Principal Component Analysis (PCA) was used for data reduction and to evaluate sperm-specific patterns during the cryopreservation process. We found that the vitality, progressive motility and sperm count from low-quality samples after cryopreservation show higher damage rates (≥40%) than in normal sperm samples. However, cytoskeleton, DNA, tail and mid-piece and acrosome display the highest cryodamage rates (∼50-99%) and are equally susceptible to cryopreservation-induced damage in both low- and normal-quality semen samples. Overall, the evaluation of these parameters provides meaningful information about different aspects of sperm functionality after cryopreservation. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Reducing barriers to energy efficiency in the German higher education sector. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Schleich, J.; Boede, U.

    2000-12-01

    This report describes the empirical research into barriers to energy efficiency in the German higher education (HE) sector. It is one of nine such reports in the BARRIERS project. The report contains description and analysis of six case studies of energy management in German universities. The results are analysed using the theoretical framework developed for the BARRIERS project (Sorrell et al., 2000). The report also provides brief recommendations on how these barriers to the rational use of energy (RUE) may be overcome and how energy efficiency within the sector may be improved. The results of the study for the higher education sector in Germany are summarised in this executive summary under the following headings: - Characterising the higher education sector; - Case studies of energy management in the German higher education sector; - Evidence of barriers in the German higher education sector; - The role of energy service companies in the higher education sector; - Policy implications. (orig.)

  20. Reducing barriers to energy efficiency in the German higher education sector. Executive summary

    Energy Technology Data Exchange (ETDEWEB)

    Schleich, J.; Boede, U.

    2000-12-01

    This report describes the empirical research into barriers to energy efficiency in the German higher education (HE) sector. It is one of nine such reports in the BARRIERS project. The report contains description and analysis of six case studies of energy management in German universities. The results are analysed using the theoretical framework developed for the BARRIERS project (Sorrell et al., 2000). The report also provides brief recommendations on how these barriers to the rational use of energy (RUE) may be overcome and how energy efficiency within the sector may be improved. The results of the study for the higher education sector in Germany are summarised in this executive summary under the following headings: - Characterising the higher education sector; - Case studies of energy management in the German higher education sector; - Evidence of barriers in the German higher education sector; - The role of energy service companies in the higher education sector; - Policy implications. (orig.)

  1. The effects of X-irradiation on ex vivo expansion of cryopreserved human hematopoietic stem/progenitor cells

    International Nuclear Information System (INIS)

    Hayashi, Naoki; Takahashi, Kenji; Kashiwakura, Ikuo

    2010-01-01

    In our previous study (Life Sciences 84: 598, 2009), we demonstrated that placental/umbilical cord blood-derived mesenchymal stem cell-like stromal cells have the effect to support the regeneration of freshly prepared X-irradiated hematopoietic stem/progenitor cells (HSPCs). Generally, HSPCs are supplied from companies, institutions, and cell banks that cryopreserve them for clinical and experimental use. In this study, the influence of cryopreservation on the responses of HSPCs to irradiation and co-culture with stromal cells is assessed. After cryopreservation with the optimal procedure, 2 Gy-irradiated HSPCs were cultured with or without stromal cells supplemented with combination of interleukin-3, stem cell factor, and thrombopoietin. The population of relatively immature CD34 + /CD38 - cells in cryopreserved cells was significantly higher than in fresh cells prior to cryopreservation; furthermore, the hematopoietic progenitor populations of CD34 + /CD45RA + cells and CD34 + /CD117 + cells in cryopreserved cells were significantly lower than that in fresh cells. However, the rate of expansion in the cryopreserved HSPCs was lower than in the fresh HSPCs. In the culture of cryopreserved cells irradiated with 2 Gy, the growth rates of CD34 + cells, CD34 + /CD38 - cells, and hematopoietic progenitors were greater than growth rates of their counter parts in the culture of fresh cells. Surprisingly, the effect to support the hematopoiesis in co-culture with stromal cells was never observed in the X-irradiated HSPCs after cryopreservation. The present results demonstrated that cryopreserving process increased the rate of immature and radio-resistant HSPCs but decreased the effects to support the hematopoiesis by stromal cells, thus suggesting that cryopreservation changes the character of HSPCs. (author)

  2. Cryopreservation of coconut (Cocos nucifera L.) zygotic embryos does not induce morphological, cytological or molecular changes in recovered seedlings.

    Science.gov (United States)

    Sisunandar; Rival, Alain; Turquay, Patricia; Samosir, Yohannes; Adkins, Steve W

    2010-07-01

    The present study aimed at exploring the fidelity of coconut (Cocos nucifera L.) plants recovered from cryopreservation. Zygotic embryos from various different cultivars were cryopreserved following four successive steps, namely: rapid dehydration, rapid freezing, rapid thawing and in vitro recovery followed by acclimatization. At the end of the acclimatization period, the seedlings were compared to counterparts of the same age, which were produced from non-cryopreserved embryos. Both series were submitted to morphological, cytological and molecular comparisons. No significant differences in terms of growth rates could be measured. In addition, no morphological variation could be detected through the measurement of shoot elongation rates, production of opened leaves, and the number and total length of primary roots. Karyotype analysis revealed the same chromosome number (2n = 32) in all studied cultivars independently of cryopreservation. No significant differences could be observed between control and cryopreserved material concerning the type of chromosomes, the length of the long and short arms, the arm length ratio and the centromeric index. However, idiogram analysis did show a greater number of black banding on chromosomes isolated from cryopreserved material. Genetic and epigenetic fidelity was assessed through microsatellite (SSR) analysis and global DNA methylation rates; no significant differences would be observed between genomic DNAs isolated from seedlings originating from cryopreserved embryos and respective controls. In conclusion, our results suggest that the method of cryopreservation under study did not induce gross morphological, genetic or epigenetic changes, thus suggesting that it is an appropriate method to efficiently preserve coconut germplasm.

  3. The economical efficiency of private investments in higher education in Russia

    Directory of Open Access Journals (Sweden)

    Elena Maksyutina

    2011-12-01

    Full Text Available The article investigates the economical efficiency of investments in higher education in modern conditions of Russia. The beginning of the article includes a characteristic of the existing empiric research concerning the efficiency of investments in human capital assets. Further the author of the article introduces the results of pay off calculation of private investments in higher education. The result of the research was that in modern conditions of Russia investments in higher education are exceedingly advantageous. High norms of higher education feedback and short period of pay off of these investments explain the reasons of continuously growing demand for it on the part of the population, especially young people. The article proves that the level of population education in Russia is quite high, however accumulated human capital asset is used insufficiently effective. Many people with higher education are forced to take jobs not requiring higher education.  Sharp shift in educational behavior of Russian people raises new demands to labor market. Graduates of higher educational institutions, appearing on a labor market, form qualitatively different demands towards it. But tempo of Russian economics development today can not provide job positions for all graduates of higher educational institutions. That is why structural change of economics is needed.

  4. CIGS cells with metallized front contact: Longer cells and higher efficiency

    NARCIS (Netherlands)

    Deelen, J. van; Frijters, C.

    2017-01-01

    We have investigated the benefit of a patterned metallization on top of a transparent conductive oxide in CIGS thin-film solar panels. It was found that cells with a grid have a higher efficiency compared to cells with only a TCO. This was observed for all cell lengths used. Furthermore, metallic

  5. Efficiency, Costs, Rankings and Heterogeneity: The Case of US Higher Education

    Science.gov (United States)

    Agasisti, Tommaso; Johnes, Geraint

    2015-01-01

    Among the major trends in the higher education (HE) sector, the development of rankings as a policy and managerial tool is of particular relevance. However, despite the diffusion of these instruments, it is still not clear how they relate with traditional performance measures, like unit costs and efficiency scores. In this paper, we estimate a…

  6. Improving extraction efficiency of the third integer resonant extraction using higher order multipoles

    Energy Technology Data Exchange (ETDEWEB)

    Brown, K. A. [Brookhaven National Lab. (BNL), Upton, NY (United States); Schoefer, V. [Brookhaven National Lab. (BNL), Upton, NY (United States); Tomizawa, M. [High Energy Accelerator Research Organization (KEK), Tsukuba (Japan)

    2017-03-09

    The new accelerator complex at J-PARC will operate with both high energy and very high intensity proton beams. With a design slow extraction efficiency of greater than 99% this facility will still be depositing significant beam power onto accelerator components [2]. To achieve even higher efficiencies requires some new ideas. The design of the extraction system and the accelerator lattice structure leaves little room for improvement using conventional techniques. In this report we will present one method for improving the slow extraction efficiency at J-PARC by adding duodecapoles or octupoles to the slow extraction system. We will review the theory of resonant extraction, describe simulation methods, and present the results of detailed simulations. From our investigations we find that we can improve extraction efficiency and thereby reduce the level of residual activation in the accelerator components and surrounding shielding.

  7. Can More Environmental Information Disclosure Lead to Higher Eco-Efficiency? Evidence from China

    Directory of Open Access Journals (Sweden)

    Yantuan Yu

    2018-02-01

    Full Text Available The present paper investigates the impact of pollution information transparency index (PITI on eco-efficiency using a novel panel dataset covering 109 key environmental protection prefecture-level cities in China over the period 2008–2015. We apply an extended data envelopment analysis (DEA model, simultaneously incorporating metafrontier, undesirable outputs and super efficiency into slack-based measure (Meta-US-SBM to estimate eco-efficiency. Then, the bootstrap Granger causality approach is utilized to test the unidirectional Granger causal relationship running from PITI to eco-efficiency. Results of DEA model show that there exist significant spatiotemporal disparities of eco-efficiency, on average, the eco-efficiency in eastern region is relative higher than those of central/western region. Estimates of ordinary least square (OLS method, quantile regression, and spatial Durbin model document that the evidence of an inverted-U-shaped relation between PITI and eco-efficiency is supported, and the turning points vary from 0.3370 to 0.4540 with different model specifications. Finally, supplementary analysis of panel threshold model also supports the robust findings. Policy implications are presented based on the empirical results.

  8. Augmented dried versus cryopreserved amniotic membrane as an ocular surface dressing.

    Directory of Open Access Journals (Sweden)

    Claire L Allen

    Full Text Available Dried amniotic membrane (AM can be a useful therapeutic adjunct in ophthalmic surgery and possesses logistical advantages over cryopreserved AM. Differences in preservation techniques can significantly influence the biochemical composition and physical properties of AM, potentially affecting clinical efficacy. This study was established to investigate the biochemical and structural effects of drying AM in the absence and presence of saccharide lyoprotectants and its biocompatibility compared to cryopreserved material.AM was cryopreserved or dried with and without pre-treatment with trehalose or raffinose and the antioxidant epigallocatechin (EGCG. Structural and visual comparisons were assessed using electron microscopy. Localisation, expression and release of AM biological factors were determined using immunoassays and immunofluorescence. The biocompatibility of the AM preparations co-cultured with corneal epithelial cell (CEC or keratocyte monolayers were assessed using cell proliferation, cytotoxicity, apoptosis and migration assays.Drying devitalised AM epithelium, but less than cryopreservation and cellular damage was reduced in dried AM pre-treated with trehalose or raffinose. Dried AM alone, and with trehalose or raffinose showed greater factor retention efficiencies and bioavailability compared to cryopreserved AM and demonstrated a more sustained biochemical factor time release in vitro. Cellular health assays showed that dried AM with trehalose or raffinose are compatible and superior substrates compared to cryopreserved AM for primary CEC expansion, with increased proliferation and reduced LDH and caspase-3 levels. This concept was supported by improved wound healing in an immortalised human CEC line (hiCEC co-cultured with dried and trehalose or raffinose membranes, compared to cryopreserved and fresh AM.Our modified preservation process and our resultant optimised dried AM has enhanced structural properties and biochemical stability

  9. Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa.

    Science.gov (United States)

    Prapaiwan, N; Tharasanit, T; Punjachaipornpol, S; Yamtang, D; Roongsitthichai, A; Moonarmart, W; Kaeoket, K; Manee-In, S

    2016-05-01

    Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL) has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05) than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05). In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.

  10. Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa

    Directory of Open Access Journals (Sweden)

    N. Prapaiwan

    2016-05-01

    Full Text Available Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05 than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05. In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.

  11. Sustainability in Chinese Higher Educational Institutions’ Social Science Research: A Performance Interface toward Efficiency

    Directory of Open Access Journals (Sweden)

    Xianmei Wang

    2017-10-01

    Full Text Available Sustainability issues in higher educational institutions’ (HEIs research, especially in the social science field, have attracted increasing levels of attention in higher education administration in recent decades as HEIs are confronted with a growing pressure worldwide to increase the efficiency of their research activities under a limited volume and relatively equitable division of public funding resources. This paper introduces a theoretical analysis framework based on a data envelopment analysis, separating the social science research process into a foundation stage and a construction stage, and then projecting each HEI into certain quadrants to form several clusters according to their overall and stage efficiencies and corresponding Malmquist Productivity Indices. Furthermore, the interfaces are formed in each cluster as feasible potential improvement directions. The empirical results in detail are demonstrated from a data set of Chinese HEIs in Jiangsu Province over the Twelfth Five-Year period as offering a closer approximation to the “China social science research best practice”.

  12. Why do French civil-law countries have higher levels of financial efficiency?

    OpenAIRE

    Asongu Simplice

    2011-01-01

    The dominance of English common-law countries in prospects for financial development in the legal-origins debate has been debunked by recent findings. Using exchange rate regimes and economic/monetary integration oriented hypotheses, this paper proposes an “inflation uncertainty theory” in providing theoretical justification and empirical validity as to why French civil-law countries have higher levels of financial allocation efficiency. Inflation uncertainty, typical of floating exchange rat...

  13. Advances in Stallion Semen Cryopreservation.

    Science.gov (United States)

    Alvarenga, Marco Antonio; Papa, Frederico Ozanam; Ramires Neto, Carlos

    2016-12-01

    The use of stallion frozen semen minimizes the spread of disease, eliminates geographic barriers, and preserves the genetic material of the animal for an unlimited time. Significant progress on the frozen thawed stallion semen process and consequently fertility has been achieved over the last decade. These improvements not only increased fertility rates but also allowed cryopreservation of semen from "poor freezers." This article reviews traditional steps and new strategies for stallion semen handling and processing that are performed to overcome the deleterious effects of semen preservation and consequently improve frozen semen quality and fertility. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Post-weaning feed efficiency decreased in progeny of higher milk yielding beef cows.

    Science.gov (United States)

    Mulliniks, J T; Edwards, S R; Hobbs, J D; McFarlane, Z D; Cope, E R

    2018-02-01

    Current trends in the beef industry focus on selecting production traits with the purpose of maximizing calf weaning weight; however, such traits may ultimately decrease overall post-weaning productivity. Therefore, the objective of this study was to evaluate the effects of actual milk yield in mature beef cows on their offspring's dry matter intake (DMI), BW, average daily gain, feed conversion ratio (FCR) and residual feed intake (RFI) during a ~75-day backgrounding feeding trial. A period of 24-h milk production was measured with a modified weigh-suckle-weigh technique using a milking machine. After milking, cows were retrospectively classified as one of three milk yield groups: Lower (6.57±1.21 kg), Moderate (9.02±0.60 kg) or Higher (11.97±1.46 kg). Calves from Moderate and Higher milk yielding dams had greater (Pfeeding phase; however, day 75 BW were not different (P=0.36) between Lower and Moderate calves. Body weight gain was greater (P=0.05) for Lower and Moderate calves from the day 0 BW to day 35 BW compared with Higher calves. Overall DMI was lower (P=0.03) in offspring from Lower and Moderate cows compared with their Higher milking counterparts. With the decreased DMI, FCR was lower (P=0.03) from day 0 to day 35 in calves from Lower and Moderate milk yielding dams. In addition, overall FCR was lower (P=0.02) in calves from Lower and Moderate milk yielding dams compared with calves from Higher milk yielding dams. However, calving of Lower milk yielding dams had an increased (P=0.04) efficiency from a negative RFI value compared with calves from Moderate and Higher milking dams. Results from this study suggest that increased milk production in beef cows decreases feed efficiency during a 75-day post-weaning, backgrounding period of progeny.

  15. Higher-fidelity yet efficient modeling of radiation energy transport through three-dimensional clouds

    International Nuclear Information System (INIS)

    Hall, M.L.; Davis, A.B.

    2005-01-01

    Accurate modeling of radiative energy transport through cloudy atmospheres is necessary for both climate modeling with GCMs (Global Climate Models) and remote sensing. Previous modeling efforts have taken advantage of extreme aspect ratios (cells that are very wide horizontally) by assuming a 1-D treatment vertically - the Independent Column Approximation (ICA). Recent attempts to resolve radiation transport through the clouds have drastically changed the aspect ratios of the cells, moving them closer to unity, such that the ICA model is no longer valid. We aim to provide a higher-fidelity atmospheric radiation transport model which increases accuracy while maintaining efficiency. To that end, this paper describes the development of an efficient 3-D-capable radiation code that can be easily integrated into cloud resolving models as an alternative to the resident 1-D model. Applications to test cases from the Intercomparison of 3-D Radiation Codes (I3RC) protocol are shown

  16. Development of methods for cryopreservation of rooster sperm from the endangered breed "Gallina Valenciana de Chulilla" using low glycerol concentrations.

    Science.gov (United States)

    Blanch, E; Tomás, C; Casares, L; Gómez, E A; Sansano, S; Giménez, I; Mocé, E

    2014-06-01

    Glycerol (11%; v:v) is the cryoprotectant most often used for the cryopreservation of rooster sperm. However, chicken breeds differ in the resistance of their sperm to the cryopreservation process and endangered or local breeds usually present low fertilizing ability when conventional sperm cryopreservation protocols are used. The objective of this study was to optimize the protocol for the cryopreservation of the sperm from the endangered breed "Gallina Valenciana de Chulilla". For this purpose, 10 pools of semen from 43 roosters of this breed were cryopreserved using 8%, 7%, 6%, or 4% glycerol, and the sperm quality was determined immediately after thawing and in the insemination doses. Lohmann Brown Classic laying hens (n = 40) were used for the insemination trials. The sperm quality after cryopreservation progressively decreased as the glycerol concentration was reduced (P roosters frozen using 4% glycerol exhibited lower sperm quality but similar fertilizing ability compared with samples processed using higher glycerol concentrations. These results may provide useful information for developing cryopreservation protocols for other breeds. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Cryopreservation: a cold look at technology for fertility preservation.

    Science.gov (United States)

    Gosden, Roger

    2011-08-01

    To outline the history of cryopreservation technology and its contributions to reproductive medicine, including fertility preservation. A search of the relevant literature using Medline and other online tools. Research and laboratory protocol development. The biology of preserving cells at low temperatures is complex and still being unraveled. Principles were first established more than half a century ago, with progress being driven empirically and often by trial and error. The protocols vary widely, and practice is still heavily dependent on operator skill, accounting for wide differences in the success rates between centers. No single protocol fits all specimen types, and differential vulnerability to cryoinjury remains a major obstacle. Nevertheless, semen cryopreservation has long been established, embryo banking is now highly effective, and vitrification appears to overcome problems with oocytes. Protocols in the future, although specific to the cell type and tissue, are likely to evolve toward generally acknowledged standards. But heterogeneity between patients and even within samples implies that each cell may have its own peculiar optimum for minimizing cryoinjury; because protocols are therefore compromises, "perfect" preservation may be unattainable. Cryopreservation has become a mainstay in the assisted reproduction laboratory and underpins fertility preservation for patients with cancer and other conditions. The practice is currently evolving from slow freezing methods toward more vitrification, and future technology is likely to reduce dependence on operator skill, which should raise success rates to higher, more uniform levels. Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  18. Cryopreservation of zebrafish (Danio rerio) oocytes by vitrification.

    Science.gov (United States)

    Guan, M; Rawson, D M; Zhang, T

    2010-01-01

    Cryopreservation of fish oocytes is challenging because these oocytes have low membrane permeability to water and cryoprotectant and are highly chilling sensitive. Vitrification is considered to be a promising approach for their cryopreservation as it involves rapid freezing and thawing of the oocytes and therefore minimising the chilling injury. In the present study, vitrification properties and the toxicity of a range of vitrification solutions containing different concentrations of Me2SO, methanol, propylene glycol and ethylene glycol were investigated. Two different base media and vitrification methods were compared. The effect of different post-thaw dilution solutions together with incubation periods on oocyte viability were also investigated. Stage III zebrafish oocytes were equilibrated in increasing concentrations of cryoprotectants for 30 min in 3 steps. Oocytes were thawed rapidly in a water bath and cryoprotectants were removed in 4 steps. Oocyte viability was assessed using trypan blue staining. The results showed that vitrification solutions V3 and V4 in KCl buffer had low toxicity and vitrified well. The survivals of oocytes after stepwise dilution using solutions containing permeable cryoprotectants were significant higher than those diluted in 0.5M glucose, and the use of CVA65 vitrification system improved oocyte survival when compared with plastic straws after 30 min at 22 degrees C post-thawing. Cryopreservation of zebrafish oocytes by vitrification is reported here for the first time, although oocyte survivals after cryopreservation assessed by trypan blue staining were relatively high shortly after thawing, they became swollen and translucent after incubation in KCl buffer. Further studies are needed to optimise the post-thaw culturing conditions.

  19. Apoptosis in fresh and cryopreserved cardiac valves of pig samples.

    Science.gov (United States)

    Rendal Vázquez, M Esther; Díaz Román, T M; Rodríguez Cabarcos, M; Zavanella Botta, C; Domenech García, N; González Cuesta, M; Sánchez Dopico, M J; Pértega Díaz, S; Andión Núñez, C

    2008-06-01

    To analyse the influence of cold ischemic time (CIT) (2-24 h) and of cryopreservation (liquid phase) on the viability of the valvular fibroblasts and in the presence of apoptosis. Cardiac valves from 10 pigs were evaluated by anatomo-pathological study of the wall, muscle and leaflet. At the same time, the presence of cellular death due to apoptosis was investigated in two ways; directly on tissue by Apodetec system and by two-colour flow cytometry assay analyzing a suspension of fibroblast from valve leaflets using Anexina V and propidium iodure (PI). We established three groups of samples to compare different experimental conditions: 2 h of ischemia (group 1), 24 h of ischemia (group 2), and a programme of cryopreservation (-1 degrees C/min) after 2 h of ischemia, followed by storage in liquid nitrogen during a week and thawing was performed (group 3). The analysis of viabilities showed slight differences between all three groups. The results indicated CIT of 24 h undergoing more structural affectation than CIT of 2 h. Flow cytometry analysis did not show important differences between groups; however cryopreserved samples (group 3) slightly less viability and a higher percentage of death by apoptosis than group 1 and 2 using flow cytometry. Apoptosis was confirmed on tissue from all valves but mainly in samples of group 2 and group 3. In summary, the viability of the valves in the case of ischemic times of 2 h, 24 h or after cryopreservation/thawing differs slightly. The death of the cells is mainly mediated by necrosis and not by apoptosis.

  20. Economic efficiency of e-learning in higher education: An industrial approach

    Directory of Open Access Journals (Sweden)

    Jordi Vilaseca

    2008-07-01

    Full Text Available Little work has been yet done to analyse if e-learning is an efficiency way in economic terms to produce higher education, especially because there are not available data in official statistics. Despite of these important constrains, this paper aims to contribute to the study of economic efficiency of e-learning through the analysis of a sample of e-learning universities during a period of time (1997-2002. We have wanted to obtain some empirical evidence to understand if e-learning is a feasible model of providing education for universities and which are the variables that allow for feasibility attainment. The main findings are: 1 that the rise of the number of students enrolled is consistent with increasing labour productivity rates; 2 that cost labour savings are explained by the improvement of universities’ economic efficiency (or total factor productivity; and 3 that improvement of total factor productivity in e-learning production is due to the attainment of scale economies, but also to two organisational innovations: outsourcing processes that leads to the increase of variable costs consistent with decreasing marginal costs, and the sharing of assets’ control and use that allow for a rise in assets rotation.

  1. Factor Analysis on Criteria Affecting Lean Retrofit for Energy Efficient Initiatives in Higher Learning Institution Buildings

    Directory of Open Access Journals (Sweden)

    Abidin Nur IzieAdiana

    2017-01-01

    Full Text Available The expansion of Higher Learning Institution (HLI is a global concerns on energy demand due to campus act like a small city. Intensive mode of operation of a building is correlated to the energy utilization. Improvement in the current energy efficiency is crucial effort to minimize the environmental effect through minimisation of energy in operation by retrofitting and upgrade the existing building system or components to be more efficient. Basically, there are three recommended steps for the improvement known as lean initiatives, green technology and clean energy in response to becoming zero energy solutions for building. The deliberation of this paper is aimed to highlight the criteria affecting in retrofitting of existing building in HLI with lean initiatives in order to achieve energy efficiency and reduction of energy comsumption. Attention is devoted to reviewing the lean energy retrofitting initiatives criteria for daylighting (side lighting, daylighting (skylight and glazing. The questionnaire survey was employed and distributed to the architects who has an expertise in green building design. Factor analysis was adopted as a method of analysis by using Principal Component with Varimax Rotation. The result is presented through summarizing the sub-criteria according to its importance with a factor loading 0.50 and above. The result found that majority of the criteria developed achieved the significant factor loading value and in accordance with the protocal of analysis. In conclusion the results from analysis of this paper assists the stakeholders in assessing the significant criteria based on the desired lean energy retrofitting initiatives and also provides a huge contribution for future planning improvement in existing buildings to become an energy efficient building.

  2. Mature Oocyte Cryopreservation for Fertility Preservation.

    Science.gov (United States)

    Liang, Tina; Motan, Tarek

    2016-01-01

    In recent decades, advances in cancer treatment have led to a dramatic improvement in long term survival. This has led to an increasing focus on quality of life after surviving cancer treatment, with fertility being an important aspect. Given the known reproductive risks of cancer therapies, there has been a growing interest in the field of fertility preservation (also referred to as oncofertility). Mature oocyte cryopreservation is no longer considered experimental and has become a realistic option for reproductive aged women prior to undergoing cancer treatment. Additionally, as cryopreservation techniques improve, mature oocyte cryopreservation is increasing being marketed to healthy women without cancer wishing to delay child bearing, also termed "social egg freezing". This chapter provides a review of the current technology, use, and outcomes of mature oocyte cryopreservation. It also outlines the ethical debate surrounding social egg freezing and directions for future research in female fertility preservation.

  3. Protectants used in the cryopreservation of microorganisms

    Czech Academy of Sciences Publication Activity Database

    Hubálek, Zdeněk

    2003-01-01

    Roč. 46, č. 3 (2003), s. 205-229 ISSN 0011-2240 Institutional research plan: CEZ:AV0Z6093917 Keywords : cryopreservation * cryoprotectants Subject RIV: EE - Microbiology, Virology Impact factor: 1.445, year: 2003

  4. Challenges in cryopreservation of regulatory T cells (Tregs) for clinical therapeutic applications.

    Science.gov (United States)

    Golab, Karolina; Leveson-Gower, Dennis; Wang, Xiao-Jun; Grzanka, Jakub; Marek-Trzonkowska, Natalia; Krzystyniak, Adam; Millis, J Michael; Trzonkowski, Piotr; Witkowski, Piotr

    2013-07-01

    Promising results of initial studies applying ex-vivo expanded regulatory T cell (Treg) as a clinical intervention have increased interest in this type of the cellular therapy and several new clinical trials involving Tregs are currently on the way. Methods of isolation and expansion of Tregs have been studied and optimized to the extent that such therapy is feasible, and allows obtaining sufficient numbers of Tregs in the laboratory following Good Manufacturing Practice (GMP) guidelines. Nevertheless, Treg therapy could even more rapidly evolve if Tregs could be efficiently cryopreserved and stored for future infusion or expansions rather than utilization of only freshly isolated and expanded cells as it is preferred now. Currently, our knowledge regarding the impact of cryopreservation on Treg recovery, viability, and functionality is still limited. Based on experience with cryopreserved peripheral blood mononuclear cells (PBMCs), cryopreservation may have a detrimental effect on Tregs, can decrease Treg viability, cause abnormal cytokine secretion, and compromise expression of surface markers essential for proper Treg function and processing. Therefore, optimal strategies and conditions for Treg cryopreservation in conjunction with cell culture, expansion, and processing for clinical application still need to be investigated and defined. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Theoretical and methodological grounds of formation of the efficient system of higher education

    Directory of Open Access Journals (Sweden)

    Raevneva Elena V.

    2013-03-01

    Full Text Available The goal of the article lies in generalisation of the modern theoretical and methodological, methodical and instrumentation provision of building of efficient system of higher education. Analysis of literature on the problems of building educational systems shows that there is a theoretical and methodological and instrumentation level of study of this issue. The article considers a theoretical and methodological level of the study and specifies theories and philosophic schools, concepts, educational paradigms and scientific approaches used during formation of the educational paradigm. The article considers models of education and models and technologies of learning as instrumental provision. In the result of the analysis the article makes a conclusion that the humanistic paradigm, which is based on the competency building approach and which assumes the use of modern (innovation technologies of learning, should be in the foundation of reformation of the system of higher education. The prospect of further studies in this directions is formation of competences of potential specialists (graduates of higher educational establishments with consideration of requirements of employers and market in general.

  6. Beyond Frontiers: Comparing the Efficiency of Higher Education Decision-Making Units across More than One Country

    Science.gov (United States)

    Agasisti, Tommaso; Johnes, Geraint

    2009-01-01

    We employ Data Envelopment Analysis to compute the technical efficiency of Italian and English higher education institutions. Our results show that, in relation to the country-specific frontier, institutions in both countries are typically very efficient. However, institutions in England are more efficient than those in Italy when we compare…

  7. Integrating Micro- or Nanoscale Encapsulation Technology with Vitreous Cryopreservation: A New Strategy to Improve Biopreservation.

    Science.gov (United States)

    Zheng, Y Y; Zhao, G

      BACKGROUND: None-uniform distributions of temperatures, limited freezing and thawing rates, and thermal stresses are three main hindering factors for successful vitreous cryopreservation of mass volume of biosamples. Micro- and nanoscale encapsulation, owning the intrinsic features to avoid those limitations introduced by the traditional approaches, has been opened up a new way for effective and high-efficiency biopreservation. This short review article summarizes recent advances in cell encapsulation technology for biopreservation by manipulating cells and biological agents in micro- or nanoscale volume droplets and microgels, and discusses its promising applications for future vitreous cryopreservation.

  8. [The destiny of cryopreserved embryos].

    Science.gov (United States)

    Karpel, L; Achour-Frydman, N; Frydman, R; Flis-Trèves, M

    2007-12-01

    To know the psychological motivations of couples who keep their embryos so long (five years and more) and do not make a decision about them. We studied 84 couples refrained from making a decision on their cryopreserved embryos for at least five years. They were invited to fill out a questionnaire focusing on three points: the reasons of the indecision, their own representation of the cryopreserved embryos and their choice for the future: donation to another couple, to research, pregnancy or no solution for the moment. Mean (S.D.) women's and men's age were respectively, 38.8 (2.5)- and 41.3 (2.5)-years old. On average, three (1-9) embryos are preserved since 7.5 (5-12) years. Most of couples are parents. Four major reasons explain their attitudes: feeling of being too aged (25%), fear of a multiple pregnancy (45%), disagreement between members of couple (20%) and fear of failure (42.5%). Multiple choices were given to the future of the embryos: 25% wanted a pregnancy, 8% wanted to give them to infertile couples, 20% to research and 27.5% did not find any solution. Twenty percent were hesitating. The representation of those embryos is more symbolic than material. Most of the time, they see them like a potential child, a hope for the future or a brother or sister of their alive children. Those embryos are symbolized. They are a proof of fertility, a hope for another child. So, whatever the legal statement, couples will be in a dilemma because it is never easy for an infertile person to renounce to embryos, and the hope for children.

  9. Achieving higher efficiency of production through knowledge management via social capital management

    Directory of Open Access Journals (Sweden)

    Jana Plchová

    2015-09-01

    Full Text Available The article shows a new approach – how to reach higher efficiency in the production from knowledge management via managing social capital through measurement, motivation and stimulation. The test in a real company on the Toyota system implementation is de-scribed. The active involvement of people is an important part of the Toyota system success. This is obvious in Japan but creates a big problem in Europe. These problems were tested in order to answer the following questions: 1. Is it possible to measure the social system level before the application of the system?, 2. Is it possible to evaluate the necessary level of the social system for successful implementation in advance?, 3. Is it possible to cultivate the social system to the desired level? We try to answer all of these questions adopting the Kopčaj Spiral Management approach. The practical results on an existing company are presented together with managerial recommendations.

  10. A rapid and simple method for cryopreservation of human liver slices

    NARCIS (Netherlands)

    de Kanter, R; Olinga, Peter; Hof, I.H; de Jager, M.H; Verwillegen, W.A; Slooff, M.JH; Meijer, D.K F; Groothuis, Geny; Koster, H

    1. Precision-cut liver slices represent a suitable and convenient in vitro preparation for studying metabolism and toxicity mechanisms of drugs and toxic chemicals. Particularly in the case of human liver slices, cryopreservation would enable more efficient utilization of this scarce and irregularly

  11. Evaluation of human platelet lysate and dimethyl sulfoxide as cryoprotectants for the cryopreservation of human adipose-derived stem cells.

    Science.gov (United States)

    Wang, Chuan; Xiao, Ran; Cao, Yi-Lin; Yin, Hong-Yu

    2017-09-09

    Cryopreservation provides an effective technique to maintain the functional properties of human adipose-derived stem cells (ASCs). Dimethylsulfoxide (DMSO) and fetal bovine serum (FBS) are frequently used as cryoprotectants for this purpose. However, the use of DMSO can result in adverse effects and toxic reactions and FBS can introduce risks of viral, prion, zoonose contaminations and evoke immune responses after injection. It is therefore crucial to reduce DMSO concentrations and use serum-free solution in the cryopreservation process. Human platelet lysate (PL) is a promising candidate for use as an alternative to DMSO and FBS. Therefore, in this study, with an aim to identify a cryoprotective agent for ASC cryopreservation, we determined the viability, proliferation potential, phenotype, and differentiation potential of fresh ASCs and ASCs cryopreserved using different combinations of three cryoprotective agents: fetal bovine serum (FBS), dimethylsulfoxide (DMSO), and human platelet lysate (PL). The viability of the ASCs cryopreserved with 90% FBS and 10% DMSO, 95% FBS and 5% DMSO, and 97% PL and 3% DMSO was >80%, and the proliferation potentials, cell phenotypes, and differentiation potentials of these groups were similar to those of fresh ASCs. Together, our findings suggest that a combination of 97% PL and 3% DMSO is an ideal cryoprotective agent for the efficient cryopreservation of human ASCs. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Cryopreservation of canine ovarian and testicular fibroblasts.

    Science.gov (United States)

    Yu, Il-Jeoung; Leibo, S P; Songsasen, Nucharin; Dresser, Betsy L; Kim, In-Shik

    2009-01-01

    To derive a practical procedure to store canine somatic cells, fibroblasts isolated from testicular or ovarian tissues were cryopreserved in 1.2 M ethylene glycol or in 1.2 M dimethylsulfoxide prepared in Dulbecco's Modified Eagle Medium as cryoprotectants, and were frozen either in plastic straws or vials. Thawed cells were cultured for 24 hr at 38.5 degree C in a humidified atmosphere of 5 percent CO2 95 percent air, and then their membrane integrity was assayed with a double fluorescent stain, Fertilight. In addition, frozen-thawed fibroblasts were cultured for 4 days, and then their functional survival was measured after staining small colonies with trypan blue. After freezing and thawing, membrane integrity of testicular fibroblasts was 55-70 percent and functional survival ranged from 20-40 percent. With frozen-thawed ovarian cells, the average membrane integrity was 55-75 percent and the average functional survival was 35-40 percent. When frozen in ethylene glycol, functional survival of ovarian fibroblasts was significantly higher than that of testicular cells (P less than 0.05). These methods should prove useful to preserve cells collected from canids in the wild.

  13. Comparing sugar type supplementation for cryopreservation of boar semen in egg yolk based extender.

    Science.gov (United States)

    Malo, C; Gil, L; Gonzalez, N; Cano, R; de Blas, I; Espinosa, E

    2010-08-01

    Cryopreservation of boar semen is still considered suboptimal due to lower fertility when compared to fresh semen. The aim of this study was to evaluate the effects of the addition of different sugars (lactose, trehalose and glucose) on boar spermatozoa cryopreserved in an egg yolk based extender. Ejaculates were collected from a boar previously selected and semen samples were processed using the straw freezing procedure. In experiment 1, subsamples of semen were frozen in three different extenders: recommended lactose egg yolk extender (LEY); trehalose egg yolk extender (TEY) and glucose egg yolk extender (GEY). Sperm quality was assessed for motility, viability, acrosome integrity and hypoosmotic swelling test response upon collection, after freezing and thawing and then every hour for 3h. Results showed that total motility at 1 and 3h, progressive motility at 3h, positive hypoosmotic response at 2 and 3h and acrosome integrity at all times were significantly improved when trehalose was added to the extender. In experiment 2, sugar influence was also demonstrated in vitro fertilization. A total of 1691 oocytes were in vitro matured and inseminated with frozen-thawed sperm at 2000:1 sperm:oocyte ratio and coincubated for 6h. Presumptive zygotes were cultured in NCSU-23 medium to assess fertilization parameters and embryo development. Both penetration and monospermy rates were significantly higher for trehalose frozen semen. A significant increase was observed in efficiency and blastocyst formation rates from TEY to the other groups. Our results demonstrated that trehalose extender enhances spermatozoa viability and its in vitro fertilization parameters in boar ejaculates with good sperm freezability. Further studies are necessary to assess the impact of sugars on the entire population. (c) 2010 Elsevier Inc. All rights reserved.

  14. Selected Private Higher Educational Institutions in Metro Manila: A DEA Efficiency Measurement

    Science.gov (United States)

    de Guzman, Maria Corazon Gwendolyn N.; Cabana, Emilyn

    2009-01-01

    This paper measures the technical efficiency of 16 selected colleges and universities in Metro Manila, Philippines, using academic data for the SY 2001-2005. Using the data envelopment analysis (DEA), on average, schools posted 0.807 index score and need additional 19.3% efficiency growth to be efficient. Overall, there are top four efficient…

  15. Effects of combined cryopreservation and decellularization on the biomechanical, structural and biochemical properties of porcine pulmonary heart valves.

    Science.gov (United States)

    Theodoridis, Karolina; Müller, Janina; Ramm, Robert; Findeisen, Katja; Andrée, Birgit; Korossis, Sotirios; Haverich, Axel; Hilfiker, Andres

    2016-10-01

    Non-fixed, decellularized allogeneic heart valve scaffolds seem to be the best choice for heart valve replacement, their availability, however, is quite limited. Cryopreservation could prolong their shelf-life, allowing for their ideal match to a recipient. In this study, porcine pulmonary valves were decellularized using detergents, either prior or after cryopreservation, and analyzed. Mechanical integrity was analyzed by uniaxial tensile testing, histoarchitecture by histological staining, and composition by DNA, collagen (hydroxyproline) and GAG (chondroitin sulfate) quantification. Residual sodium dodecyl sulfate (SDS) in the scaffold was quantified by applying a methylene blue activation assay (MBAS). Cryopreserved decellularized scaffolds (DC) and scaffolds that were decellularized after cryopreservation (CD) were compared to fresh valves (F), cryopreserved native valves (C), and decellularized only scaffolds (D). The E-modulus and tensile strength of decellularized (D) tissue showed no significant difference compared to DC and CD. The decellularization resulted in an overall reduction of DNA and GAG, with DC containing the lowest amount of GAGs. The DNA content in the valvular wall of the CD group was higher than in the D and DC groups. CD valves showed slightly more residual SDS than DC valves, which might be harmful to recipient cells. In conclusion, cryopreservation after decellularization was shown to be preferable over cryopreservation before decellularization. However, in vivo testing would be necessary to determine whether these differences are significant in biocompatibility or immunogenicity of the scaffolds. Absence of adverse effects on biomechanical stability of acellular heart valve grafts by cryopreservation, neither before nor after decellularization, allows the identification of best matching patients in a less time pressure dictated process, and therefore to an optimized use of a very limited, but best-suited heart valve prosthesis

  16. Cryopreservation of Boer goat spermatozoa: Comparison of two freezing extenders based on post-thaw sperm quality and fertility rates

    Directory of Open Access Journals (Sweden)

    Fitra Aji Pamungkas

    2014-06-01

    Full Text Available Boer goat have recently been popularly used for cross breeding with local goats. However, it is currently considered a breed at very limited number with relatively high prices . In this context, the cryopreservation of spermatozoa is important because it could be conserved for a very long period of time. Egg yolk extenders are most commonly used for cryopreservation of goat sperm. The aim of this study was to compare the ability of two extenders to maintain sperm viability after cryopreservation. Semen from three male Boer goat aged about 2-3 years old was collected using artificial vagina and frozen with Tris and Triladyl extender. The results showed that percentage of motility, viability and membrane integrity of spermatozoa with Tris and Triladyl extenders at every stage of cryopreservation showed not significantly difference (P>0.05, except the percentage of sperm motility post thawing of Triladyl was higher than Tris extender (52.00±4.47% vs 47.50±2.74%, P<0.05. Cryopreserved semen in Tris extender provided the same fertility rates after cervical insemination compared to Triladyl (62.50% vs 60.00%. In conclusion, the Tris extender has the same capabilities to Triladyl in cryopreservation of Boer goat spermatozoa as to maintain sperm quality and fertility rates.

  17. Effects of micro-encapsulation on morphology and endocrine function of cryopreserved neonatal porcine islet-like cell clusters.

    Science.gov (United States)

    Murakami, M; Satou, H; Kimura, T; Kobayashi, T; Yamaguchi, A; Nakagawara, G; Iwata, H

    2000-10-27

    For the success of clinical islets transplantation, the development of a long-term storage method is necessary. However, the structure of digested islets is scanty for culture and cryopreservation. In this study, the effect of micro-encapsulation to cryopreserved porcine islet-like cell clusters (ICCs) was investigated. The ICCs prepared from neonatal pigs by collagenase digestion and culture technique were cryopreserved and micro-encapsulated in 5% agarose membranes. After cryopreservation, ICC cultured without encapsulation (group A) and cultured with encapsulation (group B) were assessed by comparison with no cryopreserved ICC (control) both in vitro by static incubation test and in vivo in a xenotransplantation study. Micro-encapsulation was able to maintain the fine morphology and the number of ICCs of group B after 7 days of culture. There were not significant differences in insulin secretion of group B and control on day 1 and 7 of culture (1 day:11+/-0.99, 7 days: 5.30+/-1.08 microU/ICC/hr NS versus control). On day 7 of culture, the retrieval rate of group B (105.2+/-9.8%) is obviously higher compared with group A (63.0+/-6.3%). In the xenotransplatation model, the ICCs of group B showed long survival time (7.9+/-0.4 weeks) and good transplantation effect. Our study suggests that micro-encapsulation is one of the useful method for cryopreserved ICC to maintain the fine morphology and effectively recover the endocrine function.

  18. Cryopreservation of mouse embryos by ethylene glycol-based vitrification.

    Science.gov (United States)

    Mochida, Keiji; Hasegawa, Ayumi; Taguma, Kyuichi; Yoshiki, Atsushi; Ogura, Atsuo

    2011-11-18

    Cryopreservation of mouse embryos is a technological basis that supports biomedical sciences, because many strains of mice have been produced by genetic modifications and the number is consistently increasing year by year. Its technical development started with slow freezing methods in the 1970s(1), then followed by vitrification methods developed in the late 1980s(2). Generally, the latter technique is advantageous in its quickness, simplicity, and high survivability of recovered embryos. However, the cryoprotectants contained are highly toxic and may affect subsequent embryo development. Therefore, the technique was not applicable to certain strains of mice, even when the solutions are cooled to 4°C to mitigate the toxic effect during embryo handling. At the RIKEN BioResource Center, more than 5000 mouse strains with different genetic backgrounds and phenotypes are maintained(3), and therefore we have optimized a vitrification technique with which we can cryopreserve embryos from many different strains of mice, with the benefits of high embryo survival after vitrifying and thawing (or liquefying, more precisely) at the ambient temperature(4). Here, we present a vitrification method for mouse embryos that has been successfully used at our center. The cryopreservation solution contains ethylene glycol instead of DMSO to minimize the toxicity to embryos(5). It also contains Ficoll and sucrose for prevention of devitrification and osmotic adjustment, respectively. Embryos can be handled at room temperature and transferred into liquid nitrogen within 5 min. Because the original method was optimized for plastic straws as containers, we have slightly modified the protocol for cryotubes, which are more easily accessible in laboratories and more resistant to physical damages. We also describe the procedure of thawing vitrified embryos in detail because it is a critical step for efficient recovery of live mice. These methodologies would be helpful to researchers and

  19. Cryopreservation and revival of mesenchymal stromal cells

    DEFF Research Database (Denmark)

    Haack-Sørensen, Mandana; Kastrup, Jens

    2011-01-01

    initiated. As there has been a precedent for the use of bone marrow stem cells in the treatment of hematological malignancies and ischemic heart diseases through randomized clinical safety and efficacy trials, the development of new therapies based on culture-expanded human mesenchymal stromal cells (MSCs......Over the past few years, the pace of preclinical stem cell research is astonishing and adult stem cells have become the subject of intense research. Due to the presence of promising supporting preclinical data, human clinical trials for stem cell regenerative treatment of various diseases have been......) opens up new possibilities for cell therapy. To facilitate these applications, cryopreservation and long-term storage of MSCs becomes an absolute necessity. As a result, optimization of this cryopreservation protocol is absolutely critical. The major challenge during cellular cryopreservation...

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

    Science.gov (United States)

    Komatsu, Hirotake; Barriga, Alyssa; Medrano, Leonard; Omori, Keiko; Kandeel, Fouad; Mullen, Yoko

    2017-05-06

    Pancreatic islet transplantation is an effective treatment for Type 1 diabetic patients to eliminate insulin injections; however, a shortage of donor organs hinders the widespread use. Although long-term islet storage, such as cryopreservation, is considered one of the key solutions, transplantation of cryopreserved islets is still not practical due to the extensive loss during the cryopreservation-rewarming process. We have previously reported that culturing islets in a hyperoxic environment is an effective treatment to prevent islet death from the hypoxic injury during culture. In this study, we explored the effectiveness of thawing and rewarming cryopreserved islets in a hyperoxic environment. Following cryopreservation of isolated human islets, the thawing solution and culture media were prepared with or without pre-equilibration to 50% oxygen. Thawing/rewarming and the pursuant two-day culture were performed with or without oxygenation. Short-term recovery rate, defined as the volume change during cryopreservation and thawing/rewarming, was assessed. Ischemia-associated and inflammation-associated gene expressions were examined using qPCR after the initial rewarming period. Long-term recovery rate, defined as the volume change during the two-day culture after the thawing/rewarming, was also examined. Islet metabolism and function were assessed by basal oxygen consumption rate and glucose stimulated insulin secretion after long-term recovery. Oxygenated thawing/rewarming did not alter the short-term recovery rate. Inflammation-associated gene expressions were elevated by the conventional thawing/rewarming method and suppressed by the oxygenated thawing/rewarming, whereas ischemia-associated gene expressions did not change between the thawing/rewarming methods. Long-term recovery rate experiments revealed that only the combination therapy of oxygenated thawing/rewarming and oxygenated culture alleviated islet volume loss. These islets showed higher metabolism

  1. Karyotype of cryopreserved bone marrow cells

    Directory of Open Access Journals (Sweden)

    M.L.L.F. Chauffaille

    2003-07-01

    Full Text Available The analysis of chromosomal abnormalities is important for the study of hematological neoplastic disorders since it facilitates classification of the disease. The ability to perform chromosome analysis of cryopreserved malignant marrow or peripheral blast cells is important for retrospective studies. In the present study, we compared the karyotype of fresh bone marrow cells (20 metaphases to that of cells stored with a simplified cryopreservation method, evaluated the effect of the use of granulocyte-macrophage colony-stimulating factor (GM-CSF as an in vitro mitotic index stimulator, and compared the cell viability and chromosome morphology of fresh and cryopreserved cells whenever possible (sufficient metaphases for analysis. Twenty-five bone marrow samples from 24 patients with hematological disorders such as acute myeloid leukemia, acute lymphoblastic leukemia, myelodysplastic syndrome, chronic myeloid leukemia, megaloblastic anemia and lymphoma (8, 3, 3, 8, 1, and 1 patients, respectively were selected at diagnosis, at relapse or during routine follow-up and one sample was obtained from a bone marrow donor after informed consent. Average cell viability before and after freezing was 98.8 and 78.5%, respectively (P < 0.05. Cytogenetic analysis was successful in 76% of fresh cell cultures, as opposed to 52% of cryopreserved samples (P < 0.05. GM-CSF had no proliferative effect before or after freezing. The morphological aspects of the chromosomes in fresh and cryopreserved cells were subjectively the same. The present study shows that cytogenetic analysis of cryopreserved bone marrow cells can be a reliable alternative when fresh cell analysis cannot be done, notwithstanding the reduced viability and lower percent of successful analysis that are associated with freezing.

  2. Karyotype of cryopreserved bone marrow cells.

    Science.gov (United States)

    Chauffaille, M L L F; Pinheiro, R F; Stefano, J T; Kerbauy, J

    2003-07-01

    The analysis of chromosomal abnormalities is important for the study of hematological neoplastic disorders since it facilitates classification of the disease. The ability to perform chromosome analysis of cryopreserved malignant marrow or peripheral blast cells is important for retrospective studies. In the present study, we compared the karyotype of fresh bone marrow cells (20 metaphases) to that of cells stored with a simplified cryopreservation method, evaluated the effect of the use of granulocyte-macrophage colony-stimulating factor (GM-CSF) as an in vitro mitotic index stimulator, and compared the cell viability and chromosome morphology of fresh and cryopreserved cells whenever possible (sufficient metaphases for analysis). Twenty-five bone marrow samples from 24 patients with hematological disorders such as acute myeloid leukemia, acute lymphoblastic leukemia, myelodysplastic syndrome, chronic myeloid leukemia, megaloblastic anemia and lymphoma (8, 3, 3, 8, 1, and 1 patients, respectively) were selected at diagnosis, at relapse or during routine follow-up and one sample was obtained from a bone marrow donor after informed consent. Average cell viability before and after freezing was 98.8 and 78.5%, respectively (P < 0.05). Cytogenetic analysis was successful in 76% of fresh cell cultures, as opposed to 52% of cryopreserved samples (P < 0.05). GM-CSF had no proliferative effect before or after freezing. The morphological aspects of the chromosomes in fresh and cryopreserved cells were subjectively the same. The present study shows that cytogenetic analysis of cryopreserved bone marrow cells can be a reliable alternative when fresh cell analysis cannot be done, notwithstanding the reduced viability and lower percent of successful analysis that are associated with freezing.

  3. Cryopreservation of spermatozoa of black marlin, Makaira indica (Teleostei: Istiophoridae).

    Science.gov (United States)

    van der Straten, K M; Leung, L K-P; Rossini, R; Johnston, S D

    2006-01-01

    As a first step towards the development of a method for the cryopreservation of black marlin spermatozoa, this study investigated the effect of dimethylsulfoxide (DMSO) concentration and pellet size on post-thaw spermatozoal motility. Spermatozoa were recovered from the spermatic duct of testes retrieved post-mortem from four adult black marlin caught in the Coral Sea spawning grounds of Australia. Undiluted spermatozoa were stored on ice for 4 to 10 hours during transport to shore, then evaluated for motility after activation in seawater (1:10 v:v). Spermatozoa were prepared for cryopreservation in pellets by extension (1:3 v:v) in a defined fish Ringer's solution to give two final DMSO concentrations of 2.5% or 5.0%. Diluted spermatozoa were frozen directly on a dry ice block in pellet sizes of either 0.25 ml or 0.50 ml. Frozen pellets were thawed in a water bath at 40 degrees C for 60 seconds and assessed for post-thaw motility following activation in seawater. Spermatozoa recovered within 50 minutes of death and chilled on ice for 4 to 10 hours showed a mean (+/- SEM) motility immediately following activation of 91.6 +/- 7.9%. 50% of the spermatozoa remained motile for approximately 4 to 5 minutes. Following cryopreservation, mean motility declined significantly across all cryoprotectant and pellet size combinations (P < 0.001) but spermatozoa frozen in 2.5% DMSO showed higher motility than those frozen in 5.0% DMSO (P = 0.014). Pellet size had no effect on post-thaw motility (P = 0.179).

  4. Impact of procedural steps and cryopreservation agents in the cryopreservation of chlorophyte microalgae.

    Directory of Open Access Journals (Sweden)

    Tony V L Bui

    Full Text Available The maintenance of traditional microalgae collections based on liquid and solid media is labour intensive, costly and subject to contamination and genetic drift. Cryopreservation is therefore the method of choice for the maintenance of microalgae culture collections, but success is limited for many species. Although the mechanisms underlying cryopreservation are understood in general, many technical variations are present in the literature and the impact of these are not always elaborated. This study describes two-step cryopreservation processes in which 3 microalgae strains representing different cell sizes were subjected to various experimental approaches to cryopreservation, the aim being to investigate mechanistic factors affecting cell viability. Sucrose and dimethyl sulfoxide (DMSO were used as cryoprotectants. They were found to have a synergistic effect in the recovery of cryopreserved samples of many algal strains, with 6.5% being the optimum DMSO concentration. The effect of sucrose was shown to be due to improved cell survival and recovery after thawing by comparing the effect of sucrose on cell viability before or after cryopreservation. Additional factors with a beneficial effect on recovery were the elimination of centrifugation steps (minimizing cell damage, the reduction of cell concentration (which is proposed to reduce the generation of toxic cell wall components and the use of low light levels during the recovery phase (proposed to reduce photooxidative damage. The use of the best conditions for each of these variables yielded an improved protocol which allowed the recovery and subsequent improved culture viability of a further 16 randomly chosen microalgae strains. These isolates included species from Chlorellaceae, Palmellaceae, Tetrasporaceae, Palmellopsis, Scenedesmaceae and Chlamydomonadaceae that differed greatly in cell diameter (3-50 µm, a variable that can affect cryopreservation success. The collective improvement

  5. Directional freezing for the cryopreservation of adherent mammalian cells on a substrate

    Science.gov (United States)

    Braslavsky, Ido

    2018-01-01

    Successfully cryopreserving cells adhered to a substrate would facilitate the growth of a vital confluent cell culture after thawing while dramatically shortening the post-thaw culturing time. Herein we propose a controlled slow cooling method combining initial directional freezing followed by gradual cooling down to -80°C for robust preservation of cell monolayers adherent to a substrate. Using computer controlled cryostages we examined the effect of cooling rates and dimethylsulfoxide (DMSO) concentration on cell survival and established an optimal cryopreservation protocol. Experimental results show the highest post-thawing viability for directional ice growth at a speed of 30 μm/sec (equivalent to freezing rate of 3.8°C/min), followed by gradual cooling of the sample with decreasing rate of 0.5°C/min. Efficient cryopreservation of three widely used epithelial cell lines: IEC-18, HeLa, and Caco-2, provides proof-of-concept support for this new freezing protocol applied to adherent cells. This method is highly reproducible, significantly increases the post-thaw cell viability and can be readily applied for cryopreservation of cellular cultures in microfluidic devices. PMID:29447224

  6. High-throughput optimization by statistical designs: example with rat liver slices cryopreservation.

    Science.gov (United States)

    Martin, H; Bournique, B; Blanchi, B; Lerche-Langrand, C

    2003-08-01

    The purpose of this study was to optimize cryopreservation conditions of rat liver slices in a high-throughput format, with focus on reproducibility. A statistical design of 32 experiments was performed and intracellular lactate dehydrogenase (LDHi) activity and antipyrine (AP) metabolism were evaluated as biomarkers. At freezing, modified University of Wisconsin solution was better than Williams'E medium, and pure dimethyl sulfoxide was better than a cryoprotectant mixture. The best cryoprotectant concentrations were 10% for LDHi and 20% for AP metabolism. Fetal calf serum could be used at 50 or 80%, and incubation of slices with the cryoprotectant could last 10 or 20 min. At thawing, 42 degrees C was better than 22 degrees C. After thawing, 1h was better than 3h of preculture. Cryopreservation increased the interslice variability of the biomarkers. After cryopreservation, LDHi and AP metabolism levels were up to 84 and 80% of fresh values. However, these high levels were not reproducibly achieved. Two factors involved in the day-to-day variability of LDHi were identified: the incubation time with the cryoprotectant and the preculture time. In conclusion, the statistical design was very efficient to quickly determine optimized conditions by simultaneously measuring the role of numerous factors. The cryopreservation procedure developed appears suitable for qualitative metabolic profiling studies.

  7. Embryo transfer using cryopreserved Boer goat blastocysts ...

    African Journals Online (AJOL)

    The aim of this trial was to evaluate the effect of embryo cryopreservation techniques on the survivability of embryos and fertility following transfer to Boer goat does. The oestrous cycles of 27 mature recipients Boer goat does were synchronised using controlled internal drug release dispensers (CIDR's) for 16 days. At CIDR ...

  8. Changing rooster sperm membranes to facilitate cryopreservation

    Science.gov (United States)

    Cryopreservation damages rooster sperm membranes. Part of this damage is due to membrane transitioning from the fluid to the gel state as temperature is reduced. This damage may be prevented by increasing membrane fluidity at low temperatures by incorporating cholesterol or unsaturated lipids into t...

  9. Preliminary investigation of cryopreservation by encapsulation ...

    African Journals Online (AJOL)

    Protocorm-like bodies (PLBs) of Brassidium Shooting Star, a new commercial ornamental orchid hybrid, were cryopreserved by an encapsulation-dehydration technique. The effects of PLB size, various sucrose concentrations in preculture media and sodium alginate concentration for encapsulation were the main ...

  10. Cryopreservation and Cryotherapy of Citrus Cultivars

    Science.gov (United States)

    Long-term conservation of Citrus clones can be accomplished by cryopreservation. Shoot tips will survive liquid nitrogen exposure and storage when appropriately desiccated and treated with cryoprotectant solutions. In our research, vegetative Citrus budwood is shipped from Riverside to Fort Collin...

  11. Cryopreservation of medicinal plants: role of melatonin

    Science.gov (United States)

    Many useful plant species found in Canada are of conservation concern. In vitro storage and cryopreservation techniques guarantees safety of these species and have potential applications which may result in sustainable agriculture. Shoot tips of in vitro-grown plantlets of American elm, St John’s Wo...

  12. Cryopreservation of South African indigenous goat semen

    African Journals Online (AJOL)

    use

    2011-12-05

    Dec 5, 2011 ... sperm cell motility rate of 83.1%, progressive sperm cell motility of 49.3% ... cells can be stored and used after a long period of time. ... artificial insemination to enhance improvement of .... Effect of cryopreservation on semen quality (mean ± S.E) of South African .... Successful pregnancies with directional.

  13. Generation of juvenile rainbow trout derived from cryopreserved whole ovaries by intraperitoneal transplantation of ovarian germ cells.

    Science.gov (United States)

    Lee, Seungki; Katayama, Naoto; Yoshizaki, Goro

    2016-09-23

    Cryopreservation of fish sperm offers the practical applications in the selective breeding and biodiversity conservation. However, because of the lack of cryopreservation methods for fish eggs and embryos, maternally inherited cytoplasmic compartments cannot be successfully preserved. We previously developed an alternative method to derive functional eggs and sperm from cryopreserved whole testis by transplanting testicular cells into female and male recipients. However, if target fish had ovaries, the previous method employing male-derived germ cells would be ineffective. Here, we aimed to generate functional gametes from cryopreserved whole ovaries by transplanting ovarian germ cells into peritoneal cavity of sterile hatchlings. Cryopreservation conditions for rainbow trout ovaries (1.0 M DMSO, 0.1 M trehalose, and 10% egg yolk) were optimized by testing several different cryoprotective agents. Ovarian germ cells from thawed ovaries were intraperitoneally transplanted into allogeneic triploid hatchlings. Transplanted germ cells migrated toward and were incorporated into recipient gonads, where they underwent gametogenesis. Transplantation efficiency of ovarian germ cells remained stable after cryopreservation period up to 1185 days. Although all triploid recipients that did not undergo transplantation were functionally sterile, 5 of 25 female recipients and 7 of 25 male recipients reached sexual maturity at 2.5 years post-transplantation. Inseminating the resultant eggs and sperm generated viable offspring displaying the donor characteristics of orange body color, green fluorescence, and chromosome numbers. This method is thus a breakthrough tool for the conservation of endangered fish species that are crucial to cryopreserve the genetic resources of female fish. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Impact of higher energy efficiency standards on housing affordability in Alberta

    International Nuclear Information System (INIS)

    2010-07-01

    As a result of changes to provincial and national building and energy costs, the impact of increasing energy efficiency standards on housing affordability has been questioned. Determining housing affordability is a complicated process. This report presented the results of a costing analysis completed for upgrades of EnerGuide 80 levels of energy efficiency in homes in Calgary and Edmonton, Alberta. The elements of residential construction were identified. In order to better understand the cost impact of energy efficiency upgrades on a home, pricing data was obtained. Costing elements that were examined included housing price indexes; construction material price indexes; unionized trade wages; and land value. Specifically, the report presented the new housing price index analysis using material and labour costs. An analysis of energy efficiency improvement was then presented in terms of lifecycle costs (capital costs and life cycle costing results). It was concluded that although the price of labour and materials is increasing, the value of land is the primary driver for rising house prices. The price of housing is strongly correlated to the price of land and not the price of labour or materials. In addition, moving to EnerGuide 80 levels of energy efficiency for housing in Alberta made homes more affordable for homebuyers by lowering their total monthly housing costs. 4 tabs., 3 figs., 3 appendices.

  15. Cryopreservation of peach palm zygotic embryos.

    Science.gov (United States)

    Steinmacher, Douglas A; Saldanha, Cleber W; Clement, Charles R; Guerra, Miguel P

    2007-01-01

    Cryopreservation is a safe and cost-effective option for long-term germplasm conservation of non-orthodox seed species, such as peach palm (Bactris gasipaes). The objective of the present study was to establish a cryopreservation protocol for peach palm zygotic embryos based on the encapsulation-dehydration technique. After excision, zygotic embryos were encapsulated with 3 percent sodium alginate plus 2 M glycerol and 0.4 M sucrose, and pre-treated or not with 1 M sucrose during 24 h, followed by air-drying. Fresh weight water contents of beads decreased from 83 percent and 87 percent to 18 percent and 20 percent for pre-treated or non-pretreated beads, respectively, after 4 h of dehydration. Sucrose pre-treatment at 1 M caused lower zygotic embryo germination and plantlet height in contrast to non-treated beads. All the variables were statistically influenced by dehydration time. Optimal conditions for recovery of cryopreserved zygotic embryos include encapsulation and dehydration for 4 h in a forced air cabinet to 20 percent water content, followed by rapid freezing in liquid nitrogen (-196 degree C) and rapid thawing at 45 degree C. In these conditions 29 percent of the zygotic embryos germinated in vitro. However, plantlets obtained from dehydrated zygotic embryos had stunted haustoria and lower heights. Histological analysis showed that haustorium cells were large, vacuolated, with few protein bodies. In contrast, small cells with high nucleus:cytoplasm ratio formed the shoot apical meristem of the embryos, which were the cell types with favorable characteristics for survival after exposure to liquid nitrogen. Plantlets were successfully acclimatized and showed 41+/-9 percent and 88+/-4 percent survival levels after 12 weeks of acclimatization from cryopreserved and non-cryopreserved treatments, respectively.

  16. Cryopreservation of Mycobacterium bovis isolates

    Directory of Open Access Journals (Sweden)

    Cássia Yumi Ikuta

    2016-11-01

    Full Text Available Research, development of new biotechnological methods, diagnostic tests, confirmation of results, and reinvestigations are possible because of the availability of well-preserved living organisms maintained without any changes. Cryopreservation is a simpler, more reliable and long-term stable method for culture maintenance. Storage temperature and composition of the suspending vehicle are factors that affect the viability of mycobacterial strains. Three vehicles and three storage temperatures were evaluated to define a suitable cryoprotective medium for the preservation of Mycobacterium bovis strains. Colonies of sixteen M. bovis isolates were used to prepare the suspensions, which were then added to three vehicles: sterile 0.85% saline solution (SS, Middlebrook 7H9 broth (7H9, and Middlebrook 7H9 broth with sodium pyruvate (7H9p replacing glycerol. Aliquots of these suspensions were frozen by three different methods, directly in the -20°C freezer, directly in the -80°C freezer, and at -196°C by immersion in liquid nitrogen (LN. The frozen aliquots were thawed at room temperature after 45, 90 and 120 days. Mycobacterial viability was assessed by counting the living cells on plates of Stonebrink medium before and after the freezing procedure. Storage at -20°C exhibited a lower recovery of M. bovis compared to storage at -80°C (Dunn’s test, p=0.0018 and LN (Dunn’s test, p=0.0352. There was no statistically significant difference between storage at -80°C and in LN (Dunn’s test, p=0.1403, yet -80°C showed better results than LN. All three suspending vehicles showed no statistically significant difference in terms of viability (Friedman’s test, p=0.7765. Given the low loss proportion of 5% during storage at -20°C and the high cost equipment required for storage at -80°C and LN, we recommend storage at -20°C or -80°C, when this is available, for preservation of M. bovis field strains.

  17. Climbing Mount Efficiency--small steps, not giant leaps towards higher cloning success in farm animals.

    Science.gov (United States)

    Oback, Björn

    2008-07-01

    Despite more than a decade of research efforts, farm animal cloning by somatic cell nuclear transfer (SCNT) is still frustratingly inefficient. Inefficiency manifests itself at different levels, which are currently not well integrated. At the molecular level, it leads to widespread genetic, epigenetic and transcriptional aberrations in cloned embryos. At the organismal level, these genome-wide abnormalities compromise development of cloned foetuses and offspring. Specific molecular defects need to be causally linked to specific cloned phenotypes, in order to design specific treatments to correct them. Cloning efficiency depends on the ability of the nuclear donor cell to be fully reprogrammed into an embryonic state and the ability of the enucleated recipient cell to carry out the reprogramming reactions. It has been postulated that reprogrammability of the somatic donor cell epigenome is influenced by its differentiation status. However, direct comparisons between cells of divergent differentiation status within several somatic lineages have found no conclusive evidence for this. Choosing somatic stem cells as donors has not improved cloning efficiency, indicating that donor cell type may be less critical for cloning success. Different recipient cells, on the other hand, vary in their reprogramming ability. In bovine, using zygotes instead of oocytes has increased cloning success. Other improvements in livestock cloning efficiency include better coordinating donor cell type with cell cycle stage and aggregating cloned embryos. In the future, it will be important to demonstrate if these small increases at every step are cumulative, adding up to an integrated cloning protocol with greatly improved efficiency.

  18. Alternative geometry for cylindrical natural draft cooling tower with higher cooling efficiency under crosswind condition

    International Nuclear Information System (INIS)

    Goodarzi, M.; Ramezanpour, R.

    2014-01-01

    Highlights: • Alternative cross sections for natural draft cooling tower were proposed. • Numerical solution was applied to study thermal and hydraulic performances. • Thermal and hydraulic performances were assessed by comparative parameters. • Cooling tower with elliptical cross section had better thermal performance under crosswind. • It could successfully used at the regions with invariant wind direction. - Abstract: Cooling efficiency of a natural draft dry cooling tower may significantly decrease under crosswind condition. Therefore, many researchers attempted to improve the cooling efficiency under this condition by using structural or mechanical facilities. In this article, alternative shell geometry with elliptical cross section is proposed for this type of cooling tower instead of usual shell geometry with circular cross section. Thermal performance and cooling efficiency of the two types of cooling towers are numerically investigated. Numerical simulations show that cooling tower with elliptical cross section improves the cooling efficiency compared to the usual type with circular cross section under high-speed wind moving normal to the longitudinal diameter of the elliptical cooling tower

  19. Automation of analytical processes. A tool for higher efficiency and safety

    International Nuclear Information System (INIS)

    Groll, P.

    1976-01-01

    The analytical laboratory of a radiochemical facility is usually faced with the fact that numerous analyses of a similar type must be routinely carried out. Automation of such routine analytical procedures helps in increasing the efficiency and safety of the work. A review of the requirements for automation and its advantages is given and demonstrated on three examples. (author)

  20. Evaluation of amides and centrifugation temperature in boar semen cryopreservation.

    Science.gov (United States)

    Bianchi, I; Calderam, K; Maschio, E F; Madeira, E M; da Rosa Ulguim, R; Corcini, C D; Bongalhardo, D C; Corrêa, E K; Lucia, T; Deschamps, J C; Corrêa, M N

    2008-03-15

    Two experiments were conducted to evaluate the use of amides as cryoprotectants and two centrifugation temperatures (15 or 24 degrees C) in boar semen cryopreservation protocols. Semen was diluted in BTS, cooled centrifuged, added to cooling extenders, followed by the addition of various cryoprotectants. In experiment 1, mean (+/-S.E.M.) sperm motility for 5% dimethylformamide (DMF; 50.6+/-1.9%) and 5% dimethylacetamide (DMA; 53.8+/-1.7%) were superior (P0.05). In experiment 2, we tested MF, DMF, and DMA at 3, 5, and 7%. Sperm motility and membrane integrity were higher for 5% DMA (53.8+/-1.7 and 50.9+/-1.9%) and 5% DMF (50.6+/-1.9 and 47.9+/-2.1%), in comparison with 7% DMF and all MF concentrations (P0.05). In conclusion, boar semen was successfully cryopreserved by replacement of glycerol with amides (especially 5% DMA) and centrifugation at 15 degrees C, with benefits for post-thaw sperm motility and membrane integrity.

  1. Vitality of oligozoospermic semen samples is improved by both swim-up and density gradient centrifugation before cryopreservation.

    Science.gov (United States)

    Counsel, Madeleine; Bellinge, Rhys; Burton, Peter

    2004-05-01

    To ascertain whether washing sperm from oligozoospermic and normozoospermic samples before cryopreservation improves post-thaw vitality. Normozoospermic (n = 18) and oligozoospermic (n = 16) samples were divided into three aliquots. The first aliquot remained untreated and the second and third aliquots were subjected to the swim-up and discontinuous density gradient sperm washing techniques respectively. Vitality staining was performed, samples mixed with cryopreservation media and frozen. Spermatozoa were thawed, stained, and vitality quantified and expressed as the percentage of live spermatozoa present. Post-thaw vitality in untreated aliquots from normozoospermic samples (24.9% +/- 2.3; mean +/- SEM) was significantly higher (unpaired t-tests; P vitality was significantly higher after swim-up in normozoospermic samples (35.6% +/- 2.1; P vitality in oligozoospermic (22.4% +/- 1.0; P vitality in cryopreserved oligozoospermic samples was improved by both the swim-up and density gradient centrifugation washing techniques prior to freezing.

  2. Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes.

    Science.gov (United States)

    Bircher, Lea; Geirnaert, Annelies; Hammes, Frederik; Lacroix, Christophe; Schwab, Clarissa

    2018-04-17

    Strict anaerobic gut microbes have been suggested as 'next-generation probiotics' for treating several intestinal disorders. The development of preservation techniques is of major importance for therapeutic application. This study investigated cryopreservation (-80°C) and lyophilization survival and storage stability (4°C for 3 months) of the strict anaerobic gut microbes Bacteroides thetaiotaomicron, Faecalibacterium prausnitzii, Roseburia intestinalis, Anaerostipes caccae, Eubacterium hallii and Blautia obeum. To improve preservation survival, protectants sucrose and inulin (both 5% w/v) were added for lyophilization and were also combined with glycerol (15% v/v) for cryopreservation. Bacterial fitness, evaluated by maximum growth rate and lag phase, viability and membrane integrity were determined using a standardized growth assay and by flow cytometry as markers for preservation resistance. Lyophilization was more detrimental to viability and fitness than cryopreservation, but led to better storage stability. Adding sucrose and inulin enhanced viability and the proportion of intact cells during lyophilization of all strains. Viability of protectant-free B. thetaiotaomicron, A. caccae and F. prausnitzii was above 50% after cryopreservation and storage and increased to above 80% if protectants were present. The addition of glycerol, sucrose and inulin strongly enhanced the viability of B. obeum, E. hallii and R. intestinalis from 0.03-2% in protectant-free cultures to 11-37%. This is the first study that quantitatively compared the effect of cryopreservation and lyophilization and the addition of selected protectants on viability and fitness of six strict anaerobic gut microbes. Our results suggest that efficiency of protectants is process- and species-specific. © 2018 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  3. Arbiters of Effectiveness and Efficiency: The Frames and Strategies of Management Consulting Firms in US Higher Education Reform

    Science.gov (United States)

    McClure, Kevin R.

    2017-01-01

    A growing number of public colleges and universities in the United States have hired management consulting firms to help develop strategies aimed at increasing institutional effectiveness and efficiency. The purpose of this paper is to explore the frames and strategies of consultants in US public higher education reform efforts. Drawing upon a…

  4. Appraising the Cost Efficiency of Higher Technological and Vocational Education Institutions in Taiwan Using the Metafrontier Cost-Function Model

    Science.gov (United States)

    Lu, Yung-Hsiang; Chen, Ku-Hsieh

    2013-01-01

    This paper aims at appraising the cost efficiency and technology of institutions of higher technological and vocational education. Differing from conventional literature, it considers the potential influence of inherent discrepancies in output quality and characteristics of school systems for institutes of technology (ITs) and universities of…

  5. Micropropagation and cryopreservation of garlic (Allium sativum L.).

    Science.gov (United States)

    Keller, E R Joachim; Senula, Angelika

    2013-01-01

    Garlic (Allium sativum L.) is a very important medicinal and spice plant. It is conventionally propagated by daughter bulbs ("cloves") and bulbils from the flower head. Micropropagation is used for speeding up the vegetative propagation mainly using the advantage to produce higher numbers of healthy plants free of viruses, which have higher yield than infected material. Using primary explants from bulbs and/or bulbils (shoot tips) or unripe inflorescence bases, in vitro cultures are initiated on MS-based media containing auxins, e.g., naphthalene acetic acid, and cytokinins, e.g., 6-γ-γ-(dimethylallylaminopurine) (2iP). Rooting is accompanying leaf formation. It does not need special culture phases. The main micropropagation methods rely on growth of already formed meristems. Long-term storage of micropropagated material, cryopreservation, is well-developed to maintain germplasm. The main method is vitrification using the cryoprotectant mixture PVS3.

  6. Between Efficiency and Transformation: The Opinion of Deans on the Meaning of Quality in Higher Education

    Science.gov (United States)

    Olaskoaga-Larrauri, Jon; Barrenetxea-Ayesta, Miren; Cardona-Rodríguez, Antonio; Mijangos-Del Campo, Juan José; Barandiaran-Galdós, Marta

    2016-01-01

    The literature on quality management at higher education institutions has for some time been working on the basis of two issues: a) the diversity of ideas as to what "quality" means, which makes it harder to apply the principles of quality management in this context; and b) the idea that this diversity is in some way a response to the…

  7. Managing the higher risks of low-cost high-efficiency advanced power generation technologies

    International Nuclear Information System (INIS)

    Pearson, M.

    1997-01-01

    Independent power producers operate large coal-fired installations and gas turbine combined-cycle (GTCC) facilities. Combined cycle units are complex and their reliability and availability is greatly influenced by mechanical, instrumentation and control weaknesses. It was suggested that these weaknesses could be avoided by tighter specifications and more rigorous functional testing before acceptance by the owner. For the present, the difficulties of developing reliable, lower installed cost/kw, more efficient GTCC designs, pressure for lower NO x emissions with 'dry' combustors continue to be the most difficult challenges for all GT manufacturers

  8. Higher efficiency with cylinder-bank comprehensive exhaust manifold; Effizienzsteigerung durch Zylinderbank-uebergreifenden Kruemmer

    Energy Technology Data Exchange (ETDEWEB)

    Diez, Rainer; Kornherr, Heinz; Pirntke, Frank; Schmidt, Juergen [Friedrich Boysen GmbH und Co. KG, Altensteig (Germany)

    2010-05-15

    In close interdisciplinary cooperation with BMW Group, Boysen has developed an air-gap-insulated exhaust manifold that encompasses both banks of the 4.4 l V8 spark-ignition twin turbo engine of the BMW X5 M and BMW X6 M. The manifold merges the exhaust gas flow from the cylinders of the left-hand and right-hand cylinder banks in opposing pairs, thus optimising gas exchange. Due to improvements in response, torque and power characteristics of the engine, the cylinder-bank comprehensive exhaust manifold helps achieve high fuel efficiency. (orig.)

  9. Does centennial morphodynamic evolution lead to higher channel efficiency in San Pablo Bay, California?

    Science.gov (United States)

    van der Wegen, M.; Jaffe, B.E.; Barnard, P.L.; Jaffee, B.E.; Schoellhamer, D.H.

    2013-01-01

    Measured bathymetries on 30 year interval over the past 150 years show that San Pablo Bay experienced periods of considerable deposition followed by periods of net erosion. However, the main channel in San Pablo Bay has continuously narrowed. The underlying mechanisms and consequences of this tidal channel evolution are not well understood. The central question of this study is whether tidal channels evolve towards a geometry that leads to more efficient hydraulic conveyance and sediment throughput. We applied a hydrodynamic process-based, numerical model (Delft3D), which was run on 5 San Pablo Bay bathymetries measured between 1856 and 1983. Model results shows increasing energy dissipation levels for lower water flows leading to an approximately 15% lower efficiency in 1983 compared to 1856. During the same period the relative seaward sediment throughput through the San Pablo Bay main channel increased by 10%. A probable explanation is that San Pablo Bay is still affected by the excessive historic sediment supply. Sea level rise and Delta surface water area variations over 150 years have limited effect on the model results. With expected lower sediment concentrations in the watershed and less impact of wind waves due to erosion of the shallow flats, it is possible that energy dissipations levels will decrease again in future decades. Our study suggests that the morphodynamic adaptation time scale to excessive variations in sediment supply to estuaries may be on the order of centuries.

  10. An Efficient Higher-Order Quasilinearization Method for Solving Nonlinear BVPs

    Directory of Open Access Journals (Sweden)

    Eman S. Alaidarous

    2013-01-01

    Full Text Available In this research paper, we present higher-order quasilinearization methods for the boundary value problems as well as coupled boundary value problems. The construction of higher-order convergent methods depends on a decomposition method which is different from Adomain decomposition method (Motsa and Sibanda, 2013. The reported method is very general and can be extended to desired order of convergence for highly nonlinear differential equations and also computationally superior to proposed iterative method based on Adomain decomposition because our proposed iterative scheme avoids the calculations of Adomain polynomials and achieves the same computational order of convergence as authors have claimed in Motsa and Sibanda, 2013. In order to check the validity and computational performance, the constructed iterative schemes are also successfully applied to bifurcation problems to calculate the values of critical parameters. The numerical performance is also tested for one-dimension Bratu and Frank-Kamenetzkii equations.

  11. Which Are the Determinants of Online Students' Efficiency in Higher Education?

    Science.gov (United States)

    Castillo-Merino, David; Serradell-Lopez, Enric; González-González, Inés

    International literature shows that the positive effect on students performance from the adoption of innovations in the technology of teaching and learning do not affect all teaching methods and learning styles equally, as it depends on university strategy and policy towards Information and Communication Technologies (ICT) adoption, students abilities, technology uses in the educational process by teachers and students, or the selection of a methodology that matches with digital uses. This paper provides empirical answers to these questions with data from online students at the Open University of Catalonia (UOC). An empirical model based on structural equations has been defined to explain complex relationships between variables. Our results show that motivation is the main variable affecting online students' performance. It appears as a latent variable influenced by students' perception of efficiency, a driver for indirect positive and significant effect on students' performance from students' ability in ICT uses.

  12. Trophic assimilation efficiency markedly increases at higher trophic levels in four-level host-parasitoid food chain.

    Science.gov (United States)

    Sanders, Dirk; Moser, Andrea; Newton, Jason; van Veen, F J Frank

    2016-03-16

    Trophic assimilation efficiency (conversion of resource biomass into consumer biomass) is thought to be a limiting factor for food chain length in natural communities. In host-parasitoid systems, which account for the majority of terrestrial consumer interactions, a high trophic assimilation efficiency may be expected at higher trophic levels because of the close match of resource composition of host tissue and the consumer's resource requirements, which would allow for longer food chains. We measured efficiency of biomass transfer along an aphid-primary-secondary-tertiary parasitoid food chain and used stable isotope analysis to confirm trophic levels. We show high efficiency in biomass transfer along the food chain. From the third to the fourth trophic level, the proportion of host biomass transferred was 45%, 65% and 73%, respectively, for three secondary parasitoid species. For two parasitoid species that can act at the fourth and fifth trophic levels, we show markedly increased trophic assimilation efficiencies at the higher trophic level, which increased from 45 to 63% and 73 to 93%, respectively. In common with other food chains, δ(15)N increased with trophic level, with trophic discrimination factors (Δ(15)N) 1.34 and 1.49‰ from primary parasitoids to endoparasitic and ectoparasitic secondary parasitoids, respectively, and 0.78‰ from secondary to tertiary parasitoids. Owing to the extraordinarily high efficiency of hyperparasitoids, cryptic higher trophic levels may exist in host-parasitoid communities, which could alter our understanding of the dynamics and drivers of community structure of these important systems. © 2016 The Authors.

  13. Enhanced efficiency in the excitation of higher modes for atomic force microscopy and mechanical sensors operated in liquids

    Energy Technology Data Exchange (ETDEWEB)

    Penedo, M., E-mail: mapenedo@imm.cnm.csic.es; Hormeño, S.; Fernández-Martínez, I.; Luna, M.; Briones, F. [IMM-Instituto de Microelectrónica de Madrid (CNM-CSIC), Isaac Newton 8, PTM, E-28760 Tres Cantos, Madrid (Spain); Raman, A. [Birck Nanotechnology Center and School of Mechanical Engineering, Purdue University, West Lafayette, Indiana 47904 (United States)

    2014-10-27

    Recent developments in dynamic Atomic Force Microscopy where several eigenmodes are simultaneously excited in liquid media are proving to be an excellent tool in biological studies. Despite its relevance, the search for a reliable, efficient, and strong cantilever excitation method is still in progress. Herein, we present a theoretical modeling and experimental results of different actuation methods compatible with the operation of Atomic Force Microscopy in liquid environments: ideal acoustic, homogeneously distributed force, distributed applied torque (MAC Mode™), photothermal and magnetostrictive excitation. From the analysis of the results, it can be concluded that magnetostriction is the strongest and most efficient technique for higher eigenmode excitation when using soft cantilevers in liquid media.

  14. IMPROVEMENT OF THE SUPPORTING STRUCTURE OF PLATFORM CAR FOR HIGHER EFFICIENCY OF CONTAINER TRANSPORTATIONS

    Directory of Open Access Journals (Sweden)

    A. O. Lovska

    2017-02-01

    Full Text Available Purpose. The article is aimed to improve supporting structures of the platform car to increase the efficiency of container transportations. Methodology. In order to achieve the objective, the strength investigations of the universal platform car of the model 13-401 were conducted, strength reserves of the supporting elements were defined, and more optimal profiles of basic longitudinal beams of the frame in terms of the minimum material capacity were proposed. Decision correctness was confirmed by the strength calculation of the platform car supporting structure at basic loading operational modes and fatigue taking into account the research database of 107 cycles. It has been proposed to equip a platform car with swing fitting stops for fastening containers on the frame, which allows transportation of 20ft and 40ft containers. In order to improve container transportation efficiency along international transport corridors running through Ukraine, a platform car of articulated type has been designed on the base of the improved platform car structure. The mathematical simulation of dynamic loads of the platform car with containers (two 1CC containers at operational loading modes has been carried out, the maximum accelerations influencing the support structure have been defined, and their multiple values have been considered in computer simulation of the strength of the platform car of articulated type. Findings. The support structure of the platform car of articulated type on the basis of the standard platform car has been developed. Refined values of dynamic loads influencing supporting structure the platform car of articulated type with containers at operational loading modes have been obtained; the maximum equivalent stresses in the platform car support structure have been defined. Originality and practical value. A mathematical model of displacements for a platform car of articulated type with containers at operational loading modes of

  15. Agrobacterium-mediated transformation of the recalcitrant Vanda Kasem's Delight orchid with higher efficiency.

    Science.gov (United States)

    Gnasekaran, Pavallekoodi; Antony, Jessica Jeyanthi James; Uddain, Jasim; Subramaniam, Sreeramanan

    2014-01-01

    The presented study established Agrobacterium-mediated genetic transformation using protocorm-like bodies (PLBs) for the production of transgenic Vanda Kasem's Delight Tom Boykin (VKD) orchid. Several parameters such as PLB size, immersion period, level of wounding, Agrobacterium density, cocultivation period, and concentration of acetosyringone were tested and quantified using gusA gene expression to optimize the efficiency of Agrobacterium-mediated genetic transformation of VKD's PLBs. Based on the results, 3-4 mm PLBs wounded by scalpel and immersed for 30 minutes in Agrobacterium suspension of 0.8 unit at A 600 nm produced the highest GUS expression. Furthermore, cocultivating infected PLBs for 4 days in the dark on Vacin and Went cocultivation medium containing 200 μM acetosyringone enhanced the GUS expression. PCR analysis of the putative transformants selected in the presence of 250 mg/L cefotaxime and 30 mg/L geneticin proved the presence of wheatwin1, wheatwin2, and nptII genes.

  16. Cryopreserved semen in ecotoxicological bioassays: sensitivity and reliability of cryopreserved Sparus aurata spermatozoa.

    Science.gov (United States)

    Fabbrocini, Adele; D'Adamo, Raffaele; Del Prete, Francesco; Langellotti, Antonio Luca; Rinna, Francesca; Silvestri, Fausto; Sorrenti, Gerarda; Vitiello, Valentina; Sansone, Giovanni

    2012-10-01

    The aim of this study was to evaluate the feasibility of using cryopreserved S. aurata semen in spermiotoxicity tests. Cryopreservation is a biotechnology that can provide viable gametes and embryos on demand, rather than only in the spawning season, thus overcoming a limitation that has hindered the use of some species in ecotoxicological bioassays. Firstly, the sperm motility pattern of cryopreserved semen was evaluated after thawing by means of both visual and computer-assisted analyses. Motility parameters in the cryopreserved semen did not change significantly in the first hour after thawing, meaning that they were maintained for long enough to enable their use in spermiotoxicity tests. In the second phase of the research, bioassays were performed, using cadmium as the reference toxicant, in order to evaluate the sensitivity of cryopreserved S. aurata semen to ecotoxicological contamination. The sensitivity of the sperm motility parameters used as endpoints (motility percentages and velocities) proved to be comparable to what has been recorded for the fresh semen of other aquatic species (LOECs from 0.02 to 0.03 mg L(-1)). The test showed good reliability and was found to be rapid and easy to perform, requiring only a small volume of the sample. Moreover, cryopreserved semen is easy to store and transfer and makes it possible to perform bioassays in different sites or at different times with the same batch of semen. The proposed bioassay is therefore a promising starting point for the development of toxicity tests that are increasingly tailored to the needs of ecotoxicology and environmental quality evaluation strategies. Copyright © 2012 Elsevier Inc. All rights reserved.

  17. Cryopreservation of Leishmania Species in Manisa Province.

    Science.gov (United States)

    Çavuş, İbrahim; Ocak, Fulya; Kaya, Tuğba; Özbilgin, Ahmet

    2017-09-01

    It was aimed to assess the success of the cryopreservation process which is carried out in order to preserve the genetic material and the virulence of the Leishmania species that are an important health problem in our region. Leishmania tropica, L. infantum, L. major, and L. donovani strains in Novy-MacNeal-Nicolle (NNN) medium in MCBU were used. Promastigotes cultured in the NNN medium were transferred to RPMI 1640 medium; promastigotes in the logarithmic phase were washed three times with PBS, and 15% dimethylsulfoxide (DMSO) was added. Leishmania species were transferred to 12 separate tubes. The tubes were stored at -86°C for one night by placing them in Coolcell boxes. The tubes were transferred into a liquid nitrogen tank. One cryotube per Leishmania strain is thawed monthly and cultured in NNN medium. For the duration of study it was observed that each Leishmania isolate preserved 60-65% of their viability and entered the logarithmic phase on the 7th day following the inoculation in the NNN medium. Abnormalities in the structures and movements of the promastigotes were not observed in microscopic examinations. The following conclusions were made: cryopreservation is important for studies planned related to leishmaniasis and cryopreservation with DMSO is successful.

  18. Cryopreservation of Arachis pintoi (leguminosae) somatic embryos.

    Science.gov (United States)

    Rey, H Y; Faloci, M; Medina, R; Dolce, N; Engelmann, F; Mroginski, L

    2013-01-01

    In this study, we successfully cryopreserved cotyledonary somatic embryos of diploid and triploid Arachis pintoi cytotypes using the encapsulation-dehydration technique. The highest survival rates were obtained when somatic embryos were encapsulated in calcium alginate beads and precultured in agitated (80 rpm) liquid establishment medium (EM) with daily increasing sucrose concentration (0.50, 0.75, and 1.0 M). The encapsulated somatic embryos were then dehydrated with silica gel for 5 h to 20% moisture content (fresh weight basis) and cooled either rapidly (direct immersion in liquid nitrogen, LN) or slowly (1 degree C per min from 25 degree C to -30 degree C followed by immersion in LN). Beads were kept in LN for a minimum of 1 h and then were rapidly rewarmed in a 30 degree C water-bath for 2 min. Finally, encapsulated somatic embryos were post-cultured in agitated (80 rpm) liquid EM with daily decreasing sucrose concentration (0.75 and 0.5 M) and transferred to solidified EM. Using this protocol, we obtained 26% and 30% plant regeneration from cryopreserved somatic embryos of diploid and triploid cytotypes. No morphological abnormalities were observed in any of the plants regenerated from cryopreserved embryos and their genetic stability was confirmed with 10 isozyme systems and nine RAPD profiles.

  19. Increasing the efficiency of sulphur dioxide in wine by using of saturated higher fatty acids

    Directory of Open Access Journals (Sweden)

    Petra Bábíková

    2012-01-01

    Full Text Available This work is aimed on stopping of alcoholic fermentation to leave residual sugar and the possibility of sulfur dioxide reduction in wine technology and storage. As a very good opportunity showed mixture of higher saturated fatty acids with a reduced dose of sulfur dioxide. Experiments have confirmed that the concentration of viable yeasts in 1 ml of wine for variants treated with a mixture of fatty acids is significantly lower than in variants treated with sulfur dioxide alone. Then was monitored the influence of fatty acids on stored wine with residual sugar. At this point a dramatically prolongation of interval to secondary fermentation (depreciation of wine in the bottle was confirmed. Finally, attention was paid to influence on the organoleptic characteristics of wine treated this way. In this case, it is possible to consider the recommended concentration of fatty acid below the threshold of susceptibility.

  20. Cryotops versus open-pulled straws (OPS) as carriers for the cryopreservation of bovine oocytes: effects on spindle and chromosome configuration and embryo development.

    Science.gov (United States)

    Morató, Roser; Izquierdo, Dolors; Paramio, Maria Teresa; Mogas, Teresa

    2008-10-01

    Two experiments were designed to assess the effectiveness of cryopreserving bovine MII oocytes using cryotops as the carrier system for vitrification. In the first experiment, we examined the developmental competence of oocytes after: (i) vitrification in open-pulled straws (OPS method); or (ii) vitrification in plastic handle (Cryotop method). In the second experiment, warmed oocytes that had been vitrified in OPS or cryotops were fixed to analyze spindle and chromosome configuration. In all experiments both cow and calf oocytes were used. Significantly different fertilization rates were observed between the vitrification groups: 31.5% and 20.2% for the cow and calf oocytes vitrified in OPS, respectively, versus 46.1% and 46.4% for the oocytes vitrified using cryotops. After in vitro fertilization, 3.8% of the calf oocytes and 5.3% of the cow oocytes developed to the blastocyst stage. All blastocysts from vitrified oocytes resulted from the Cryotop method. A significantly lower percentage of the OPS-vitrified calf oocytes showed a normal spindle configuration (37.8%) compared to control fresh oocytes (69.9%), while normal spindle and chromosome configurations were observed in a significantly higher proportion of the cryotop-vitrified calf oocytes (60.2%). For the cow oocytes, 60.6% in the OPS group and 60.3% in the Cryotop group exhibited a normal morphology after warming. These findings suggest the cryotop system is a more efficient carrier for vitrification than OPS for the cryopreservation of bovine oocytes.

  1. Ocular Fluid As a Replacement for Serum in Cell Cryopreservation Media.

    Directory of Open Access Journals (Sweden)

    Vivek Phani Varma

    Full Text Available Cryostorage is of immense interest in biomedical research, especially for stem cell-based therapies and fertility preservation. Several protocols have been developed for efficient cryopreservation of cells and tissues, and a combination of dimethyl sulfoxide (DMSO and fetal bovine serum (FBS is commonly used. However, there is a need for an alternative to FBS because of ethical reasons, high cost, and risk of contamination with blood-borne diseases. The objective of the present study was to examine the possibility of using buffalo (Bubalus bubalis ocular fluid (BuOF to replace FBS in cryomedia. Frozen-thawed cells, which were cryopreserved in a cryomedia with BuOF, were assessed for viability, early and late apoptosis, and proliferation. Three cell lines (CHO, HEK, and C18-4, mouse embryonic stem (mES cells, and primary cells, such as mouse embryonic fibroblast (MEF cells, human peripheral blood mononuclear cells (hPBMCs, and mouse bone marrow cells (mBMCs, were cryopreserved in cryomedia containing 10% DMSO (D10 with 20% FBS (D10S20 or D10 with 20% BuOF (D10O20. For all three cell lines and mES cells cryopreserved in either D10S20 or D10O20, thawed cells showed no difference in cell viability or cell recovery. Western blot analysis of frozen-thawed-cultured cells revealed that the expression of Annexin V and proliferating cell nuclear antigen (PCNA proteins, and the ratio of BAX/BCL2 proteins were similar in all three cell lines, mES cells, and hPBMCs cryopreserved in D10S20 and D10O20. However, initial cell viability, cell recovery after culture, and PCNA expression were significantly lower in MEF cells, and the BAX/BCL2 protein ratio was elevated in mBMCs cryopreserved in D10O20. Biochemical and proteomic analysis of BuOF showed the presence of several components that may have roles in imparting the cryoprotective property of BuOF. These results encourage further research to develop an efficient serum-free cryomedia for several cell types

  2. Increasing emitter efficiency in 3.3-kV enhanced trench IGBTs for higher short-circuit capability

    DEFF Research Database (Denmark)

    Reigosa, Paula Diaz; Iannuzzo, Francesco; Rahimo, Munaf

    2018-01-01

    In this paper, a 3.3-kV Enhanced Trench IGBT has been designed with a high emitter efficiency, for improving its short-circuit robustness. The carrier distribution profile has been shaped in a way that it is possible to increase the electric field at the surface of the IGBT, and thereby, counteract...... the Kirk Effect onset. This design approach is beneficial for mitigating high-frequency oscillations, typically observed in IGBTs under short-circuit conditions. The effectiveness of the proposed design rule is validated by means of mixed-mode device simulations. Then, two IGBTs have been fabricated...... with different emitter efficiencies and tested under short circuit, validating that the high-frequency oscillations can be mitigated, with higher emitter efficiency IGBT designs....

  3. Towards a higher energy efficiency and lower carbon society the European approach and experience

    International Nuclear Information System (INIS)

    Hein, Klaus R.G.

    2010-01-01

    The use of natural energy sources and their conversion to secondary forms of energy are a crucial base for the development of our society with its continuous change of requirements due to an increase in population and the broadening of the needs in our modern life. As a consequence the consumption of primary energy resources rose drastically worldwide during the last 5 decades in particular in the industrialized regions such as Europe. Parallel in time the increasing awareness of negative effects of fuel dependent pollution on the environment and the introduction of stringent emission control regulations about 3 decades ago initiated extensive development and retrofit activities resulting in the today applied high level state of the art. As an additional challenge the worldwide debate about the potential effects of the emission of the s.c green house gases on the global climate in particular carbon dioxide from the use of predominantly fossil fuels have initiated in the European Union extensive efforts in efficiency improvements of large scale fuel conversion and more recently in the development of processes for CO 2 capture and its subsequent storage. Furthermore the use of the s.c CO 2 neutral fuels such as biomass and organic wastes as well as the non carbonaceous options water wind and solar power are promoted towards a growing role in a future primary energy mix. In addition the majority of the member states of the European Union support the minimization of energetic losses by the provision of incentives for energy savings and for switching to lower energy consuming alternatives. In fact all the above mentioned instruments have to be applied in order to fulfil the long term requirements e. g. a reduction of the greenhouse gas emission by at least 20% by 2020 and further alleviations for the time beyond. The presentation will start with summarizing the development of the demand and supply of energy in Europe during the recent decades. After an outline of the

  4. Partial protection of baboons against Schistosoma mansoni using radiation-attenuated cryopreserved schistosomula

    International Nuclear Information System (INIS)

    James, E.R.; Dobinson, A.R.; Otieno, M.; Monorei, J.; Else, J.G.

    1986-01-01

    Three groups of five baboons were vaccinated in Kenya using three doses of 10,000 viable cryopreserved schistosomula attenuated with either 10, 20 or 60 krad 60 co-irradiation. The results from perfusion indicated reductions in worm burdens in the 10, 20 and 60 krad vaccinated groups of 18%, 23% and 20% respectively, none of which was statistically significant. No stunting of adult worms could be demonstrated in any of the groups. Mean tissue egg burdens were higher in all vaccinated groups and consequently egg production per worm pair was also higher than in the challenge controls. The logistics of preparing and delivering a cryopreserved radiation-attenuated vaccine were amply demonstrated; however, in this study the levels of protection achieved were not statistically significant; possible reasons for this are discussed. (author)

  5. Generation of Platelet Microparticles after Cryopreservation of Apheresis Platelet Concentrates Contributes to Hemostatic Activity

    Directory of Open Access Journals (Sweden)

    İbrahim Eker

    2017-03-01

    Full Text Available Objective: In the last decade, substantial evidence has accumulated about the use of cryopreserved platelet concentrates, especially in trauma. However, little reference has been made in these studies to the morphological and functional changes of platelets. Recently platelets have been shown to be activated by cryopreservation processes and to undergo procoagulant membrane changes resulting in the generation of platelet-derived microparticles (PMPs, platelet degranulation, and release of platelet-derived growth factors (PDGFs. We assessed the viabilities and the PMP and PDGF levels of cryopreserved platelets, and their relation with thrombin generation. Materials and Methods: Apheresis platelet concentrates (APCs from 20 donors were stored for 1 day and cryopreserved with 6% dimethyl sulfoxide. Cryopreserved APCs were kept at -80 °C for 1 day. Thawed APCs (100 mL were diluted with 20 mL of autologous plasma and specimens were analyzed for viabilities and PMPs by flow cytometry, for thrombin generation by calibrated automated thrombogram, and for PDGFs by enzyme-linked immunosorbent assay testing. Results: The mean PMP and PDGF levels in freeze-thawed APCs were significantly higher (2763±399.4/μL vs. 319.9±80.5/μL, p<0.001 and 550.9±73.6 pg/mL vs. 96.5±49 pg/mL, p<0.001, respectively, but the viability rates were significantly lower (68.2±13.7% vs. 94±7.5%, p<0.001 than those of fresh APCs. The mean endogenous thrombin potential (ETP of freeze-thawed APCs was significantly higher than that of the fresh APCs (3406.1±430.4 nM.min vs. 2757.6±485.7 nM.min, p<0.001. Moreover, there was a significant positive poor correlation between ETP levels and PMP levels (r=0.192, p=0.014. Conclusion: Our results showed that, after cryopreservation, while levels of PMPs were increasing, significantly higher and earlier thrombin formation was occurring in the samples analyzed despite the significant decrease in viability. Considering the damage caused

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

    Science.gov (United States)

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

    2017-02-01

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

  7. Cryopreservation of donkey sperm using non-permeable cryoprotectants.

    Science.gov (United States)

    Diaz-Jimenez, M; Dorado, J; Ortiz, I; Consuegra, C; Pereira, B; Gonzalez-De Cara, C A; Aguilera, R; Mari, G; Mislei, B; Love, C C; Hidalgo, M

    2018-02-01

    The aim of this study was to evaluate the effect of different concentrations of sucrose combined with bovine serum albumin (BSA), as non-permeable cryoprotectants, on donkey sperm parameters after cryopreservation, in comparison to a control extender containing glycerol. Semen from five Andalusian donkeys (n = 12) were centrifuged and resuspended with a commercial extender for equine sperm (Gent A, Minitube) adding 1% BSA and different concentrations (M, mol/l) of water-diluted sucrose: 0.05, 0.1, 0.25, 0.35 and 0.45. Thereafter, semen (n = 24) were diluted in the same base extender containing 0.25 M sucrose (S25) or glycerol (GLY, Gent B). Sperm were slowly cooled, filled in 0.5 ml straws and frozen in nitrogen vapours. Post-thaw samples were assessed for sperm motility, plasma membrane and DNA integrity and results were compared by ANOVA. In Experiment 1, sperm motility was significantly higher (P < 0.001) for S25 than the remaining treatments, and no differences were found for plasma membrane or DNA integrity. In Experiment 2, no differences were found between S25 or GLY for sperm motility and DNA integrity but plasma membrane integrity was significantly higher (P < 0.05) for S25. In conclusion, the extender with sucrose 0.25 M combined with BSA can be considered as an alternative to conventional extenders with glycerol for donkey sperm cryopreservation. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Microencapsulation Technology: A Powerful Tool for Integrating Expansion and Cryopreservation of Human Embryonic Stem Cells

    Science.gov (United States)

    Malpique, Rita; Brito, Catarina; Jensen, Janne; Bjorquist, Petter; Carrondo, Manuel J. T.; Alves, Paula M.

    2011-01-01

    The successful implementation of human embryonic stem cells (hESCs)-based technologies requires the production of relevant numbers of well-characterized cells and their efficient long-term storage. In this study, cells were microencapsulated in alginate to develop an integrated bioprocess for expansion and cryopreservation of pluripotent hESCs. Different three-dimensional (3D) culture strategies were evaluated and compared, specifically, microencapsulation of hESCs as: i) single cells, ii) aggregates and iii) immobilized on microcarriers. In order to establish a scalable bioprocess, hESC-microcapsules were cultured in stirred tank bioreactors. The combination of microencapsulation and microcarrier technology resulted in a highly efficient protocol for the production and storage of pluripotent hESCs. This strategy ensured high expansion ratios (an approximately twenty-fold increase in cell concentration) and high cell recovery yields (>70%) after cryopreservation. When compared with non-encapsulated cells, cell survival post-thawing demonstrated a three-fold improvement without compromising hESC characteristics. Microencapsulation also improved the culture of hESC aggregates by protecting cells from hydrodynamic shear stress, controlling aggregate size and maintaining cell pluripotency for two weeks. This work establishes that microencapsulation technology may prove a powerful tool for integrating the expansion and cryopreservation of pluripotent hESCs. The 3D culture strategy developed herein represents a significant breakthrough towards the implementation of hESCs in clinical and industrial applications. PMID:21850261

  9. Medium-term cryopreservation of rabies virus samples

    Directory of Open Access Journals (Sweden)

    Tereza D'avila de Freitas Aguiar

    2013-12-01

    Full Text Available Introduction The cryopreservation of rabies virus has been described in detail in the literature. To date, little information is available on the use of cryoprotective agents for cold preservation of this virus, and the available data focus only on short-term virus preservation. In this study, we investigated the medium-term cryopreservation of samples of rabies virus using different cryopreservation protocols. Methods The cryopreservation protocols for the rabies virus samples were performed at -20°C and were divided according to the variables of time and cryoprotectant type used. The laboratory tests (intracerebral inoculation of mice, viral titration and direct immunofluorescence were performed at regular intervals (360 and 720 days to assess the viability of the viral samples according to the different preservation techniques used. Results After 1 year of cryopreservation, the fluorescence intensity of intracellular corpuscles of the rabies virus and the median survival time of the mice differed between the positive controls and the treatments with the cryoprotectants. After 2 years, most of the samples subjected to the cryopreservation protocols (including the controls did not produce fluorescence. However, the virus samples exposed to the cryoprotectant sucrose (68% solution responded positively in the direct immunofluorescence assay and in the intracerebral inoculation of the mice. Conclusions Medium-term cryopreservation of the rabies virus inactivates the viral sample. However, the cryoprotectant agent sucrose (68% produces a preservative effect in cryopreserved rabies virus samples.

  10. Ovarian tissue cryopreservation and transplantation among alternatives for fertility preservation in the Nordic countries

    DEFF Research Database (Denmark)

    Rodriguez-Wallberg, Kenny A; Tanbo, Tom; Tinkanen, Helena

    2016-01-01

    cryopreservation to be experimental. In Iceland, embryo cryopreservation is the only option for fertility preservation. Most centers use slow-freezing methods for ovarian tissue cryopreservation. Most patients selected for ovarian tissue cryopreservation were newly diagnosed with cancer and the tissue...

  11. Cryopreservation of Embryos and Oocytes in Human Assisted Reproduction

    Directory of Open Access Journals (Sweden)

    János Konc

    2014-01-01

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

  12. Cryopreservation of embryos and oocytes in human assisted reproduction.

    Science.gov (United States)

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

    2014-01-01

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

  13. First attempts to cryopreserve red abalone (Haliotis rufescens oocytes

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    Ramírez Torrez, A.

    2015-01-01

    Full Text Available Overall, few advances in the cryopreservation of complex cells such as oocytes, embryo or tissue have been registered and in less quantity have been reported for aquatic species. Abalone has high economic interest worldwide and the conservation of abalone germplasm may help to enhance its culture and develop repopulation programs. In this work, we reported the cytotoxic effect of two concentration of trehalose (0.2 and 0.4 M on red abalone oocytes incubated for 10, 15 and 20 min. Also, we reported the cryopreservation of red abalone oocytes using a 3-steps cryopreservation protocol and 5 thawing protocols. Significant differences on cytotoxic effect were found (p<0.01. However, none of the cryoprotectant was optimum to cryopreserve red abalone oocyte. In conclusion, it is necessary to find an appropriate method to dehydrate or make the cryoprotectant penetrate on the abalone oocyte before proceeding to cryopreservation.

  14. A comparison of cryopreservation methods: Slow-cooling vs. rapid-cooling based on cell viability, oxidative stress, apoptosis, and CD34+ enumeration of human umbilical cord blood mononucleated cells

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    Sandra Ferry

    2011-09-01

    Full Text Available Abstract Background The finding of human umbilical cord blood as one of the most likely sources of hematopoietic stem cells offers a less invasive alternative for the need of hematopoietic stem cell transplantation. Due to the once-in-a-life time chance of collecting it, an optimum cryopreservation method that can preserve the life and function of the cells contained is critically needed. Methods Until now, slow-cooling has been the routine method of cryopreservation; however, rapid-cooling offers a simple, efficient, and harmless method for preserving the life and function of the desired cells. Therefore, this study was conducted to compare the effectiveness of slow- and rapid-cooling to preserve umbilical cord blood of mononucleated cells suspected of containing hematopoietic stem cells. The parameters used in this study were differences in cell viability, malondialdehyde content, and apoptosis level. The identification of hematopoietic stem cells themselves was carried out by enumerating CD34+ in a flow cytometer. Results Our results showed that mononucleated cell viability after rapid-cooling (91.9% was significantly higher than that after slow-cooling (75.5%, with a p value = 0.003. Interestingly, the malondialdehyde level in the mononucleated cell population after rapid-cooling (56.45 μM was also significantly higher than that after slow-cooling (33.25 μM, with a p value p value = 0.138. However, CD34+ enumeration was much higher in the population that underwent slow-cooling (23.32 cell/μl than in the one that underwent rapid-cooling (2.47 cell/μl, with a p value = 0.001. Conclusions Rapid-cooling is a potential cryopreservation method to be used to preserve the umbilical cord blood of mononucleated cells, although further optimization of the number of CD34+ cells after rapid-cooling is critically needed.

  15. Exploring the Possibility of Cryopreservation of Feline and Canine Erythrocytes by Rapid Freezing with Penetrating and Non-Penetrating Cryoprotectants.

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    Denys Pogozhykh

    Full Text Available Efficient application of veterinary blood transfusion approaches for small companion animals requires readily available supply of the donor material. This can be achieved by developing of effective biobanking technologies allowing long-term storage of donor blood components via cryopreservation. Transfusion of an erythrocyte concentrate allows the successful correction of various hematological pathologies, severe bleeding, and etc. While in the past there were several approaches to cryopreserve red blood cells of dogs, to our knowledge there is virtually no data on cryopreservation of feline erythrocytes. In this paper, we performed a comprehensive parameter optimization for low temperature storage of RBCs of both species. Here, the efficiency of single-component and multicomponent cryoprotective media as well as necessary time of pre-incubation with penetrating and non-penetrating cryoprotectants prior to rapid freezing is analyzed. This study showed that glycerol was not sufficient for cryopreservation of red blood cells of the studied species under the investigated conditions. Application of 10% (v/v ME2SO allowed for a significant reduction of canine and feline erythrocytes hemolysis after thawing. 17.5% hydroxyethyl starch demonstrated the highest cryoprotective activity for both species. It was found that dog RBCs should be incubated in cryoprotective media for 30 min at 22°C prior to freezing, while for cat RBCs 20 min is sufficient. Combination of CPAs was less effective. Presented data may be considered in further studies in veterinary transfusion and blood banking optimization.

  16. Effect of Previous Chemotherapy on the Quality of Cryopreserved Human Ovarian Tissue In Vitro.

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    Babak Asadi Azarbaijani

    Full Text Available Cryopreservation of ovarian tissue has been widely accepted as an option for fertility preservation among cancer patients. Some patients are exposed to chemotherapy prior to ovarian tissue cryopreservation. Consequently, assessment of the developmental capacity of human ovarian tissue after chemotherapy is of primary importance.In order to study the impact of previous chemotherapy on in vitro development and viability of ovarian follicles, quality control samples from 34 female cancer patients at median age of 15 years (range 1‒35, cryopreserved for fertility preservation before (n = 14 or after (n = 20 initiation of chemotherapy, were thawed and cultured for 7 days. The morphology and developmental stages of ovarian follicles were studied by light microscopy before and after culture. Possible associations between follicular densities, age and exposure to alkylating agents, expressed as cyclophosphamide equivalent dose (CED were tested.Exposure to chemotherapy significantly impaired the survival and development of ovarian follicles in culture. After seven days, significantly higher densities of intermediary, primary and secondary follicles and lower densities of atretic follicles was detected in the samples collected before chemotherapy. Increasing dose of alkylating agents was identified by multivariate linear regression analysis as an independent predictor of a higher density of atretic follicles, whereas increasing age of the patient predicted a better outcome with less follicle atresia and a higher density of maturing follicles.This study provides quantitative in vitro evidence of the impact of chemotherapy on developmental capacity of cryopreserved human ovarian tissue. The results indicate that fertility preservation should be carried out, if possible, before initiation of alkylating agents in order to guarantee better in vitro survival of ovarian follicles. In addition, ovarian samples from younger girls show lower viability and fewer

  17. Characterization of cryopreserved primary human corneal endothelial cells cultured in human serum-supplemented media

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    Lucas Monferrari Monteiro Vianna

    2016-02-01

    Full Text Available ABSTRACT Purpose: To compare cryopreserved human corneal endothelial cells (HCECs grown in human serum-supplemented media (HS-SM with cryopreserved HCECs grown in fetal bovine serum-supplemented media (FBS-SM. Methods: Three pairs of human corneas from donors aged 8, 28, and 31 years were obtained from the eye bank. From each pair, one cornea was used to start a HCEC culture using HS-SM; the other cornea was grown in FBS-SM. On reaching confluence, the six cell populations were frozen using 10% dimethyl sulfoxidecontaining medium. Thawed cells grown in HS-SM were compared with those grown in FBS-SM with respect to morphology, growth curves, immunohistochemistry, real time-reverse transcriptase polymerase chain reaction (RT-PCR for endothelial cell markers, and detachment time. Results: No difference in morphology was observed for cells grown in the two media before or after cryopreservation. By growth curves, cell counts after thawing were similar in both media, with a slight trend toward higher cell counts in FBS-SM. Cells grown in both the media demonstrated a similar expression of endothelial cell markers when assessed by immunohistochemistry, although HCEC marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM as assessed by RT-PCR. With FBS-SM, there was a tendency of longer detachment time and lower cell passages. Conclusions: HS-SM was similar to FBS-SM for cryopreservation of cultured HCECs as assessed by analysis of cell morphology, proliferation, and protein expression, although marker gene expression was higher in cells grown in HS-SM than in those grown in FBS-SM. Detachment time was longer with FBS-SM and in lower passages.

  18. Effect of Cryopreservation and Post-Cryopreservation Somatic Embryogenesis on the Epigenetic Fidelity of Cocoa (Theobroma cacao L.).

    Science.gov (United States)

    Adu-Gyamfi, Raphael; Wetten, Andy; Marcelino Rodríguez López, Carlos

    2016-01-01

    While cocoa plants regenerated from cryopreserved somatic embryos can demonstrate high levels of phenotypic variability, little is known about the sources of the observed variability. Previous studies have shown that the encapsulation-dehydration cryopreservation methodology imposes no significant extra mutational load since embryos carrying high levels of genetic variability are selected against during protracted culture. Also, the use of secondary rather than primary somatic embryos has been shown to further reduce the incidence of genetic somaclonal variation. Here, the effect of in vitro conservation, cryopreservation and post-cryopreservation generation of somatic embryos on the appearance of epigenetic somaclonal variation were comparatively assessed. To achieve this we compared the epigenetic profiles, generated using Methylation Sensitive Amplified Polymorphisms, of leaves collected from the ortet tree and from cocoa somatic embryos derived from three in vitro conditions: somatic embryos, somatic embryos cryopreserved in liquid nitrogen and somatic embryos generated from cryoproserved somatic embryos. Somatic embryos accumulated epigenetic changes but these were less extensive than in those regenerated after storage in LN. Furthermore, the passage of cryopreserved embryos through another embryogenic stage led to further increase in variation. Interestingly, this detected variability appears to be in some measure reversible. The outcome of this study indicates that the cryopreservation induced phenotypic variability could be, at least partially, due to DNA methylation changes. Phenotypic variability observed in cryostored cocoa somatic-embryos is epigenetic in nature. This variability is partially reversible, not stochastic in nature but a directed response to the in-vitro culture and cryopreservation.

  19. Cryopreservation of spores of Dicksonia sellowiana: an endangered tree fern indigenous to South and Central America.

    Science.gov (United States)

    Rogge, G D; Viana, A M; Randi, A M

    2000-01-01

    Spores of Dicksonia sellowiana (Presl.) Hook., an endangered tree fern, were stored in liquid nitrogen. Surface sterilized spores were placed in 1 ml sterile polypropylene cryotubes and were plunged into liquid nitrogen cryo-cans for 15 minutes, 15 days, 1 month and 3 months. In all, of the treatments the percentage of germination was higher than the control (fresh spores). Germination in Dyer and MS media supplement with 10 (-7) M and 5 x 10(-7) M BA was also promoted as comparing to control. There was no difference between the germination of spores thawed rapidly in a water bath at 45 degree C during 5 minutes or slowly at room temperature. Cryopreservation seems to promote germination of some dormant spores of D. sellowiana. The pre-treatment in cryoprotective solution of dimethyl sulphoxide 15%(v/v) in 1 M glycerol inhibited the germination of cryopreserved spores

  20. Towards gene banking amphibian maternal germ lines: short-term incubation, cryoprotectant tolerance and cryopreservation of embryonic cells of the frog, Limnodynastes peronii.

    Science.gov (United States)

    Lawson, Bianca; Clulow, Simon; Mahony, Michael J; Clulow, John

    2013-01-01

    Gene banking is arguably the best method available to prevent the loss of genetic diversity caused by declines in wild populations, when the causes of decline cannot be halted or reversed. For one of the most impacted vertebrate groups, the amphibians, gene banking technologies have advanced considerably, and gametes from the male line can be banked successfully for many species. However, cryopreserving the female germ line remains challenging, with attempts at cryopreserving oocytes unsuccessful due to their large size and yolk content. One possible solution is to target cryopreservation of early embryos that contain the maternal germ line, but consist of smaller cells. Here, we investigate the short term incubation, cryoprotectant tolerance, and cryopreservation of dissociated early embryonic cells from gastrulae and neurulae of the Striped Marsh Frog, Limnodynastes peronii. Embryos were dissociated and cells were incubated for up to 24 hours in various media. Viability of both gastrula and neurula cells remained high (means up to 40-60%) over 24 hours of incubation in all media, although viability was maintained at a higher level in Ca(2+)-free Simplified Amphibian Ringer; low speed centrifugation did not reduce cell viability. Tolerance of dissociated embryonic cells was tested for two cryoprotectants, glycerol and dimethyl sulphoxide; dissociated cells of both gastrulae and neurulae were highly tolerant to both-indeed, cell viability over 24 hours was higher in media containing low-to-medium concentrations than in equivalent cryoprotectant-free media. Viability over 24 hours was lower in concentrations of cryoprotectant higher than 10%. Live cells were recovered following cryopreservation of both gastrula and neurula cells, but only at low rates. Optimal cryodiluents were identified for gastrula and neurula cells. This is the first report of a slow cooling protocol for cryopreservation of amphibian embryonic cells, and sets future research directions for

  1. Towards gene banking amphibian maternal germ lines: short-term incubation, cryoprotectant tolerance and cryopreservation of embryonic cells of the frog, Limnodynastes peronii.

    Directory of Open Access Journals (Sweden)

    Bianca Lawson

    Full Text Available Gene banking is arguably the best method available to prevent the loss of genetic diversity caused by declines in wild populations, when the causes of decline cannot be halted or reversed. For one of the most impacted vertebrate groups, the amphibians, gene banking technologies have advanced considerably, and gametes from the male line can be banked successfully for many species. However, cryopreserving the female germ line remains challenging, with attempts at cryopreserving oocytes unsuccessful due to their large size and yolk content. One possible solution is to target cryopreservation of early embryos that contain the maternal germ line, but consist of smaller cells. Here, we investigate the short term incubation, cryoprotectant tolerance, and cryopreservation of dissociated early embryonic cells from gastrulae and neurulae of the Striped Marsh Frog, Limnodynastes peronii. Embryos were dissociated and cells were incubated for up to 24 hours in various media. Viability of both gastrula and neurula cells remained high (means up to 40-60% over 24 hours of incubation in all media, although viability was maintained at a higher level in Ca(2+-free Simplified Amphibian Ringer; low speed centrifugation did not reduce cell viability. Tolerance of dissociated embryonic cells was tested for two cryoprotectants, glycerol and dimethyl sulphoxide; dissociated cells of both gastrulae and neurulae were highly tolerant to both-indeed, cell viability over 24 hours was higher in media containing low-to-medium concentrations than in equivalent cryoprotectant-free media. Viability over 24 hours was lower in concentrations of cryoprotectant higher than 10%. Live cells were recovered following cryopreservation of both gastrula and neurula cells, but only at low rates. Optimal cryodiluents were identified for gastrula and neurula cells. This is the first report of a slow cooling protocol for cryopreservation of amphibian embryonic cells, and sets future research

  2. Whole ovary cryopreservation with vascular transplantation – A future development in female oncofertility

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    Mats Brännström

    2010-07-01

    Full Text Available Preservation of female fertility in female cancer victims is gaining more importance in the light of the excellent survival rates today after treatment for the types of cancer, which are common in women during childhood and fertile age. In this article we discuss the risks of infertility after various cancer treatments and in what patient categories fertility preservation should be discussed. The modes of female fertility preservation used today as clinically established methods or as experimental methods that are used in the human female are reviewed. The advantages and disadvantages of the methods are also identified. Whole ovary cryopreservation has been suggested as a possible future more efficient method of fertility preservation. The research on this method in different animal models is discussed in detail and unsolved questions are identified. To date there has not been any attempt to transplant a cryopreserved whole human ovary but human ovaries have already been cryopreserved for future fertility preservation purposes, when cryobiological and microsurgical techniques have been further refined.

  3. Successful Oocyte Cryopreservation in Reproductive-Aged Cancer Survivors.

    Science.gov (United States)

    Druckenmiller, Sarah; Goldman, Kara N; Labella, Patty A; Fino, M Elizabeth; Bazzocchi, Antonia; Noyes, Nicole

    2016-03-01

    To demonstrate that oocyte cryopreservation is a feasible reproductive option for patients with cancer of childbearing age who require gonadotoxic therapies. This study is a university-based retrospective review of reproductive-aged cancer patient treatment cycles that included ovarian stimulation, transvaginal oocyte retrieval, oocyte cryopreservation, and, in some cases, subsequent oocyte thaw, in vitro fertilization, and embryo transfer. Outcome measures included ovarian stimulation response, number of oocytes retrieved, cryopreserved, and thawed, and pregnancy data. From 2005 to 2014, 176 reproductive-aged patients with cancer (median age 31 years, interquartile range 24-36) completed 182 oocyte cryopreservation cycles. Median time between consult request and oocyte retrieval was 12 days (interquartile range 10-14). Median peak stimulation estradiol was 1,446 pg/mL (interquartile range 730-2,687); 15 (interquartile range 9-23) oocytes were retrieved and 10 (interquartile range 5-18) metaphase II oocytes were cryopreserved per cycle. Ten patients (11 cycles) have returned to attempt pregnancy with their cryopreserved oocytes. Among thawed oocytes, the cryopreservation survival rate was 86% (confidence interval [CI] 78-94%). Nine of 11 thaw cycles resulted in embryos suitable for transfer. The embryo implantation rate was 27% (CI 8-46%) and the live birth rate was 44% (CI 12-77%) per embryo transfer. Chance for live birth with embryos created from cryopreserved oocytes was similar between the patients with cancer in this study and noncancer patients who underwent the same treatment at our center (44% [CI 12-77%] compared with 33% [CI 22-44%] per embryo transfer). Oocyte cryopreservation is now a feasible fertility preservation option for reproductive-aged patients with cancer who require gonadotoxic therapies.

  4. Cryopreservation Maintains Functionality of Human iPSC Dopamine Neurons and Rescues Parkinsonian Phenotypes In Vivo

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    Dustin R. Wakeman

    2017-07-01

    Full Text Available A major challenge for clinical application of pluripotent stem cell therapy for Parkinson's disease (PD is large-scale manufacturing and cryopreservation of neurons that can be efficiently prepared with minimal manipulation. To address this obstacle, midbrain dopamine neurons were derived from human induced pluripotent stem cells (iPSC-mDA and cryopreserved in large production lots for biochemical and transplantation studies. Cryopreserved, post-mitotic iPSC-mDA neurons retained high viability with gene, protein, and electrophysiological signatures consistent with midbrain floor-plate lineage. To test therapeutic efficacy, cryopreserved iPSC-mDA neurons were transplanted without subculturing into the 6-OHDA-lesioned rat and MPTP-lesioned non-human-primate models of PD. Grafted neurons retained midbrain lineage with extensive fiber innervation in both rodents and monkeys. Behavioral assessment in 6-OHDA-lesioned rats demonstrated significant reversal in functional deficits up to 6 months post transplantation with reinnervation of the host striatum and no aberrant growth, supporting the translational development of pluripotent cell-based therapies in PD.

  5. Cryopreservation of tissue engineered constructs for bone.

    Science.gov (United States)

    Kofron, Michelle D; Opsitnick, Natalie C; Attawia, Mohamed A; Laurencin, Cato T

    2003-11-01

    The large-scale clinical use of tissue engineered constructs will require provisions for its mass availability and accessibility. Therefore, it is imperative to understand the effects of low temperature (-196 degrees C) on the tissue engineered biological system. Initial studies used samples of the osteoblast-like cell line (SaOS-2) adhered to a two-dimensional poly(lactide-co-glycolide) thin film (2D-PLAGA) or a three-dimensional poly(lactide-co-glycolide) sintered microsphere matrix (3D-PLAGA) designed for bone tissue engineering. Experimental samples were tested for their ability to maintain cell viability, following low temperature banking for one week, in solutions of the penetrating cryoprotective agents, dimethylsulfoxide (DMSO), ethylene glycol, and glycerol. Results indicated the DMSO solution yielded the greatest percent cell survival for SaOS-2 cells adhered to both the 2D- and 3D-PLAGA scaffolds; therefore, DMSO was used to cryopreserve mineralizing primary rabbit osteoblasts cells adhered to 2D-PLAGA matrices for 35 days. Results indicated retention of the extracellular matrix architecture as no statistically significant difference in the pre- and post-thaw mineralized structures was measured. Percent cell viability of the mineralized constructs following low temperature storage was approximately 50%. These are the first studies to address the issue of preservation techniques for tissue engineered constructs. The ability to successfully cryopreserve mineralized tissue engineered matrices for bone may offer an unlimited and readily available source of bone-like materials for orthopaedic applications.

  6. Recent Progress in Cryopreservation of Bovine Oocytes

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    In-Sul Hwang

    2014-01-01

    Full Text Available Principle of oocyte cryoinjury is first overviewed and then research history of cryopreservation using bovine oocytes is summarized for the last two decades with a few special references to recent progresses. Various types of cryodevices have been developed to accelerate the cooling rate and applied to the oocytes from large domestic species enriched with cytoplasmic lipid droplets. Two recent approaches include the qualitative improvement of IVM oocytes prior to the vitrification and the short-term recovery culture of vitrified-warmed oocytes prior to the subsequent IVF. Supplementation of L-carnitine to IVM medium of bovine oocytes has been reported to reduce the amount of cytoplasmic lipid droplets and improve the cryotolerance of the oocytes, but it is still controversial whether the positive effect of L-carnitine is reproducible. Incidence of multiple aster formation, a possible cause for low developmental potential of vitrified-warmed bovine oocytes, was inhibited by a short-term culture of the postwarm oocytes in the presence of Rho-associated coiled-coil kinase (ROCK inhibitor. Use of an antioxidant α-tocopherol, instead of the ROCK inhibitor, also supported the revivability of the postwarm bovine oocytes. Further improvements of the vitrification procedure, combined with pre- and postvitrification chemical treatment, would overcome the high sensitivity of bovine oocytes to cryopreservation.

  7. Nucleons II: cryopreservation and metabolic activity.

    Science.gov (United States)

    Reyes, R; Flores-Alonso, J C; Rodríguez-Hernández, H M; Merchant-Larios, H M; Delgado, N M

    2001-01-01

    The establishment of intracytoplasmatic sperm injection (ICSI) as a routine procedure in assisted fertilization has been used in the treatment of male infertility. The major technical problem that has arisen with the use of immotile sperm for ICSI has been differentiating between live and dead cells. Nucleons from human, pig, hamster, mouse, rat, and bull have been able to induce their chromatin decondensation by the action of heparin/GSH. Cryopreservation is deleterious to sperm function, killing more than 50% of the spermatozoa during the process. Nucleon cryostorage was performed at 5 and -5 degrees C and analyzed for total area (mu2), perimeter (mu), width (mu), and length (mu), using Metamorph Imaging System software. On the other hand, fluorescein diacetate (FDA) is hydrolyzed by intracellular estereases to produce fluorescein, which exhibits green fluorescence when excited by blue light. This fact is a striking result since the presence of this metabolic activity opens the possibility to select the nucleons for ICSI. In the present study, the authors decided to search for a suitable metabolic test, which might reflect the metabolism and viability of these chromatin structures. This is a simple cryostorage technique that after months of cryopreservation, allow the use of nucleons for ICSI with suitable fertilization and pregnancies rates.

  8. Partial dehydration and cryopreservation of Citrus seeds.

    Science.gov (United States)

    Graiver, Natalia; Califano, Alicia; Zaritzky, Noemí

    2011-11-01

    Three categories of seed storage behavior are generally recognized among plant species: orthodox, intermediate and recalcitrant. Intermediate seeds cannot be stored in liquid nitrogen (LN) without a previous partial dehydration process. The water content (WC) of the seeds at the moment of immersion in LN must be regarded as the most critical factor in cryopreservation. The purpose of this study was to investigate the basis of the optimal hydration status for cryopreservation of Citrus seeds: C. sinensis (sweet orange), C. paradisi (grapefruit), C. reticulata (mandarin) in LN. To study the tolerance to dehydration and LN exposure, seeds were desiccated by equilibration at relative humidities between 11 and 95%. Sorption isotherms were determined and modeled; lipid content of the seeds was measured. Seed desiccation sensitivity was quantified by the quantal response model. Differential scanning calorimetry (DSC) thermograms were determined on cotyledon tissue at different moisture contents to measure ice melting enthalpies and unfrozen WC. Samples of total seed lipid extract were also analyzed by DSC to identify lipid transitions in the thermograms. The limit of hydration for LN Citrus seeds treatment corresponded to the unfrozen WC in the tissue, confirming that seed survival strictly depended on avoidance of intracellular ice formation. Copyright © 2011 Society of Chemical Industry.

  9. Evaluation of the damage in fish spermatozoa cryopreservation

    Science.gov (United States)

    Li, Jun; Liu, Qinghua; Zhang, Shicui

    2006-12-01

    Cryodamages occur during sperm cryopreservation. Cryopreservation of fish sperm usually results in marked decrease in sperm quality, such as swelling or disruption of the plasma membrane, mitochondrial dysfunction, diminished sperm motility, impaired velocity, shorter motility period, denaturation, and release of some enzymes from spermatozoa. In this paper, damages in morphology, physiology, biochemistry and metabolism, and genetic integrity of fish semen after cryopreservation are discussed. New approaches in assessment of fish thawed sperm quality such as computer assisted sperm analysis, flow cytometic analysis combined with fluorescent probes and single cell gel electrophoresis are also briefly reviewed.

  10. Higher energy efficient homes are associated with increased risk of doctor diagnosed asthma in a UK subpopulation.

    Science.gov (United States)

    Sharpe, Richard A; Thornton, Christopher R; Nikolaou, Vasilis; Osborne, Nicholas J

    2015-02-01

    The United Kingdom (UK) has one of the highest prevalence of asthma in the world, which represents a significant economic and societal burden. Reduced ventilation resulting from increased energy efficiency measures acts as a modifier for mould contamination and risk of allergic diseases. To our knowledge no previous study has combined detailed asset management property and health data together to assess the impact of household energy efficiency (using the UK Government's Standard Assessment Procedure) on asthma outcomes in an adult population residing in social housing. Postal questionnaires were sent to 3867 social housing properties to collect demographic, health and environmental information on all occupants. Detailed property data, residency periods, indices of multiple deprivation (IMD) and household energy efficiency ratings were also investigated. Logistic regression was used to calculate odds ratios and confidence intervals while allowing for clustering of individuals coming from the same location. Eighteen percent of our target social housing population were recruited into our study. Adults had a mean age of 59 (SD±17.3) years and there was a higher percentage of female (59%) and single occupancy (58%) respondents. Housing demographic characteristics were representative of the target homes. A unit increase in household Standard Assessment Procedure (SAP) rating was associated with a 2% increased risk of current asthma, with the greatest risk in homes with SAP >71. We assessed exposure to mould and found that the presence of a mouldy/musty odour was associated with a two-fold increased risk of asthma (OR 2.2 95%; CI 1.3-3.8). A unit increase in SAP led to a 4-5% reduction in the risk of visible mould growth and a mouldy/musty odour. In contrast to previous research, we report that residing in energy efficient homes may increase the risk of adult asthma. We report that mould contamination increased the risk of asthma, which is in agreement with existing

  11. Successful use of the Cryolock device for cryopreservation of scarce human ejaculate and testicular spermatozoa.

    Science.gov (United States)

    Stein, A; Shufaro, Y; Hadar, S; Fisch, B; Pinkas, H

    2015-03-01

    The existing methods for cryopreservation of very low count sperm samples are complex and sub-optimal for individual spermatozoa. Our purpose is to establish an effective simple method for cryoprotecting individual spermatozoa. Samples from patients with OTA were mixed with TYB or HEPES-buffered salt solution with glycerol + glucose and placed on a Cryolock that was plunged directly into liquid nitrogen or exposed to its vapors. Thawing was performed by direct immersion into a drop of warmed medium. The favorable method was tested on diluted samples (10-50 cells) and leftover TESE specimens from patients with azoospermia. Cryopreservation was considered successful if >30 spermatozoa, (>3 motile), or >5 spermatozoa (>1 motile) in diluted and TESE samples, were detected post-thawing. A significantly higher survival rate of seminal spermatozoa was obtained when using the Cryolock with TYB and freezing with liquid nitrogen vapor, compared to HEPES glycerol-glucose (95 vs. 35% respectively). Plunging the Cryolock into liquid nitrogen was detrimental. Cryolock combined with TYB cryoprotection and liquid nitrogen vapor freezing was highly effective for cryopreservation of individual spermatozoa in diluted and TESE samples. The Cryolock may serve for freezing very low-count sperm samples and individual spermatozoa. This method offers simplicity, efficacy, use of available materials, without requiring micromanipulation equipment or skills. © 2015 American Society of Andrology and European Academy of Andrology.

  12. The effect of Curcuma longa extracted (curcumin) on the quality of cryopreserved boar semen.

    Science.gov (United States)

    Chanapiwat, Panida; Kaeoket, Kampon

    2015-09-01

    The aim of this study was to determine the optimal concentration of curcumin needed for cryopreservation of boar semen. Semen samples (n = 9) were collected from nine Duroc boars which having proven fertility were used for routine artificial insemination. Semen samples were collected and divided into six groups (groups A-F) according to various concentrations of curcumin in freezing extender (i.e. 0, 0.125, 0.25, 0.50, 0.75 and 1.0 mmol/L, respectively). The semen was frozen by traditional liquid nitrogen vapor method and stored at -196°C in the liquid nitrogen tank. After storage, frozen semen samples were thawed at 50°C for 12 s and evaluated for progressive motility, viability and acrosome integrity. The present results indicated that the addition of curcumin at 0.25 (group C) or 0.50 mmol/L curcumin (group D) yielded the higher percentage of progressive motility (33.3 and 36.1%, respectively) (P curcumin during cryopreservation at a concentration of 0.25 or 0.50 mmol/L is the optimal concentration of curcumin for improving the quality (i.e. increased progressive motility and acrosome integrity) of cryopreserved boar semen. © 2015 Japanese Society of Animal Science.

  13. Soybean lecithin-based extender preserves spermatozoa membrane integrity and fertilizing potential during goat semen cryopreservation.

    Science.gov (United States)

    Chelucci, Sara; Pasciu, Valeria; Succu, Sara; Addis, Daniela; Leoni, Giovanni G; Manca, Maria E; Naitana, Salvatore; Berlinguer, Fiammetta

    2015-04-01

    Soybean lecithin may represent a suitable alternative to egg yolk for semen cryopreservation in livestock species. However, additional studies are needed to elucidate its effects on spermatozoa functional properties. Semen collected from five Sarda bucks was cryopreserved in Tris-based extender and glycerol (4% v:v) with different supplementations. In a preliminary experiment, different soybean lecithin concentrations were tested (1%-6% wt/vol) and results in terms of viability, percentages of progressive motile and rapid spermatozoa, and DNA integrity after thawing showed that the most effective concentration was 1%. In the second experiment, semen was frozen in a Tris-based extender with no supplementation (EXT), with 1% lecithin (EXT LC), and 20% egg yolk (EXT EY). The effectiveness of these extenders was also compared with a commercial extender. The EXT EY led to the highest viability and motility parameters after freezing and thawing (P lecithin can be considered as a suitable alternative to egg yolk in goat semen cryopreservation, because it ensures higher fertilization rates and a better protection from membrane damage by cold shock. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Ovarian tissue cryopreservation by stepped vitrification and monitored by X-ray computed tomography.

    Science.gov (United States)

    Corral, Ariadna; Clavero, Macarena; Gallardo, Miguel; Balcerzyk, Marcin; Amorim, Christiani A; Parrado-Gallego, Ángel; Dolmans, Marie-Madeleine; Paulini, Fernanda; Morris, John; Risco, Ramón

    2018-04-01

    Ovarian tissue cryopreservation is, in most cases, the only fertility preservation option available for female patients soon to undergo gonadotoxic treatment. To date, cryopreservation of ovarian tissue has been carried out by both traditional slow freezing method and vitrification, but even with the best techniques, there is still a considerable loss of follicle viability. In this report, we investigated a stepped cryopreservation procedure which combines features of slow cooling and vitrification (hereafter called stepped vitrification). Bovine ovarian tissue was used as a tissue model. Stepwise increments of the Me 2 SO concentration coupled with stepwise drops-in temperature in a device specifically designed for this purpose and X-ray computed tomography were combined to investigate loading times at each step, by monitoring the attenuation of the radiation proportional to Me 2 SO permeation. Viability analysis was performed in warmed tissues by immunohistochemistry. Although further viability tests should be conducted after transplantation, preliminary results are very promising. Four protocols were explored. Two of them showed a poor permeation of the vitrification solution (P1 and P2). The other two (P3 and P4), with higher permeation, were studied in deeper detail. Out of these two protocols, P4, with a longer permeation time at -40 °C, showed the same histological integrity after warming as fresh controls. Copyright © 2018 Elsevier Inc. All rights reserved.

  15. Supplemental effect of varying L-cysteine concentrations on the quality of cryopreserved boar semen

    Science.gov (United States)

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

    2010-01-01

    Cryopreservation is associated with the production of reactive oxygen species, which leads to lipid peroxidation of the sperm membrane and consequently a reduction in sperm motility and decreased fertility potential. The aim of this study was to determine the optimal concentration of L-cysteine needed for cryopreservation of boar semen. Twelve boars provided semen of proven motility and morphology for this study. The semen was divided into four portions in which the lactose-egg yolk (LEY) extender used to resuspend the centrifuged sperm pellet was supplemented with various concentrations of L-cysteine to reach 0 mmol L−1 (group I, control), 5 mmol L−1 (group II), 10 mmol L−1 (group III) and 15 mmol L−1 (group IV). Semen suspensions were loaded in straws (0.5 mL) and placed in a controlled-rate freezer. After cryopreservation, frozen semen samples were thawed and investigated for progressive motility, viability using SYBR-14/EthD-1 staining and acrosome integrity using FITC-PNA/EthD-1 staining. There was a significantly higher (P extender for improving the quality of frozen–thawed boar semen. PMID:20601963

  16. Modified A{sup ++}-standard refrigerator with 30% reduction of energy consumption. Higher efficiency by compressor modification - Final report

    Energy Technology Data Exchange (ETDEWEB)

    Siegrist, M.; Stahl, S.; Ganz, J.

    2010-06-15

    This final report for the Swiss Federal Office of Energy (SFOE) takes a look at how a modified refrigerator can be given a higher efficiency by modifying the compressor. The modified refrigerator was fitted with a variable-speed compressor. This compressor could be run at much lower speeds so that it was in operation for up to 90% of the time. It was shown that less electricity was consumed the more the compressor ran. The report discusses the aims of the work and presents details on the standard refrigerator used for the tests. The compressor normally used and the variable-speed compressor used in the test are described. Systems for temperature control and data acquisition during the tests are described. The results obtained are examined and the influence of various factors is discussed.

  17. Characterization of proacrosin/acrosin system after liquid storage and cryopreservation of turkey semen (Meleagris gallopavo).

    Science.gov (United States)

    Słowińska, M; Liszewska, E; Dietrich, G J; Ciereszko, A

    2012-09-15

    This study was designed to identify the effect of liquid storage at 4 °C for 48 h and cryopreservation on the proacrosin/acrosin system of turkey spermatozoa. Anti-acrosin I antibodies were produced and used to demonstrate Western blot analysis profile of the proacrosin/acrosin system of sperm and seminal plasma and possible changes in the proacrosin/acrosin system of turkey sperm stored for 2.5, 24, and 48 h or cryopreserved. At the same time acrosin-like activity was examined by the measurement of amidase activity of sperm extracts, sperm suspension, and seminal plasma of turkey semen. A computer-assisted sperm analysis system was used to monitor the sperm motility characteristics of turkey sperm stored for 48 h or cryopreserved. Different profiles of the sperm proacrosin/acrosin system were observed regarding the presence or absence of inhibitors (p-nitrophenyl-p'-guanidine benzoate [NPGB] and Kazal family inhibitor) during the extraction process. When NPGB was present three main bands were observed with the molecular weight ranging from 66 to 35 kDa. Bands corresponding to acrosin I and II were not observed. In sperm extract without NPGB, three or four bands were observed with the molecular weight ranging from 41 to 30 kDa. The bands corresponding to acrosin I and II were observed. During liquid storage a decrease in sperm motility and an increase in sperm-extracted amidase activity were observed. After 24 and 48 h of storage, extracted amidase activity was higher than at 2.5 h by 24% and 31%, respectively. However, no changes in the Western blot analysis profiles of sperm extract and seminal plasma were visible during liquid storage. After cryopreservation a decrease in sperm motility and all sperm motility parameters were observed. In contrast to liquid storage, cryopreservation did not increase extracted amidase activity. However, changes in Western blot analysis profiles were visible in sperm extract and seminal plasma after cryopreservation. After

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

    Science.gov (United States)

    Van Den Abbeel, E; Van Steirteghem, A

    2000-02-01

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

  19. Chapter 1 Historical Background on Gamete and Embryo Cryopreservation.

    Science.gov (United States)

    Ali, Jaffar; AlHarbi, Naif H; Ali, Nafisa

    2017-01-01

    This chapter describes the development of the science of cryopreservation of gametes and embryos of various species including human. It attempts to record in brief the main contributions of workers in their attempts to cryopreserve gametes and embryos. The initial difficulties faced and subsequent developments and triumphs leading to present-day state of the art are given in a concise manner. The main players and their contributions are mentioned and the authors' aim is to do justice to them. This work also attempts to ensure that credit is correctly attributed for significant advances in gamete and embryo cryopreservation. In general this chapter has tried to describe the historical development of the science of cryopreservation of gametes and embryos as accurately as possible without bias or partiality.

  20. Cryopreservation studies on the marine diatom Navicula subinflata Grun

    Digital Repository Service at National Institute of Oceanography (India)

    Redekar, P.D.; Wagh, A.B.

    of cryoprotectant decreased the growth. These experiments after cryopreservation revealed that the maximum growth was recorded in Ethylene glycol treated cells for 0 day sample and 7 days and 30 days liquid nitrogen preserved samples. No growth was observed...

  1. Vitrification versus slow freezing for women undergoing oocyte cryopreservation

    NARCIS (Netherlands)

    Glujovsky, Demián; Riestra, Barbara; Sueldo, Carlos; Fiszbajn, Gabriel; Repping, Sjoerd; Nodar, Florencia; Papier, Sergio; Ciapponi, Agustín

    2014-01-01

    Oocyte cryopreservation is a technique with considerable potential in reproductive medicine, including fertility preservation, as a way of delaying childbearing and as part of oocyte donation programs. Although the technique was relatively ineffective at first more recently numerous modifications

  2. Cryopreservation of coconut (Cocos nucifera L.) zygotic embryos by vitrification.

    Science.gov (United States)

    Sajini, K K; Karun, A; Amamath, C H; Engelmann, F

    2011-01-01

    The present study investigates the effect of preculture conditions, vitrification and unloading solutions on survival and regeneration of coconut zygotic embryos after cryopreservation. Among the seven plant vitrification solutions tested, PVS3 was found to be the most effective for regeneration of cryopreserved embryos. The optimal protocol involved preculture of embryos for 3 days on medium with 0.6 M sucrose, PVS3 treatment for 16 h, rapid cooling and rewarming and unloading in 1.2 M sucrose liquid medium for 1.5 h. Under these conditions, 70-80 survival (corresponding to size enlargement and weight gain) was observed with cryopreserved embryos and 20-25 percent of the plants regenerated (showing normal shoot and root growth) from cryopreserved embryos were established in pots.

  3. Early investigation on cryopreservation of Dendrobium sonia-28 ...

    African Journals Online (AJOL)

    User

    2011-05-02

    May 2, 2011 ... This study was conducted to determine the potential of cryostoring and regenerating Dendrobium ... Key words: Orchid, protocorm-like bodies, cryopreservation. ... technology is now taken as an ideal alternative propa-.

  4. Magnetic induction heating of superparamagnetic nanoparticles during rewarming augments the recovery of hUCM-MSCs cryopreserved by vitrification.

    Science.gov (United States)

    Wang, Jianye; Zhao, Gang; Zhang, Zhengliang; Xu, Xiaoliang; He, Xiaoming

    2016-03-01

    Cryopreservation by vitrification has been recognized as a promising strategy for long-term banking of living cells. However, the difficulty to generate a fast enough heating rate to minimize devitrification and recrystallization-induced intracellular ice formation during rewarming is one of the major obstacles to successful vitrification. We propose to overcome this hurdle by utilizing magnetic induction heating (MIH) of magnetic nanoparticles to enhance rewarming. In this study, superparamagnetic (SPM) Fe3O4 nanoparticles were synthesized by a chemical coprecipitation method. We successfully applied the MIH of Fe3O4 nanoparticles for rewarming human umbilical cord matrix mesenchymal stem cells (hUCM-MSCs) cryopreserved by vitrification. Our results show that extracellular Fe3O4 nanoparticles with MIH may efficiently suppress devitrification and/or recrystallization during rewarming and significantly improve the survival of vitrified cells. We further optimized the concentration of Fe3O4 nanoparticles and the current of an alternating current (AC) magnetic field for generating the MIH to maximize cell viability. Our results indicate that MIH in an AC magnetic field with 0.05% (w/v) Fe3O4 nanoparticles significantly facilitates rewarming and improves the cryopreservation outcome of hUCM-MSCs by vitrification. The application of MIH of SPM nanoparticles to achieve rapid and spatially homogeneous heating is a promising strategy for enhanced cryopreservation of stem cells by vitrification. Here we report the successful synthesis and application of Fe3O4 nanoparticles for magnetic induction heating (MIH) to enhance rewarming of vitrification-cryopreserved human umbilical cord matrix mesenchymal stem cells (hUCM-MSCs). We found that MIH-enhanced rewarming greatly improves the survival of vitrification-cryopreserved hUCM-MSCs. Moreover, the hUCM-MSCs retain their intact stemness and multilineage potential of differentiation post cryopreservation by vitrification with the

  5. Comparison of the Developmental Potential and Clinical Results of In Vivo Matured Oocytes Cryopreserved with Different Vitrification Media

    Directory of Open Access Journals (Sweden)

    Mei Li

    2015-01-01

    Full Text Available Background: Oocyte vitrification is widely used throughout the world, but its clinical efficacy is inconsistent and depends on the vitrification media. This study compared the developmental potential and clinical results of in vivo matured oocytes cryopreserved with different vitrification media. Methods: This retrospective study involved vitrified-warmed oocytes at one in vitro fertilization laboratory. Vitrification media kits comprised the MC kit (ethylene glycol [EG] plus 1,2-propanediol [PROH], the KT kit (EG plus dimethyl sulphoxide [DMSO], and the Modified kit (EG plus DMSO and PROH kit. Rates of oocyte survival and subsequent developmental potential were recorded and analyzed. The t-test and the Chi-square test were used to evaluate each method′s efficacy. Results: Oocyte survival rate was significantly higher for the Modified kit (92.0% than for the MC kit (88.2% (P 0.05. The high-quality embryo rate per warmed oocyte was significantly higher (23.4% in the Modified kit group than in the other groups (P 0.05. Conclusions: Modified vitrification media are efficient for oocyte vitrification and, with further verification, may be able to replace commercially available media in future clinical applications.

  6. Efficacy and prospects for cryopreserved thyroid gland transplantation as a method for treatment of different forms of primary hypothyroidism including postradiation

    International Nuclear Information System (INIS)

    Karachentsev, Yu.I.; Lyakh, I.A.; Khasiev, V.V.

    1999-01-01

    It was established that auto-and allo transplantation of cryopreserved thyroid is a physiologically well grounded technique of treatment and can be an alternative to substitution hormonotherapy of hypothyroidism and can be a main method of treatment in cases when hormone treatment is not effective or is contraindicated.Talking into account the efficiency of cryopreserved thyroid transplantation in patients with primary hypothyroidism,it is possible to predict its positive effect for c postradiation hypothyroidism treatment,if substitution hormonotherapy is not effective

  7. Impact of cryopreservation on tetramer, cytokine flow cytometry, and ELISPOT

    Directory of Open Access Journals (Sweden)

    Morse Michael A

    2005-07-01

    Full Text Available Abstract Background Cryopreservation of PBMC and/or overnight shipping of samples are required for many clinical trials, despite their potentially adverse effects upon immune monitoring assays such as MHC-peptide tetramer staining, cytokine flow cytometry (CFC, and ELISPOT. In this study, we compared the performance of these assays on leukapheresed PBMC shipped overnight in medium versus cryopreserved PBMC from matched donors. Results Using CMV pp65 peptide pool stimulation or pp65 HLA-A2 tetramer staining, there was significant correlation between shipped and cryopreserved samples for each assay (p ≤ 0.001. The differences in response magnitude between cryopreserved and shipped PBMC specimens were not significant for most antigens and assays. There was significant correlation between CFC and ELISPOT assay using pp65 peptide pool stimulation, in both shipped and cryopreserved samples (p ≤ 0.001. Strong correlation was observed between CFC (using HLA-A2-restricted pp65 peptide stimulation and tetramer staining (p Conclusion We conclude that all three assays show concordant results on shipped versus cryopreserved specimens, when using a peptide-based readout. The assays are also concordant with each other in pair wise comparisons using equivalent antigen systems.

  8. Respiratory analysis of coupled mitochondria in cryopreserved liver biopsies

    Directory of Open Access Journals (Sweden)

    Mercedes García-Roche

    2018-07-01

    Full Text Available The aim of this work was to develop a cryopreservation method of small liver biopsies for in situ mitochondrial function assessment. Herein we describe a detailed protocol for tissue collection, cryopreservation, high-resolution respirometry using complex I and II substrates, calculation and interpretation of respiratory parameters. Liver biopsies from cow and rat were sequentially frozen in a medium containing dimethylsulfoxide as cryoprotectant and stored for up to 3 months at −80 °C. Oxygen consumption rate studies of fresh and cryopreserved samples revealed that most respiratory parameters remained unchanged. Additionally, outer mitochondrial membrane integrity was assessed adding cytochrome c, proving that our cryopreservation method does not harm mitochondrial structure. In sum, we present a reliable way to cryopreserve small liver biopsies without affecting mitochondrial function. Our protocol will enable the transport and storage of samples, extending and facilitating mitochondrial function analysis of liver biopsies. Keywords: Cryopreservation, Mitochondria, Biopsy, Oxygen consumption rate, High-resolution respirometry, Mitochondrial function

  9. First production of larvae using cryopreserved sperm: Effects of preservation temperature and cryopreservation on European eel sperm fertilization capacity

    DEFF Research Database (Denmark)

    Asturiano, J.F.; Sørensen, Sune Riis; Perez, L.

    2016-01-01

    Sperm cryopreservation is a useful tool in captive fish reproduction management, that is to synchronize gamete production, especially in the case of species as the European eel, where the time of female spawning readiness is unpredictable. Several protocols to cryopreserve sperm of this species....... Fertilization of two egg batches was attempted. Diluted sperm caused a similar percentage of fertilized eggs and a similar number of embryos and larvae, independently of storage temperature (4 or 20°C). The cryopreserved sperm resulted in a lower percentage of fertilized eggs, but embryos developed and a few...... larvae ('cryolarvae') were obtained 55 h after fertilization in one of the two egg batches. This result evidences that the tested cryopreservation protocol is applicable for eel reproduction management, although improvements will be required to enhance fertilization success...

  10. Bentall Procedure Using Cryopreserved Valved Aortic Homografts

    Science.gov (United States)

    Christenson, Jan T.; Sierra, Jorge; Trindade, Pedro T.; Didier, Dominique; Kalangos, Afksendiyos

    2004-01-01

    The Bentall procedure is the standard operation for patients who have lesions of the ascending aorta associated with aortic valve disease. In many cases, however, mechanical prosthetic conduits are not suitable. There are few reports in the English-language medical literature concerning the mid- to long-term outcome of Bentall operations with cryopreserved homografts. Therefore, we reviewed our experience with this procedure and valved homografts. From January 1997 through December 2002, 21 patients underwent a Bentall operation with cryopreserved homografts at our institution. There were 14 males and 7 females; the mean age was 36 ± 21 years (range, 15–74 years). Eleven patients had undergone previous aortic valve surgery. All patients had aortic dilatation or aneurysms involving the ascending aorta. Indications for surgery included aortic valve stenosis or insufficiency, and aortic valve endocarditis (native valve or prosthetic). One patient had Takayasu's arteritis and 3 had Marfan syndrome. There was 1 hospital death (due to sepsis), but no other major postoperative complications. The mean hospital stay was 14 ± 7 days. Follow-up echocardiographic and computed tomographic scans were performed yearly. The mean follow-up was 34 months (6–72 months). Follow-up imaging revealed no calcifications or degenerative processes related to the homograft. Four patients had minimal valve regurgitation. Two patients died during follow-up. The 3-year actuarial survival rate was 85.7%. Our data suggest that the Bentall procedure with a valved homograft conduit is a safe procedure with excellent mid- to long-term results, comparable to results reported with aortic valve replacement with a homograft. PMID:15745290

  11. Cryopreservation of ectomycorrhizal fungi has minor effects on root colonization of Pinus sylvestris plantlets and their subsequent nutrient uptake capacity.

    Science.gov (United States)

    Crahay, Charlotte; Wevers, Jan; Munaut, Françoise; Colpaert, Jan V; Declerck, Stéphane

    2013-08-01

    The use of ectomycorrhizal (ECM) fungi for afforestation, bioremediation, and timber production requires their maintenance over long periods under conditions that preserve their genetic, phenotypic, and physiological stability. Cryopreservation is nowadays considered as the most suitable method to maintain the phenotypic and genetic stability of a large number of filamentous fungi including the ECM fungi. Here, we compared the ability of eight ECM fungal isolates to colonize Pinus sylvestris roots and to transport inorganic phosphate (Pi) and NH4 (+) from the substrate to the plant after cryopreservation for 6 months at -130 °C or after storage at 4 °C. Overall, the mode of preservation had no significant effect on the colonization rates of P. sylvestris, the concentrations of ergosterol in the roots and substrate, and the uptake of Pi and NH4 (+). Comparing the isolates, differences were sometimes observed with one or the other method of preservation. Suillus bovinus exhibited a reduced ability to form mycorrhizas and to take up Pi following cryopreservation, while one Suillus luteus isolate exhibited a decreased ability to take up NH4 (+). Conversely, Hebeloma crustuliniforme, Laccaria bicolor, Paxillus involutus, and Pisolithus tinctorius exhibited a reduced ability to form mycorrhizas after storage at 4 °C, although this did not result in a reduced uptake of Pi and NH4 (+). Cryopreservation appeared as a reliable method to maintain important phenotypic characteristics (i.e., root colonization and nutrient acquisition) of most of the ECM fungal isolates studied. For 50 % of the ECM fungal isolates, the colonization rate was even higher with the cultures cryopreserved at -130 °C as compared to those stored at 4 °C.

  12. Modification of trout sperm membranes associated with activation and cryopreservation. Implications for fertilizing potential

    Science.gov (United States)

    Abstract We investigated the effects of two trout sperm activation solutions on sperm physiology and membrane organization prior to and following cryopreservation using flow cytometry and investigated their impact on in vitro fertility. Cryopreservation caused greater phospholipid disorder (high pl...

  13. Determination of metabolic stability using cryopreserved hepatocytes from rainbow trout (Oncorhynchus mykiss)

    Science.gov (United States)

    Standard protocols for isolating, cryopreserving, and thawing rainbow trout hepatocytes are described, along with procedures for using fresh or cryopreserved hepatocytes to assess chemical metabolic stability in fish by means of a substrate depletion approach. Variations on thes...

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

    Science.gov (United States)

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

    2014-04-01

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

  15. Cryopreservation of Indian red jungle fowl (Gallus gallus murghi) semen.

    Science.gov (United States)

    Rakha, B A; Ansari, M S; Akhter, S; Hussain, I; Blesbois, E

    2016-11-01

    The population of red jungle fowl is declining and needs special attention for its conservation with suitable approaches. For ex situ in vitro conservation of Indian red jungle fowl, establishment of semen cryobank is an appropriate option, for which an extender with adequate retrieval capacity for functional spermatozoa is required. Therefore, studies were designed to evaluate a wide range of extenders for cryopreservation of Indian red jungle fowl (Gallus gallus murghi) sperm to achieve maximal post-thawed semen quality and fertility. For this purpose, semen from eight mature cocks were collected, initially evaluated (percent sperm motility, volume and concentration), pooled, assessed for motility, plasma membrane integrity, viability and acrosome integrity, and divided into six aliquots for dilution (1:5; 37°C) in Beltsville poultry, red fowl extender, Lake, EK, Tselutin poultry and chicken semen extenders. Diluted semen was cooled from 37°C to 4°C @ -0.275°C/min. Glycerol (20%) was added to chilled semen, equilibrated for 10min, filled in 0.5mL French straws, kept over LN 2 vapours for 10min and plunged into LN 2 and stored at -196°C. Percentages of motility, plasma membrane integrity, viability and acrosome integrity were higher (Psemen extender. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Migration of fresh and cryopreserved human spermatozoa in polyacrylamide gel.

    Science.gov (United States)

    Goldstein, M C; Wix, L S; Foote, R H; Feldschuh, R; Feldschuh, J

    1982-05-01

    The ability of freshly collected and frozen human spermatozoa to migrate in round capillary tubes containing specially formulated polyacrylamide gel was investigated, using 33 ejaculates from 27 donors. Each semen sample was divided; one portion was left undiluted, and the other portion was diluted to 50 x 10(6) sperm/ml. Glycerol was used as the cryoprotectant. The percentage of motile sperm cells was determined before and after freezing. Fresh semen contained a higher percentage of motile cells, which migrated farther than those of cryopreserved-thawed semen. Various correlations between the percentage of motile sperm and migration distance ranged from 0.57 to 0.62. There was a low positive correlation of migration distance with sperm cell concentration per milliliter, r = 0.25 to 0.34; and thus adjusting semen samples to a standard sperm concentration improved the accuracy of the test only slightly. The regression coefficient of migration distance on the percentage of motile sperm in fresh semen was 0.65, indicating that for each 10% increase in sperm motility, migration distance is predicted to increase 6.5 mm. Five batches of polyacrylamide gel gave uniform results, and the application of this stable gel to fertility investigations is discussed.

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

    Directory of Open Access Journals (Sweden)

    So-Min Hwang

    2015-03-01

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

  18. Cryopreservation of citrus seed via dehydration followed by immersion in liquid nitrogen

    Science.gov (United States)

    An important method for plant germplasm conservation is offered by a biotechnology-based approach of cryopreservation. Cryopreservation refers to the storage of plant material at ultralow temperatures in liquid nitrogen. A procedure for cryopreservation of polyembryonic seeds was improved for select...

  19. Towards higher transparency and efficiency in energy taxation. Energy taxation and environmental policy in a small open economy

    International Nuclear Information System (INIS)

    Normann, Goeran

    2002-01-01

    The accrual of energy taxation has led to a complex structure of taxes and charges that are characterized by instability and low efficiency. Other reasons for analyzing the system is the pressure from our contractual responsibilities within the European Union and the raised ambitions in the environmental policy. The report leads to the conclusion that it would be motivated to separate fiscal energy taxation from measures to internalize environmental costs that the market does not register. This separation would make it possible to create a more transparent and rational energy taxation. The fiscal energy taxation ought to be a broad, value-based tax, equal for all energy sources. Value-based means, besides the energy content in kWh, also properties such as conversion and distribution costs. Two alternatives are suggested for the fiscal energy taxation: A separate consumption tax on energy. Such a tax would amount to 48% to produce the same income as the fiscal elements of today's energy taxes. Another alternative would be to include the fiscal energy tax in the value added tax. This would raise the standard VAT level to 30%, if the lower VAT levels are kept unchanged. With this model, consumption of energy would be treated as any other consumption. The environmental policy measures against greenhouse gases should be delt with through a system with international trade with emission quotas for such gases. Measures against other external effects from energy use are not suggested in this report, except for the opinion that economic incentives are preferable to regulations. The initial allocation of quotas ought to be done through an auction, since this method would give lower national costs than the alternatives. The system should cover all greenhouse gases and (almost) all sources which indicates that an upstream solution would be best with low administrative costs. A safety vent should be considered, so that extreme costs for CO 2 -emissions are avoided, if e.g. the

  20. Development of a low-cost biogas filtration system to achieve higher-power efficient AC generator

    Science.gov (United States)

    Mojica, Edison E.; Ardaniel, Ar-Ar S.; Leguid, Jeanlou G.; Loyola, Andrea T.

    2018-02-01

    The paper focuses on the development of a low-cost biogas filtration system for alternating current generator to achieve higher efficiency in terms of power production. A raw biogas energy comprises of 57% combustible element and 43% non-combustible elements containing carbon dioxide (36%), water vapor (5%), hydrogen sulfide (0.5%), nitrogen (1%), oxygen (0 - 2%), and ammonia (0 - 1%). The filtration system composes of six stages: stage 1 is the water scrubber filter intended to remove the carbon dioxide and traces of hydrogen sulfide; stage 2 is the silica gel filter intended to reduce the water vapor; stage 3 is the iron sponge filter intended to remove the remaining hydrogen sulfide; stage 4 is the sodium hydroxide solution filter intended to remove the elemental sulfur formed during the interaction of the hydrogen sulfide and the iron sponge and for further removal of carbon dioxide; stage 5 is the silica gel filter intended to further eliminate the water vapor gained in stage 4; and, stage 6 is the activated carbon filter intended to remove the carbon dioxide. The filtration system was able to lower the non-combustible elements by 72% and thus, increasing the combustible element by 54.38%. The unfiltered biogas is capable of generating 16.3 kW while the filtered biogas is capable of generating 18.6 kW. The increased in methane concentration resulted to 14.11% increase in the power output. The outcome resulted to better engine performance in the generation of electricity.

  1. Cryopreservation of canine semen - new challenges.

    Science.gov (United States)

    Farstad, W

    2009-07-01

    Egg yolk (EY) protects cell membranes against cold shock, and it prevents or restores the loss of phospholipids from the membrane. EY has been widely used in semen extenders. It has been added to Tris-Glucose buffer and has been widely used for cooling and cryopreservation of canine semen. EY is not a defined entity, but a complex biological compound containing proteins, vitamins, phospholipids, glucose and antioxidants which are all potentially useful for cell membrane integrity. Unfortunately, it also is a biologically hazardous compound. Hence, whole EY needs to be replaced by other chemically defined components for semen processing in dogs. Freezing poor semen does not improve its quality, so attention must be focused on how to cope with dogs whose semen does not freeze well, and on designing individual freezing extenders for semen from such males. Furthermore, differences have been found among canid species in the ability of their spermatozoa to withstand freezing. There are differences in sperm membrane fatty acid composition among species, which may explain part of these differences. If the presence of long-chained polyunsaturated fatty acids contributes to increased membrane fluidity, this relationship may be biphasic, i.e. either too much membrane fluidity, or too little, could compromise successful sperm cryopreservation. An increase in fluidity of the outer leaflet of the plasma membrane has been shown in frozen thawed dog spermatozoa. The protective effect of exogenous lipids may lie in close association with the membrane rather than in modification or rearrangement of the membrane. This also points at lipids as an important, if not entirely new group of substances, which may substitute standard EY-based diluents in preserving sperm survival during freezing. EY-derived phospholipids or lecithin could be used to replace whole EY. Vegetable lecithin is currently investigated to avoid using substances of animal origin. EY also contains antioxidants which

  2. Novel and potential application of cryopreservation to plant genetic transformation.

    Science.gov (United States)

    Wang, Biao; Zhang, Zhibo; Yin, Zhenfang; Feng, Chaohong; Wang, Qiaochun

    2012-01-01

    The world population now is 6.7 billion and is predicted to reach 9 billion by 2050. Such a rapid growing population has tremendously increased the challenge for food security. Obviously, it is impossible for traditional agriculture to ensure the food security, while plant biotechnology offers considerable potential to realize this goal. Over the last 15 years, great benefits have been brought to sustainable agriculture by commercial cultivation of genetically modified (GM) crops. Further development of new GM crops will with no doubt contribute to meeting the requirements for food by the increasing population. The present article provides updated comprehensive information on novel and potential application of cryopreservation to genetic transformation. The major progresses that have been achieved in this subject include (1), long-term storage of a large number of valuable plant genes, which offers a good potential for further development of novel cultivars by genetic transformation; (2), retention of regenerative capacity of embryogenic tissues and protoplasts, which ensures efficient plant regeneration system for genetic transformation; (3), improvement of transformation efficiency and plant regeneration of transformed cells; (4), long-term preservation of transgenic materials with stable expression of transgenes and productive ability of recombinant proteins, which allows transgenic materials to be stored in a safe manner before being analyzed and evaluated, and allows establishment of stable seed stocks for commercial production of homologous proteins. Data provided in this article clearly demonstrate that cryo-technique has an important role to play in the whole chain of genetic transformation. Further studies coupling cryotechnique and genetic transformation are expected to significantly improve development of new GM crops. Copyright © 2011 Elsevier Inc. All rights reserved.

  3. Cryopreservation of human colorectal carcinomas prior to xenografting

    International Nuclear Information System (INIS)

    Linnebacher, Michael; Maletzki, Claudia; Ostwald, Christiane; Klier, Ulrike; Krohn, Mathias; Klar, Ernst; Prall, Friedrich

    2010-01-01

    Molecular heterogeneity of colorectal carcinoma (CRC) is well recognized, forming the rationale for molecular tests required before administration of some of the novel targeted therapies that now are rapidly entering the clinics. For clinical research at least, but possibly even for future individualized tumor treatment on a routine basis, propagation of patients' CRC tissue may be highly desirable for detailed molecular, biochemical or functional analyses. However, complex logistics requiring close liaison between surgery, pathology, laboratory researchers and animal care facilities are a major drawback in this. We here describe and evaluate a very simple cryopreservation procedure for colorectal carcinoma tissue prior to xenografting that will considerably reduce this logistic complexity. Fourty-eight CRC collected ad hoc were xenografted subcutaneously into immunodeficient mice either fresh from surgery (N = 23) or after cryopreservation (N = 31; up to 643 days). Take rates after cryopreservation were satisfactory (71%) though somewhat lower than with tumor tissues fresh from surgery (74%), but this difference was not statistically significant. Re-transplantation of cryopreserved established xenografts (N = 11) was always successful. Of note, in this series, all of the major molecular types of CRC were xenografted successfully, even after cryopreservation. Our procedure facilitates collection, long-time storage and propagation of clinical CRC specimens (even from different centres) for (pre)clinical studies of novel therapies or for basic research

  4. Cryopreservation of GABAergic Neuronal Precursors for Cell-Based Therapy.

    Directory of Open Access Journals (Sweden)

    Daniel Rodríguez-Martínez

    Full Text Available Cryopreservation protocols are essential for stem cells storage in order to apply them in the clinic. Here we describe a new standardized cryopreservation protocol for GABAergic neural precursors derived from the medial glanglionic eminence (MGE, a promising source of GABAergic neuronal progenitors for cell therapy against interneuron-related pathologies. We used 10% Me2SO as cryoprotectant and assessed the effects of cell culture amplification and cellular organization, as in toto explants, neurospheres, or individualized cells, on post-thaw cell viability and retrieval. We confirmed that in toto cryopreservation of MGE explants is an optimal preservation system to keep intact the interneuron precursor properties for cell transplantation, together with a high cell viability (>80% and yield (>70%. Post-thaw proliferation and self-renewal of the cryopreserved precursors were tested in vitro. In addition, their migration capacity, acquisition of mature neuronal morphology, and potency to differentiate into multiple interneuron subtypes were also confirmed in vivo after transplantation. The results show that the cryopreserved precursor features remained intact and were similar to those immediately transplanted after their dissection from the MGE. We hope this protocol will facilitate the generation of biobanks to obtain a permanent and reliable source of GABAergic precursors for clinical application in cell-based therapies against interneuronopathies.

  5. Social oocyte cryopreservation: a portrayal of Brazilian women.

    Science.gov (United States)

    Santo, Elisangela V Espirito; Dieamant, Felipe; Petersen, Claudia G; Mauri, Ana L; Vagnini, Laura D; Renzi, Adriana; Zamara, Camila; Oliveira, João Batista A; Baruffi, Ricardo L R; Franco, José G

    2017-06-01

    This study aimed to determine what Brazilian childless women of reproductive age think about oocyte cryopreservation to postpone pregnancy and their reasons for performing or not performing this procedure. Women of reproductive age were randomly selected from the general population using different e-mail lists and were invited to participate in the study by completing an online web survey regarding social oocyte cryopreservation. The survey was also distributed through social media to women of reproductive age. Although most of the responders had a partner (86.9%) and had already planned the pregnancy of their first child (69.6%), 85.4% (379) considered the potential of social oocyte freezing to improve their chances of giving birth later in life. Those that had already planned pregnancy were two times more likely to intend to freeze their oocytes (p=0.03). The most important barrier for not undergoing oocyte cryopreservation was cost. The women who indicated that they could not currently undergo the procedure now because of cost were two times (p=0.03) more likely to intend to cryopreserve their oocytes than women who thought that they would not need to delay pregnancy. Brazilian women who think that they are not ready to have a family are discovering the option of oocyte cryopreservation. Most participants considered safeguarding their reproductive potential. Making the procedure more accessible could give women the opportunity to make proactive decisions about the future of their fertility.

  6. The effect of the melatonin on cryopreserved mouse testicular cells

    Directory of Open Access Journals (Sweden)

    Ghasem Saki

    2016-01-01

    Full Text Available Background: After improvements in various cancer treatments, life expectancy has been raised, but success in treatment causes loss of fertility in many of the survived young men. Cryopreservation of immature testicular tissues or cells introduced as the only way to preserve fertility. However, freezing has some harmful effects. Melatonin, a pineal gland hormone, has receptors in reproductive systems of different species. It is assumed that melatonin has free radical scavenger properties. Objective: The aim of this study was to evaluate the effects of melatonin on the cryopreserved testicular cells in mouse. Materials and Methods: Cells from 7- 10 days old NMRI mice testes were isolated using two step enzymatic digestion. The testicular cells were divided into two groups randomly and cryopreserved in two different freezing media with and without the addition of 100 μm melatonin. Finally, apoptosis of the cells was assayed by flow cytometry. Also, lactate dehydrogenase activity test was performed to assess the cytotoxicity. Results: The results of lactate dehydrogenase showed the nearly cytotoxic effect of melatonin. The results of flow cytometry showed increase in apoptosis in the cryopreserved cells in the media containing melatonin compared to the control group. Conclusion: The present study shows that melatonin has an apoptotic effect on cryopreserved mouse testicular cells.

  7. Thixotropic injectable hydrogel using a polyampholyte and nanosilicate prepared directly after cryopreservation

    Energy Technology Data Exchange (ETDEWEB)

    Jain, Minkle; Matsumura, Kazuaki, E-mail: mkazuaki@jaist.ac.jp

    2016-12-01

    Success of tissue engineering applications in regenerative medicine requires the preservation of tissue-engineered products at a low temperature. This can be successfully achieved by the use of cryoprotective agent (CPA). In this study, we formulated a unique injectable hydrogel for the purpose of cell delivery after cryopreservation by using polyampholyte CPA. The polyampholyte showed excellent post-thaw cell survival, and after thawing, the polymeric CPA did not have to be removed because of its low cytotoxicity. The polyampholyte could be transformed into a hydrogel by mixing with nanosilicates. Previously, nanosilicates were used to improve mechanical properties, but this is the first report of the use of a nanosilicate together with CPA to formulate hydrogels. Inclusion of the nanosilicate led to the formation of thixotropic hydrogels, which can be injected using fine needles. These gels with tunable mechanical properties can be injected into defect sites to form scaffolds for cell growth and tissue repair, and they do not require any separate seeding of cells before injection, thus eliminating the need for cell harvesting and cell maintenance. This is a distinct system in which cells can be cryopreserved until before usage; when required, the cells in the polyampholyte can be revived to their original state and the thixotropic hydrogel can be formed. The combination of thixotropy and cytocompatibility of the gels could enable a wide range of biomedical applications such as cell delivery and orthopedic repair. - Graphical abstract: A novel thixotropic, cytocompatible and injectable nanocomposite hydrogel with tunable mechanical properties directly after cryopreservation was formulated using an efficient polymer cryoprotectant and laponite. This is an efficient system in which cells remain viable and can proliferate even after one week. The combined use of thixotropy and cytocompatibility enables this system to be used for cell delivery applications

  8. Novel glyceryl glucoside is a low toxic alternative for cryopreservation agent

    Energy Technology Data Exchange (ETDEWEB)

    Su, Cathy; Allum, Allison J. [Department of Environmental & Radiological Health Sciences, Colorado State University, 1618 Campus Delivery, Fort Collins, CO 80523 (United States); Aizawa, Yasushi [Research and Development Group, Toyo Sugar Refining Co. Ltd., Tokyo 103-0046 (Japan); Kato, Takamitsu A., E-mail: Takamitsu.Kato@Colostate.edu [Department of Environmental & Radiological Health Sciences, Colorado State University, 1618 Campus Delivery, Fort Collins, CO 80523 (United States)

    2016-08-05

    Glyceryl glucoside (GG, α-D-glucosyglycerol) is a natural glycerol derivative found in alcoholic drinks. Recently GG has been used as an alternative for glycerol in cosmetic products. However, the safety of using GG is still unclear. Currently, dimethyl sulfoxide (DMSO) and glycerol are wildly used in cryopreservation. Despite GG being a derivative of glycerol, the ability of GG in cryopreservation is still unknown. By using a system of Chinese Hamster Ovary cells (CHO), A549 cells and AG1522 cells, the study examined the cryoprotective effects of DMSO, glycerol and GG. Cytotoxic and genotoxic responses induced by the three chemicals were also investigated with CHO to determine the safety of GG for cosmetic products. Our data suggests that GG has great cryopresearvation ability in the concentration of 30%–40% (v/v). For cytotoxic studies, DMSO showed the highest cytotoxicity above 3% (v/v) in cell doubling time delay among three chemicals. For the acute cytotoxicity with trypan blue dye exclusion assay, GG showed stronger cell killing effect within 24 h above 4% (v/v). For the continuous cytotoxicity with colony formation assay for 7 days, DMSO showed significantly reduced clonogenic ability above 2%. In genotoxicity studies, CHO treated with glycerol at 2% concentration induced three times higher frequencies of sister chromatid exchange (SCE) than background levels. GG did not induce significant amounts of SCE compared to background. Micronuclei formation was equally observed in the 2% and above concentrations of glycerol and GG. Our data showed that GG has significant effects on cryopreservation compared to DMSO. Glycerol and GG have similar cytotoxicity effects to CHO, but glycerol induced genotoxic responses in the same concentration. Therefore, we conclude that GG may be a safer alternative compound to glycerol in cosmetic products and safer alternative to DMSO in cryopreservation. -- Highlights: •Glyceryl Glucoside is low cytotoxicity and genotoxicity

  9. Cryopreservation of Tetraclinis articulata (vahl.) Masters.

    Science.gov (United States)

    Serrano-Martinez, Francisco; Casas, José Luis

    2011-01-01

    Tetraclinis articulata shoot tips excised from in vitro grown shoots were cryopreserved using a modified PVS2-based vitrification protocol. Preliminary experiments with non-cryostored shoot tips showed that the high concentrations of sucrose in loading (LS), vitrification (PVS2) and unloading (US) solutions employed in the protocol were very toxic for the explants. Replacement of sucrose by sorbitol in equal molar concentration in all these solutions enhanced survival of shoot tips after all treatments. However, cold-hardening of donor shoots before shoot tip excision was strictly required to obtain post-rewarming survival. Therefore, the protocol was outlined as follows: pre-conditioning of explants at 4 degree C for 3 weeks in the dark; excision of 1 mm long shoot tips; loading for 20 min in modified LS at room temperature; dehydration in modified PVS2 at 0 degree C for 60 min; immersion in liquid nitrogen (LN); rewarming at 40 degree C for 2 min and subsequent transfer of shoot tips in modified US for 20 min.

  10. Double perovskites overtaking the single perovskites: A set of new solar harvesting materials with much higher stability and efficiency

    Science.gov (United States)

    Kangsabanik, Jiban; Sugathan, Vipinraj; Yadav, Anuradha; Yella, Aswani; Alam, Aftab

    2018-05-01

    Solar energy plays an important role in substituting the ever declining source of fossil fuel energy. Finding novel materials for solar cell applications is an integral part of photovoltaic research. Hybrid lead halide perovskites are one of, if not the most, well sought material in the photovoltaic research community. Its unique intrinsic properties, flexible synthesis techniques, and device fabrication architecture made the community highly buoyant over the past few years. Yet, there are two fundamental issues that still remain a concern, i.e., the stability in external environment and the toxicity due to Pb. This led to a search for alternative materials. More recently, double perovskite [A2B B'X6 (X =Cl, Br, I)] materials have emerged as a promising choice. Few experimental synthesis and high throughput computational studies have been carried out to check for promising candidates of this class. The main outcome from these studies, however, can essentially be summarized into two categories: (i) either they have an indirect band gap or (ii) a direct but large optical band gap, which is not suitable for solar devices. Here we propose a large set of stable double perovskite materials, Cs2B B 'X6 (X =Cl, Br, I), which show indirect to direct band gap transition via small Pb+2 doping at both B and B'sites. This is done by careful band (orbital) engineering using first-principles calculations. This kind of doping has helped to change the topology of the band structure, triggering an indirect to direct transition that is optically allowed. It also reduces the band gap significantly, bringing it well into the visible region. We also simulated the optical absorption spectra of these systems and found a comparable/higher absorption coefficient and solar efficiency with respect to the state of the art photovoltaic absorber material CH3NH3PbI3 . A number of materials Cs2(B0.75Pb0.25) (B0.75'Pb0.25) X6 (for various combinations of B ,B ', and X ) are found to be promising

  11. Eggs on Ice. Imaginaries on Eggs and Cryopreservation in Denmark

    DEFF Research Database (Denmark)

    Herrmann, Janne Rothmar; Kroløkke, Charlotte

    2018-01-01

    While Denmark is widely known as a global exporter of cryopreserved sperm, Danish women’s eggs follow very different trajectories. This paper combines legal and rhetorical analyses with the concept of sociotechnical imaginaries (Jasanoff, 2015). In establishing the genealogy of the sociotechnical...... imaginaries that shaped the Danish regulation on the cryopreservation of eggs, we analyze the relevant Acts, Bills, preparatory work and readings in Parliament along with the concurrent public and ethical debates that in time relaxed the legal limit for the cryopreservation of eggs to the current 5 years...... and today continue to ignite discussions on elective egg freezing. We rely on welfare state perspectives to discuss why reproduction, in the Danish context, is seen as a legitimate and appropriate sphere to regulate and we turn to feminist theorizing to discuss their gendered implications captured...

  12. Proximate composition analysis posterior to the cryopreservation of Chaetoceros calcitrans

    Directory of Open Access Journals (Sweden)

    Joan Salas-Leiva

    2015-12-01

    Full Text Available Objective. The effect of cryopreservation on the proximate composition of microalgae Chaetoceros calcitrans was evaluated. Materials and methods. C. calcitrans was cultured and cryopreserved using 5% (v/v dimethylsulfoxide as cryoprotectant. The freezing was controlled at a rate of 3°C/min, up to -60°C and then the microalgae were immersed in liquid nitrogen (-196°C. After storage in liquid nitrogen, microalgae were rapidly thawed out and subcultured. The percentage of proteins, lipids and carbohydrates was analyzed using absorption spectrophotometry and the organic matter was studied by gravimetric analysis. Results. There was no significant variation between the proximate composition of C. calcitrans cryopreserved and the controls (p>0.05. Conclusion. Our results show that, despite low cell recovery after the preservation of this organism at low temperatures, there is no apparent loss of nutritional characteristics caused by the storing process at low temperatures.

  13. Cryopreservation of roe deer abomasal nematodes for morphological identification.

    Science.gov (United States)

    Beraldo, Paola; Pascotto, Ernesto

    2014-02-01

    Conventional methods to preserve adult nematodes for taxonomic purposes involve the use of fixative or clearing solutions (alcohol, formaldehyde, AFA and lactophenol), which cause morphological alterations and are toxic. The aim of this study is to propose an alternative method based on glycerol-cryopreservation of nematodes for their subsequent identification. Adults of trichostrongylid nematodes from the abomasum of roe deer (Capreolus capreolus Linnaeus) were glycerol-cryopreserved and compared with those fixed in formaldehyde, fresh and frozen without cryoprotectans. Morphology, transparency and elasticity of the anterior and posterior portion of male nematodes were compared, especially the caudal cuticular bursa and genital accessories. The method presented is quick and easy to use, and the quality of nematode specimens is better than that of nematodes fixed by previously used fixatives. Moreover, glycerol cryopreserved nematodes can be stored for a long time at -20 degrees C in perfect condition and they could be suitable for further analyses, such as histological or ultrastructural examinations.

  14. Fertility preservation for girls and young women with cancer: population-based validation of criteria for ovarian tissue cryopreservation.

    Science.gov (United States)

    Wallace, W Hamish B; Smith, Alice Grove; Kelsey, Thomas W; Edgar, Angela E; Anderson, Richard A

    2014-09-01

    assessable patients who had declined the procedure, one had developed premature ovarian insufficiency. Assessment of ovarian function was possible for 141 of the 376 patients who were not offered cryopreservation; one of these patients had developed premature ovarian insufficiency. The cumulative probability of developing premature ovarian insufficiency after treatment was completed was significantly higher for patients who met the criteria for ovarian tissue cryopreservation than for those who did not (15-year probability 35% [95% CI 10-53] vs 1% [0-2]; pgirls and young women who will develop premature ovarian insufficiency, and validate their use for selection of patients for ovarian tissue cryopreservation. Further follow-up of this cohort of patients is likely to allow refinement of the criteria for this experimental procedure in girls and young women with cancer. UK Medical Research Council. Copyright © 2014 Wallace et al. Open Access article distributed under the terms of CC BY-NC-ND. Published by Elsevier Ltd. All rights reserved.

  15. Fertility in cancer patients after cryopreservation of one ovary

    DEFF Research Database (Denmark)

    Schmidt, K T; Andersen, Anders Nyboe; Greve, T

    2013-01-01

    This questionnaire study describes the fertility and ovarian function in 143 adult female cancer survivors with only one ovary due to cryopreservation of the other. The women were asked about their ovarian function (as defined by the presence of a spontaneous menstrual cycle), pregnancies...... and their outcome. The mean follow-up time was 58months after cryopreservation (range 24-129months). The risk of premature ovarian failure was high in the group of patients with leukaemia (13/15; 87%) but low in the breast cancer group (5/54; 9%). Fifty-seven women had actively tried to become pregnant after end...

  16. The Effect of L-Carnitine, Hypotaurine, and Taurine Supplementation on the Quality of Cryopreserved Chicken Semen.

    Science.gov (United States)

    Partyka, Agnieszka; Rodak, Olga; Bajzert, Joanna; Kochan, Joanna; Niżański, Wojciech

    2017-01-01

    The objective of this study was to investigate the effect of L-carnitine (LC), hypotaurine (HT), and taurine (T) on the quality of frozen-thawed chicken semen. Pooled semen samples were divided into seven aliquots (control, 1 mM LC, 5 mM LC, 1 mM HT, 10 mM HT, 1 mM T, and 10 mM T) and subjected to cryopreservation. Postthaw sperm motility was determined by IVOS system and sperm characteristics were assessed with fluorochromes and flow cytometry. The highest sperm motility and the highest percentage of viable sperm were in the HT1 group ( P < 0.01 and P < 0.05) following cryopreservation. After thawing, we observed a higher percentage of sperm without apoptosis and membrane reorganization changes in the LC1 and T1 group when compared to the control ( P < 0.05). There was a higher percentage of live sperm without lipid peroxidation (LPO) in all treatments ( P < 0.01; P < 0.05), when compared to the control group. The percentage of sperm with high mitochondrial potential significantly increased with LC1, T1, and T10 ( P < 0.05). Supplementation of the diluent with LC1, LC5, and T1 significantly ( P < 0.05) reduced DNA susceptibility to fragmentation, compared to the control and HT1 groups. These results indicate that the addition of examined antioxidants improves the quality of cryopreserved chicken semen.

  17. Thermomechanical Stress in Cryopreservation Via Vitrification With Nanoparticle Heating as a Stress-Moderating Effect.

    Science.gov (United States)

    Eisenberg, David P; Bischof, John C; Rabin, Yoed

    2016-01-01

    This study focuses on thermomechanical effects in cryopreservation associated with a novel approach of volumetric heating by means on nanoparticles in an alternating electromagnetic field. This approach is studied for the application of cryopreservation by vitrification, where the crystalline phase is completely avoided-the cornerstone of cryoinjury. Vitrification can be achieved by quickly cooling the material to cryogenic storage, where ice cannot form. Vitrification can be maintained at the end of the cryogenic protocol by quickly rewarming the material back to room temperature. The magnitude of the rewarming rates necessary to maintain vitrification is much higher than the magnitude of the cooling rates that are required to achieve it in the first place. The most common approach to achieve the required cooling and rewarming rates is by exposing the specimen's surface to a temperature-controlled environment. Due to the underlying principles of heat transfer, there is a size limit in the case of surface heating beyond which crystallization cannot be prevented at the center of the specimen. Furthermore, due to the underlying principles of solid mechanics, there is a size limit beyond which thermal expansion in the specimen can lead to structural damage and fractures. Volumetric heating during the rewarming phase of the cryogenic protocol can alleviate these size limitations. This study suggests that volumetric heating can reduce thermomechanical stress, when combined with an appropriate design of the thermal protocol. Without such design, this study suggests that the level of stress may still lead to structural damage even when volumetric heating is applied. This study proposes strategies to harness nanoparticles heating in order to reduce thermomechanical stress in cryopreservation by vitrification.

  18. A Matching Activity When Entering Higher Education: Ongoing Guidance for the Students or Efficiency Instrument for the School?

    Science.gov (United States)

    Mittendorff, Kariene; Faber, Marike; Staman, Laura

    2017-01-01

    In order to lower dropout rates and stimulate student success in higher education, the Dutch government implemented a new law demanding that every higher education institute offer a matching activity to applying students. This article evaluates how students and teachers experience this matching activity. Data were collected in a Dutch university…

  19. Cryopreservation of yamú (Brycon amazonicus) sperm for large scale fertilization

    DEFF Research Database (Denmark)

    Velasco-Santamaría, Yohana M.; Medina-Robles, Mauricio; Cruz-Casallas, Pablo E.

    2006-01-01

      To determine the effect of straw size and thawing temperature on cryopreserved sperm quality of yamú (Brycon amazonicus), ovulation and spermiation were induced in sexually mature broodstock using Carp Pituitary Extract. Sperm quality was evaluated by motility, activation time and fertility...... assays consisted of 40 g eggs inseminated with approximately 5.0 mL (ca. 75,000 motile spermatozoa/egg) of cryopreserved sperm in large straws thawed at 35 °C. The fertilization rate was estimated 6 h post-insemination. In all straws, postthaw motility was significantly lower than for fresh sperm (pb0.......05) to sperm frozen in 0.5-mL straws (48±2%, 51±2%, 52±2% and 54±3%, respectively). In large scale fertilization trials, fresh sperm showed a higher (pb0.05) fertilization rate (83±1%) than frozen-thawed sperm (68±1%). Although the fertility percentage with fresh sperm was significantly higher than with frozen...

  20. Lipid content and composition of oocytes from five coral species: potential implications for future cryopreservation efforts.

    Directory of Open Access Journals (Sweden)

    Chiahsin Lin

    Full Text Available Given the previously documented importance of lipid concentration and composition in the successful cryopreservation of gorgonian corals, these parameters were assessed in oocytes of five species of scleractinian coral; Platygyra daedalea, Echinopora gemmacea, Echinophyllia aspera, Oxypora lacera and Astreopora expansa. Wax esters, phosphatidylethanolamine, phosphatidylcholine, and fatty acids were all measured at detectable levels, and the latter were produced at significantly elevated quantities in E. gemmacea, E. aspera, and O. lacera. On the other hand, phosphatidylethanolamine, phosphatidylcholine, and wax ester were found at significantly higher concentrations in A. expansa oocytes. Triacylglycerol was not present in any species. Interestingly, the total lipid content of oocytes from all five scleractinians was significantly lower than that of oocytes of two gorgonian species, Junceella juncea and Junceella fragilis. As higher total lipid concentrations may be correlated with greater degrees of cellular membrane fluidity at lower temperatures, it stands to reason that gorgonian coral oocytes may be more likely to survive the cryopreservation process than oocytes of scleractinian corals.

  1. Higher energy efficiency in administrative and office buildings. Optimisation of primary energy consumption and economic efficiency; Energieeffiziente Buero- und Verwaltungsgebaeude. Hinweise zur primaerenergetischen und wirtschaftlichen Optimierung

    Energy Technology Data Exchange (ETDEWEB)

    Knissel, J.

    1999-12-15

    The study investigates measures to reduce primary energy consumption in administrative and office buildings and their effects in terms of economic efficiency. An exemplary office building is modernised step by step while recording the changes in the primary energy consumption coefficient. [German] In der vorliegenden Studie wird untersucht, wie weit und mit welchen Massnahmen der Primaerenergiebedarf von Buero- und Verwaltungsgebaeuden gesenkt werden kann und welche Auswirkungen dies auf die Wirtschaftlichkeit hat. Hierzu wird die energetische Ausfuehrungsqualitaet eines einfachen Beispielgebaeudes schrittweise verbessert und die Veraenderung des Primaerenergiekennwertes ermittelt. (orig.)

  2. Anti-oxidant supplementation improves boar sperm characteristics and fertility after cryopreservation: comparison between cysteine and rosemary (Rosmarinus officinalis).

    Science.gov (United States)

    Malo, C; Gil, L; Gonzalez, N; Martínez, F; Cano, R; de Blas, I; Espinosa, E

    2010-08-01

    Anti-oxidants partially ameliorated the detrimental effects of reactive oxidative substances produced during cryopreservation. The objective of the study was to determine the effect of anti-oxidant addition to the freezing extender on boar semen qualities and fertility capacity. Ejaculates were collected from a previously selected boar and semen samples were processed using the straw freezing procedure. In experiment 1, semen samples were cryopreserved in lactose-egg yolk solution supplemented with various concentrations of cysteine (0, 5 and 10mM) to determinate a cysteine concentration capable of producing a protective effect during cryopreservation. Semen quality (total motility, progressive motility, viability, acrosome integrity and hypoosmotic swelling test) was evaluated after freezing and thawing and then every hour for 3h. In experiment 2, ejaculates were cryopreserved with lactose-egg yolk extender with or without the following anti-oxidants: cysteine, rosemary (Rosmarinus officinalis) and cysteine plus rosemary. Semen quality was evaluated. In the experiment 3, fertility capacity of semen frozen in anti-oxidant supplementation extenders was examined in vitro. A total of 2232 oocytes were in vitro matured and inseminated with frozen-thawed sperm. In summary: (i) the effective concentration of cysteine in freezing extender was 10mM; (ii) the addition of exogenous rosemary or cysteine to the freezing extender positively affected post-thawed viability and acrosome integrity. Only rosemary supplementation improved total motility at 3h and progressive motility at any time; (iii) the inclusion of rosemary into the extender was effective in penetration and cleavage rate and also in the efficiency of the fertilization system. (c) 2010 Elsevier Inc. All rights reserved.

  3. A method for generation of bone marrow-derived macrophages from cryopreserved mouse bone marrow cells.

    Directory of Open Access Journals (Sweden)

    Fernanda M Marim

    Full Text Available The broad use of transgenic and gene-targeted mice has established bone marrow-derived macrophages (BMDM as important mammalian host cells for investigation of the macrophages biology. Over the last decade, extensive research has been done to determine how to freeze and store viable hematopoietic human cells; however, there is no information regarding generation of BMDM from frozen murine bone marrow (BM cells. Here, we establish a highly efficient protocol to freeze murine BM cells and further generate BMDM. Cryopreserved murine BM cells maintain their potential for BMDM differentiation for more than 6 years. We compared BMDM obtained from fresh and frozen BM cells and found that both are similarly able to trigger the expression of CD80 and CD86 in response to LPS or infection with the intracellular bacteria Legionella pneumophila. Additionally, BMDM obtained from fresh or frozen BM cells equally restrict or support the intracellular multiplication of pathogens such as L. pneumophila and the protozoan parasite Leishmania (L. amazonensis. Although further investigation are required to support the use of the method for generation of dendritic cells, preliminary experiments indicate that bone marrow-derived dendritic cells can also be generated from cryopreserved BM cells. Overall, the method described and validated herein represents a technical advance as it allows ready and easy generation of BMDM from a stock of frozen BM cells.

  4. SPECIFIC FEATURES OF HIGHER EFFICIENCY IN FUNCTIONING OF ROAD-TRANSPORT COMPLEX IN THE REPUBLIC OF BELARUS

    Directory of Open Access Journals (Sweden)

    M. G. Solodkaya

    2015-01-01

    Full Text Available The road-transport complex objectively reflects the essence of efficient transportation process which is carried out by transport facilities along the highways. The complex  emphasizes an equivalent contribution of transport facilities and highways in a unified transportation process. Efficiency of the state economy rigidly depends on availability of the developed and well-functioning network of highways. Countries with the developed economy which have generally finished creation of national highway networks continue to invest money in public road systems that stimulates development of industrial sectors, agriculture and trade, etc. Their progress and efficient functioning is possible only with the balanced, overall development of the road-transport complex of the country. Functioning of the road-transport complex is inextricable connected with the operation of automotive transport and road infrastructure. Interaction of these two components of the unified economic system is determined by technical characteristics of the automotive transport and transport and operational indices of  the highways. Development of methods for optimum organization of management for functioning of the road complex is considered as an important problem of the national economy while forming market economy mechanisms. Further growth of capital expenditures including  investments will be needed in order to ensure such road conditions that meet the requirements of modern and perspective road traffic. Management of the highway network conditions presupposes a selection of such set of regulatory impacts on road conditions which will allow to minimize expenses in the road-transport complex. Elaboration and realization of the most efficient repair measures serve as such regulatory impact. The purpose is achieved while solving the problem pertaining to minimization of expenses on  transportations in the road-transport complex in the process of the realization of the most

  5. Development of a higher-efficiency tubular cavity receiver for direct steam generation on a dish concentrator

    Science.gov (United States)

    Pye, John; Hughes, Graham; Abbasi, Ehsan; Asselineau, Charles-Alexis; Burgess, Greg; Coventry, Joe; Logie, Will; Venn, Felix; Zapata, José

    2016-05-01

    An integrated model for an axisymmetric helical-coil tubular cavity receiver is presented, incorporating optical ray-tracing for incident solar flux, radiosity analysis for thermal emissions, computational fluid dynamics for external convection, and a one-dimensional hydrodynamic model for internal flow-boiling of water. A receiver efficiency of 98.7% is calculated, for an inlet/outlet temperature range of 60-500 °C, which is the ratio of fluid heating to receiver incident irradiance. The high-efficiency design makes effective use of non-uniform flux in its non-isothermal layout, matching lower temperature regions to areas of lower flux. Full-scale testing of the design will occur in late 2015.

  6. Development and application of VISAR probe with higher signal-collecting efficiency and adjusted depth of field

    International Nuclear Information System (INIS)

    Zhao Jianheng; Sun Chengwei; Ma Ruchao

    2001-01-01

    The new design of optical fiber VISAR probes has been described. It consists of optical fibers and two lenses, and has simpler structure and lower cost than others. If the size of the image near the end face of collecting fiber is larger than the diameter of the core fiber, the distance between the probe and the target can be decreased. Requirement for the precision in the design is lower in this way, and is easier to build up. At the same time, the signal-collecting efficiency is improved in some degree. During the process of designing the probe, the technique for manufacturing the lens of plexiglass is developed. The lens of plexiglass is used to replace the lens of glass, which can reduce the cost of the probe. The factors affecting the collecting efficiency are analyzed

  7. Seasonal and cryopreservation impacts on semen quality in boars

    Science.gov (United States)

    Seasonal boar infertility occurs worldwide and contributes to economic loss to the pork industry. The current study evaluated cooled vs cryopreserved semen quality of 11 Duroc boars collected in June (cool season) and August 2014 (warm season). Semen was cooled to 16°C (cooled) or frozen over liquid...

  8. Effect of season on fresh and cryopreserved stallion semen

    Science.gov (United States)

    The objective of this study was to determine the effect of season on sperm quality parameters, expression of the fertility-related protein SP22 and selected mRNA transcripts infresh and cryopreserved stallion sperm. Four stallions were collected in each of the four seasons: summ...

  9. Sperm cryopreservation before cancer treatment: a 15-year monocentric experience.

    Science.gov (United States)

    Bizet, P; Saias-Magnan, J; Jouve, E; Grillo, J M; Karsenty, G; Metzler-Guillemain, C; Perrin, J

    2012-03-01

    Sperm banking is an important procedure to preserve fertility before cancer therapy. The aim of this study was to comprehensively analyse cryopreservation activity retrospectively for 1080 patients referred to the sperm bank for sperm cryopreservation before cancer treatment. This study included 1007 patients diagnosed with testicular cancer (TC) (41.7%), lymphoma (26%), other haematological cancers (9.4%) or other types of cancer (22.8%); of these, 29 patients did not produce any semen sample and cryopreservation was impossible for 67 patients. Semen characteristics before treatment were within normal ranges, except moderate asthenospermia. Sperm concentration was significantly lower in TC than in non-TC. Straws from 57 patients (6.3%) were used in assisted reproductive technologies, which led to a 46.8% cumulative birth rate. Straws were destroyed for 170 patients (18.7%) and 140 patients performed semen analyses after cancer therapy. After an average delay of 22.5 months after the end of therapy, 43 patients (30.7%) exhibited azoospermia. This study of a large population of cancer patients revealed a high level of successful sperm storage. Utilization of cryopreserved spermatozoa led to good chances of fatherhood. Nevertheless, sperm banks should be aware of the low rates of straw use and straw destruction by cancer patients. Copyright © 2011 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  10. Cryopreservation of human skeletal muscle impairs mitochondrial function

    DEFF Research Database (Denmark)

    Larsen, Steen; Wright-Paradis, C; Gnaiger, E

    2012-01-01

    functionality after long term cryopreservation (1 year). Skeletal muscle samples were preserved in dimethyl sulfoxide (DMSO) for later analysis. Human skeletal muscle fibres were thawed and permeabilised with saponin, and mitochondrial respiration was measured by high-resolution respirometry. The capacity...

  11. Development of a cryopreservation protocol for type A spermatogonia

    NARCIS (Netherlands)

    Izadyar, Fariborz; Matthijs-Rijsenbilt, J. J.; den Ouden, Krista; Creemers, Laura B.; Woelders, Henri; de Rooij, Dirk G.

    2002-01-01

    The aim of this study was to develop a cryopreservation protocol for type A spermatogonia. Testes from 5- to 7-month-old calves were collected, and type A spermatogonia were isolated using two-step enzymatic digestion and Percoll separation. Cells were resuspended in minimum essential medium (MEM)

  12. Fertility rate of daily collected and cryopreserved fowl semen

    NARCIS (Netherlands)

    Voorst, van A.; Leenstra, F.R.

    1995-01-01

    Semen was collected for 4 consecutive d individually from experimental broiler breeder males that had not been massaged for 7 d. The semen was mixed and diluted with the Beltsville Poultry Semen Extender with dimethylsulfoxide (DMSO) as cryoprotectant and cryopreserved. After thawing of the semen,

  13. Cryopreservation of in vitro -grown shoot tips of apricot ( Prunus ...

    African Journals Online (AJOL)

    In vitro grown apricot (Prunus armeniaca L.) cv. El-Hamawey shoot tips were successfully cryopreserved using an encapsulation-dehydration procedure. Shoot tips were encapsulated in calcium-alginate beads before preculture on woody plant (WP) medium supplemented with different sucrose concentrations; 0.1, 0.3, 0.5, ...

  14. Midterm Results of Aortic Valve Replacement with Cryopreserved Homografts

    Directory of Open Access Journals (Sweden)

    Emre Özker

    2012-06-01

    Full Text Available Objective: The aim of this study was to analyze the midterm clinical results of aortic valve replacement with cryopreserved homografts.Materials and Methods: Aortic valve replacement was performed in 40 patients with cryopreserved homograft. The indications were aortic valve endocarditis in 20 patients (50%, truncus arteriosus in 6 patients (15%, and re-stenosis or regurtitation after aortic valve reconstruction in 14 (35% patients. The valve sizes ranged from 10 to 27mm. A full root replacement technique was used for homograft replacement in all patients.Results: The 30-day postoperative mortality rate was 12.5% (5 patients. There were four late deaths. Only one of them was related to cardiac events. Overall mortality was 22.5%. Thirty-three patients were followed up for 67±26 months. Two patients needed reoperation due to aortic aneurysm caused by endocarditis. The mean transvalvular gradient significantly decreased after valve replacement (p<0.003. The last follow up showed that the 27 (82% patients had a normal left ventricular function.Conclusion: Cryopreserved homografts are safe alternatives to mechanical valves that can be used when there are proper indications. Although it has a high perioperative mortality rate, cryopreserved homograft implantation is an alternative for valve replacement, particularly in younger patients and for complex surgical problems such as endocarditis that must be minimalized.

  15. The Need for a Higher Fuel Efficiency of the Electricity Sector - An Analysis of Opportunities and Barriers

    Energy Technology Data Exchange (ETDEWEB)

    Klimstra, J.

    2007-07-01

    The electricity sector is the single largest user of primary energy in the world. The issues of fuel prices, security of supply and greenhouse gas emissions are therefore closely connected with electricity generation. The total energy efficiency of the electricity sector is only 32.5% so that quick improvements are required. However, the uncertainty over fuel prices and technology preferences is such that most investors are hesitant. The life of existing, often low-efficiency, power plants is therefore extended. At the same time, the demand for electricity is rapidly increasing and the gap between capacity and demand decreases. This paper intends to bring more clarity into the economic and environmental boundary conditions of power plants. The goal is to find an attractive way for rapid efficiency improvement with an even better system reliability without increasing the costs. The paper discusses fuel price developments and the costs of generating technologies in connection with the typical demand pattern of electricity. Ultimately, it appears that local generation, preferably coupled with cogeneration, can be an important part of the solution. (auth)

  16. Energy conservation in industrial buildings. Higher energy efficiency with smart control systems; Energieeinsparung im Gewerbebau. Hoehere Energieeffizienz durch 'intelligente' Regeltechnik

    Energy Technology Data Exchange (ETDEWEB)

    Mayer, Edgar [CentraLine c/o Honeywell GmbH, Schoenaich (Germany)

    2009-07-01

    With smart control systems, the energy conservation potential of industrial buildings could be fully utilized. This means, e.g., that classic control algorithms must be replaced by new solutions. New methods will ensure higher energy efficiency with maximum comfort; they will also prolong the service life and the inspection intervals of the technical facilities. (orig.)

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

    Directory of Open Access Journals (Sweden)

    Balint Bela

    2006-01-01

    Full Text Available Background/Aim. Thermodynamical and cryobiological parameters responsible for cell damages during cryopreservation (cryoinjuries have not yet been completely explained. Thus, freezing procedures should be revised, exactly optimized to obtain an enhanced structural and functional recovery of frozen- thawed cells. The aim of this study was to compare microprocessor- controlled (controlled-rate with the compensation of the released fusion heat and “dump-freezing” (uncontrolled- rate of the platelet and lymphocyte cryopreservation efficacy. Methods. Platelet quantitative recovery (post-thaw vs. unfrozen cell count, viability (using hypotonic shock response - HSR, morphological score (PMS, ultrastructural (electron microscopy properties and expression of different surface antigens were investigated. In lymphocyte setting, cell recovery and viability (using trypan blue exclusion test as well as functionality (by plant mitogens were determined. Controlled- rate freezing and uncontrolled-rate cryopreservation were combined with 6% (platelets and 10% (lymphocytes dimethyl sulfoxide (DMSO. Results. Platelet recovery and functionality were superior in the controlled-rate system. The majority of surface antigen expression was reduced in both freezing groups vs. unfrozen cells, but GP140/CD62p was significantly higher in controlled-rate vs. uncontrolled-rate setting. Controlled- rate freezing resulted with better lymphocyte recovery and viability (trypan blue-negative cell percentage. In mitogen-induced lymphocyte proliferative response no significant intergroup difference (controlled-rate vs. uncontrolled-rate were found. Conclusion. The data obtained in this study showned the dependence of cell response on the cryopreservation type. Controlled-rate freezing provided a superior platelet quantitative and functional recovery. Lymphocyte recovery and viability were better in the controlled-rate group, although only a minor intergroup difference for cell

  18. Comparison of glycerolisation with cryopreservation methods on HIV-1 inactivation

    International Nuclear Information System (INIS)

    Van Baare, J.; Pagnon, J.; Cameron, P.; Vardaxis, N.; Middlekoop, E.; Crowe, S.

    1999-01-01

    Cryopreservation and glycerolisation are two successful long-term preservation methods for human cadaveric donor skin, which is used in the treatment of bum patients. High concentrations of glycerol has been shown to be antibacterial and virucidal. Because fear of possible transmission of HIV-1 following allograft transplantation, this study was undertaken to investigate whether HIV can be effectively eliminated from skin explants. HIV-1 Ba-L, which has been shown to infect monocytes in skin explants and also dendritic cells, was. For the experiments we used cell-free virus, exogenously HIV infected peripheral blood mononuclear cells (PBMCs) and exogenously HIV infected cadaver split skin. Different concentrations of glycerol at various temperatures and the glycerolisation procedure as used by the Euro Skin Bank were used to determine the effects on HIV-1 Ba-L infectivity. For the cryopreservation technique we used 10% DMSO and a controlled rate freezer. HIV-1 Ba-L transfer was determined by adding uninfected PBMCs to the infected material and reverse transcriptase was measured. Cell-free HIV-1 Ba-L was not inactivated by 50% glycerol but was effectively inactivated within 30 minutes by 70% and 85% glycerol at 4 degree C, room temperature and 37 degree C. In contrast, cell-free HIV-1 Ba-L was not inactivated by cryopreservation. Most importantly, we have shown that HIV-1 Ba-L present in split skin is inactivated by incubating skin in 70% glycerol for three hours at 37-C. HIV in exogenously infected skin was not inactivated by cryopreservation. High concentrations of glycerol effectively inactivates free HIV-1 Ba-L and intracellular HIV-1 Ba-L. Also the current glycerolisation procedure carried out by the Euro Skin Bank effectively inactivates infectious virus. However, the cryopreservation technique did not show any reduction in HIV-1 Ba-L infectivity

  19. Cryopreservation of preimplantation embryos of cattle, sheep, and goats.

    Science.gov (United States)

    Youngs, Curtis R

    2011-08-05

    Preimplantation embryos from cattle, sheep, and goats may be cryopreserved for short- or long-term storage. Preimplantation embryos consist predominantly of water, and the avoidance of intracellular ice crystal formation during the cryopreservation process is of paramount importance to maintain embryo viability. Embryos are placed into a hypertonic solution (1.4 - 1.5 M) of a cryoprotective agent (CPA) such as ethylene glycol (EG) or glycerol (GLYC) to create an osmotic gradient that facilitates cellular dehydration. After embryos reach osmotic equilibrium in the CPA solution, they are individually loaded in the hypertonic CPA solution into 0.25 ml plastic straws for freezing. Embryos are placed into a controlled rate freezer at a temperature of -6°C. Ice crystal formation is induced in the CPA solution surrounding the embryo, and crystallization causes an increase in the concentration of CPA outside of the embryo, causing further cellular dehydration. Embryos are cooled at a rate of 0.5°C/min, enabling further dehydration, to a temperature of -34°C before being plunged into liquid nitrogen (-196°C). Cryopreserved embryos must be thawed prior to transfer to a recipient (surrogate) female. Straws containing the embryos are removed from the liquid nitrogen dewar, held in room temperature air for 3 to 5 sec, and placed into a 37°C water bath for 25 to 30 sec. Embryos cryopreserved in GLYC are placed into a 1 M solution of sucrose for 10 min for removal of the CPA before transfer to a recipient (surrogate) female. Embryos cryopreserved in EG, however, may be directly transferred to the uterus of a recipient.

  20. The ratio of nurse consultation and physician efficiency index of senior rheumatologists is significantly higher than junior physicians in rheumatology residency training

    DEFF Research Database (Denmark)

    Emamifar, Amir; van Bui Hansen, Morten Hai; Jensen Hansen, Inger Marie

    2017-01-01

    To elucidate the difference between ratios of nurse consultation sought by senior rheumatologists and junior physicians in rheumatology residency training, and also to evaluate physician efficiency index respecting patients with rheumatoid arthritis (RA). Data regarding outpatient visits for RA...... patients between November 2013 and 2015 were extracted. The mean interval (day) between consultations, the nurse/physician visits ratio, and physician efficiency index (nurse/physician visits ratio × mean interval) for each senior and junior physicians were calculated. Disease Activity Score in 28 joints....../physician visits ratio (P = .01) and mean efficiency index (P = .04) of senior rheumatologists were significantly higher than that of junior physicians. Regression analysis showed a positive correlation between physician postgraduate experience and physician efficiency index adjusted for DAS28 at baseline...

  1. Funding System of Full-Time Higher Education and Technical Efficiency: Case of the University of Ljubljana

    Science.gov (United States)

    Tajnikar, Maks; Debevec, Jasmina

    2008-01-01

    The present paper tackles the issue of the higher education funding system in Slovenia. Its main attribute is that institutions are classified into study groups according to their fields of education, and funds granted by the state are based on their weights or study group factors (SGF). Analysis conducted using data envelopment analysis tested…

  2. Carbon Isotope discrimination in acacia auriculiformis - can it be used to select for higher water-use-efficiency in trees?

    International Nuclear Information System (INIS)

    Montagu, K.D.; Woo, K.C.; Puangchit, L.

    1999-01-01

    Full text: Determining the water-use-efficiency of trees in relation lo wood production is problematic due to the sheer size of the plant and the number of years taken to produce the wood. Indirect measures of water-use-efficiency, such as carbon isotope discrimination (Δ), are therefore attractive to tree breeders wishing to select for increased water-use-efficiency. To begin to evaluate the usefulness of Δ as a selection parameter for the tropical tree Acacia auriculiformis we addressed the following questions: 1. Within the tree canopy, how variable is Δ? 2. Is there any genotypic variation in Δ? and 3. Does water availability affect genotypic variation? To address these questions we sampled foliage from pot trials and field trials of A. auriculiformis ranging in age from 3 months lo 8 years in Australia and Thailand. In 16-18m high 8-year-old trees, canopy variation in Δ was large (P>0.01). Foliage Δ values increased down the tree from 22.0 %o at the top to 24.7 %o at the base. The decrease was rapid in the lop 3 m of the canopy thus considerable care must be taken to sampling foliage from the same position in the canopy. Genotype variations in Δ was observed in seedlings and 2 year-old trees (P>0.01) but not in 8 year-old trees (P=0.60). Where genotypic variation were observed the differences between the lowest and highest values were 2.2 - 3.6 %o. Reduced water availability decreased Δ values in both pot and field studies but not in a consistent way across seedlots. Thus it would appear that the Δ of trees grown under favourable conditions does not give an indication of the Δ value which will be obtained under water-limited conditions. This complicates the use of Δ as a screening method. We have clearly shown that genotype variation occurs in A. auriculiformis in both seedlings and young field-grown trees. Considerable care is required when sampling large trees, as variation in Δ within the tree can be as large as between genotypes. The challenge

  3. In vitro assessment of soybean lecithin and egg yolk based diluents for cryopreservation of goat semen.

    Science.gov (United States)

    Salmani, Hossein; Towhidi, Armin; Zhandi, Mahdi; Bahreini, Majid; Sharafi, Mohsen

    2014-04-01

    Soybean lecithin is a suitable plant-based cryoprotectant for freezing ruminant sperm. Optimum level of lecithin was not clear for goat semen cryopreservation. The objective of this study was to investigate the effects of different levels of soybean lecithin in semen extender on post-thaw sperm quality including CASA-motion parameters, viability, plasma membrane integrity and lipid peroxidation. Semen samples were collected from 4 Mahabadi bucks using an artificial vagina. Different concentrations of soy lecithin (SL, 0.5%, 1%, 1.5%, 2% and 2.5% w/v) were compared to 15% (v/v) egg yolk-based extender (TR-EY). No significant difference was observed for sperm progressive motility, viability or plasma membrane integrity in 1.5% SL media (33.8%, 66%, and 62.7%, respectively) and TR-EY medium (35.4%, 67.2%, and 64.9%, respectively). Sperm motion characteristics (VAP, VSL, VCL, ALH and LIN) and rapid spermatozoa were improved with extender containing 1% and 1.5% SL, compared to TR-EY extender. Furthermore, egg yolk produced significantly higher malondialdehyde (4.02±0.21) than other groups. Results suggest that the optimal lecithin concentration in the semen extender was 1.5% and also soy lecithin can substitute for egg yolk during cryopreservation for caprine sperm. Copyright © 2014 Elsevier Inc. All rights reserved.

  4. Application of Pectin From Rauvolfia serpentina (L.) Benth to the Cryopreservation of Human Leucocyte Cell Suspensions.

    Science.gov (United States)

    Zaitseva, O O; PoleZhaeva, T V; Khudvakov, A N; Solomina, O N; Golovchenko, V V

      BACKGROUND: Due to their valuable medicinal properties and high physiological activity, plant polysaccharides are currently being extensively studied. The present study aims to investigate rauwolfian (pectin for Rauvolfia serpentina) supplementation on human leukocytes cryopreservation. We determined the сharacteristics of leukocytes undergoing freezing with pectin at different temperatures. Donor leukocytes were frozen under the protection of comprehensive cryoprotectant solution and stored in electrical freezers (-20C, -40C, -80C). A regular decrease of all values starting from a higher temperature (-20С) through to the lower temperature (-80С) was identified. The study showed that pectin rauwolfian stimulated both the oxygen-independent and the oxygen-dependent killer response. We also found that the oxygen-dependent neutrophil killer effect was reduced as the storage temperature was lowered. It was determined that the LPO levels in the cells with added pectin-containing solutions remained the same before freezing, while their antioxidant activity positively increased, which is beneficial for neutrophils for their further freezing to -20C, -40C and -80C. The results of the study make it possible to assume that rauwolfian, a pectin extracted from Rauvolfia serpentina, has an exocellular protectant effect as part of cryopreservative solution on human white blood cells stored at different low temperatures. The versatility of the substance is probably due to the degree of the macromolecule branching, in particular, the structure of carbohydrate side chains, which contain a large number of hydroxyl groups.

  5. Determination of Intracellular Vitrification Temperatures for Unicellular Micro Organisms under Conditions Relevant for Cryopreservation.

    Science.gov (United States)

    Fonseca, Fernanda; Meneghel, Julie; Cenard, Stéphanie; Passot, Stéphanie; Morris, G John

    2016-01-01

    During cryopreservation ice nucleation and crystal growth may occur within cells or the intracellular compartment may vitrify. Whilst previous literature describes intracellular vitrification in a qualitative manner, here we measure the intracellular vitrification temperature of bacteria and yeasts under conditions relevant to cryopreservation, including the addition of high levels of permeating and nonpermeating additives and the application of rapid rates of cooling. The effects of growth conditions that are known to modify cellular freezing resistance on the intracellular vitrification temperature are also examined. For bacteria a plot of the activity on thawing against intracellular glass transition of the maximally freeze-concentrated matrix (Tg') shows that cells with the lowest value of intracellular Tg' survive the freezing process better than cells with a higher intracellular Tg'. This paper demonstrates the role of the physical state of the intracellular environment in determining the response of microbial cells to preservation and could be a powerful tool to be manipulated to allow the optimization of methods for the preservation of microorganisms.

  6. Effect of different monosaccharides and disaccharides on boar sperm quality after cryopreservation.

    Science.gov (United States)

    Gómez-Fernández, José; Gómez-Izquierdo, Emilio; Tomás, Cristina; Mocé, Eva; de Mercado, Eduardo

    2012-07-01

    The aim of the present study was to evaluate the cryoprotectant effect of different non-permeating sugars for boar sperm. Pooled semen from three boars was used for the experiments. In the first experiment, the sperm quality of boar sperm cryopreserved with an egg-yolk based extender supplemented with different monosaccharides (glucose, galactose or fructose) was compared to a control cryopreserved in lactose-egg yolk extender. In the second experiment, the effect of five disaccharides (lactose, sucrose, lactulose, trehalose or melibiose) on boar sperm cryosurvival was studied. Several sperm quality parameters were assessed by flow cytometry in samples incubated for 30 and 150 min at 37°C after thawing: percentages of sperm with intact plasma membrane (SIPM), sperm presenting high plasma membrane fluidity (HPMF), sperm with intracellular reactive oxygen substances production (IROSP) and apoptotic sperm (AS). In addition, the percentages of total motile (TMS) and progressively motile sperm (PMS) were assessed at the same incubation times with a computer-assisted sperm analysis system. Freezing extenders supplemented with each of the monosaccharide presented smaller cryoprotective effect than the control extender supplemented with lactose (Pextender supplemented with lactulose exhibited in general the lowest sperm quality, except for the percentage of capacitated sperm, which was highest (Pextender. Our results suggest that disaccharides have higher cryoprotective effect than monosaccharides, although the monosaccharide composition of the disaccharides is also important, since the best results were obtained with those disaccharides presenting glucose in their composition. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. A novel efficient coupled polynomial field interpolation scheme for higher order piezoelectric extension mode beam finite elements

    International Nuclear Information System (INIS)

    Sulbhewar, Litesh N; Raveendranath, P

    2014-01-01

    An efficient piezoelectric smart beam finite element based on Reddy’s third-order displacement field and layerwise linear potential is presented here. The present formulation is based on the coupled polynomial field interpolation of variables, unlike conventional piezoelectric beam formulations that use independent polynomials. Governing equations derived using a variational formulation are used to establish the relationship between field variables. The resulting expressions are used to formulate coupled shape functions. Starting with an assumed cubic polynomial for transverse displacement (w) and a linear polynomial for electric potential (φ), coupled polynomials for axial displacement (u) and section rotation (θ) are found. This leads to a coupled quadratic polynomial representation for axial displacement (u) and section rotation (θ). The formulation allows accommodation of extension–bending, shear–bending and electromechanical couplings at the interpolation level itself, in a variationally consistent manner. The proposed interpolation scheme is shown to eliminate the locking effects exhibited by conventional independent polynomial field interpolations and improve the convergence characteristics of HSDT based piezoelectric beam elements. Also, the present coupled formulation uses only three mechanical degrees of freedom per node, one less than the conventional formulations. Results from numerical test problems prove the accuracy and efficiency of the present formulation. (paper)

  8. Achieving 99.9% proton spin-flip efficiency at higher energy with a small rf dipole

    CERN Document Server

    Leonova, M A; Gebel, R; Hinterberger, F; Krisch, A D; Lehrach, A; Lorentz, B; Maier, R; Morozov, V S; Prasuhn, D; Raymond, R S; Schnase, A; Stockhorst, H; Ulbrich, K; Wong, V K; 10.1103/PhysRevLett.93.224801

    2004-01-01

    We recently used a new ferrite rf dipole to study spin flipping of a 2.1 GeV/c vertically polarized proton beam stored in the COSY Cooler Synchrotron in Julich, Germany. We swept the rf dipole's frequency through an rf-induced spin resonance to flip the beam's polarization direction. After determining the resonance's frequency, we varied the frequency range, frequency ramp time, and number of flips. At the rf dipole's maximum strength and optimum frequency range and ramp time, we measured a spin-flip efficiency of 99.92+or-0.04%. This result, along with a similar 0.49 GeV/c IUCF result, indicates that, due to the Lorentz invariance of an rf dipole's transverse integral Bdl and the weak energy dependence of its spin-resonance strength, an only 35% stronger rf dipole should allow efficient spin flipping in the 100 GeV BNL RHIC Collider or even the 7 TeV CERN Large Hadron Collider.

  9. Achieving Lower Nitrogen Balance and Higher Nitrogen Recovery Efficiency Reduces Nitrous Oxide Emissions in North America's Maize Cropping Systems

    Directory of Open Access Journals (Sweden)

    Rex A. Omonode

    2017-06-01

    Full Text Available Few studies have assessed the common, yet unproven, hypothesis that an increase of plant nitrogen (N uptake and/or recovery efficiency (NRE will reduce nitrous oxide (N2O emission during crop production. Understanding the relationships between N2O emissions and crop N uptake and use efficiency parameters can help inform crop N management recommendations for both efficiency and environmental goals. Analyses were conducted to determine which of several commonly used crop N uptake-derived parameters related most strongly to growing season N2O emissions under varying N management practices in North American maize systems. Nitrogen uptake-derived variables included total aboveground N uptake (TNU, grain N uptake (GNU, N recovery efficiency (NRE, net N balance (NNB in relation to GNU [NNB(GNU] and TNU [NNB(TNU], and surplus N (SN. The relationship between N2O and N application rate was sigmoidal with relatively small emissions for N rates <130 kg ha−1, and a sharp increase for N rates from 130 to 220 kg ha−1; on average, N2O increased linearly by about 5 g N per kg of N applied for rates up to 220 kg ha−1. Fairly strong and significant negative relationships existed between N2O and NRE when management focused on N application rate (r2 = 0.52 or rate and timing combinations (r2 = 0.65. For every percentage point increase, N2O decreased by 13 g N ha−1 in response to N rates, and by 20 g N ha−1 for NRE changes in response to rate-by-timing treatments. However, more consistent positive relationships (R2 = 0.73–0.77 existed between N2O and NNB(TNU, NNB(GNU, and SN, regardless of rate and timing of N application; on average N2O emission increased by about 5, 7, and 8 g N, respectively, per kg increase of NNB(GNU, NNB(TNU, and SN. Neither N source nor placement influenced the relationship between N2O and NRE. Overall, our analysis indicated that a careful selection of appropriate N rate applied at the right time can both increase NRE and reduce N

  10. MEGA. Higher energy-efficiency through systematic user information - Final report; MEGA. Mehr Energieeffizienz durch gezielte Anwenderinformationen - Schlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Fercu, M.; Kistler, R. [Hochschule Luzern Technik und Architektur, CEESAR - iHomeLab, Horw (Switzerland); Egli, A. [Hochschule Luzern Technik und Architektur, ISIS, Horw (Switzerland); Gallati, J. [Hochschule Luzern Technik und Architektur, Wirtschaft, Horw (Switzerland)

    2010-09-15

    Individuals are empowered to conserve energy and natural resources when provided with motivational and personalized information on its use. By presenting information about the energy consumption from the home's energy meters along with recommended actions, the residential customer becomes aware of how in/efficiently energy is consumed within his home and can decide on how to act to conserve. This information can provide an accurate metric of how effective a conservation action is even to inhabitants that are not yet knowledgeable about or self-motivated by the monetary and ecologic rewards of conserving. This project was designed to build knowledge on technically and economically feasible ways to create an awareness of energy (especially electricity) for the sake of conservation. Specifically, it implements an exemplar prototype of a highly effective energy feedback system that is an interactive, real-time, in-home display. Toward this goal, four system architecture configuration proposals, a set of system requirements, and ideal system features are synthesized; they are based on the results of the research that evaluates effectiveness of existing energy-efficiency and -conservation methods and studies related technologies. Three of the four systems proposed represent energy technologies expected to be available within the next decade. The fourth system proposal is a demonstration prototype designed for implementation in the iHomeLab. This prototype is an open, modular, robust, cross-platform software framework that collects data, processes, and presents it interactively and visually on hardware available in most households. The results of this project both indicate that the creation of such energy feedback systems appear beneficial and also provide guidelines for their design. However, further development of infrastructure and elaboration of design is foreseen as necessary for this system to be suitable for mass deployment. (author)

  11. MEGA. Higher energy-efficiency through systematic user information - Final report; MEGA. Mehr Energieeffizienz durch gezielte Anwenderinformationen - Schlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Fercu, M.; Kistler, R. [Hochschule Luzern Technik und Architektur, CEESAR - iHomeLab, Horw (Switzerland); Egli, A. [Hochschule Luzern Technik und Architektur, ISIS, Horw (Switzerland); Gallati, J. [Hochschule Luzern Technik und Architektur, Wirtschaft, Horw (Switzerland)

    2010-09-15

    Individuals are empowered to conserve energy and natural resources when provided with motivational and personalized information on its use. By presenting information about the energy consumption from the home's energy meters along with recommended actions, the residential customer becomes aware of how in/efficiently energy is consumed within his home and can decide on how to act to conserve. This information can provide an accurate metric of how effective a conservation action is even to inhabitants that are not yet knowledgeable about or self-motivated by the monetary and ecologic rewards of conserving. This project was designed to build knowledge on technically and economically feasible ways to create an awareness of energy (especially electricity) for the sake of conservation. Specifically, it implements an exemplar prototype of a highly effective energy feedback system that is an interactive, real-time, in-home display. Toward this goal, four system architecture configuration proposals, a set of system requirements, and ideal system features are synthesized; they are based on the results of the research that evaluates effectiveness of existing energy-efficiency and -conservation methods and studies related technologies. Three of the four systems proposed represent energy technologies expected to be available within the next decade. The fourth system proposal is a demonstration prototype designed for implementation in the iHomeLab. This prototype is an open, modular, robust, cross-platform software framework that collects data, processes, and presents it interactively and visually on hardware available in most households. The results of this project both indicate that the creation of such energy feedback systems appear beneficial and also provide guidelines for their design. However, further development of infrastructure and elaboration of design is foreseen as necessary for this system to be suitable for mass deployment. (author)

  12. New aeration systems for higher efficiency in mine water treatment in the Lausitz region; Einsatz neuer Belueftungssysteme zur Effizienzsteigerung bei der Grubenwasserbehandlung in der Lausitz

    Energy Technology Data Exchange (ETDEWEB)

    Janneck, E.; Glombitza, F. [G.E.O.S. Freiberg Ingenieurgesellschaft mbH, Freiberg (Germany); Schlee, K.; Arnold, I. [Vattenfall Europe Mining AG, Cottbus (Germany)

    2006-07-01

    The article presents experiences and results of the application of new aerator-systems in the mine water treatment. The processes of ferrous iron oxidation and sludge removal became more stable and efficiently by the application of the aerators. For the first time, spiral aerators were used in the Lower Lusatia lignite mining district to clean ferrous iron containing mine water. These devices lead to an enhanced iron oxidation rate under the existing conditions, where the oxygen diffusion is the rate determining step. Furthermore, the application caused increased throughput, optimal lime utilisation and better sludge thickening, which led to a higher efficiency of the mine water treatment. (orig.)

  13. Hatchery-scale trials using cryopreserved spermatozoa of black-lip pearl oyster, Pinctada margaritifera

    OpenAIRE

    Hui, Belinda; Vonau, Vincent; Moriceau, Jacques; Tetumu, Roger; Vanaa, Vincent; Demoy-schneider, Marina; Suquet, Marc; Le Moullac, Gilles

    2011-01-01

    Cryopreservation is a valuable tool for genetic improvement programs. Several bivalve mollusc species have already been the subject of such programs and the Tahitian black pearl oyster industry is now planning the development of selective breeding for desirable traits in Pinctada margaritifera. The ability to cryopreserve spermatozoa would, therefore, offer significant benefits to the cultured black pearl industry. Spermatozoa were cryopreserved with CPA 0.7 M trehalose in 0.8 M Me2SO and...

  14. Data on antioxidant activity in grapevine (Vitis vinifera L.) following cryopreservation by vitrification

    OpenAIRE

    María Fernanda Lazo-Javalera; Martín Ernesto Tiznado-Hernández; Irasema Vargas-Arispuro; Elisa Valenzuela-Soto; María del Carmen Rocha-Granados; Marcos Edel Martínez-Montero; Marisela Rivera-Domínguez

    2015-01-01

    Cryopreservation is used for the long-term conservation of plant genetic resources. This technique very often induces lethal injury or tissue damage. In this study, we measured indicators of viability and cell damage following cryopreservation and vitrification-cryopreservation in Vitis vinifera L. axillary buds cv. ?Flame seedless? stored in liquid nitrogen (LN) for: three seconds, one hour, one day, one week and one month; after LN thawed at 38??C for three minutes. The enzymatic activity o...

  15. The hybrid methylene blue-zeolite system: a higher efficient photo catalyst for photo inactivation of pathogenic microorganisms

    International Nuclear Information System (INIS)

    Smolinska, M.; Cik, G.; Sersen, F.; Caplovicova, M.; Takacova, A.; Kopani, M.

    2015-01-01

    The composite system can be prepared by incorporation of methylene blue into the channels of zeolite and by adsorption on the surface of the crystals. The composite photo sensitizer effectively absorbs the red light (kmax = 648 nm) and upon illumination with light-emitting diode at a fluence rate of 1.02 mW cm-2 generates effectively reactive singlet oxygen in aqueous solution, which was proved by EPR spectroscopy. To test efficiency for inactivation of pathogenic microorganisms, we measured photo killing of bacteria Escherichia coli and Staphylococcus aureus and yeasts Candida albicans. We found out that after the microorganisms have been adsorbed at the surface of such modified zeolite, the photo generated singlet oxygen quickly penetrates their cell walls, bringing about their effective photo inactivation. The growth inhibition reached almost 50 % at 200 and 400 mg modified zeolite in 1 ml of medium in E. coli and C. albicans, respectively. On the other hand, the growth inhibition of S. aureus reached 50 % at far smaller amount of photo catalyst (30 lg per 1 ml of medium). These results demonstrate differences in sensitivities of bacteria and yeast growth. The comparison revealed that concentration required for IC50 was in case of C. albicans several orders of magnitude lower for a zeolite-immobilized dye than it was for a freely dissolved dye. In S. aureus, this concentration was even lower by four orders of magnitude. Thus, our work suggested a new possibility to exploitation of zeolite and methylene blue in the protection of biologically contaminated environment, and in photodynamic therapy.

  16. Measures for improving the adoption of higher efficiency appliances in Indonesian households: An analysis of lifetime use and decision-making in the purchase of electrical appliances

    International Nuclear Information System (INIS)

    Wijaya, Muhammad Ery; Tezuka, Tetsuo

    2013-01-01

    Highlights: ► We observe human psychosocial variables regarding purchase of electrical appliances. ► Two cities with different cultures are subject of this study – Bandung and Yogyakarta. ► Differences in the lifetime of appliances can be attributed to the cultural. ► Ads and store’s staff have the greatest impact on people’s choice of appliances. ► Adoption of higher-efficiency appliances could be implemented based on each culture. - Abstract: One approach to decreasing electricity consumption is to facilitate the replacement of older appliances with new, higher-efficiency. The objectives of this paper are to compare and analyse the replacement of appliances in two cities of Indonesia – Yogyakarta and Bandung – that are characterised by different cultural backgrounds, ethnicities, and decision-making processes in the household purchase of electrical appliances. A questionnaire survey method was employed to obtain information on behavioural economics and human psychosocial variables such as attitudes, beliefs and perceived benefits regarding the replacement and purchase of electrical appliances. The results show that refrigerators in Yogyakarta have a longer lifetime than in Bandung. However, in Bandung, air conditioners, electric fans, rice cookers, and water pumps have a longer lifetime than in Yogyakarta. These differences in the lifetime of appliances can be attributed to the cultural differences within the two cities that are reflected in the manner in which people use electrical appliances as well as to their lack of knowledge regarding appliance operation. An analysis of the factors influencing the purchase of appliances indicated that people in Yogyakarta show a greater awareness of the benefits of adopting higher-efficiency appliances than do persons in Bandung. The following suggestions could be implemented to improve the strategy of encouraging the adoption of higher-efficiency appliances: (1) in Yogyakarta, energy labelling could be

  17. The Effect of Phyto-Lecithin on Preservation and Cryopreservation of Semen: A Review

    Directory of Open Access Journals (Sweden)

    AS Aku

    2007-01-01

    Full Text Available Artificial insemination represents one of technologies in livestock reproduction that can be applied to cattle, sheep, goats and other livestock. Application of livestock reproduction technology includes artificial insemination to increase reproductive efficiency. Semen processing is one critical phase in an artificial insemination program. The use of animal origin ingredient for semen extenders, such as egg yolk and milk, presents a risk of microbial contamination, which lead to the search for alternatives. To increase standard of quality, researchers exploits phyto-lesitin for semen extender and the results showed no significant differences in motility, viability, and acrosomal status of spermatozoa with phyto-lesitin extender when compared to tris-egg yolk-containing extenders. (Animal Production 9(1: 49-52 (2007 Key Words : Phyto-Lechitin, preservation, cryopreservation, semen

  18. A new strategy and system for the ex vivo ovary perfusion and cryopreservation: An innovation.

    Science.gov (United States)

    Ali Mohamed, Mohamed Shehata

    2017-06-01

    Children and young adults, who suffer from cancer, receive gonadotoxic therapy, which destroys their fertile abilities after survival. Ovarian cryopreservation and transplantation provide the promising solution to this problem, where the ovary can be removed before the gonadotoxic therapy and reimplanted after patient's survival, where the ovary is to be cryopreserved during the period of the therapy. However, cryopreservation of the whole ovary is still facing great obstacles, namely the ischemic reperfusion injury and the defective cryopreservation related to the defective ability to universally deliver the cryopreservation/warming solutions through the ovarian vascular bed. Meanwhile, the currently applied technique of ovarian tissue cryopreservation provides limited follicular recovery because many follicles are lost until the development of revascularization post-transplantation. To solve the problems, an innovative system has been developed to insure immediate and universal delivery of the cryopreservation/warming solutions to the graft, in addition to keeping the graft under continuous perfusion before and after cryopreservation, minimizing any chance for microthrombi formation or ischemia-reperfusion. This innovative system can be applied in the following surgical and clinical interventions: 1) Allogeneic ovarian transplantation; 2) Preservation of fertility after systemic chemotherapy or bone marrow transplantation in young females, where the ovaries could be removed before the therapy and exposed to the adequate cryopreservation provided by the system till re-implantation after the patient's survival; 3) The system is also suitable for the corresponding applications on the testicles.

  19. Efficacy of postal communication with patients who have cryopreserved pre-embryos.

    Science.gov (United States)

    Brzyski, R G

    1998-11-01

    To compare the characteristics of patients who did and did not respond to a request for information regarding their cryopreserved pre-embryos. Mail survey. Academic-assisted reproductive technology program. One hundred thirty-six patients with cryopreserved pre-embryos. Patients were surveyed by first-class mail regarding their plans for their cryopreserved pre-embryos and their interest in embryo donation. Age, number of stored pre-embryos, and duration of storage of responders and nonresponders at 6 weeks after mailing. Eighty-three patients (62%) did not respond to the survey. Compared with responders, nonresponders were significantly older at the time of embryo cryopreservation, had fewer pre-embryos cryopreserved, and had the pre-embryos cryopreserved for a longer duration. Five responders (9%) expressed an interest in embryo donation. Three patients requested disposal of pre-embryos. Sixteen surveys (12%) were returned as undeliverable. As a group, these patients had the fewest pre-embryos cryopreserved and had the longest duration of storage. A disturbing number of patients with cryopreserved pre-embryos ignored efforts by our program to maintain contact. Older patients with few cryopreserved pre-embryos may require special attention to avoid abandonment.

  20. A questionnaire survey on attitude toward sperm cryopreservation among hematologists in Japan.

    Science.gov (United States)

    Kobayashi, Tomohiro; Shin, Takeshi; Nishio, Kojiro; Shimomura, Yukihito; Iwahata, Toshiyuki; Suzuki, Keisuke; Miyata, Akane; Kobori, Yoshitomo; Arai, Gaku; Okada, Hiroshi

    2017-03-01

    Advances in multimodal treatment have led to dramatic improvement in cancer treatment outcomes. It is now necessary to consider cancer patients' holistic quality of life. Fertility preservation is the top concern for cancer survivors of reproductive age. Sperm cryopreservation before treatment is recommended for postpubescent men, but many patients lose fertility without having been informed about options for fertility preservation. To determine how sperm cryopreservation is perceived and practiced in Japan, we surveyed hematologists who often treat young males. A questionnaire about sperm cryopreservation was sent to 45 major hematology institutions. A total of 22 institutions responded before the deadline. All institutions but one responded that they felt sperm cryopreservation is necessary. Only 15 institutions responded that they inform patients about sperm cryopreservation, and 12 institutions responded that they perform sperm cryopreservation before chemotherapy. A total of 213 young males started their first course of chemotherapy during the survey period, of whom 61 (28.6%) had their sperm cryopreserved. Although almost all hematologists stated that sperm cryopreservation is necessary for fertility preservation, not all institutions informed patients about it. Our findings indicate that, to promote fertility preservation in Japan, it will be necessary to systematize sperm cryopreservation and build inter-hospital networks.

  1. Seasonal Thermal-Energy Storage: A Critical Review on BTES Systems, Modeling, and System Design for Higher System Efficiency

    Directory of Open Access Journals (Sweden)

    Michael Lanahan

    2017-05-01

    Full Text Available Buildings consume approximately ¾ of the total electricity generated in the United States, contributing significantly to fossil fuel emissions. Sustainable and renewable energy production can reduce fossil fuel use, but necessitates storage for energy reliability in order to compensate for the intermittency of renewable energy generation. Energy storage is critical for success in developing a sustainable energy grid because it facilitates higher renewable energy penetration by mitigating the gap between energy generation and demand. This review analyzes recent case studies—numerical and field experiments—seen by borehole thermal energy storage (BTES in space heating and domestic hot water capacities, coupled with solar thermal energy. System design, model development, and working principle(s are the primary focus of this analysis. A synopsis of the current efforts to effectively model BTES is presented as well. The literature review reveals that: (1 energy storage is most effective when diurnal and seasonal storage are used in conjunction; (2 no established link exists between BTES computational fluid dynamics (CFD models integrated with whole building energy analysis tools, rather than parameter-fit component models; (3 BTES has less geographical limitations than Aquifer Thermal Energy Storage (ATES and lower installation cost scale than hot water tanks and (4 BTES is more often used for heating than for cooling applications.

  2. Combined effects of cooled EGR and a higher geometric compression ratio on thermal efficiency improvement of a downsized boosted spark-ignition direct-injection engine

    International Nuclear Information System (INIS)

    Su, Jianye; Xu, Min; Li, Tie; Gao, Yi; Wang, Jiasheng

    2014-01-01

    Highlights: • Experiments for the effects of cooled EGR and two compression ratios (CR) on fuel efficiency were conducted. • The mechanism for the observed fuel efficiency behaviors by cooled EGR and high CR was clarified. • Cooled EGR offers more fuel efficiency improvement than elevating CR from 9.3 to 10.9. • Combining 18–25% cooled EGR with 10.9 CR lead to 2.1–3.5% brake thermal efficiency improvements. - Abstract: The downsized boosted spark-ignition direct-injection (SIDI) engine has proven to be one of the most promising concepts to improve vehicle fuel economy. However, the boosted engine is typically designed at a lower geometric compression ratio (CR) due to the increased knock tendency in comparison to naturally aspirated engines, limiting the potential of improving fuel economy. On the other hand, cooled exhaust gas recirculation (EGR) has drawn attention due to the potential to suppress knock and improve fuel economy. Combing the effects of boosting, increased CR and cooled EGR to further improve fuel economy within acceptable knock tolerance has been investigated using a 2.0 L downsized boosted SIDI engine over a wide range of engine operating conditions from 1000 rpm to 3000 rpm at low to high loads. To clarify the mechanism of this complicated effects, the first law of thermodynamics analysis was conducted with the inputs from GT-Power® engine simulation. Experiment results indicate that cooled EGR provides more brake thermal efficiency improvement than increasing geometric CR from 9.3 to 10.9. The benefit of brake thermal efficiency from the higher CR is limited to low load conditions. The attributes for improving brake thermal efficiency by cooled EGR include reduced heat transfer loss, reduced pumping work and increased ratio of specific heats for all the engine operating conditions, as well as higher degree of constant volume heat release only for the knock-limited high load conditions. The combined effects of 18–25% cooled EGR

  3. Random Start Ovarian Stimulation for Oocyte or Embryo Cryopreservation in Women Desiring Fertility Preservation Prior to Gonadotoxic Cancer Therapy.

    Science.gov (United States)

    Danis, Rachel B; Pereira, Nigel; Elias, Rony T

    2017-11-10

    Women of reproductive age diagnosed with cancer are often interested in preserving gametes or reproductive tissue that would allow for future genetic parenthood. Preservation of fertility is often accomplished in young cancer patients via ovarian stimulation followed by oocyte or embryo cryopreservation. Conventional stimulation protocols, however, require 2-4 weeks to complete ovarian stimulation, oocyte retrieval and possible fertilization. Such a strategy may not be feasible in patients requiring urgent cancer treatment. Recent studies have highlighted that random start ovarian stimulation can be initiated irrespective of the phase of the menstrual cycle and is an attractive alternative to conventional ovarian stimulation. The primary aim of the current review is to discuss the feasibility and success of random start ovarian stimulation for oocyte or embryo cryopreservation in women desiring fertility preservation prior to gonadotoxic cancer therapy. We performed a systematic review of medical literature published between January 2000 to June 2017 reporting the utility of random start ovarian stimulation for fertility preservation. Search terms included "fertility preservation," "cancer," "ovarian stimulation," "random-start ovarian stimulation," "embryo cryopreservation, and" "oocyte cryopreservation." Publications were included in this review only if patients underwent random start ovarian stimulation prior to cancer therapy. Nineteen publications were identified and perused by the authors. Most publications described the utility of random start ovarian stimulation in the setting of breast cancer. Radom-start stimulation was associated with a reduced time interval between ovarian stimulation initiation and oocyte or embryo cryopreservation. The yield of mature oocytes and their developmental potential into embryos was comparable between conventional and random-start protocols, albeit with higher gonadotropin doses in the latter. The current review suggests

  4. Numerical Optimization of a Bifacial Bi-Glass Thin-Film a-Si:H Solar Cell for Higher Conversion Efficiency

    Science.gov (United States)

    Berrian, Djaber; Fathi, Mohamed; Kechouane, Mohamed

    2018-02-01

    Bifacial solar cells that maximize the energy output per a square meter have become a new fashion in the field of photovoltaic cells. However, the application of thin-film material on bifacial solar cells, viz., thin-film amorphous hydrogenated silicon ( a- Si:H), is extremely rare. Therefore, this paper presents the optimization and influence of the band gap, thickness and doping on the performance of a glass/glass thin-film a- Si:H ( n- i- p) bifacial solar cell, using a computer-aided simulation tool, Automat for simulation of hetero-structures (AFORS-HET). It is worth mentioning that the thickness and the band gap of the i-layer are the key parameters in achieving higher efficiency and hence it has to be handled carefully during the fabrication process. Furthermore, an efficient thin-film a- Si:H bifacial solar cell requires thinner and heavily doped n and p emitter layers. On the other hand, the band gap of the p-layer showed a dramatic reduction of the efficiency at 2.3 eV. Moreover, a high bifaciality factor of more than 92% is attained, and top efficiency of 10.9% is revealed under p side illumination. These optimizations demonstrate significant enhancements of the recent experimental work on thin-film a- Si:H bifacial solar cells and would also be useful for future experimental investigations on an efficient a- Si:H thin-film bifacial solar cell.

  5. Germination, carbohydrate composition and vigor of cryopreserved Caesalpinia echinata seeds

    Directory of Open Access Journals (Sweden)

    Rafael Fonsêca Zanotti

    2012-10-01

    Full Text Available The present study investigated the germination and vigor of Caesalpinia echinata (Brazilwood seeds stored at negative temperatures. Recently harvested seeds were cryopreserved at -18º or -196ºC and periodically evaluated for germination, seed vigor and carbohydrate composition. The temperatures did not influence the germination percentages or vigor. The germination percentage decreased from 88% in recently harvested seeds to 60% after 730 days of storage. The different temperature and storage times tested did not affect the vigor seed germination as indicated by the measures of plant growth and survival. The different temperatures used did not cause changes in the carbohydrate composition. The tegument cell walls were rich in lignin, arabinose and xylose. The cytoplasm of the cotyledons and embryos had high levels of glucose, fructose, and sucrose. The cryopreservation technique here presented was effective in the conservation of Brazilwood seeds for the medium term.

  6. Efficiency of assisted hatching of the cryopreserved–melted embryos

    Directory of Open Access Journals (Sweden)

    V. A. Pitko

    2018-04-01

    Full Text Available Purpose. To measure outcomes of clinical research of efficiency of assisted hatching of cryopreserved embryos. Materials and methods. Patients who had un successful cycles IVF/ICSI with transfer of fresh embryos have been selected for participation in the research between 2014 and 2016 years. Patients were distributed in a random way for participation in the experiment and control groups. Results of embryos transfer of one or two cryopreserved and melted embryos were considered only. Embryos were cryopreserved at a stage of blastocyst, 5 days after extraction of oocytes by method of vitrification. Melting procedure was conducted in the morning of a day of embryos transfer following the instructions of the vitrification medium producer Cryotech (Japan. Assisted hatching was conducted with use of micropipettes of Holding Pipette Cook Medical (Australia and Assisted Hatching/Zona Drilling Pipette Cook Medical (Australia. The treated embryos were cultivated up to a repeated estimation of morphology of embryos before transfer. Transfer of embryos has been conducted by a standard method with the use of catheter for non-invasive transfer of embryo Sydney IVF Cook Medical (Australia. The quantity of the transferred embryos varied from one to two. Results. 100 cryopreserved embryos were transferred which have been distributed in a random way either to the group with the assisted hatching or to the control group (without assisted hatching. A number of parameters of patients from both groups was analyzed, i.e. age of the patient at the time of melting of embryos, duration of infertility, causes of infertility, quantity of previous unsuccessful cycles IVF/ICSI. Any essential differences between patients within two groups based on the aforementioned parameters were not revealed. Also, there were no essential differences in number of the melted embryos, survival rate of embryos, quantity of the embryos transferred to patients. However, at the same time

  7. Fertilization and Embryo Development of Fresh and Cryopreserved Sibling Oocytes

    Directory of Open Access Journals (Sweden)

    Robert F. Casper

    2010-01-01

    Full Text Available Background: Oocyte cryopreservation is potentially the best way to preserve female fertility forunmarried women or young girls at risk of losing ovarian function. The aim of this study was tocompare fertilization and embryo development in frozen-thawed oocytes to their fresh siblings inwomen undergoing in vitro fertilization (IVF and embryo transfer (ET.Materials and Methods: Eleven infertile women undergoing infertility treatment, between theages of 24 to 37 years (mean ± SD = 31.6 ± 3.5, were included in this study. Mature oocytesfrom each patient were randomized into cryopreserved and fresh groups prior to intracytoplasmicsperm injection (ICSI. One hundred and thirty nine oocytes were retrieved, of which 105 were atmetaphase II (MII. Forty- five fresh MII oocytes were kept in culture whereas their sibling 60 MIIoocytes were cryopreserved using a slow cooling protocol. The frozen oocytes remained in LN2for 2 hours before thawing. ICSI was performed 1-2 hours after thawing for frozen oocytes and 4-5hours after retrieval for fresh oocytes. Fertilization and embryo development were compared.Results: Following thawing, 31 oocytes (51.6 % survived and 22 fertilized (79% while 32 freshoocytes fertilized upon ICSI (71%. The mean ± SE scores for embryos developing from frozenthawedoocytes were significantly lower at 48 and 72 hours post-ICSI than for embryos resultingfrom fresh oocytes (p<0.05.Conclusion: Our data demonstrated that oocyte freezing resulted in acceptable survival ratesfollowing cryopreservation, and similar fertilization rates following ICSI as compared to the freshsibling oocytes. However the number of blastomeres and the embryo quality on day three wassuperior in embryos from fresh oocytes when compared to the frozen oocytes.

  8. Effective Condition for Whole Testis Cryopreservation of Endangered Miho Spine Loach (Cobitis choii) Through the Optimization of Mud Loach (Misgurnus mizolepis) Whole Testis Cryopreservation Condition.

    Science.gov (United States)

    Kim, J J; Nam, Y K; Bang, I C; Gong, S P

      BACKGROUND: Miho spine loach (Cobitis choii) is an endangered Korean endemic fish. Whole testis cryopreservation is a good way for species preservation, but needs to the sacrifice of a large number of fish to optimize the freezing condition. Considering this limitation, a surrogate fish species was used for the protocol development. This study was to establish the effective condition for Miho spine loach whole testis cryopreservation by optimizing the conditions for whole testis cryopreservation in an allied species, mud loach (Misgurnus mizolepis). The condition for whole testis cryopreservation was optimized in mud loach first, and then the optimal condition was applied to Miho spine loach testes. The optimal condition for mud loach testis cryopreservation consists of the freezing medium containing 1.3 M dimethyl sulfoxide, 6% fetal bovine serum and 0.3 M trehalose, -1 C/min cooling rate and 26 degree C thawing temperature, which also permits effective cryopreservation of Miho spine loach testes. An effective cryopreservation condition for whole testis of the endangered Miho spine loach has been established by using mud loach as a surrogate fish.

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

    Science.gov (United States)

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

    2006-07-01

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

  10. Cryopreservation of cocoa (Theobroma cacao L.) somatic embryos by vitrification.

    Science.gov (United States)

    Adu-Gyamfi, Raphael; Wetten, Andy

    2012-01-01

    Losses of cultivated cocoa (Theobroma cacao L.) due to diseases and continued depletion of forests that harbour the wild progenitors of the crop make ex situ conservation of cocoa germplasm of paramount importance. In order to enhance security of in situ germplasm collections, 2-3 mm floral-derived secondary somatic embryos were cryopreserved by vitrification. This work demonstrates the most uncomplicated clonal cocoa cryopreservation. Optimal post-cryostorage survival (74.5 percent) was achieved by 5 d preculture of SSEs on 0.5 M sucrose medium followed by 60 min dehydration in cold PVS2. To minimise free radical related cryo-injury, cation sources were removed from the embryo development solution and/or the recovery medium, the former treatment resulting in a significant benefit. After optimisation with cocoa genotype AMAZ 15, the same protocol was effective across all five additional cocoa genotypes tested. For the multiplication of clones, embryos regenerated following cryopreservation were used as explant sources, and vitrification was found to maintain their embryogenic potential.

  11. Cryopreservation of in vitro shoot apices of Oxalis tuberosa Mol.

    Science.gov (United States)

    Gonzalez-Benito, M E; Mendoza-Condori, V H; Molina-Garcia, A D

    2007-01-01

    Oca (Oxalis tuberosa Mol.) is an under-utilized tuber crop from the Andean region. Cryopreservation would allow the safe and long-term preservation of the genetic resources of this crop. A protocol for the cryopreservation of in vitro grown shoots has been developed using the vitrification solution PVS2. Two genotypes were studied (G1 and G27). Nodal segments were cultured on MS medium and incubated at 10 degree C with 16 h photoperiod and 10 mol per square meter per second irradiance, for two weeks. Apices were then excised and cultured on MS+0.15 M sucrose for 3 days at 5 degree C in darkness. Subsequently, apices were immersed in a loading solution (liquid MS medium+2 M glycerol+0.4 M sucrose), and then treated with the vitrification solution PVS2 for 0 to 40 minutes. Cryovials were then immersed in liquid nitrogen. Four weeks after rewarming and culture on recovery medium, genotype G1 showed approximately 60 percent recovery (normal growth) with 20 min PVS2 treatment. Genotype G27 showed lower recovery (30 percent). Differential scanning calorimetry yielded a Tg midpoint for PSV2 solution of ca. -120 degree C. Calorimetric studies on apices at different stages of the cryopreservation protocol showed a change in calorimetric parameters consistent with a decrease in the amount of frozen water as the protocol advanced.

  12. [Prospects for Application of Gases and Gas Hydrates to Cryopreservation].

    Science.gov (United States)

    Shishova, N V; Fesenko, E E

    2015-01-01

    In the present review, we tried to evaluate the known properties of gas hydrates and gases participating in the formation of gas hydrates from the point of view of the mechanisms of cryoinjury and cryoprotection, to consider the papers on freezing biological materials in the presence of inert gases, and to analyze the perspectives for the development of this direction. For the purpose, we searched for the information on the physical properties of gases and gas hydrates, compared processes occured during the formation of gas hydrates and water ice, analyzed the influence of the formation and growth of gas hydrates on the structure of biological objects. We prepared a short review on the biological effects of xenon, krypton, argon, carbon dioxide, hydrogen sulfide, and carbon monoxide especially on hypothermal conditions and probable application of these properties in cryopreservation technologies. The description of the existing experiments on cryopreservation of biological objects with the use of gases was analyzed. On the basis of the information we found, the most perspective directions of work in the field of cryopreservation of biological objects with the use of gases were outlined. An attempt was made to forecast the potential problems in this field.

  13. Development of a sperm cryopreservation protocol for redside dace, Clinostomus elongatus: implications for genome resource banking

    DEFF Research Database (Denmark)

    Butts, Ian A.E.; Mokdad, A.; Trippel, E.A.

    2013-01-01

    of extirpation.We developed cryopreservation protocols by testing the effects of diluent (buffered sperm motility-inhibiting saline solution [BSMIS]; BSMIS + glycine; sucrose; and Hanks’ balanced salt solution [HBSS]), cryoprotectant (dimethyl sulfoxide [DMSO]; propylene glycol [PG]; N,N-dimethylacetamide [DMA...... rate was not significant for BSMIS + glycine or for HBSS, but an effect was detected for sucrose, with sperm frozen at 5◦C/min or 10◦C/min having higher motility than sperm frozen at 1◦C/min. The effect of extender was not significant at 1◦C/min or 5◦C/min, but an effect was detected at 10◦C...

  14. Efficiency of Photosynthesis in a Chl d-Utilizing Cyanobacterium is Comparable to or Higher than that in Chl a-Utilizing Oxygenic Species

    Science.gov (United States)

    Mielke, S. P.; Kiang, N. Y.; Blankenship, R. E.; Gunner, M. R.; Mauzerall, D.

    2011-01-01

    The cyanobacterium Acaryochloris marina uses chlorophyll d to carry out oxygenic photosynthesis in environments depleted in visible and enhanced in lower-energy, far-red light. However, the extent to which low photon energies limit the efficiency of oxygenic photochemistry in A. marina is not known. Here, we report the first direct measurements of the energy-storage efficiency of the photosynthetic light reactions in A. marina whole cells,and find it is comparable to or higher than that in typical, chlorophyll a-utilizing oxygenic species. This finding indicates that oxygenic photosynthesis is not fundamentally limited at the photon energies employed by A. marina, and therefore is potentially viable in even longer-wavelength light environments.

  15. Higher photosynthesis, nutrient- and energy-use efficiencies contribute to invasiveness of exotic plants in a nutrient poor habitat in northeast China.

    Science.gov (United States)

    Liu, Ming-Chao; Kong, De-Liang; Lu, Xiu-Rong; Huang, Kai; Wang, Shuo; Wang, Wei-Bin; Qu, Bo; Feng, Yu-Long

    2017-08-01

    The roles of photosynthesis-related traits in invasiveness of introduced plant species are still not well elucidated, especially in nutrient-poor habitats. In addition, little effort has been made to determine the physiological causes and consequences of the difference in these traits between invasive and native plants. To address these problems, we compared the differences in 16 leaf functional traits related to light-saturated photosynthetic rate (P max ) between 22 invasive and native plants in a nutrient-poor habitat in northeast China. The invasive plants had significantly higher P max , photosynthetic nitrogen- (PNUE), phosphorus- (PPUE), potassium- (PKUE) and energy-use efficiencies (PEUE) than the co-occurring natives, while leaf nutrient concentrations, construction cost (CC) and specific leaf area were not significantly different between the invasive and native plants. The higher PNUE contributed to higher P max for the invasive plants, which in turn contributed to higher PPUE, PKUE and PEUE. CC changed independently with other traits such as P max , PNUE, PPUE, PKUE and PEUE, showing two trait dimensions, which may facilitate acclimation to multifarious niche dimensions. Our results indicate that the invasive plants have a superior resource-use strategy, i.e. higher photosynthesis under similar resource investments, contributing to invasion success in the barren habitat. © 2017 Scandinavian Plant Physiology Society.

  16. A HIGHER EFFICIENCY OF CONVERTING GAS TO STARS PUSHES GALAXIES AT z ∼ 1.6 WELL ABOVE THE STAR-FORMING MAIN SEQUENCE

    Energy Technology Data Exchange (ETDEWEB)

    Silverman, J. D.; Rujopakarn, W. [Kavli Institute for the Physics and Mathematics of the Universe (WPI), The University of Tokyo Institutes for Advanced Study, The University of Tokyo, Kashiwa, Chiba 277-8583 (Japan); Daddi, E.; Liu, D. [Laboratoire AIM, CEA/DSM-CNRS-Universite Paris Diderot, Irfu/Service d’Astrophysique, CEA Saclay (France); Rodighiero, G. [Dipartimento di Fisica e Astronomia, Universita di Padova, vicolo Osservatorio, 3, I-35122 Padova (Italy); Sargent, M. [Astronomy Centre, Department of Physics and Astronomy, University of Sussex, Brighton BN1 9QH (United Kingdom); Renzini, A. [Instituto Nazionale de Astrofisica, Osservatorio Astronomico di Padova, v.co dell’Osservatorio 5, I-35122 Padova (Italy); Feruglio, C. [IRAM—Institut de RadioAstronomie Millimétrique, 300 rue de la Piscine, F-38406 Saint Martin d’Hères (France); Kashino, D. [Division of Particle and Astrophysical Science, Graduate School of Science, Nagoya University, Nagoya 464-8602 (Japan); Sanders, D. [Institute for Astronomy, University of Hawaii, 2680 Woodlawn Drive, Honolulu, HI 96822 (United States); Kartaltepe, J. [National Optical Astronomy Observatory, 950 N. Cherry Avenue, Tucson, AZ 85719 (United States); Nagao, T. [Graduate School of Science and Engineering, Ehime University, 2-5 Bunkyo-cho, Matsuyama 790-8577 (Japan); Arimoto, N. [Subaru Telescope, 650 North A’ohoku Place, Hilo, HI-96720 (United States); Berta, S.; Lutz, D. [Max-Planck-Institut für extraterrestrische Physik, D-84571 Garching (Germany); Béthermin, M. [European Southern Observatory, Karl-Schwarzschild-Strasse 2, D-85748 Garching (Germany); Koekemoer, A., E-mail: john.silverman@ipmu.jp [Space Telescope Science Institute, 3700 San Martin Drive, Baltimore, MD, 21218 (United States); and others

    2015-10-20

    Local starbursts have a higher efficiency of converting gas into stars, as compared to typical star-forming galaxies at a given stellar mass, possibly indicative of different modes of star formation. With the peak epoch of galaxy formation occurring at z > 1, it remains to be established whether such an efficient mode of star formation is occurring at high redshift. To address this issue, we measure the molecular gas content of seven high-redshift (z ∼ 1.6) starburst galaxies with the Atacama Large Millimeter/submillimeter Array and IRAM/Plateau de Bure Interferometer. Our targets are selected from the sample of Herschel far-infrared-detected galaxies having star formation rates (∼300–800 M{sub ⊙} yr{sup −1}) elevated (≳4×) above the star-forming main sequence (MS) and included in the FMOS-COSMOS near-infrared spectroscopic survey of star-forming galaxies at z ∼ 1.6 with Subaru. We detect CO emission in all cases at high levels of significance, indicative of high gas fractions (∼30%–50%). Even more compelling, we firmly establish with a clean and systematic selection that starbursts, identified as MS outliers, at high redshift generally have a lower ratio of CO to total infrared luminosity as compared to typical MS star-forming galaxies, although with a smaller offset than expected based on past studies of local starbursts. We put forward a hypothesis that there exists a continuous increase in star formation efficiency with elevation from the MS with galaxy mergers as a possible physical driver. Along with a heightened star formation efficiency, our high-redshift sample is similar in other respects to local starbursts, such as being metal rich and having a higher ionization state of the interstellar medium.

  17. Sperm cryopreservation in endangered felids: developing linkage of in situ-ex situ populations.

    Science.gov (United States)

    Swanson, W F; Magarey, G M; Herrick, J R

    2007-01-01

    Many of the world's cat species face growing threats to their continued survival in nature. For some species, managed captive populations may provide a reservoir for future reintroduction or genetic augmentation. Because most zoo populations are derived from small founder sizes and are subject to loss of genetic variation over time, periodic infusion of founder alleles is necessary to avoid the dire consequences of inbreeding. Collection and freezing of semen from free-living nondomestic felids offers a viable option for introducing founder genes into captive populations without removal of animals from the wild. The effective application of this strategy requires established protocols for safely capturing and anaesthetising wild cats coupled with suitable methods of semen recovery, processing and cryopreservation under field conditions. In small-sized non-domestic felids, the general feasibility of this approach is being explored in two studies of black-footed cats and Pallas' cats. Two factors - relatively low sperm numbers per ejaculate and compromised status of frozen-thawed cat spermatozoa - suggest that in vitro fertilisation (IVF) and embryo transfer present the most efficient use of this limiting resource in small-sized cats. Our studies with captive felids have explored alternative methods of sperm cryopreservation that are adaptable to field situations and shown that frozen-thawed spermatozoa from Pallas' cats, ocelots, and fishing cats exhibit adequate function to fertilise heterologous and/or homologous oocytes in vitro. Most recently, we investigated the fertilising capacity of frozen-thawed spermatozoa obtained from wild Pallas' cats in Mongolia. Combined with improved methods for embryo culture and transfer in small cat species, sperm banking in situ will facilitate introduction of new founders into captive populations without causing further depletion of their wild counterparts. As one component of holistic conservation programs, including ongoing

  18. Utilization and quality of cryopreserved red blood cells in transfusion medicine

    NARCIS (Netherlands)

    Henkelman, S.; Noorman, F.; Badloe, J. F.; Lagerberg, J. W. M.

    Cryopreserved (frozen) red blood cells have been used in transfusion medicine since the Vietnam war. The main method to freeze the red blood cells is by usage of glycerol. Although the usage of cryopreserved red blood cells was promising due to the prolonged storage time and the limited cellular

  19. Cryopreservation of human embryos and its contribution to in vitro fertilization success rates

    NARCIS (Netherlands)

    Wong, Kai Mee; Mastenbroek, Sebastiaan; Repping, Sjoerd

    2014-01-01

    Cryopreservation of human embryos is now a routine procedure in assisted reproductive technologies laboratories. There is no consensus on the superiority of any protocol, and substantial differences exist among centers in day of embryo cryopreservation, freezing method, selection criteria for which

  20. Cryopreservation of in vitro-grown shoot tips of apricot (Prunus ...

    African Journals Online (AJOL)

    akpobome

    2013-03-20

    Mar 20, 2013 ... Thus, they can be preserved in such a state for a long period (Sakai, 1995). Many new cryopreservation ..... formation of intracellular ice crystals during rapid cooling in LN (Wilkinson et al., 1998). Effect preculture and ..... Cryopreservation of protocorm-like bodies. (PLBs) of Phalaenopsis bellina. (Rchb.f.) ...

  1. Cryopreservation techniques and their application in vegetatively propagated crop plants in Finland

    Directory of Open Access Journals (Sweden)

    A. NUKARI

    2008-12-01

    Full Text Available Cryopreservation protocols have been introduced as techniques for germplasm preservation of vegetatively propagated horticultural and staple food crops. In Finland, cryopreservation has been studied since 1990’s, beginning with cryopreservation of forest tree breeding material and since 2004 on cryopreservation of genetic resources of horticultural plants and potato. Priority was given to cryopreservation of raspberry (Rubus ideaus L., strawberry (Fragaria x ananassa Duch. and potato (Solanum tuberosum L. and the possibility to use cryotherapy in eradication of raspberry bushy dwarf virus (RBDV from in vitro cultures were studied on raspberry. Modified droplet vitrification cryopreservation protocols were designed for raspberry and strawberry and cryotherapy combined with thermotherapy was proven to be a successful application to eliminate RBDV from infected raspberries. Cryotherapy method can be applied for a large scale elimination of viruses from plant germplasm and from candidate nuclear stock in a certified plant production scheme. Routine use of cryotechniques in germplasm preservation of vegetatively propagated horticultural plants was started. Besides for long term germplasm preservation, cryopreservation techniques can be applied also for maintenance of mother stocks in certified plant production schemes and in commercial plant production. Cryopreservation of potato shoot tips needs additional detailed research to obtain sufficient recovery and regrowth rates.;

  2. Parental desire and acceptability of spermatogonial stem cell cryopreservation in boys with cancer

    NARCIS (Netherlands)

    van den Berg, H.; Repping, S.; van der Veen, F.

    2007-01-01

    BACKGROUND: In the near future, a substantial proportion of adults will be childhood cancer survivors. The cryopreservation and transplantation of spermatogonial stem cells (SSCs) is currently successful in animals; application in humans seems likely in the near future. Cryopreserving SSCs might

  3. The Effect of L-Carnitine, Hypotaurine, and Taurine Supplementation on the Quality of Cryopreserved Chicken Semen

    Directory of Open Access Journals (Sweden)

    Agnieszka Partyka

    2017-01-01

    Full Text Available The objective of this study was to investigate the effect of L-carnitine (LC, hypotaurine (HT, and taurine (T on the quality of frozen-thawed chicken semen. Pooled semen samples were divided into seven aliquots (control, 1 mM LC, 5 mM LC, 1 mM HT, 10 mM HT, 1 mM T, and 10 mM T and subjected to cryopreservation. Postthaw sperm motility was determined by IVOS system and sperm characteristics were assessed with fluorochromes and flow cytometry. The highest sperm motility and the highest percentage of viable sperm were in the HT1 group (P<0.01 and P<0.05 following cryopreservation. After thawing, we observed a higher percentage of sperm without apoptosis and membrane reorganization changes in the LC1 and T1 group when compared to the control (P<0.05. There was a higher percentage of live sperm without lipid peroxidation (LPO in all treatments (P<0.01; P<0.05, when compared to the control group. The percentage of sperm with high mitochondrial potential significantly increased with LC1, T1, and T10 (P<0.05. Supplementation of the diluent with LC1, LC5, and T1 significantly (P<0.05 reduced DNA susceptibility to fragmentation, compared to the control and HT1 groups. These results indicate that the addition of examined antioxidants improves the quality of cryopreserved chicken semen.

  4. Banking cryopreserved heart valves in Europe: assessment of a 5-year operation in an international tissue bank in Brussels.

    Science.gov (United States)

    Goffin, Y; Grandmougin, D; Van Hoeck, B

    1996-01-01

    The heart valve bank of the European Homograft Bank has been set up in 1988 to meet the growing demand of cardiac surgeons for various sized and quality controlled cryopreserved homografts. Heart valve donors less than 60 years of age were classified in 3 categories: multiorgan donors with non transplantable hearts, recipients of cardiac transplantation and non beating heart cadavers with a warm ischemic time of less than 6 hours. Past history and biology were checked for transmissible diseases. Preparation, progressive freezing and storage in liquid nitrogen vapors, and quality control were according to the standards of the Belgian Ministry of Health. From end January 1989 to end May 1994, 989 homograft valves were cryopreserved (514 pulmonary, 475 aortic and 3 mitral) whereas 962 valves were discarded. The first cause of rejection being a major macroscopic lesion (41.48%). 138 hearts accepted at inspection were contaminated and 43 cases remained so after antibiotics. 38 cases were positive for hepatitis B or C. Complication at distribution and thawing included 10 instances of bag rupture and 15 of transversal fracture through the wall of the conduit. 477 aortic, 474 pulmonary valves as well as one mitral were implanted between May 1989 and May 1994, either for left or right ventricular outflow tract reconstruction. In the left ventricular outflow tract series 111 aortic and 23 pulmonary homograft valves were used in cases of native endocarditis, prosthetic endocarditis or recurrent endocarditis after homograft implantation. 9.6% of the requests could no be satisfied. Regular follow up information was available from 382 implants-40.1% only. The assessment of 5 years operation of the heart valve bank indicates: 1) the efficiency of selecting, cryopreserving and allocating quality controlled homograft valves from a large pool of donor hearts provided by a network of hospitals; 2) the difficulty of obtaining regular follow up information on the implants.

  5. Cryopreservation of Indian red jungle fowl (Gallus gallus murghi) semen with polyvinylpyrrolidone.

    Science.gov (United States)

    Rakha, Bushra Allah; Ansari, Muhammad Sajjad; Akhter, Shamim; Zafar, Zartasha; Hussain, Iftikhar; Santiago-Moreno, Julian; Blesbois, Elisabeth

    2017-10-01

    The Indian red jungle fowl is a sub-species of the genus Gallus native to South Asia; facing high risk of extinction in its native habitat. During cryopreservation, permeable cryoprotectants like glycerol are usually employed and we previously showed encouraging results with 20% glycerol. Because bird spermatozoa contain very little intracellular water, the possibility of replacing an internal cryoprotectant by an external one is opened. In the present study, we tested the replacement of internal cryoprotectant glycerol by the external cryoprotectant Polyvinylpyrrolidone (PVP). PVP is a non-permeable cryoprotectant and keeps the sperm in glassy state both in cooling and warming stages without making ice crystallization within the sperm cell. We evaluated the effect of various levels of polyvinylpyrrolidone (PVP) on Indian red jungle fowl semen quality and fertility outcomes. The qualifying semen ejaculates collected from eight mature cocks were pooled, divided into five aliquots, diluted (37 °C) with red fowl semen extender having PVP [0% (control) 4% (w/v), 6% (w/v), 8% (w/v) and 10% (w/v)]. Diluted semen was cryopreserved and stored in liquid nitrogen. The whole experiment was repeated/replicated for five times independently. Sperm motility, plasma membrane integrity, viability and acrosome integrity were recorded highest (P < 0.05) with 6% PVP at post-dilution, cooling, equilibration and freeze-thawing. Higher (P < 0.05) no. of fertile eggs, fertility, no. of hatched chicks, percent hatch and hatchability was recorded with 6% PVP compared to control. It is concluded that 6% PVP maintained better post-taw quality and fertility of Indian red jungle fowl spermatozoa than glycerol and can be used in routine practice avoiding the contraceptive effects of glycerol. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Comparison of two different extenders for cryopreservation of epididymal dog sperm.

    Science.gov (United States)

    Martins, Mim; Justino, R C; Sant'anna, M C; Trautwein, L G C; Souza, F F

    2012-12-01

    The collection of epididymal sperm is an option for preservation of germplasm of genetically superior animals that need to be orchiectomized or have died. The extender type used to freeze sperm is important to avoid spermatozoal membrane damage and to preserve semen quality after cryopreservation. The objective of this study was to verify the effects of a commercial bovine extender (Bovimix(®); Nutricell, Campinas) and a traditional TRIS-citric acid-glucose-egg yolk-7% glycerol extender on cryopreservation of canine epididymal sperm. The testes of 13 adult dogs were kept at 5 °C for 24 h in saline solution, and epididymal sperm was recovered in Ringers solution without lactate and were evaluated for motility. Samples with ≥ 80% motility were pooled and then divided before dilution and packaging in 0.5 ml plastic straws, equilibration at 4 °C for 1 h, freezing in nitrogen vapour for 20 min and storing at -196 °C. The straws were thawed at 56 °C for 10 s and were evaluated for motility by computer assisted analysis (CASA). The semen parameters, sperm movement index, linearity, total motility and rapid progressive motility were statistically higher in Bovimix(®) than TRIS. In contrast, amplitude of lateral head displacement, slow sperm and static sperm were lower in Bovimix(®). Despite the high percentage of sperm defects in epididymal cells, regardless of the extender, we concluded that Bovimix(®) is a viable alternative for the freezing of canine epididymal sperm. © 2012 Blackwell Verlag GmbH.

  7. USE OF DIFFERENT EXTENDERS TO CRYOPRESERVATION OF MANGALARGA MARCHADOR SPERM

    Directory of Open Access Journals (Sweden)

    Jessica Neri Nascimento

    2015-07-01

    Full Text Available The techniques applied to animal reproduction such as artificial insemination, transfer and in vitro production of embryos, heat synchronization and induction, and gametes cryopreservation have been more utilized in veterinary practice each day. Nevertheless, some techniques have not achieved their full technical capacity within the equine reproduction field, such as semen cryopreservation. The aim of this study was to evaluate the characteristics of post-thawing semen (total motility, strength, plasmatic and acrosomal membrane integrity of Mangalarga Marchador breed stallions, using three different extenders (Botucrio, FR5 and FR6. Ejaculates from five stallions were collected and the gel-free semen was diluted in a 1:1 dilution in skim milk extender, and centrifuged at 600 g for 10 minutes. After the centrifugation the supernatant was removed and sperm pellet was divided and re-suspended using three different extenders to a concentration of 200 x 106 cells/mL. The samples were packed into 0.5 mL straws, placed in a stainless steel support and kept inside the refrigerator (5 oC for one hour. Subsequently, these straws were kept at a height of 6 cm from liquid nitrogen for 15 minutes in an isotherm box and, after that, plunged into liquid nitrogen (-196 oC and stored in a liquid nitrogen holding tank. There were no differences in the parameters evaluated when extenders using mixed amides and glycerol (Botucrio and FR6 were used (P > 0.05. All parameters evaluated were lower for the extender containing only glycerol (P<0.05. The use of cryoprotectants (methylformamide and dimethylformamide in association with glycerol concentrations around 1 to 2% is an alternative for semen cryopreservation of Mangalarga Marchador breed stallions.

  8. Two-piece cryopreserved tracheal allotransplantation: an experimental study.

    Science.gov (United States)

    Iyikesici, Tuncel; Tuncozgur, Bulent; Sanli, Maruf; Isik, Ahmet Feridun; Meteroglu, Fatih; Elbeyli, Levent

    2009-10-01

    For successful reconstruction with tracheal allotransplants following long tracheal resections, problems related to the preservation and vascularisation of the tracheal graft have to be solved. In this study, instead of using a long-segment single-piece graft, we used a graft that has been split into two. The aim was to use this graft after cryopreservation in order to ease neo-vascularisation and to maintain tracheal integrity by transplanting it to two separate regions of the dog cervical trachea. This experimental study was conducted in animal laboratories of the medical school on 11 half-blood dogs. The trachea obtained from the first dog was 8 cm in length; it was split into two pieces of 4 cm each and stored in the preservation solution at -80 degrees C for 4 weeks. Following this, the dog was sacrificed. Two 2 cm portions of cervical trachea were excised from the second dog. These parts were then reconstructed with two tracheal grafts of the same length as the cryopreserved ones. Ten dogs that were grouped into five groups of two dogs each underwent the same procedure. The subjects had a bronchoscopic evaluation on the third postoperative week. Anastomosis regions of the test tracheas were resected to be examined histopathologically. Seven subjects were found to have third-degree obstructions during bronchoscopy; two had close to fourth-degree obstructions. In the histopathological examination, contrary to the findings of the bronchoscopies, 75% of the anastomoses had intact epithelium. The cartilage was seen to have well-preserved structural characteristics in all the anastomoses. Twelve anastomoses had moderate, seven mild and one had severe inflammation. All anastomoses had either good or very good level of vascularisation. The integrity of the tracheal epithelium can be maintained with cryopreservation and split anastomosis technique. The cartilage preserves its structural characteristics despite losing its viability, thereby offering an advantage to

  9. Effects of scaffold surface morphology on cell adhesion and survival rate in vitreous cryopreservation of tenocyte-scaffold constructs

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Zhi [State Key Laboratory of Biotherapy, West China Hospital of Sichuan University, Chengdu 610041 (China); Department of Bone and Joint Surgery, The affiliated hospital of Luzhou Medical College, Luzhou 646000 (China); Qing, Quan [Sichuan College of Traditional Chinese Medicine, Mianyang 621000 (China); Regenerative Medicine Research Center, West China Hospital of Sichuan University, Chengdu 610041 (China); Chen, Xi; Liu, Cheng-Jun; Luo, Jing-Cong [State Key Laboratory of Biotherapy, West China Hospital of Sichuan University, Chengdu 610041 (China); Hu, Jin-Lian [Institute of Textiles and Clothing, The Hong Kong Polytechnic University, Hong Kong (China); Qin, Ting-Wu, E-mail: tingwuqin@hotmail.com [State Key Laboratory of Biotherapy, West China Hospital of Sichuan University, Chengdu 610041 (China)

    2016-12-01

    Highlights: • The shapes of tenocytes varied when seeded on different surface of scaffolds. • Tenocytes were flat on smooth surface and spindle on micro-grooved surface. • Tenocytes were ellipse or spindle on porous surface. • Tenocytes got varying adhesion shape and elongation index on varying surfaces. • The tenocyte survival on porous surface was superior to the other two groups. - Abstract: The purpose of this study was to investigate the effects of scaffold surface morphology on cell adhesion and survival rate in vitreous cryopreservation of tenocyte-scaffold constructs. Tenocytes were obtained from tail tendons of rats. Polydimethylsiloxane (PDMS) was used to fabricate three types of scaffolds with varying surface morphological characteristics, i.e., smooth, micro-grooved, and porous surfaces, respectively. The tenocytes were seeded on the surfaces of the scaffolds to form tenocyte-scaffold constructs. The constructs were cryopreserved in a vitreous cryoprotectant (CPA) with a multi-step protocol. The cell adhesion to scaffolds was observed with electronic scanning microscopy (SEM). The elongation index of the living tenocytes and ratio of live/dead cell number were examined based on a live/dead dual fluorescent staining technique, and the survival rate of tenocytes was studied with flow cytometry (FC). The results showed the shapes of tenocytes varied between the different groups: flat or polygonal (on smooth surface), spindle (on micro-grooved surface), and spindle or ellipse (on porous surface). After thawing, the porous surface got the most living tenocytes and a higher survival rate, suggesting its potential application for vitreous cryopreservation of engineered tendon constructs.

  10. Effects of three different types of antifreeze proteins on mouse ovarian tissue cryopreservation and transplantation.

    Directory of Open Access Journals (Sweden)

    Jaewang Lee

    Full Text Available Ovarian tissue (OT cryopreservation is effective in preserving fertility in cancer patients who have concerns about fertility loss due to cancer treatment. However, the damage incurred at different steps during the cryopreservation procedure may cause follicular depletion; hence, preventing chilling injury would help maintain ovarian function.This study was designed to investigate the beneficial effects of different antifreeze proteins (AFPs on mouse ovarian tissue cryopreservation and transplantation.Ovaries were obtained from 5-week-old B6D2F1 mice, and each ovary was cryopreserved using two-step vitrification and four-step warming procedures. In Experiment I, ovaries were randomly allocated into fresh, vitrification control, and nine experimental groups according to the AFP type (FfIBP, LeIBP, type III and concentration (0.1, 1, 10 mg/mL used. After vitrification and warming, 5,790 ovarian follicles were evaluated using histology and TUNEL assays, and immunofluorescence for τH2AX and Rad51 was used to detect DNA double-strand breaks (DSBs and repair (DDR, respectively. In Experiment II, 20 mice were randomly divided into two groups: one where the vitrification and warming media were supplemented with 10 mg/mL LeIBP, and the other where media alone were used (control. Ovaries were then autotransplanted under both kidney capsules 7 days after vitrification together with the addition of 10 mg/mL LeIBP in the vitrification-warming media. After transplantation, the ovarian follicles, the percentage of apoptotic follicles, the extent of the CD31-positive area, and the serum FSH levels of the transplanted groups were compared.In Experiment I, the percentage of total grade 1 follicles was significantly higher in the 10 mg/mL LeIBP group than in the vitrification control, while all AFP-treated groups had significantly improved grade 1 primordial follicle numbers compared with those of the vitrification control. The number of apoptotic (TUNEL

  11. Cryopreservation at -75 °C of Agaricus subrufescens on wheat grains with sucrose

    Directory of Open Access Journals (Sweden)

    Lienine Luiz Zaghi Júnior

    Full Text Available Abstract Agaricus subrufescens is a basidiomycete which is studied because of its medicinal and gastronomic importance; however, less attention has been paid to its preservation. This study aimed to evaluate the effect of sucrose addition to substrate and cryotube on the viability of Agaricus subrufescens cryopreserved at -20 °C and at -75 °C for one and two years. Zero, 10% or 20% sucrose was added to potato dextrose agar or wheat grain. The mycelia were cryopreserved in the absence of cryoprotectant or with sucrose solutions at 15%, 30% or 45%. After one or two years at -75 °C or at -20 °C, mycelia were thawed and evaluated about viability, initial time of growth, colony diameter and genomic stability. Cryopreservation at -20 °C is not effective to keep mycelial viability of this fungus. Cryopreservation at -75 °C is effective when sucrose is used in substrates and/or cryotubes. Without sucrose, cryopreservation at -75 °C is effective only when wheat grains are used. Physiological characteristic as mycelial colony diameter is negatively affected when potato dextrose agar is used and unaffected when wheat grain is used after two-year cryopreservation at -75 °C. The fungus genome does not show alteration after two-year cryopreservation at -75 °C.

  12. Cryopreservation of somatic embryogenic cultures of Pinus pinaster: effects on regrowth and embryo maturation.

    Science.gov (United States)

    Álvarez, José M; Cortizo, Millán; Ordás, Ricardo J

    2012-01-01

    Pinus pinaster is one of the most economically important conifers in the world. Somatic embryogenesis is a powerful tool in breeding programmes because it allows the generation of a great number of different clonal lines from seeds of superior genotypes. Unfortunately, embryogenic competence decreases with the age of cultures. Therefore, it is necessary to have a cryopreservation protocol that ensures a continuous supply of juvenile mass while allowing good maturation and conversion rates into vigorously growing plants. In this work we studied the influence of several cryopreservation parameters, such as cryoprotectant solution and pre-cooling temperature, on embryogenic culture regrowth and embryo maturation. Recovery of rewarmed samples after cryopreservation in a -150 degree C freezer depended on the cooling temperature reached prior to plunging the tubes into liquid nitrogen. As a result, we present an optimised cryopreservation protocol that ensures high recovery and embryo maturation rates. The protocol presented is a simple and fast alternative and enabled successful cryopreservation and recovery of 100 percent of the lines tested. Cryopreserved lines presented the same maturation rates as non-cryopreserved controls.

  13. Recovery and cryopreservation of epididymal sperm from agouti (Dasiprocta aguti) using powdered coconut water (ACP-109c) and Tris extenders.

    Science.gov (United States)

    Silva, M A; Peixoto, G C X; Santos, E A A; Castelo, T S; Oliveira, M F; Silva, A R

    2011-10-01

    The objective was to compare the use of powdered coconut water (ACP-109c; ACP Biotecnologia, Fortaleza, CE, Brazil) and Tris extenders for recovery and cryopreservation of epididymal sperm from agouti. The caudae epididymus and proximal ductus deferens from 10 sexually mature agoutis were subjected to retrograde washing using ACP-109c (ACP Biotecnologia) or Tris. Epididymal sperm were evaluated for motility, vigor, sperm viability, membrane integrity, and morphology. Samples were centrifuged, and extended in the same diluents plus egg yolk (20%) and glycerol (6%), frozen in liquid nitrogen, and subsequently thawed at 37°C for 1 min, followed by re-evaluation of sperm characteristics. The two extenders were similarly efficient for epididymal recovery, with regard to the number and quality of sperm recovered. However, for both extenders, sperm quality decreased (P Biotecnologia) group, which was significantly better than 9.7 ± 2.6% motile sperm with 1.2 ± 0.3 vigor in Tris. In conclusion, agouti epididymal sperm were successfully recovered using either ACP-109c (ACP Biotecnologia) or Tris extenders; however, ACP-109c (ACP Biotecnologia) was a significantly better extender for processing and cryopreserving these sperm. Copyright © 2011 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

    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

  15. A TLBO based gradient descent learning-functional link higher order ANN: An efficient model for learning from non-linear data

    Directory of Open Access Journals (Sweden)

    Bighnaraj Naik

    2018-01-01

    Full Text Available All the higher order ANNs (HONNs including functional link ANN (FLANN are sensitive to random initialization of weight and rely on the learning algorithms adopted. Although a selection of efficient learning algorithms for HONNs helps to improve the performance, on the other hand, initialization of weights with optimized weights rather than random weights also play important roles on its efficiency. In this paper, the problem solving approach of the teaching learning based optimization (TLBO along with learning ability of the gradient descent learning (GDL is used to obtain the optimal set of weight of FLANN learning model. TLBO does not require any specific parameters rather it requires only some of the common independent parameters like number of populations, number of iterations and stopping criteria, thereby eliminating the intricacy in selection of algorithmic parameters for adjusting the set of weights of FLANN model. The proposed TLBO-FLANN is implemented in MATLAB and compared with GA-FLANN, PSO-FLANN and HS-FLANN. The TLBO-FLANN is tested on various 5-fold cross validated benchmark data sets from UCI machine learning repository and analyzed under the null-hypothesis by using Friedman test, Holm’s procedure and post hoc ANOVA statistical analysis (Tukey test & Dunnett test.

  16. Cryopreserved irradiated tracheal homograft reconstruction for subglottic-tracheal stenosis

    International Nuclear Information System (INIS)

    Somyos Kunachak; Yongyudh Vajaradul; Boonchu Kulapaditharom

    1999-01-01

    Subglottic-tracheal stenosis is a common clinical entity. Handling on severe case is often problematic. Various tracheal replacement techniques have been used with varying degree of success and dispute. In this study we worked on cryopreserved irradiated tracheal homograft, of which its use in human has not been reported. The tracheas were harvested from donor cadavers within 24 hours of death in a sterile condition. After 1-2 weeks of preservation at -70 degree C, the grafts were irradiated at 25 kGy, then stored at -70 degree C until used. Four patients, 2 males and 2 females (aged 2-40 years, mean 16 years) with severe subglottic-tracheal stenosis underwent segmental tracheal graft reconstruction using this graft. Immunosuppressant was not given in any patient. The follow up period ranged from 11-1 5 months. Three patients were successfully decapulated, 1 patient developed local infection and dislodgement of intraluminal stent with subsequent restenosis. Postoperative tracheal lumen appeared near normal with histologic evidence of normal respiratory epithelium at the grafted site. In conclusion, cryopreserved irradiated tracheal homograft is a valuable alternative for tracheal transplant or reconstruction, without the need of immunosuppression

  17. Fertility in cancer patients after cryopreservation of one ovary.

    Science.gov (United States)

    Schmidt, K T; Nyboe Andersen, A; Greve, T; Ernst, E; Loft, A; Yding Andersen, C

    2013-03-01

    This questionnaire study describes the fertility and ovarian function in 143 adult female cancer survivors with only one ovary due to cryopreservation of the other. The women were asked about their ovarian function (as defined by the presence of a spontaneous menstrual cycle), pregnancies and their outcome. The mean follow-up time was 58months after cryopreservation (range 24-129months). The risk of premature ovarian failure was high in the group of patients with leukaemia (13/15; 87%) but low in the breast cancer group (5/54; 9%). Fifty-seven women had actively tried to become pregnant after end of treatment; of these, 41 women obtained a total of 68 pregnancies resulting in 45 live births and five ongoing pregnancies, 15 spontaneous abortions, one ectopic pregnancy and two elective abortions. In the remaining 86 women without a pregnancy wish, there had been five elective abortions. Ninety-three per cent of the pregnancies were after natural conception and only four cases were a result of fertility treatment. The overall risk of premature ovarian failure was low (22%). Patients who retain their ovarian function after treatment of a malignant disease have a good chance of becoming pregnant. Copyright © 2013 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  18. NEW BIOTECHNOLOGICAL METHODS FOR CRYOPRESERVATION OF REPRODUCTIVE CELLS OF STURGEON

    Directory of Open Access Journals (Sweden)

    E. N. Ponomareva

    2016-01-01

    Full Text Available The aim of the research is to increase the survivability of reproductive cells of sturgeon at cryopreservation and developing reliable technology suitable for use on an industrial scale.Methods. We have used standard methods of freezing, thawing reproductive cells, fertilization and incubation of eggs and larval rearing of sturgeon. Fundamentally new is cryoprotective composition: for sperm we have adjusted the composition of cryoprotective medium (for beluga 3% of dimethyl sulfoxide, for Russian sturgeon 4% of dimethyl sulfoxide; for freezing the eggs we have used cryoprotective mixture of unrefined vegetable and animal oils.Results. Survivability of defrosted sperm sturgeon has been increased: for Beluga it is up to 20%, for Russian sturgeon - 47%. At insemination of cryopreserved eggs of Russian sturgeon with native sperm the fertilization rate has made 41%.Main conclusions. The research proves the effectiveness of reducing the toxic effect of cryoprotective substances, thus leading to increased survivability of reproductive cells of sturgeon. During the insemination of eggs, stored in liquid nitrogen, the resulting offspring were viable and by the reactivity of the central nervous system and receptor complex it does not differ from the young obtained by conventional technology.

  19. Predictors of sperm recovery after cryopreservation in testicular cancer

    Directory of Open Access Journals (Sweden)

    James M Hotaling

    2016-01-01

    Full Text Available Our objective was to identify predictors of improved postthaw semen quality in men with testicular cancer banking sperm for fertility preservation. We reviewed 173 individual semen samples provided by 67 men with testicular germ cell tumor (TGCT who cryopreserved sperm before gonadotoxic treatment between 1994 and 2010 at our tertiary university medical center. Our main outcomes measures were independent predictors for the greater postthaw total motile count (TMC in men with TGCT. Men with NSGCT were more likely to be younger (P median fresh TMC each had increased odds of a postthaw TMC greater than median postthaw TMC. Interestingly, age, advanced cancer stage (II or III, rapid freezing protocol, and motility enhancer did not show increased odds of improved postthaw TMC in our models. In conclusion, men with TGCT or poor fresh TMC should consider preserving additional vials (at least 15 vials before oncologic treatment. Density gradient purification should be routinely used to optimize postthaw TMC in men with TGCT. Larger, randomized studies evaluating cancer stage and various cryopreservation techniques are needed to assist in counseling men with TGCT regarding fertility preservation and optimizing cryosurvival.

  20. Data on antioxidant activity in grapevine (Vitis vinifera L.) following cryopreservation by vitrification.

    Science.gov (United States)

    Lazo-Javalera, María Fernanda; Tiznado-Hernández, Martín Ernesto; Vargas-Arispuro, Irasema; Valenzuela-Soto, Elisa; Rocha-Granados, María Del Carmen; Martínez-Montero, Marcos Edel; Rivera-Domínguez, Marisela

    2015-12-01

    Cryopreservation is used for the long-term conservation of plant genetic resources. This technique very often induces lethal injury or tissue damage. In this study, we measured indicators of viability and cell damage following cryopreservation and vitrification-cryopreservation in Vitis vinifera L. axillary buds cv. "Flame seedless" stored in liquid nitrogen (LN) for: three seconds, one hour, one day, one week and one month; after LN thawed at 38 °C for three minutes. The enzymatic activity of catalase (CAT) and superoxide dismutase (SOD), as well as the amount of malondialdehyde (MDA), total protein and viability were assayed.

  1. Data on antioxidant activity in grapevine (Vitis vinifera L. following cryopreservation by vitrification

    Directory of Open Access Journals (Sweden)

    María Fernanda Lazo-Javalera

    2015-12-01

    Full Text Available Cryopreservation is used for the long-term conservation of plant genetic resources. This technique very often induces lethal injury or tissue damage. In this study, we measured indicators of viability and cell damage following cryopreservation and vitrification-cryopreservation in Vitis vinifera L. axillary buds cv. “Flame seedless” stored in liquid nitrogen (LN for: three seconds, one hour, one day, one week and one month; after LN thawed at 38 °C for three minutes. The enzymatic activity of catalase (CAT and superoxide dismutase (SOD, as well as the amount of malondialdehyde (MDA, total protein and viability were assayed.

  2. Assessment of basic seminal characteristics, sperm cryopreservation and heterologous in vitro fertilisation in the fishing cat (Prionailurus viverrinus).

    Science.gov (United States)

    Thiangtum, Khongsak; Swanson, William F; Howard, JoGayle; Tunwattana, Wanchai; Tongthainan, Dakara; Wichasilpa, Wisid; Patumrattanathan, Pornchai; Pinyopoommintr, Tanu

    2006-01-01

    Conservation of the fishing cat, a threatened south-east Asian felid, could benefit from effective ex situ genetic management and breeding programmes, including the use of assisted reproduction. The aims of the present study were to: (1) characterise basal seminal traits of fishing cats in Thailand zoos; and (2) investigate the effect of cryopreservation on sperm motility, acrosomal integrity and in vitro function. Seminal traits were evaluated in electroejaculates collected from eight males. Spermatozoa were diluted in n-tris(hydroxymethyl)-methyl-2-aminoethanesulfonic acid Tris (TEST)-yolk buffer (TYB) without glycerol, then diluted further with TYB with glycerol (4% final concentration) at either 25 degrees C or after slow cooling to 5 degrees C and frozen in straws over liquid nitrogen vapour. After thawing, sperm function was assessed by insemination of viable domestic cat oocytes. Fishing cat ejaculates averaged (+/- s.e.m.) 43.6 +/- 14.2 x 10(6) motile spermatozoa with 33.5 +/- 6.8% normal sperm morphology. Semen processing had a negligible effect (P > 0.05) on sperm motility and acrosomal integrity, but values were reduced (P cat oocytes, with 62.1% (36/58) of mature oocytes cleaving. Glycerol addition at 5 degrees C resulted in higher (P fishing cats and their spermatozoa exhibit adequate function after cryopreservation for in vitro fertilisation procedures.

  3. Cryopreserved embryo transfer: adjacent or non-adjacent to failed fresh long GnRH-agonist protocol IVF cycle.

    Science.gov (United States)

    Volodarsky-Perel, Alexander; Eldar-Geva, Talia; Holzer, Hananel E G; Schonberger, Oshrat; Reichman, Orna; Gal, Michael

    2017-03-01

    The optimal time to perform cryopreserved embryo transfer (CET) after a failed oocyte retrieval-embryo transfer (OR-ET) cycle is unknown. Similar clinical pregnancy rates were recently reported in immediate and delayed CET, performed after failed fresh OR-ET, in cycles with the gonadotrophin-releasing hormone (GnRH) antagonist protocol. This study compared outcomes of CET performed adjacently (<50 days, n = 67) and non-adjacently (≥50 to 120 days, n = 62) to the last OR-day of cycles with the GnRH agonist down-regulation protocol. Additional inclusion criteria were patients' age 20-38 years, the transfer of only 1-2 cryopreserved embryos, one treatment cycle per patient and artificial preparation for CET. Significantly higher implantation, clinical pregnancy and live birth rates were found in the non-adjacent group than in the adjacent group: 30.5% versus 11.3% (P = 0.001), 41.9% versus 17.9% (P = 0.003) and 32.3% versus 13.4% (P = 0.01), respectively. These results support the postponement of CET after a failed OR-ET for at least one menstrual cycle, when a preceding long GnRH-agonist protocol is used. Copyright © 2016 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  4. Expanded cryopreserved mesenchymal stromal cells as an optimal source for graft-versus-host disease treatment

    Czech Academy of Sciences Publication Activity Database

    Holubová, M.; Lysák, D.; Vlas, T.; Vannucci, Luca; Jindra, P.

    2014-01-01

    Roč. 42, č. 3 (2014), s. 139-144 ISSN 1045-1056 Institutional support: RVO:61388971 Keywords : Mesenchymal stromal cells * Cryopreservation * Immunomodulation Subject RIV: EC - Immunology Impact factor: 1.209, year: 2014

  5. Improved droplet-vitrification and histological studies of cryopreserved shoot tips of cultivated Jerusalem artichoke genotypes

    Science.gov (United States)

    Germplasm conservation of Jerusalem artichoke (Helianthus tuberosus L.) is crucial to preserve genetic diversity and to secure materials for genetic improvement. Long-term conservation is accomplished through cryopreservation, storing cells or tissues at an ultralow temperature in liquid nitrogen (-...

  6. Cryopreservation of common carp (Cyprinus carpio L.) sperm induces protein phosphorylation in tyrosine and threonine residues

    Czech Academy of Sciences Publication Activity Database

    Li, P.; Hulák, M.; Li, Z.; H.; Šulc, Miroslav; Pšenička, M.; Rodina, M.; Gela, D.; Linhart, O.

    2013-01-01

    Roč. 80, č. 2 (2013), s. 84-89 ISSN 0093-691X Institutional support: RVO:61388971 Keywords : Cryopreservation * Sperm * Phosphorylation Subject RIV: CE - Biochemistry Impact factor: 1.845, year: 2013

  7. Cryopreservation of sperm bundles (spermatozeugmata) from endangered livebearing goodeids.

    Science.gov (United States)

    Liu, Yue; Torres, Leticia; Tiersch, Terrence R

    2018-04-14

    More than half of fishes in the family Goodeidae are considered to be endangered, threatened, or vulnerable. Sperm cryopreservation is an effective tool for conserving genetic resources of imperiled populations, but development of protocols with livebearing fishes faces numerous challenges including the natural packaging of sperm into bundles. In this study the cryopreservation of sperm bundles (spermatozeugmata) of three goodeids species was evaluated. Sperm quality was evaluated by activation with NaCl-NaOH solution (at 300 mOsmol/kg and pH 11.8), and analysis of dissociable bundles and dissociation duration. Using Redtail Splitfin (Xenotoca eiseni) as a model, the effects of cryoprotectants (dimethyl sulfoxide, methanol, and glycerol) with different concentrations (5-15% v/v %), equilibration exposure times (1-60 min), cooling rates (5-40 °C/min), concentrations (4 × 10 4 -4 × 10 6 bundles/ml), buffers (HBSS, PBS and NaCl), and buffer osmolalities (200-400 mOsmol/kg) were investigated. After cooling and thawing, sperm bundles maintained their packed form. A specific protocol was developed (10% dimethyl sulfoxide, 20-min equilibration, 10 °C/min cooling rate, 4 × 10 6 bundles/ml, and 300 mOsmol/kg HBSS). This protocol yielded 89 ± 5% of post-thaw dissociable bundles with 209 ± 10 s of dissociation duration for X. eiseni, 96 ± 9% with 814 ± 14 s for Blackfin Goodea (Goodea atripinni), and 66 ± 2% with 726 ± 25 s for Striped Goodeid (Ataeniobius toweri). This is the first study of cryopreservation of sperm within bundles for livebearing fishes and provides a basis for establishment of germplasm repositories for goodeids and other livebearers. Copyright © 2018 Elsevier Inc. All rights reserved.

  8. Cryopreservation of Embryos of the Mediterranean Fruit Fly Ceratitis capitata Vienna 8 Genetic Sexing Strain.

    Directory of Open Access Journals (Sweden)

    Antonios A Augustinos

    Full Text Available The Mediterranean fruit fly, Ceratitis capitata, is one of the most serious pests of fruit crops world-wide. During the last decades, area-wide pest management (AW-IPM approaches with a sterile insect technique (SIT component have been used to control populations of this pest in an effective and environment-friendly manner. The development of genetic sexing strains (GSS, such as the Vienna 8 strain, has been played a major role in increasing the efficacy and reducing the cost of SIT programs. However, mass rearing, extensive inbreeding, possible bottleneck phenomena and hitch-hiking effects might pose major risks for deterioration and loss of important genetic characteristics of domesticated insect. In the present study, we present a modified procedure to cryopreserve the embryos of the medfly Vienna 8 GSS based on vitrification and used this strain as insect model to assess the impact of the cryopreservation process on the genetic structure of the cryopreserved insects. Forty-eight hours old embryos, incubated at 24°C, were found to be the most suitable developmental stage for cryopreservation treatment for high production of acceptable hatch rate (38%. Our data suggest the absence of any negative impact of the cryopreservation process on egg hatch rate, pupation rates, adult emergence rates and stability of the temperature sensitive lethal (tsl character on two established cryopreserved lines (flies emerged from cryopreserved embryos, named V8-118 and V8-228. Taken together, our study provides an optimized procedure to cryopreserve the medfly Vienna 8 GSS and documents the absence of any negative impact on the genetic structure and quality of the strain. Benefits and sceneries for utilization of this technology to support operational SIT projects are discussed in this paper.

  9. Cryopreservation of Embryos of the Mediterranean Fruit Fly Ceratitis capitata Vienna 8 Genetic Sexing Strain.

    Science.gov (United States)

    Augustinos, Antonios A; Rajamohan, Arun; Kyritsis, Georgios A; Zacharopoulou, Antigone; Haq, Ihsan Ul; Targovska, Asya; Caceres, Carlos; Bourtzis, Kostas; Abd-Alla, Adly M M

    2016-01-01

    The Mediterranean fruit fly, Ceratitis capitata, is one of the most serious pests of fruit crops world-wide. During the last decades, area-wide pest management (AW-IPM) approaches with a sterile insect technique (SIT) component have been used to control populations of this pest in an effective and environment-friendly manner. The development of genetic sexing strains (GSS), such as the Vienna 8 strain, has been played a major role in increasing the efficacy and reducing the cost of SIT programs. However, mass rearing, extensive inbreeding, possible bottleneck phenomena and hitch-hiking effects might pose major risks for deterioration and loss of important genetic characteristics of domesticated insect. In the present study, we present a modified procedure to cryopreserve the embryos of the medfly Vienna 8 GSS based on vitrification and used this strain as insect model to assess the impact of the cryopreservation process on the genetic structure of the cryopreserved insects. Forty-eight hours old embryos, incubated at 24°C, were found to be the most suitable developmental stage for cryopreservation treatment for high production of acceptable hatch rate (38%). Our data suggest the absence of any negative impact of the cryopreservation process on egg hatch rate, pupation rates, adult emergence rates and stability of the temperature sensitive lethal (tsl) character on two established cryopreserved lines (flies emerged from cryopreserved embryos), named V8-118 and V8-228. Taken together, our study provides an optimized procedure to cryopreserve the medfly Vienna 8 GSS and documents the absence of any negative impact on the genetic structure and quality of the strain. Benefits and sceneries for utilization of this technology to support operational SIT projects are discussed in this paper.

  10. Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa

    OpenAIRE

    N. Prapaiwan; T. Tharasanit; S. Punjachaipornpol; D. Yamtang; A. Roongsitthichai; W. Moonarmart; K. Kaeoket; S. Manee-in

    2016-01-01

    Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL) has been reported to have a cryoprotective property for sperm cryopreservation. Howeve...

  11. A simple improvement of the conventional cryopreservation for human ES and iPS cells

    OpenAIRE

    sprotocols

    2014-01-01

    Authors: Midori Ozawa, Yutaka Ozawa, Masashi Iemura, Arihiro Kohara, Kana Yanagihara & Miho K Furue ### Abstract In this study, a simple method for the cryopreservation of human embryonic stem (ES) and induced pluripotent stem (iPS) cells is proposed. It is based on the conventional slow-freezing method with 10% DMSO and modified mainly in a thawing protocol without specific equipment or reagents. Recovery rate of the cells cryopreserved by this method was equally high, which is compa...

  12. Effects of Droplet-Vitrification Cryopreservation Based on Physiological and Antioxidant Enzyme Activities of Brassidium Shooting Star Orchid

    Directory of Open Access Journals (Sweden)

    Safrina Rahmah

    2015-01-01

    Full Text Available Protocorm-like bodies (PLBs of Brassidium Shooting Star orchid were successfully cryopreserved using droplet-vitrification method. Vitrification based cryopreservation protocol is comprised of preculture, osmoprotection, cryoprotection, cooling, rewarming, and growth recovery and each and every step contributes to the achievement of successful cryopreservation. In order to reveal the lethal and nonlethal damage produced by cryopreservation, histological observation, scanning electron microscopy (SEM, and biochemical analysis were carried out in both cryopreserved and noncryopreserved PLBs of Brassidium Shooting Star orchid comparing with the control PLBs stock culture. Histological and scanning electron microscopy analyses displayed structural changes in cryopreserved PLBs due to the impact of cryoinjury during exposure to liquid nitrogen. Total soluble protein significantly increased throughout the dehydration process and the highest value was achieved when PLBs were stored in liquid nitrogen. Ascorbate peroxidase (APX and catalase (CAT showed the highest enzyme activities in both dehydration and cryostorage treatments indicating that stress level of PLBs was high during these stages.

  13. Advanced CFD modelling of air and recycled flue gas staging in a waste wood-fired grate boiler for higher combustion efficiency and greater environmental benefits.

    Science.gov (United States)

    Rajh, Boštjan; Yin, Chungen; Samec, Niko; Hriberšek, Matjaž; Kokalj, Filip; Zadravec, Matej

    2018-07-15

    Grate-fired boilers are commonly used to burn biomass/wastes for heat and power production. In spite of the recent breakthrough in integration of advanced secondary air systems in grate boilers, grate-firing technology needs to be advanced for higher efficiency and lower emissions. In this paper, innovative staging of combustion air and recycled flue gas in a 13 MW th waste wood-fired grate boiler is comprehensively studied based on a numerical model that has been previously validated. In particular, the effects of the jet momentum, position and orientation of the combustion air and recycled flue gas streams on in-furnace mixing, combustion and pollutant emissions from the boiler are examined. It is found that the optimized air and recycled flue gas jets remarkably enhance mixing and heat transfer, result in a more uniform temperature and velocity distribution, extend the residence time of the combustibles in the hot zone and improve burnout in the boiler. Optimizing the air and recycled flue gas jet configuration can reduce carbon monoxide emission from the boiler by up to 86%, from the current 41.0 ppm to 5.7 ppm. The findings of this study can serve as useful guidelines for novel design and optimization of the combustion air supply and flue gas recycling for grate boilers of this type. Copyright © 2018 Elsevier Ltd. All rights reserved.

  14. The use of cryopreserved sea urchin embryos (Paracentrotus lividus) in marine quality assessment.

    Science.gov (United States)

    Paredes, E; Bellas, J

    2015-06-01

    We have established for first time an ecotoxicological bioassay using cryopreserved sea urchin embryos (Paracentotus lividus) and provided a comparison to the already standardized sea urchin embryo-larval bioassay, using selected (organic and inorganic) pollutants and sediment elutriates from 4 different locations from Ria de Vigo harbour (Galicia, NW Iberian Peninsula). A cryopreservation protocol was designed in order to enable the successful cryopreservation and cryobanking of gametes and embryos to be used for marine quality assessment and ensure the accessibility to high quality reproductive material all year round, as an option to conditioning adults for out of season reproduction. The calculated EC50 using the cryopreserved blastula was 53.7 μg L(-1) for copper, 81.0 μg L(-1) for lead, 300.6 μg L(-1) for BP-3 and 300.6 μg L(-1) for 4-MBC. The sensitivity of the classic sea urchin embryo-larval bioassay was compared with the bioassay conducted with cryopreserved blastula. The results showed that the use of cryopreserved blastula bioassay allows detecting lower concentrations of pollutants in comparison with the classic bioassay. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Sperm Cryopreservation before Testicular Cancer Treatment and Its Subsequent Utilization for the Treatment of Infertility

    Directory of Open Access Journals (Sweden)

    Jana Žáková

    2014-01-01

    Full Text Available Aims. In this study we report our results with storage of cryopreserved semen intended for preservation and subsequent infertility treatment in men with testicular cancer during the last 18 years. Methods. Cryopreserved semen of 523 men with testicular cancer was collected between October 1995 and the end of December 2012. Semen of 34 men (6.5% was used for fertilization of their partners. They underwent 57 treatment cycles with cryopreserved, fresh, and/or donor sperm. Results. A total of 557 men have decided to freeze their semen before cancer treatment. Azoospermia was diagnosed in 34 men (6.1%, and semen was cryopreserved in 532 patients. Seminoma was diagnosed in 283 men (54.1% and nonseminomatous germ cell tumors in 240 men (45.9%. 34 patients who returned for infertility treatment underwent 46 treatment cycles with cryopreserved sperm. Totally 16 pregnancies were achieved, that is, 34.8% pregnancy rate. Conclusion. The testicular cancer survivors have a good chance of fathering a child by using sperm cryopreserved prior to the oncology treatment, even when it contains only limited number of spermatozoa.

  16. Recent Advances in Boar Sperm Cryopreservation: State of the Art and Current Perspectives.

    Science.gov (United States)

    Yeste, M

    2015-07-01

    While sperm cryopreservation is the best technology to store boar semen for long-term periods, only 1% of all artificial inseminations (AI) conducted worldwide are made using frozen-thawed boar sperm. With the emergence of long-term extenders for liquid storage, the use of cryopreserved sperm in routine AI is less required. However, banks of boar semen contain cryopreserved sperm and planning inseminations in AI centres may benefit from the use of frozen-thawed semen. Therefore, there is an interest in the use of this technology to preserve boar sperm. In this regard, although the first attempts to cryopreserve boar semen date back to the seventies and this technology is still considered as optimal, some relevant improvements have been made in the last decade. After giving a general picture about boar sperm cryodamage, the present review seeks to shed light on these recent cryopreservation advances. These contributions regard to protein markers for predicting ejaculate freezability, sperm selection prior to start cryopreservation procedures, additives to freezing and thawing extenders, relevance of the AI-technique and insemination-to-ovulation interval. In conclusion, most of these progresses have allowed counteracting better boar sperm cryodamage and are thus considered as forward steps for this storage method. It is also worth noting that, despite being lower than fresh/extended semen, reproductive performance outcomes following AI with frozen-thawed boar sperm are currently acceptable. © 2015 Blackwell Verlag GmbH.

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

    Science.gov (United States)

    Chanapiwat, Panida; Kaeoket, Kampon; Tummaruk, Padet

    2012-03-01

    The present study determined the effect of different types of sugars (lactose, fructose, glucose and sorbitol) used in egg yolk-based extender on the post-thawed boar semen quality. Twenty-two ejaculates from 6 fertility-proven Yorkshire boars were cryopreserved by liquid nitrogen vapor method. Sperm motility, viability, acrosome integrity and intact functional plasma membrane were determined at 0, 2 and 4 hr after thawing. It was found that the lactose-based extender resulted in a higher percentage of post-thawed sperm motility, viability, intact acrosome and functional plasma membrane than sorbitol-based extender (Pextender yielded a higher post-thawed sperm motility and viability than sorbitol-based extender (Pboar semen.

  18. In Vitro and Cryopreservation Techniques for Conservation of Snow Mountain Garlic.

    Science.gov (United States)

    Mahajan, Ritu

    2016-01-01

    Garlic is an important medicinal herb of culinary value by imparting its flavors and odors to the food. Allicin, a notable flavonoid in garlic, is a powerful antibiotic and antifungal compound. Due to poor bioavailability, garlic is of limited use for oral human consumption. Being sexually sterile, propagation of garlic is done by individual cloves from a bulb which increases the chances of transfer of viral diseases. In this chapter, an efficient and improved regeneration protocol for explant establishment and shoot multiplication under in vitro conditions is described. A high rate of shoot multiplication is obtained on MS medium supplemented with 0.5 mg/l BAP, 1.0 mg/l KN, and 2.0 mg/l GA3. Addition of 1.0 mg/l NAA to MS medium resulted in rooting at the shoot bases. A detailed method for encapsulation of explant in sodium alginate beads and their cryopreservation using encapsulation-dehydration is also described.

  19. Reproductive experiences of women who cryopreserved oocytes for non-medical reasons.

    Science.gov (United States)

    Hammarberg, Karin; Kirkman, Maggie; Pritchard, Natasha; Hickey, Martha; Peate, Michelle; McBain, John; Agresta, Franca; Bayly, Chris; Fisher, Jane

    2017-03-01

    What are the reproductive experiences of women who cryopreserve oocytes for non-medical reasons? One in three women had been pregnant at some stage in their lives and while most still wanted to have a child or another child, very few had used their stored oocytes, predominantly because they did not want to be single parents. The number of healthy women who freeze oocytes to avoid age-related infertility is increasing. Evidence about reproductive outcomes after oocyte cryopreservation for non-medical reasons is needed to help women make informed decisions. A cross-sectional survey was carried out. Study packs which included a self-administered questionnaire were mailed by clinic staff to 193 eligible women. Women who had stored oocytes for non-medical reasons at Melbourne IVF, a private ART clinic, between 1999 and 2014 were identified from medical records and invited to complete an anonymous questionnaire about their reproductive histories and experience of oocyte cryopreservation. A total of 10 survey packs were returned to the clinic marked 'address unknown'. Of the 183 potential respondents, 96 (53%) returned the questionnaire. One respondent provided only free-text comments, thus data from 95 respondents were compiled. The mean age at the time of freezing oocytes was 37.1 years (SD ± 2.6, range: 27-42) and the average number of oocytes stored was 14.2 (SD ± 7.9, range: 0-42); 2% had attempted to store oocytes but had none suitable for freezing, 24% had stored 23 oocytes. About one-third of respondents (34%) had been pregnant at some point in their lives. Six women (6%) had used their stored oocytes and three of them had given birth as a result. The main reason for not using stored oocytes was not wanting to be a single parent. Of the 87 (91%) women who still had oocytes stored, 21% intended to use them while 69% indicated that their circumstances would determine usage. The mean number of children respondents would ideally have liked to have was significantly

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

    Science.gov (United States)

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

    2015-01-01

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

  1. Cryopreservation of canine sperm using egg yolk and soy bean based extenders.

    Science.gov (United States)

    Sánchez-Calabuig, María Jesús; Maillo, Verónica; Beltrán-Breña, Paula; de la Fuente Martínez, Julio; Galera-Carrillo, Silvestre; Pérez-Gutiérrez, José Félix; Pérez-Cerezales, Serafín

    2017-09-01

    Animal protein-based extenders are widely used despite being a potential source of bacterial or mycoplasma contamination. Its replacement with vegetal protein-based extenders could represent an interesting alternative for dog sperm cryopreservation. This technique could be further improved by the addition of Tris-Glucose-Citric acid (TGC) that could physically protect the spermatozoa and improve its homeostasis. The aim of this study was to evaluate a cryopreservation protocol for dog spermatozoa using a soybean-based extender (LP1 ℗ ) as well as the effects of the addition of (TGC) immediately after the semen collection. Eleven ejaculates from purebred adult dogs were collected, centrifuged in the absence or presence of TGC and processed as fresh or cryopreserved spermatozoa with: egg yolk-based extender (CaniPRO) or LP1 ℗ . Freezing the spermatozoa in LP1 ℗ reduced the amplitude of the lateral head displacement, the percentage of spermatozoa that showed the intact acrosome and the mitochondrial function (P<0.05). These samples also showed a trend towards increased percentage of apoptotic spermatozoa (P<0.05). The addition of TGC before centrifugation did not improve the seminal parameters and adversely affected motility (P<0.05) in the spermatozoa cryopreserved in CaniPRO. However, TGC did not affect motility and increased (P<0.05) the percentage of intact acrosomes in the spermatozoa cryopreserved in LP1 ℗ , reaching similar values than those cryopreserved in CaniPRO. In conclusion, LP1 ® plus TGC provide the same level of protection to dog spermatozoa cryopreservation than the egg yolk based extender CaniPRO when comparing standard post-thaw sperm quality parameters. Copyright © 2017. Published by Elsevier Urban & Partner Sp. z o.o.

  2. Safety and efficacy of cryopreserved autologous platelet concentrates in HLA-alloimmunized patients with hematologic malignancies.

    Science.gov (United States)

    Gerber, Bernhard; Alberio, Lorenzo; Rochat, Sophie; Stenner, Frank; Manz, Markus G; Buser, Andy; Schanz, Urs; Stussi, Georg

    2016-10-01

    Curative chemotherapy approaches in patients with malignancies and platelet (PLT) transfusion refractoriness due to alloimmunization may be hampered by the lack of suitable PLT donors. For these patients, transfusion of cryopreserved autologous PLTs is an option, but is time- and resource-consuming. We aimed at further simplifying this process. A retrospective single-center analysis was conducted on the transfusion of cryopreserved autologous PLTs in nine female alloimmunized, PLT transfusion-refractory patients treated for acute leukemia (n = 8) and non-Hodgkin's lymphoma (n = 1). No additional processing was used before transfusion, and most notably, washing and centrifugation steps were omitted. Clinical efficacy and safety, as well as a flow cytometric assessment of structural and functional PLT changes, were analyzed. A total of 40 autologous PLT concentrates were thawed at bedside and transfused a median of 32 (range, 9 to 994) days after cryopreservation. No major bleeds and no severe dimethyl sulfoxide toxicity were observed. The median PLT count increments did not differ 1 and 18 to 24 hours after transfusion and reached 6 × 10 9 /L (interquartile range [IQR], 3 × 10 9 -7.5 × 10 9 /L) and 6 × 10 9 /L (IQR, 2.5 × 10 9 -9.5 × 10 9 /L), respectively. Cryopreservation resulted in partial activation of one-third of the PLTs. In vitro stimulation with strong agonists induced additional full activation of cryopreserved PLTs: median, 55% (IQR, 42%-60%) after thrombin and 39% (IQR, 36%-39%) after convulxin. The transfusion of cryopreserved autologous PLTs is feasible and safe. Despite the cryopreservation process, PLT functionality is partially maintained. © 2016 AABB.

  3. Cryopreservation of dermal fibroblasts and keratinocytes in hydroxyethyl starch-based cryoprotectants.

    Science.gov (United States)

    Naaldijk, Yahaira; Johnson, Adiv A; Friedrich-Stöckigt, Annett; Stolzing, Alexandra

    2016-12-01

    Preservation of human skin fibroblasts and keratinocytes is essential for the creation of skin tissue banks. For successful cryopreservation of cells, selection of an appropriate cryoprotectant agent (CPA) is imperative. The aim of this study was to identify CPAs that minimize toxic effects and allow for the preservation of human fibroblasts and keratinocytes in suspension and in monolayers. We cryopreserved human fibroblasts and keratinocytes with different CPAs and compared them to fresh, unfrozen cells. Cells were frozen in the presence and absence of hydroxyethyl starch (HES) or dimethyl sulfoxide (DMSO), the latter of which is a commonly used CPA known to exert toxic effects on cells. Cell numbers were counted immediately post-thaw as well as three days after thawing. Cellular structures were analyzed and counted by labeling nuclei, mitochondria, and actin filaments. We found that successful cryopreservation of suspended or adherent keratinocytes can be accomplished with a 10% HES or a 5% HES, 5% DMSO solution. Cell viability of fibroblasts cryopreserved in suspension was maintained with 10% HES or 5% HES, 5% DMSO solutions. Adherent, cryopreserved fibroblasts were successfully maintained with a 5% HES, 5% DMSO solution. We conclude that skin tissue cells can be effectively cryopreserved by substituting all or a portion of DMSO with HES. Given that DMSO is the most commonly used CPA and is believed to be more toxic than HES, these findings are of clinical significance for tissue-based replacement therapies. Therapies that require the use of keratinocyte and fibroblast cells, such as those aimed at treating skin wounds or skin burns, may be optimized by substituting a portion or all of DMSO with HES during cryopreservation protocols.

  4. Functional integrity of Colossoma macropomum (Cuvier, 1816 sperm cryopreserved with enriched extender solutions

    Directory of Open Access Journals (Sweden)

    Raycon Roberto Freitas Garcia

    Full Text Available Cryoprotectant solutions are used to protect the sperm from alterations caused by the low temperature in the cryopreservation process. We evaluated the quality of Colossoma macropomum semen after freezing, using dimethyl sulfoxide (DMSO as a cryoprotectant, combined with two extender solutions (T1 - Solution 1: Glucose 90.0 g/L, Sodium Citrate 6.0 g/L, EDTA 1.5 g/L, Sodium Bicarbonate 1.5 g/L, Potassium Chloride 0.8 g/L, Gentamycin Sulphate 0.2 g/L, and T2 - Solution 2: Glucose 90.0 g/L, ACP(r-104 10.0 g/L. Motility rate and motility time did not differ between T1 and T2 and were lower than fresh semen. The number of normal sperm was significantly different in treatments T1 (15.1% and T2 (21.9%, and both showed a reduction in the percentage of normal sperm compared to fresh semen (57.4%. The values found for the rates of fertilization and hatching, mitochondrial functionality and sperm DNA, did not differ between the treatments (T1 and T2. Regarding membrane integrity, there was a higher percentage of spermatozoa with intact membranes in T1 (53.4% than T2 (43.7%. The extender solutions, combined with 10% DMSO, maintained the sperm DNA intact in almost all the C. macropomum sperm cells, however there was a loss in their functionality.

  5. Supplementing rooster sperm with Cholesterol-Loaded-Cyclodextrin improves fertility after cryopreservation.

    Science.gov (United States)

    Chuaychu-Noo, Napapach; Thananurak, Pachara; Chankitisakul, Vibantita; Vongpralub, Thevin

    2017-02-01

    Little is known about the effects of Cholesterol-Loaded Cyclodextrin (CLC) on post-thaw semen quality in chicken. The aim of the present study is to investigate the efficacy of CLC levels (0, 1, 2 and 3 mg/mL Schramm diluent) on post-thawed semen quality and fertility in two breeds of chicken Pradu Hang Dum (native chicken) and Rhode Island Red. Semen samples of each breed were pooled, divided into 4 aliquots and diluted with Schramm diluents, cooled to 5 °C when DMF was added (6% of final volume). Semen straws were subjected to cryopreservation using the liquid nitrogen vapor method. Post-thawed sperm motility, viability, acrosome integrity, mitochondrial function, and the Malondialdehyde (MDA) level were determined. The fertility of frozen semen was tested by inseminating laying hens. Post-thaw motility between Pradu Hang Dum and Rhode Island Red was no different; but Rhode Island Red had a higher semen viability and live cell intact acrosomes than Pradu Hang Dum (P rooster semen. There was no interaction among breeds and CLC on post-thaw semen quality and fertility. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. A novel micro-straw for cryopreservation of small number of human spermatozoon

    Directory of Open Access Journals (Sweden)

    Feng Liu

    2017-01-01

    Full Text Available Cryopreservation of few spermatozoa is still a major challenge for male fertility preservation. This study reports use a new micro-straw (LSL straw for freezing few spermatozoa for intracytoplasmic sperm injection (ICSI. Semen samples from 22 fertile donors were collected, and each semen sample was diluted and mixed with cryoprotectant in a ratio of 1:1, and then frozen using three different straws such as LSL straw (50-100 μl, traditional 0.25 ml and 0.5 ml straws. For freezing, all straws were fumigated with liquid nitrogen, with temperature directly reducing to −130-−140°C. Sperm concentration, progressive motility, morphology, acrosome integrity, and DNA fragmentation index were evaluated before and after freezing. After freezing-thawing, LSL straw group had significantly higher percentage of sperm motility than traditional 0.25 ml and 0.5 ml straw groups (38.5% vs 27.4% and 25.6%, P 0.05. As LSL straws were thinner and hold very small volume, the freezing rate of LSL straw was obviously faster than 0.25 ml straw and 0.5 ml straws. In conclusion, LSL micro-straws may be useful to store few motile spermatozoa with good recovery of motility for patients undergoing ICSI treatment.

  7. Milt cryopreservation for rheophilic fish threatened by extinction in the Rio Grande, Brazil.

    Science.gov (United States)

    de Andrade, Estefânia Souza; Paula, Daniella Aparecida de Jesus; Felizardo, Viviane de Oliveira; Murgas, Luis David Solis; Veras, Galileu Crovatto; Vieira e Rosa, Priscila

    2014-01-01

    Specific protocols for milt cryopreservation have been established for some freshwater fish species. However, cryopreservation reduces sperm quality, giving unsatisfactory results in reproduction. The objective of this work was to evaluate the effect of different cryoprotectants on the quality of Prochilodus lineatus, Brycon orbignyanus and Piaractus mesopotamicus milt after cryopreservation. The milt was diluted in different cryoprotectant solutions containing 10% methanol, dimethyl sulfoxide, glycerol, propylene glycol or ethylene glycol combined with the Beltsville Thawing Solution extender (5%), then placed in the vapour of a liquid nitrogen (LN) storage tank for 24 h, after which they were immersed in LN. After rewarming, the rate (%) and duration (s) of milt motility and abnormal morphology were evaluated. All of cryoprotectant solutions tested used maintained the viability of P. lineatus and P. mesopotamicus milt. However, in P. lineatus, glycerol ensured a lower percentage of abnormal morphology. In case of P. mesopotamicus, all of the cryoprotectant solutions tested may be used in the cryopreservation process, with the exception of those containing glycerol. For B. orbignyanus, cryoprotectant solutions containing methanol and ethylene glycol are recommended for use in the cryopreservation process, although they reduced the quality of sperm post-rewarming.

  8. Cryopreservation of PLBs of Brassidium Fly Away Using Encapsulation-Dehydration Technique

    Directory of Open Access Journals (Sweden)

    Arulvilee Rajasegar

    2015-12-01

    Full Text Available In vitro grown protocorm-like bodies (PLBs of Brassidium Fly Away orchid hybrid were cryopreserved using encapsulation- dehydration technique. The viability of the cryopreserved cells was determined by 2,3,5-triphenyltetrazolium chloride (TTC assay. For the preculture treatment, the PLBs were excised into two standard sizes of 1-2 and 4-5 mm and were precultured on half-strength Murashige and Skoog (MS semi solid medium supplemented with diff erent concentrations of sucrose (0, 0.2, 0.4, 0.6, 0.8 and 1.0M. The PLBs size 4-5 mm and 0.6 M sucrose concentration was selected based on highest viability obtained in TTC assay. The PLBs were encapsulated for 30 minutes using 3% (w/v liquid s odium alginate medium supplemented with 0.4M sucrose and 0.1M calcium chl oride and osmoprotected in 0.75M sucrose solution for 24 hours at 25°C. Th e beads were then dehydrated using 50g heat-sterilised silica gel for four hours , cryopreserved for 24 hours, thawed in a 40±2°C water bath for 90 seconds, and r egenerated in semi-solid half-strength. Biochemical analyses were conducted and th e cryopreserved PLBs had produced lower content of chlorophyll while the highest specifi c peroxidase activity was observed in cryopreserved PLBs

  9. Sperm harvesting and cryopreservation during vasectomy reversal is not cost effective.

    Science.gov (United States)

    Boyle, Karen E; Thomas, Anthony J; Marmar, Joel L; Hirshberg, Steven; Belker, Arnold M; Jarow, Jonathan P

    2006-04-01

    To determine whether sperm harvesting and cryopreservation at the time of vasectomy reversal is cost-effective. Model of actual costs and results at five institutions. Multicenter study comprising five centers, including university hospitals and private practices. Men undergoing vasectomy reversal. We established two models for vasectomy reversal. The first model was sperm harvesting and cryopreservation at the time of vasectomy reversal. The second model was sperm harvesting at the time of IVF only if the patient remained azoospermic after vasectomy reversal. Vasectomy reversal procedures modeled included bilateral vasovasostomy and bilateral epididymovasostomy. The costs for each procedure at the five institutions were collated and median costs determined. Median cost of procedure and calculated financial comparisons. The median cost of testicular sperm extraction/cryopreservation performed at the time of bilateral vasovasostomy was $1,765 (range, $1,025-$2,800). The median cost of microsurgical epididymal sperm aspiration or testicular sperm extraction with cryopreservation performed at the time of epididymovasostomy was $1,209 (range, $905-$2,488). The average of the median costs for percutaneous sperm aspiration or testicular sperm aspiration for those patients with a failed vasectomy reversal was $725 (range, $400-$1,455). Sperm retrieval with cryopreservation at the time of vasectomy reversal is not a cost-effective management strategy.

  10. A validation study of new cryopreservation bags for implementation in a blood and marrow transplant laboratory.

    Science.gov (United States)

    Pomper, Gregory J; Wilson, Emily; Isom, Scott; Hurd, David D

    2011-06-01

    A new cryopreservation bag for hematopoietic cell transplantation requires validation as a safe alternative to the bag currently being used in the laboratory. The new bag was validated using both laboratory and clinical criteria. Laboratory validation proceeded using paired samples of mononuclear cells processed using standard procedures. Cells cryopreserved in the new and old bags were compared for viability, cell counts, CD34 enumeration, colony-forming unit assays, and bag integrity. After completion of laboratory investigations, engraftment with the new bags was followed and compared to historical engraftment using the old bags. There were no significant differences between the old and new bags detected using laboratory studies. Bag integrity was equivalent. The validation data suggested impaired cell function after cryopreservation in the new bags, but there were no significant differences in engraftment potential using either material. Days to engraftment was longer using the new bags, but statistical analysis revealed an association with CD34 dose and not with cryopreservation bag type. The new bags were noninferior to the old bags. A change in cryopreservation bag type may appear to affect cell function and potentially affect engraftment. Multiple analyses may be needed to understand the effect of cell processing changes. © 2010 American Association of Blood Banks.

  11. Replacement of serum with ocular fluid for cryopreservation of immature testes.

    Science.gov (United States)

    Pothana, Lavanya; Devi, Lalitha; Venna, Naresh Kumar; Pentakota, Niharika; Varma, Vivek Phani; Jose, Jedy; Goel, Sandeep

    2016-12-01

    Cryopreservation of immature testis is a feasible approach for germplasm preservation of male animals. Combinations of dimethyl sulfoxide (DMSO) and foetal bovine serum (FBS) are used for testis cryopreservation. However, an alternative to FBS is needed, because FBS is expensive. Buffalo ocular fluid (BuOF), a slaughter house by-product, could be an economical option. The objective of the present study was to assess whether BuOF can replace FBS for cryopreservation of immature mouse (Mus musculus), rat (Rattus norvegicus), and buffalo (Bubalus bubalis) testes. Results showed that rodent and buffalo testes frozen in DMSO (10% for rodents and 20% for buffalo) with 20% FBS or BuOF had similar numbers of viable and DNA-damaged cells (P > 0.05). The expression of cell proliferation- (PCNA) and apoptosis-specific proteins (Annexin V and BAX/BCL2 ratio) were also comparable in mouse and buffalo testes frozen in DMSO with FBS or BuOF (P > 0.05). Interestingly, rat testis frozen in DMSO with BuOF had lower expression of Annexin V protein than testis frozen in DMSO with FBS (P  0.05). These findings provide evidence that BuOF has potential to replace FBS for cryopreservation of immature rodent and buffalo testis. Further investigation is needed to explore whether BuOF can replace FBS for testis cryopreservation of other species. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Cryopreservation of Populus trichocarpa and Salix dormant buds with recovery by grafting or direct rooting.

    Science.gov (United States)

    Bonnart, Remi; Waddell, John; Haiby, Kathy; Widrlenchner, Mark P; Volk, Gayle M

    2014-01-01

    Methods are needed for the conservation of clonally maintained trees of Populus and Salix. In this work, Populus trichocarpa and Salix genetic resources were cryopreserved using dormant scions as the source explant. We quantified the recovery of cryopreserved materials that originated from diverse field environments by using either direct sprouting or grafting. Scions (either at their original moisture content of 48 to 60% or dried to 30%) were slowly cooled to -35 degree C, transferred to the vapor phase of liquid nitrogen (LNV, -160 degree C), and warmed before determining survival. Dormant buds from P. trichocarpa clones from Westport and Boardman, OR had regrowth levels between 42 and 100%. Direct rooting of cryopreserved P. trichocarpa was also possible. Ten of 11 cryopreserved Salix accessions, representing 10 different species, exhibited at least 40% bud growth and rooting after 6 weeks when a bottom-heated rooting system was implemented. We demonstrate that dormant buds of P. trichocarpa and Salix accessions can be cryopreserved and successfully regenerated without the use of tissue culture.

  13. Cryopreservation of in vitro grown shoot tips and apical meristems of the forage legume Arachis pintoi.

    Science.gov (United States)

    Rey, Hebe Y; Faloci, Mirta; Medina, Ricardo; Dolce, Natalia; Mroginski, Luis; Engelmann, Florent

    2009-01-01

    A cryopreservation protocol using the encapsulation-dehydration procedure was established for shoot tips (2-3 mm in length) and meristems (0.3-0.5 mm) sampled from in vitro plantlets of diploid and triploid cytotypes of Arachis pintoi. The optimal protocol was the following: after dissection, explants were precultured for 24 h on establishment medium (EM), encapsulated in calcium alginate beads and pretreated in liquid EM medium with daily increasing sucrose concentration (0.5, 0.75, 1.0 M) and desiccated to 22-23 percent moisture content (fresh weight basis). Explants were frozen using slow cooling (1 C per min from 25C to -30C followed by direct immersion in liquid nitrogen), thawed rapidly and post-cultured in liquid EM medium enriched with daily decreasing sucrose concentrations (0.75, 0.50, 0.1 M). Explants were then transferred to solid EM medium in order to achieve shoot regeneration, then on Murashige and Skoog medium supplemented with 0.05 microM naphthalene acetic acid to induce rooting of shoots. With this procedure, 53 percent and 56 percent of cryopreserved shoot tips of the diploid and triploid cytotypes, respectively, survived and formed plants. However, only 16 percent of cryopreserved meristems of both cytotypes regenerated plants. Using ten isozyme systems and seven RAPD profiles, no modification induced by cryopreservation could be detected in plantlets regenerated from cryopreserved material.

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

    Science.gov (United States)

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

  15. Assessment of some critical factors in the freezing technique for the cryopreservation of precision-cut rat liver slices

    NARCIS (Netherlands)

    Maas, W.J.M.; Graaf, I.A.M. de; Schoen, E.D.; Koster, H.J.; Sandt, J.J.M. van de; Groten, J.P.

    2000-01-01

    A number of studies on the cryopreservation of precision-cut liver slices using various techniques have been reported. However, the identification of important factors that determine cell viability following cryopreservation is difficult because of large differences between the various methods

  16. Recovery patterns, histological observations and genetic integrity in Malus shoot tips cryopreserved using droplet vitrification and encapsulation-dehydration procedures

    Science.gov (United States)

    A droplet-vitrification procedure is described for cryopreservation of Malus shoot tips. Survival patterns, recovery types, histological observations, and genetic integrity were compared for Malus shoot tips cryopreserved using this droplet-vitrification procedure and an encapsulation-dehydration pr...

  17. Isolation and Characterization of Human Dental Pulp Stem Cells from Cryopreserved Pulp Tissues Obtained from Teeth with Irreversible Pulpitis.

    Science.gov (United States)

    Malekfar, Azin; Valli, Kusum S; Kanafi, Mohammad Mahboob; Bhonde, Ramesh R

    2016-01-01

    Human dental pulp stem cells (DPSCs) are becoming an attractive target for therapeutic purposes because of their neural crest origin and propensity. Although DPSCs can be successfully cryopreserved, there are hardly any reports on cryopreservation of dental pulp tissues obtained from teeth diagnosed with symptomatic irreversible pulpitis during endodontic treatment and isolation and characterization of DPSCs from such cryopreserved pulp. The aim of this study was to cryopreserve the said pulp tissues to propagate and characterize isolated DPSCs. A medium consisting of 90% fetal bovine serum and 10% dimethyl sulfoxide was used for cryopreservation of pulp tissues. DPSCs were isolated from fresh and cryopreserved pulp tissues using an enzymatic method. Cell viability and proliferation were determined using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. DPSC migration and interaction were analyzed with the wound healing assay. Mesenchymal characteristics of DPSCs were verified by flow cytometric analysis of cell surface CD markers. The osteogenic and adipogenic potential of DPSCs was shown by von Kossa and oil red O staining methods, respectively, and the polymerase chain reaction method. We found no significant difference in CD marker expression and osteogenic and adipogenic differentiation potential of DPSCs obtained from fresh and cryopreserved dental pulp tissue. Our study shows that dental pulp can be successfully cryopreserved without losing normal characteristics and differentiation potential of their DPSCs, thus making them suitable for dental banking and future therapeutic purposes. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  18. Effects of chilled storage and cryopreservation on sperm characteristics, antioxidant enzyme activities, and lipid peroxidation in Pacific cod Gadus microcephalus

    Science.gov (United States)

    Wang, Xueying; Shi, Xuehui; Liu, Yifan; Yu, Daode; Guan, Shuguang; Liu, Qinghua; Li, Jun

    2016-07-01

    The present study evaluated the effects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm motility and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (Gr), and lipid peroxidation (measured via malondialdehyde (MDA) content) were determined after the milt was stored at 4°C for 12 h, cryopreserved without cryoprotectant in 12% propylene glycol (PG), cryopreserved in 12% PG+0.1 mol/L trehalose, or cryopreserved in 12% PG spermatozoa but centrifuged to decant the supernatant prior to cryopreservation (only sperm cells were cryopreserved). After chilled storage or cryopreservation, the SOD, CAT and GPx activities were reduced in sperm cells and increased in seminal plasma in almost all treatments; sperm motility parameters were also decreased. However, the addition of trehalose into the cryoprotectant could significantly improve the postthaw sperm quality as revealed by the sperm average path velocity. This improvement might be attributed to the function of trehalose in scavenging reactive oxygen species. Chilled storage and cryopreservation had significant effects on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod.

  19. Effects of cryopreservation on excretory function, cellular adhesion molecules and vessel lumen formation in human umbilical vein endothelial cells.

    Science.gov (United States)

    Cai, Guoping; Lai, Binbin; Hong, Huaxing; Lin, Peng; Chen, Weifu; Zhu, Zhong; Chen, Haixiao

    2017-07-01

    Cryopreservation is widely used in regenerative medicine for tissue preservation. In the present study, the effects of cryopreservation on excretory function, cellular adhesion molecules and vessel lumen formation in human umbilical vein endothelial cells (HUVECs) were investigated. After 0, 4, 8, 12 or 24 weeks of cryopreservation in liquid nitrogen, the HUVECs were thawed. The excretory functions markers (endothelin‑1, prostaglandin E1, von Willebrand factor and nitric oxide) of HUVECs were measured by ELISA assay. The expression of intercellular adhesion molecule‑1 (ICAM‑1) in HUVECs was analyzed using flow cytometry. An angiogenesis assay was used to determine the angiogeneic capabilities of the thawed HUVECs. The results demonstrated that cryopreserved/thawed and recultivated HUVECs were unsuitable for tissue‑engineered microvascular construction. Specifically, the excretory function of the cells was significantly decreased in the post‑cryopreserved HUVECs at 24 weeks. In addition, the level of ICAM‑1 in HUVECs was significantly upregulated from the fourth week of cryopreservation. Furthermore, the tube‑like structure‑forming potential was weakened with increasing cryopreservation duration, and the numbers of lumen and the length of the pipeline were decreased in the thawed HUVECs, in a time‑dependent manner. In conclusion, the results of the present study revealed that prolonged cryopreservation may lead to HUVEC dysfunction and did not create stable cell lines for tissue‑engineered microvascular construction.

  20. Autotransplantation of cryopreserved ovarian tissue in 12 women with chemotherapy-induced premature ovarian failure: the Danish experience

    DEFF Research Database (Denmark)

    Schmidt, Kirsten Tryde; Rosendahl, Mikkel; Ernst, Erik

    2011-01-01

    To describe a cohort of 12 Danish women who received autotransplantation of frozen-thawed cryopreserved ovarian tissue because of premature ovarian failure after cancer treatment.......To describe a cohort of 12 Danish women who received autotransplantation of frozen-thawed cryopreserved ovarian tissue because of premature ovarian failure after cancer treatment....

  1. Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples.

    Science.gov (United States)

    Roque, M; Valle, M; Marques, F; Sampaio, M; Geber, S

    2016-04-01

    Intracytoplasmic sperm injection (ICSI) may be performed with testicular frozen-thawed spermatozoa in patients with nonobstructive azoospermia (NOA). Sperm retrieval can be performed in advance of oocyte aspiration, as it may avoid the possibility of no recovery of spermatozoa on the day of oocyte pickup. There are few studies available in the literature concerning the use of frozen-thawed spermatozoa obtained from testicular sperm aspiration (TESA). To evaluate the effects and the outcomes of ICSI with frozen-thawed spermatozoa obtained by TESA, we performed a retrospective analysis of 43 ICSI cycles using frozen-thawed TESA. We obtained acceptable results with a fertilisation rate of 67.9%, an implantation rate (IR) of 17.1%, and clinical and ongoing pregnancy rates of 41.9% and 37.2% respectively. The results of this study suggest that performing ICSI using cryopreserved frozen-thawed testicular spermatozoa with TESA as a first option is a viable, safe, economic and effective method for patients with NOA. © 2015 Blackwell Verlag GmbH.

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

    Science.gov (United States)

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

    2016-04-01

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

  3. Cryopreservation of in vitro grown nodal segments of Rauvolfia serpentina by PVS2 vitrification.

    Science.gov (United States)

    Ray, Avik; Bhattacharya, Sabita

    2008-01-01

    This paper describes the cryopreservation by PVS2 vitrification of Rauvolfia serpentina (L.) Benth ex kurz, an important tropical medicinal plant. The effects of type and size of explants, sucrose preculture (duration and concentration) and vitrification treatment were tested. Preliminary experiments with PVS1, 2 and 3 produced shoot growth only for PVS2. When optimizing the PVS2 vitrification of nodal segments, those of 0.31 - 0.39 cm in size were better than other nodal sizes and or apices. Sucrose preculture had a positive role in survival and subsequent regrowth of the cryopreserved explants. Seven days on 0.5 M sucrose solution significantly improved the viability of nodal segments. PVS2 incubation for 45 minutes combined with a 7-day preculture gave the optimum result of 66 percent. Plantlets derived after cryopreservation resumed growth and regenerated normally.

  4. Triplet pregnancy after intracytoplasmic sperm injection of cryopreserved oocytes: case report.

    Science.gov (United States)

    Young, E; Kenny, A; Puigdomenech, E; Van Thillo, G; Tiverón, M; Piazza, A

    1998-08-01

    To report a triplet pregnancy that occurred after intracytoplasmic injection of sperm into cryopreserved oocytes. Case report. Instituto de Ginecología y Fertilidad (IFER), Buenos Aires, Argentina. A 36-year-old infertile patient with premature ovarian failure and a previous term pregnancy with fresh donated oocytes. We administered leuprolide acetate for pituitary down-regulation followed by E2 valerianate in incremental doses until an endometrial lining of >8 mm was observed by ultrasound. Thawing of frozen donated oocytes, intracytoplasmic sperm injection (ICSI), and translaparoscopic fallopian tube ET also were performed. Natural micronized progesterone was administered intravaginally (600 mg/d) before ET. Ultrasound at the 8th week of gestation revealed a triplet pregnancy with active fetal heartbeats. A triple intrauterine gestation was achieved with the use of microinjection into cryopreserved oocytes. This case illustrates the feasibility of oocyte cryopreservation for clinical use in the era of ICSI.

  5. Optimizing human semen cryopreservation by reducing test vial volume and repetitive test vial sampling

    DEFF Research Database (Denmark)

    Jensen, Christian F S; Ohl, Dana A; Parker, Walter R

    2015-01-01

    OBJECTIVE: To investigate optimal test vial (TV) volume, utility and reliability of TVs, intermediate temperature exposure (-88°C to -93°C) before cryostorage, cryostorage in nitrogen vapor (VN2) and liquid nitrogen (LN2), and long-term stability of VN2 cryostorage of human semen. DESIGN......: Prospective clinical laboratory study. SETTING: University assisted reproductive technology (ART) laboratory. PATIENT(S): A total of 594 patients undergoing semen analysis and cryopreservation. INTERVENTION(S): Semen analysis, cryopreservation with different intermediate steps and in different volumes (50......-1,000 μL), and long-term storage in LN2 or VN2. MAIN OUTCOME MEASURE(S): Optimal TV volume, prediction of cryosurvival (CS) in ART procedure vials (ARTVs) with pre-freeze semen parameters and TV CS, post-thaw motility after two- or three-step semen cryopreservation and cryostorage in VN2 and LN2. RESULT...

  6. Cryopreservation of ram semen with antioxidant supplemented soybean lecithin-based extenders and impacts on incubation resilience.

    Science.gov (United States)

    Toker, M Berk; Alcay, Selim; Gokce, Elif; Ustuner, Burcu

    2016-06-01

    The scope of this study was investigation the affects of various antioxidants on 1% soybean lecithin-based semen extenders for ram semen cryopreservation. Ejaculates, collected via electrically stimulated ejaculation, that have a thick consistency, rapid wave motion (3-5 on a 0-5 scale) and >75% initial motility were pooled. The pooled samples were split into four equal aliquots as 5 mM Methionine, 5 mM Cysteamine, 1 mM Cysteine and a sample of antioxidant-free control group. Each sample group was diluted to a ratio of 1/5 (semen/extender, v/v) as final concentration and two step dilution method was used for cryopreservation. Extender groups were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Semen samples also incubated for 6 h in humidified air with 5% CO2 at 39 °C to evaluate post-thaw incubation resilience of semen characteristics. The results showed that freezing and thawing procedures had negative effects on motility (P < 0.05), plasma membrane integrity (P < 0.05) and acrosomal integrity (P < 0.05). After 6 h of incubation time, the Cysteine supplemented extender group yielded significantly higher results than other extender groups in terms of spermatological parameters. Furthermore MDA levels in the antioxidant groups were lower than control group (P < 0.05). Nevertheless, there were no significant differences among antioxidant groups. Copyright © 2016 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    2012-01-01

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

  8. Testicular tissue cryopreservation in prepubertal male children: an analysis of parental decision-making.

    Science.gov (United States)

    Ginsberg, Jill P; Li, Yimei; Carlson, Claire A; Gracia, Clarisa R; Hobbie, Wendy L; Miller, Victoria A; Mulhall, John; Shnorhavorian, Margarett; Brinster, Ralph L; Kolon, Thomas F

    2014-09-01

    Infertility is an unfortunate treatment-related consequence for some pediatric malignancies as well as some non-malignant conditions treated with stem cell transplant. Unlike pubertal males, prepubertal males cannot produce semen for cryopreservation. This manuscript reports on the acceptability and safety of a multi-institutional protocol for offering testicular tissue cryopreservation to families of prepubertal male children at highest risk for infertility. Data on decision influences, decision-making control, and emotional state when considering this option are described. Prepubertal males facing gonadotoxic therapy were offered testicular cryopreservation. Post-biopsy, patients were followed for acute side effects. In addition, parents and patients were asked to complete questionnaires, whether or not they chose to cryopreserve tissue. Seventy-four prepubertal male children were approached. Fifty-seven families (77%) consented to the testicular biopsy; 48 of 57 underwent the procedure. There was one post-operative side effect. Parents who agreed to testicular cryopreservation and those that did not felt in control of this decision. Parents who consented to the biopsy and refusers were not deterred by the experimental nature of the protocol. An important decision-making influence was the risk of the biopsy. Biopsy and cryopreservation of testicular tissue from prepubertal male children was performed successfully and safely at three institutions. Parents faced with this option at diagnosis can make an informed decision and weigh carefully the risks and benefits. Although asked to make a decision soon after they were given a difficult diagnosis, parents uniformly felt in control of this decision. © 2014 Wiley Periodicals, Inc.

  9. Circulating angiogenic cells can be derived from cryopreserved peripheral blood mononuclear cells.

    Directory of Open Access Journals (Sweden)

    Tanja Sofrenovic

    Full Text Available Cell transplantation for regenerative medicine has become an appealing therapeutic method; however, stem and progenitor cells are not always freshly available. Cryopreservation offers a way to freeze cells as they are generated, for storage and transport until required for therapy. This study was performed to assess the feasibility of cryopreserving peripheral blood mononuclear cells (PBMCs for the subsequent in vitro generation of their derived therapeutic population, circulating angiogenic cells (CACs.PBMCs were isolated from healthy human donors. Freshly isolated cells were either analyzed immediately or cryopreserved in media containing 6% plasma serum and 5% dimethyl sulfoxide. PBMCs were thawed after being frozen for 1 (early thaw or 28 (late thaw days and analyzed, or cultured for 4 days to generate CACs. Analysis of the cells consisted of flow cytometry for viability and phenotype, as well as functional assays for their adhesion and migration potential, cytokine secretion, and in vivo angiogenic potential.The viability of PBMCs and CACs as well as their adhesion and migration properties did not differ greatly after cryopreservation. Phenotypic changes did occur in PBMCs and to a lesser extent in CACs after freezing; however the potent CD34(+VEGFR2(+CD133(+ population remained unaffected. The derived CACs, while exhibiting changes in inflammatory cytokine secretion, showed no changes in the secretion of important regenerative and chemotactic cytokines, nor in their ability to restore perfusion in ischemic muscle.Overall, it appears that changes do occur in cryopreserved PBMCs and their generated CACs; however, the CD34(+VEGFR2(+CD133(+ progenitor population, the secretion of pro-vasculogenic factors, and the in vivo angiogenic potential of CACs remain unaffected by cryopreservation.

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

    Science.gov (United States)

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

    2017-01-01

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

  11. Slow cryopreservation is not superior to vitrification in human spermatozoa; an experimental controlled study

    Directory of Open Access Journals (Sweden)

    Mohamed Shehata Ali Mohamed

    2015-10-01

    Full Text Available Background: Spermatozoa cryopreservation is used for the management of infertility and some other medical conditions. The routinely applied cryopreservation technique depends on permeating cryoprotectants, whose toxic effects have raised the attention towards permeating cryoprotectants-free vitrification technique. Objective: To compare between the application of slow cryopreservation and vitrification on human spermatozoa. Materials and Methods: This was an experimental controlled study involving 33 human semen samples, where each sample was divided into three equal parts; fresh control, conventional slow freezing, and permeating cryoprotectants-free vitrification. Viability and mitochondrial membrane potential (MMP of control and post-thawing spermatozoa were assessed with the sperm viability kit and the JC-1 kit, respectively, using fluorescence-activated cell sorting analysis. Results: Significant reduction of the progressive motility, viability and MMP was observed by the procedure of freezing and thawing, while there was not any significant difference between both cryopreservation techniques. Cryopreservation resulted in 48% reduction of the percentage of viable spermatozoa and 54.5% rise in the percentage of dead spermatozoa. In addition, high MMP was reduced by 24% and low MMP was increased by 34.75% in response to freezing and thawing. Progressive motility of spermatozoa was correlated significantly positive with high MMP and significantly negative with low MMP in control as well as post-thawing specimens (r=0.8881/ -0.8412, 0.7461/ -0.7510 and 0.7603/ -0.7839 for control, slow and vitrification respectively, p=0.0001. Conclusion: Although both cryopreservation techniques have similar results, vitrification is faster, easier and associated with less toxicity and costs. Thus, vitrification is recommended for the clinical application.

  12. Evaluation of the Efficiency of Two Different Freezing Media and Two Different Protocols to Preserve Human Spermatozoa from Cryoinjury

    Directory of Open Access Journals (Sweden)

    Gemma Fabozzi

    2016-01-01

    Full Text Available It is universally recognized that cryopreservation impairs sperm quality. In order to improve postthawing sperm survival and motility, media of different composition and different protocols have been proposed. However, no clear evidence is available to understand which are the most efficient protocol and medium for sperm cryopreservation. The present study evaluates the efficiency of two different cryopreservation protocols and two common freezing media (FM containing different cryoprotectants (CPs, TEST Yolk Buffer (TYB and Sperm Freeze (SF, to preserve human sperm quality. Our data suggest that TYB is better than SF both in terms of postthaw viability and in terms of progressive motility, while the direct addition of FM to the sperm sample resulted in the most efficient protocol in terms of postthaw viability but not in terms of progressive motility.

  13. Cryopreservation of ovarian tissue for a decade in Denmark: a view of the technique

    DEFF Research Database (Denmark)

    Rosendahl, Mikkel; Schmidt, Kirsten Louise Tryde; Ernst, Erik

    2011-01-01

    evaluation of mouse and human ovarian tissue after freezing with four different combinations of cryoprotectants. Viability was confirmed by transplantation of frozen-thawed human ovarian tissue (n = 49) to oophorectomized Nude mice. Viability after transport of fresh tissue 4-5 h prior to freezing had...... previously been validated. Overnight transport of fresh ovarian tissue prior to cryopreservation was evaluated when human ovarian tissue was kept on ice for 20 h and then cryopreserved. The thawed ovarian tissue was transplanted to an oophorectomized Nude mouse and histology confirmed viability. In Denmark...

  14. Transplantation of cryopreserved allogeneic bone marrow after its long-term storage to lethally irradiated dogs

    International Nuclear Information System (INIS)

    Novikova, N.N.; Fedotenkov, A.G.; Sukyasyan, G.V.; Timakova, L.A.

    1982-01-01

    The study of the dog bone marrow preserved at -196 deg C during 6-12 years has shown that in the body of lethally irradiated animals (8Gy), due to the antigenic difference in the tissues of the donor and the irradiated recipients, the cells of cryopreserved allogeneic bone marrow were differentiated by the lymphoid type similar to that observed in transplantation of freshly prepared myelocaryocytes. However, their proliferative activity in the period of active lymphocyte transformation was quantitatively less manifest than in freshly transplanted cells. The results of the study evidence that the bone marrow cells cryopreserved during 6-12 years retain their functional activity

  15. Energy and climate protection management, the key to higher energy efficiency in communities; Energie- und Klimaschutzmanagement. Der Schluessel zu mehr Energieeffizienz in Kommunen

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2011-07-15

    The brochure explains the dena energy and climate protection management concepts and presents tools for long-term reduction of energy consumption in communities. It presents valuable information for better organization of internal processes in community administrations and for the management of energy efficiency measures. The dena energy and climate protection management concept is developed in cooperation with model communities of different sizes since 2010. All interested communities can use this brochure as a guide for initiating effective climate protection measures.

  16. Polymyxin B effects on motility parameters of cryopreserved bull semen

    Directory of Open Access Journals (Sweden)

    Mojtaba Rashedi

    2017-01-01

    Full Text Available Objective: To evaluate the effect of adding different values of polymyxin B (PMB to bull semen on various motility parameters of post-thawed semen such as total motility, progressive motility and velocity parameters using kinetic parameters of sperm by Computer Assisted Sperm Analysis.Methods: Gram negative bacteria release lipopolysaccharide, which induces the apoptotic pathway. Antibiotics are added to semen in order to prevent bacterial contaminations in bovine semen. These antibiotics kill the bacteria especially gram negative bacteria. Therefore, their endotoxins are released during bacteriolysis and bind to the head region and midpiece of sperm. PMB is a bactericidal antibiotic against multidrug resistant gram-negative bacteria and is able to neutralize the toxic effects of the released endotoxin. This study was performed on 3-year old Taleshi bulls.Results: The results showed both positive and negative significant effects of PMB on semen quality. Total motility and progressive motility were significantly increased (P<0.000 1 by 100 μg per mL of PMB (55.2% and 48.8% respectively against the control groups (43.5% and 37.7%, respectively. Moreover, they were significantly decreased (P<0.000 1 by 1 000 μg per mL of PMB (35.2% and 28.8% respectively against the control groups (43.5% and 37.7% respectively in above-mentioned parameters. In Computer Assisted Semen Analyzer, parameter VAP was significantly decreased (P<0.04 in 1 000 μg (69.6 μm/s against the control group (78.7 μm/s. Finally, using PMB in processing cryopreserved bull semen is advised, but before using it, the rate of endotoxins must be measured.Conclusions: We advise using PMB after measuring endotoxin concentration; In vitro, in vivo and in field fertilization, adding other sperm evaluation factors such as acrosomal integrity, DNA integrity, mitochondrial function to PMB treated semen.

  17. The Lust for Efficiency: A Downhome Story or The Implications of Zero-Based Budgeting for Institutions of Higher Education as Seen from the State of Georgia's Experience.

    Science.gov (United States)

    Fincher, Cameron

    Reactions to zero-based budgeting in the State of Georgia as it pertains to institutions of higher education are discussed. Major advantages and disadvantages of zero-based budgeting as reported by budget analysts and selected department heads in state agencies were examined by George Minmier and Roger Hermanson (1976). Zero-based budgeting was…

  18. Cryopreservation and other assisted reproductive technologies for the conservation of threatened amphibians and reptiles: bringing the ARTs up to speed.

    Science.gov (United States)

    Clulow, John; Clulow, Simon

    2016-06-01

    Amphibians and reptiles are experiencing serious declines, with the number of threatened species and extinctions growing rapidly as the modern biodiversity crisis unfolds. For amphibians, the panzootic of chytridiomycosis is a major driver. For reptiles, habitat loss and harvesting from the wild are key threats. Cryopreservation and other assisted reproductive technologies (ARTs) could play a role in slowing the loss of amphibian and reptile biodiversity and managing threatened populations through genome storage and the production of live animals from stored material. These vertebrate classes are at different stages of development in cryopreservation and other ARTs, and each class faces different technical challenges arising from the separate evolutionary end-points of their reproductive biology. For amphibians, the generation of live offspring from cryopreserved spermatozoa has been achieved, but the cryopreservation of oocytes and embryos remains elusive. With reptiles, spermatozoa have been cryopreserved in a few species, but no offspring from cryopreserved spermatozoa have been reported, and the generation of live young from AI has only occurred in a small number of species. Cryopreservation and ARTs are more developed and advanced for amphibians than reptiles. Future work on both groups needs to concentrate on achieving proof of concept examples that demonstrate the use of genome storage and ARTs in successfully recovering threatened species to increase awareness and support for this approach to conservation.

  19. Effect of seminal plasma removal before cryopreservation of bovine semen obtained by electroejaculation on semen quality and in vitro fertility.

    Science.gov (United States)

    Campanholi, Suzane Peres; Monteiro, Fabio Morato; Ribeiro Dias, Erika Aline; Mercadante, Maria Eugênia Zerlotti; de Paz, Claudia Cristina Paro; Dell'Aqua Junior, José Antonio; Papa, Frederico Ozanam; Dell'Aqua, Camila de Paula Freitas; Vantini, Roberta; Garcia, Joaquim Mansano

    2017-02-01

    Cryopreservation of bull semen is a common biotechnology procedure in cattle breeding. However, when the ejaculate is obtained by electroejaculation, wide variation is observed in the sperm/seminal plasma (SP) ratio that can affect the freezability of semen in this species. The removal of SP may improve the quality of frozen bull semen. The objective of this study was to evaluate the effect of SP removal from the ejaculate on the cryopreservation of semen from 38 Nellore bulls collected by electroejaculation. After collection, the ejaculate was divided into three aliquots: (1) control (N) diluted to a concentration of 60 × 10 6 spermatozoa/mL and frozen with SP; (2) centrifugation (C) at ×600g for 10 minutes and the pellet resuspended and frozen at the same concentration as N; and (3) filtration (F) through SpermFilter and sperm recovered and frozen at the same concentration as N. After thawing, sperm kinetics, plasma and acrosome membrane integrity, mitochondrial membrane potential, oxidative stress, and in vitro fertility were evaluated. Statistical analysis was performed using the SAS 9.2 package, and differences were considered significant when P semen freezing reduced the rate of in vitro-produced embryos, whereas filtration of prefrozen semen was found to be an efficient alternative in terms of semen freezability and in vitro production of bovine embryos. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. LONG-TERM PRECONDITIONING OF PLANTLETS: A PRACTICAL METHOD FOR ENHANCING SURVIVAL OF PINEAPPLE (Ananas comosus Merr.) SHOOT TIPS CRYOPRESERVED USING VITRIFICATION.

    Science.gov (United States)

    Hu, W H; Liu, S F; Liaw, S I

    2015-01-01

    The purpose of this study was to develop an efficient cryopreservation protocol for pineapple (Ananas comosus Merr.) shoot tips. The optimal state of pineapple plantlets was investigated by using sucrose preconditioning to enhance survival after cryostorage. To achieve a suitable state of plantlets before cryopreservation, 0.2 M to 0.4 M sucrose concentrations combined with short- (0-7 days), medium- (15-30 days), and long-term (75-150 days) preconditioning periods were compared. The highest survival (100 %) was achieved using the following procedure: intact plantlets underwent long-term preconditioning with 0.2 M sucrose for 135 days, dissected shoot tips were treated with a loading solution containing 2.0 M glycerol + 0.4 M sucrose for 60 min at 25 degree and the shoot tips were dehydrated in PVS2 for 2h at 0 degree C before being plunged in liquid nitrogen. Rewarming was conducted in a water-bath for 30 s at 40 degree C and PVS2 was replaced with a 1.2 M sucrose solution for 30 min at 25 degree C. The shoot tips were transferred on semisolid medium and left in the dark for 1 week, then in dim light for 3 weeks.

  1. Characteristics of seminal plasma and cryopreservation of anoa (Bubalus sp. semen obtained by electroejaculation

    Directory of Open Access Journals (Sweden)

    Yudi

    2011-03-01

    Full Text Available The population of anoa, which is an endemic fauna to Indonesia, was getting decrease caused by the illegal hunting and deforestation. Anoa is included in endangered species by IUCN, and Appendix I by CITES. The experiment aimed to characterize the seminal plasma contents and to cryopreserve the anoa semen for artificial insemination application in captivity. The experiment was carried out in Taman Safari Indonesia (Bogor. Semen was collected from 2 anesthetized males (4-10 years by electroejaculation. Seminal plasma gained by centrifugation of ejaculate (3000 rpm, 20 minutes, and then was evaluated the biochemical contents. Other ejaculates were evaluated macroscopically and microscopically, and then extended in Tris and Na-citrate media to a total concentration of 100 billion cells mL-1. Extended semen was stored at 4oC, and evaluated the motility and viability every 12 h. Frozen semen was made in Tris medium added with 5% of glycerol. The seminal plasma of anoa contained total lipid, Na, Ca and Mg higher than the buffalo, but its total protein, K and Cl were lower. Electrophoresis of seminal plasma using by SDS-PAGE method showed 10 bands of proteins (17-148 kDa. The motility and viability of chilled-extended semen in Tris and Na-citrate media were not significantly different (P > 0.05 during 72 h of evaluation. Extended semen in both of media may applicable for AI program for 24-48 h. Post thawing motility of frozen semen was still low, 26.00 ± 9.62%. Therefore, it is necessary to improve each stages of semen processing, so the motility will increased and resulted high pregnancy in AI program.

  2. Effect of rosemary (Rosmarinus officinalis) extracts and glutathione antioxidants on bull semen quality after cryopreservation

    Energy Technology Data Exchange (ETDEWEB)

    Daghigh-Kia, H.; Olfati-Karaji, R.; Hoseinkhani, A.; Ashrafi, I.

    2014-06-01

    The present study determined the effects of the addition of rosemary extract (ROM), glutathione (GSH), and their combination (ROM + GSH) to freezing extender on the quality of bull semen after cryopreservation. Before cryoperservation, the samples were diluted in a tris-egg yolk (TEY) extender containing 5 mM GSH (treatment I), 5 or 10 g L{sup -}1 ROM (treatments II and III), and ROM with GSH (5 mM GSH with 5 or 10 g L{sup -}1 of ROM) (treatments IV and V). An extender containing no antioxidants (non-ROM/GSH-treated) served as control group. Kinematic parameters were evaluated by means of a computer-assisted semen analysis (CASA). The viability and membrane integrity of the sperm were assessed using eosin-nigrosin stain and the hypo-osmotic swelling test (HOST) at 0 and 2 h after freezethawing. Lipooxidative parameters, superoxide dismutase, and glutathione peroxidase (GPx) activity were assessed after thawing. Treatment III showed positive effects for total motility (TM) (p < 0.01), average path velocity (VAP) (p < 0.001), viability (p < 0.01) and HOST (p < 0.01); however, lipid peroxidation (LPO) decreased (p < 0.05) and GPx activity increased (p < 0.05) immediately after thawing compared to the control. The TM (p < 0.01), VAP (p < 0.01), viability (p < 0.01), HOST (p < 0.01) decreased in LPO (p < 0.01) and GPx activity (p < 0.05) for treatment V and the viability and GPx activity (p < 0.05) for treatment I were significantly higher than for the control group at 2 h after thawing. It was concluded that the inclusion of ROM and its combination with GSH improves the post-thaw quality of bull semen. (Author)

  3. Siebel eEnergy ensures higher efficiency at HEW; Siebel eEnergy sorgt bei HEW fuer mehr Effizienz. Vertriebsvertraege und Service managen

    Energy Technology Data Exchange (ETDEWEB)

    Ritter, U. [EMEA, Siebel Systems, Ismaning (Germany)

    2003-01-27

    HEW, a leading German energy provider, uses Siebel eEnergy to create a single, comprehensive view of customer information across multiple channels and integrating with the company's back office applications. Siebel eEnergy provides improved management of energy contracts and enables HEW to increase sales efficiency, accelerate the service delivery process, and enhance customer satisfaction and loyalty. (orig.) [German] HEW, eines der fuehrenden deutschen Energieversorgungsunternehmen, nutzt Siebel eEnergy, um ueber verschiedene Kanaele hinweg umfassende Kundenprofile zu erstellen und sie mit den Back-Office-Anwendungen zu integrieren. Siebel eEnergy bietet verbessertes Management der Energieversorgungs-Vertraege und ermoeglicht HEW dadurch, die Effizienz im Vertrieb zu steigern, die Bereitstellung und Lieferung von Energie zu beschleunigen und so Kundenzufriedenheit und Kundentreue zu erhoehen. (orig.)

  4. How private housholds can use environment - higher energy consumption despite efficiency increasement; Wie private Haushalte die Umwelt nutzen - hoeherer Energieverbrauch trotz Effizienzsteigerung. Hintergrundpapier

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2006-11-15

    Energy saving potentials are not being used satisfactorily to date, especially as regards the use of electricity. No-load losses caused by electrical devices such as television sets and computers amount to some 17 billion kilowatt hours per year for private households alone. This makes 3.3. billion euros in electricity costs. Having an easily accessible mains switch that disconnects electrical devices from the mains can already help to avoid no-load losses. White goods such as fridge-freezers, washing machines or dish washers also offer considerable saving potentials. Efficient technologies could contribute annual savings from this product sector in the order of seven billion kilowatt hours, equivalent to 5.8% of the total electricity consumption of private households.

  5. Cryopreservation of third-stage larvae of Strongylus vulgaris (large strongyle of horses).

    Science.gov (United States)

    Titoy, G A; Van Rensburg, L J

    1997-06-01

    A technique for the cryopreservation of third-stage larvae of Strongylus vulgaris is described. Infective larvae of S. vulgaris were exsheathed in a 0.16% sodium hypochlorite solution and then transferred into cryotubes containing 0.09% saline. The samples were stored in the gas phase of liquid nitrogen.

  6. Cryopreservation of porcine fetal ventral mesencephalic tissue for intrastriatal transplantation in Parkinson's disease

    NARCIS (Netherlands)

    Koopmans, J.; Hogenesch, I.; Copray, S.; Middel, B.; van Dijk, H.; Go, K-G.; Staal, M.

    2001-01-01

    In this study we examined the efficacy of cryopreserving porcine fetal mesencephalic tissue. After microscopical dissection of the ventral mesencephalon (VM) from E28 pig fetuses, the collection of explants was randomly divided into two equal parts. One part was directly prepared as cell suspension.

  7. Perfusion bioreactor-based cryopreservation of 3D human mesenchymal stromal cell tissue grafts

    Czech Academy of Sciences Publication Activity Database

    Petrenko, Yuriy; Petrenko, A.; Martin, I.; Wendt, D.

    2017-01-01

    Roč. 76, jun. (2017), s. 150-153 ISSN 0011-2240 Institutional support: RVO:68378041 Keywords : cryopreservation * tissue engineering * mesenchymal stromal cells Subject RIV: FP - Other Medical Disciplines OBOR OECD: Cell biology Impact factor: 1.996, year: 2016

  8. CRYOPRESERVATION OF RAM SPERM FROM AUTOCHTHONOUS BREEDS DURING A NON-MATING SEASON

    Directory of Open Access Journals (Sweden)

    Milko SABEV

    2007-07-01

    Full Text Available It is possible to collect and successfully cryopreserve ejaculates in a non-mating season from rams of the autochthonous breeds Karakachan, Cooper-red Shumen and Karnobat-local, raised in Bulgaria. Studies are in progress aiming the elaboration of optimal cryoprotective extenders and freezing technology.

  9. Heat and mass transfer during the cryopreservation of a bioartificial liver device: a computational model.

    Science.gov (United States)

    Balasubramanian, Saravana K; Coger, Robin N

    2005-01-01

    Bioartificial liver devices (BALs) have proven to be an effective bridge to transplantation for cases of acute liver failure. Enabling the long-term storage of these devices using a method such as cryopreservation will ensure their easy off the shelf availability. To date, cryopreservation of liver cells has been attempted for both single cells and sandwich cultures. This study presents the potential of using computational modeling to help develop a cryopreservation protocol for storing the three dimensional BAL: Hepatassist. The focus is upon determining the thermal and concentration profiles as the BAL is cooled from 37 degrees C-100 degrees C, and is completed in two steps: a cryoprotectant loading step and a phase change step. The results indicate that, for the loading step, mass transfer controls the duration of the protocol, whereas for the phase change step, when mass transfer is assumed negligible, the latent heat released during freezing is the control factor. The cryoprotocol that is ultimately proposed considers time, cooling rate, and the temperature gradients that the cellular space is exposed to during cooling. To our knowledge, this study is the first reported effort toward designing an effective protocol for the cryopreservation of a three-dimensional BAL device.

  10. Rooster Semen Cryopreservation: Effect of Pedigree Line and Male Age on Post-Thaw Sperm Function

    Science.gov (United States)

    The fertility rates of cryopreserved poultry semen are highly variable and not reliable for use in preservation of commercial genetic stocks. Our objective was to evaluate the cryosurvival of semen from 8 pedigreed layer lines at the onset and end of production. Semen from 160 roosters (20/line) was...

  11. Effects of alternative cryoprotectants, diluents, straw size and cholesterol addition on cryopreserved rooster sperm

    Science.gov (United States)

    Rooster sperm do not cryopreserve well. This is due in part to osmotic changes that sperm undergo during addition and removal of cryoprotectant (CPA), as well as membrane damage during the membrane phase transition from fluid at 37 degrees C to gel at low temperatures. When a CPA is removed from spe...

  12. Schistosoma mansoni: interactive effects of irradiation and cryopreservation on parasite maturation and immunization of mice

    International Nuclear Information System (INIS)

    James, E.R.; Dobinson, A.R.

    1984-01-01

    Mechanically transformed schistosomula of Schistosoma mansoni were irradiated with levels of 60Co irradiation between 2.5 and 54 krad, cryopreserved by the two-step addition of ethanediol and rapid cooling technique, and were injected intramuscularly into groups of mice which were perfused 40 days later. The schistosomula were either irradiated and then cryopreserved (IC) or cryopreserved and then irradiated in the frozen state (CI). Development into adult worms was prevented with 4 krad for IC schistosomula, but for CI schistosomula a small number of worms (1.6%) was recovered using 8.8 krad. A dose of 4 krad was sufficient to prevent development of unfrozen controls (I), but for schistosomula irradiated while exposed to ethanediol (EI), a dose of 7 krad was required. Using the different protocols, the peak levels of protection against a challenge infection were achieved with 9 (IC) and 16 krad (CI), compared to 20 krad for unfrozen schistosomula (I) reported previously. The highest level of protection (65%) was achieved with CI schistosomula. Possible interactions between the radioprotective and damaging effects of cryopreservation are discussed

  13. Cryopreservation of Precision-cut Tissue Slices for Application in Drug Metabolism Research

    NARCIS (Netherlands)

    Graaf, Inge Anne Maria de

    2002-01-01

    The research described in this thesis had two important aims. The first was to determine whether tissue slices could be used as an in vitro tool to predict the in vivo metabolism of new drugs. The second aim was to find a manner to store tissue slices for longer time periods by cryopreservation.

  14. Comparison of the efficacy of conventional slow freezing and rapid cryopreservation methods for bovine embryos

    NARCIS (Netherlands)

    Wagtendonk-de Leeuw, van A.M.; Daas, den J.H.; Kruip, T.A.; Rail, W.F.

    1995-01-01

    Day 7 bovine morulae and early blastocysts were randomly assigned to one of four cryopreservation methods: (i) a modified conventional controlled slow freezing and stepwise dilution after thawing; and three methods which enable direct transfer of the embryo into the recipient upon thawing: (ii)

  15. The potential of cryopreservation and reproductive technologies for animal genetic resources conservation strategies

    NARCIS (Netherlands)

    Hiemstra, S.J.; Lende, van der T.; Woelders, H.

    2006-01-01

    This chapter focuses on ex situ conservation. An overview of the state of the art cryopreservation and reproductive technology for farm animals and fish is followed by a discussion on the implications of ex situ conservation strategies. Ex situ conservation of genetic material from livestock and

  16. Melatonin enhances the recovery of cryopreserved shoot tips of American elm (Ulmus Americana L.)

    Science.gov (United States)

    Climate change and the global migrations of people and goods have exposed trees to new diseases and abiotic challenges that threaten the survival of species. In vitro germplasm storage via cryopreservation is an effective tool to ensure conservation of tree species, but plant cells and tissues are e...

  17. Cryopreservation of Populus trichocarpa and Salix using dormant buds with recovery by grafting or direct rooting

    Science.gov (United States)

    Populus trichocarpa and Salix can be successfully cryopreserved by using dormant scions as the source explants. These scions (either at their original moisture content of 48 to 60% or dried to 30%) were slowly cooled to –35 degree Celsius, transferred to the vapor phase of liquid nitrogen (LNV,-160...

  18. Crosstalk between Substrates and Rho-Associated Kinase Inhibitors in Cryopreservation of Tissue-Engineered Constructs

    Directory of Open Access Journals (Sweden)

    Arindam Bit

    2017-01-01

    Full Text Available It is documented that human mesenchymal stem cells (hMSCs can be differentiated into various types of cells to present a tool for tissue engineering and regenerative medicine. Thus, the preservation of stem cells is a crucial factor for their effective long-term storage that further facilitates their continuous supply and transportation for application in regenerative medicine. Cryopreservation is the most important, practicable, and the only established mechanism for long-term preservation of cells, tissues, and organs, and engineered tissues; thus, it is the key step for the improvement of tissue engineering. A significant portion of MSCs loses cellular viability while freeze-thawing, which represents an important technical limitation to achieving sufficient viable cell numbers for maximum efficacy. Several natural and synthetic materials are extensively used as substrates for tissue engineering constructs and cryopreservation because they promote cell attachment and proliferation. Rho-associated kinase (ROCK inhibitors can improve the physiological function and postthaw viability of cryopreserved MSCs. This review proposes a crosstalk between substrate topology and interaction of cells with ROCK inhibitors. It is shown that incorporation of ionic nanoparticles in the presence of an external electrical field improves the generation of ROCK inhibitors to safeguard cellular viability for the enhanced cryopreservation of engineered tissues.

  19. STABILITY IN REAL TIME OF SOME CRYOPRESERVED MICROBIAL STRAINS WITH REFERENCE TO GENETICALLY MODIFIED MICROORGANISMS

    Directory of Open Access Journals (Sweden)

    DANIELA VINTILĂ

    2007-05-01

    Full Text Available The aim of this work is to analyze the viability of microorganisms from Collection of Industrial Microorganisms from Faculty of Animal Science and Biotechnology – Timisoara, during freezing and thawing as part of cryopreservation technique. The stability in real time of 19 strains cryopreserved in 16% glycerol was evaluated during a 6-months period. The strains studied were: Escherichia coli, Lactobacillus acidophilus, Rhizobium meliloti, Saccharomyces cerevisiae, Aspergillus oryzae, Aspergillus niger, Trichoderma viride, Bacillus globigii, Bacillus licheniformis, and 9 strains of Bacillus subtilis. The strains cryopreserved at -20oC and -70oC were activated using the fast thawing protocol. A better cell recovery was achieved with the -70oC protocol reaching an average viability for E. coli of 86,3%, comparing with 78,6% in -20oC protocol. The cell recovery percentages for the other strains were: 92,4% for L. acidophilus, 93,9% for A.niger, 89% for A. oryzae, 86,7% for T. viride, 94,2% for R. meliloti, 82,1% for S. cerevisiae, 89,9% for B. licheniformis. Regarding the viability of genetically modified microorganisms, the values shows a good recovering after freezing and thawing, even after 180 days of cryopreservation. With the -20oC protocol lower viability was observed due probably to the formation of eutectic mixtures and recrystalization processes.

  20. Droplet vitrification technique for cryopreservation of different pineapple (Ananas comosus L. Merrill) accessions

    Science.gov (United States)

    Germplasm conservation of pineapple is crucial to secure the genetic variability of the genus for breeding programs and supporting new research. Long-term conservation is done through cryopreservation, by storing cells or tissues at ultra-low temperature in liquid nitrogen (LN; -196°C) or in the LN ...

  1. Droplet-vitrification and morphohistological studies of cryopreserved shoot tips of cultivated and wild pineapple genotypes

    Science.gov (United States)

    Germplasm conservation of pineapple [Ananas comosus (L.) Mer.] is crucial to preserve the genus’ genetic diversity to secure material for genetic improvement and to support innovative and new research. Long-term conservation is accomplished through cryopreservation that is done by storing cells or t...

  2. Collection, analysis and cryopreservation of semen from Malayan gaur (Bos gaurus hubbacki: A preliminary study

    Directory of Open Access Journals (Sweden)

    M.S. Khairiah

    2012-10-01

    Full Text Available The Malayan gaur (Bos gaurus hubbacki or Seladang is classified as vulnerable by the International Union for Conservation of Nature and Natural Resources (IUCN. The Malayan gaur is mainly distributed in the tropical woodlands of Peninsular Malaysia and Southern Thailand. The aim of this study was to collect, analyze and cryopreserve the semen of wild Malayan gaur. Transrectal massage (TM and electroejaculation (EEJ technique was applied in semen collection of the Malayan gaur. The semen was then cryopreserved in liquid nitrogen using slow freezing technique. Makler counting chamber was used to evaluate sperm concentration and motility, while the sperm viability and morphology of fresh and post-thaw sperm was determined using eosin-nigrosin staining protocol. As a result, we have successfully collected the Malayan gaur semen using EEJ technique. Sperm motility, viability and morphological changes of the post-thaw semen of Malayan gaur were found undesirable due to the complication of the cryopreservation process. On the basis of current study it can be concluded that Malayan gaur bulls semen can be obtain by EEJ with no evidence of rectal trauma. Optimization of the process of cryopreservation for Malayan gaur sperm is needed to maintain the cryoviability of the good sperm quality. The data generated in this study would be useful in conservation of genetic diversity program for Malayan gaur.

  3. CRYOPRESERVATION OF FRESHLY ISOLATED SYNAPTOSOMES PREPARED FROM THE CEREBRAL-CORTEX OF RATS

    NARCIS (Netherlands)

    GLEITZ, J; BEILE, A; WILFFERT, B; TEGTMEIER, F

    In the present study, we established a cryopreservation method for freshly isolated synaptosomes prepared from the cerebral cortex of rats. Freshly prepared synaptosomes were either shock-frozen or frozen under temperature-controlled conditions using a programmable temperature controller. Each group

  4. The Different Calcium+2 Intensity Profile and Quality of Oocyte and Goat Sperms after Cryopreservation

    Science.gov (United States)

    Ciptadi, G.; Rahayu, S.; Fatchiyah; Wahyuningsih, S.; Budiarto, A.; Nasich, M.; Putri, A. R. I.; Mudawamah, M.; Ihsan, M. N.

    2018-02-01

    This research aims were to study the effect of the oocyte and sperms cryopreservation of Indonesian local goat on the post-thawing quality and profile or characters of Calcium+2 intensity in relating with their fertility capacity. A study was conducted to test the freezing method and post-thawing viability both stock cells stored in the deep freezer and liquid nitrogen (-80°C of vs -196°C). A fertility test of sperms has been conducted through in vitro of sperm quality, while the oocytes cryopreserved test was done by in vitro maturation (IVM) rate (%). The profile of Calcium 2+ was performed and analysis by Confocal Laser Scanned Microscope (CLSM). The result showed that IVM rate of goat oocyte is considered lower when cryopreserved in -80°C than in -196°C. Meanwhile, sperm is considered having a good quality in 2 methods of cryopreservation with post-thawing motility > 40 % (SNI 2014). There is an important difference between Calcium intensity of fresh and post-thawing both for oocyte and spermatozoa. Calcium +2 profiles is varied individually on the peak of intensity, but it considered expressed the same profile of each fresh and post-thawing cell. In vitro fertilization test need to be performed to complete the viability and fertility competence of these sperm and oocyte freezing stocks.

  5. Effects of seed cryopreservation, stratification and scarification on germination for five rare species of pitcher plants.

    Science.gov (United States)

    Khanna, Sruti; Jenkins, Heather; Bucalo, Kylie; Determann, Ron O; Cruse-Sanders, Jennifer M; Pullman, Gerald S

    2014-01-01

    Habitat loss and over collection have caused North American pitcher plants to become rare, including U.S. federally endangered Sarracenia alabamensis and S. oreophila, and S. leucophylla, S. psittacina and S. purpurea spp. venosa, endangered in several states. To develop reliable seed cryopreservation protocols for endangered Sarracenia species enabling similar germination percentages before and after storage in liquid nitrogen (LN) either in vivo or using in vitro tools. Seed germination pre- and post-cryopreservation were compared following seed drying with germination in soil, aseptic environment with wet filter paper or enriched medium, and using scarification or stratification for dormancy removal. After cryostorage, germination in vitro (1/6- or 1/3-strength MS medium) increased compared to germination on peat moss. Germination pre- and post-cryopreservation was similar for S. alabamensis and S. oreophila when seeds were stratified and grown in vitro. S. leucophylla and S. psittacina also showed high germination after cryopreservation when germinated on medium following stratification. Rapid liquid nitrogen exposure and rewarming induced seed coat cracking that damaged seeds, likely allowing internal damage during acid scarification and microbial entry during germination in non-sterile environments.

  6. Shoot regeneration and embryogenesis in lily shoot tips cryopreserved by droplet vitrification

    Science.gov (United States)

    Shoot regeneration and embryogenesis were, for the first time, achieved directly in shoot tips of Lilium Oriental hybrid ‘Siberia’ following cryopreservation by droplet-vitrification. Shoot tips (2 mm in length) including 2-3 leaf primordia were excised from 4-week-old adventitious shoots directly r...

  7. The effect of Tribulus terrestris extract on motility and viability of human sperms after cryopreservation.

    Science.gov (United States)

    Asadmobini, Atefeh; Bakhtiari, Mitra; Khaleghi, Sara; Esmaeili, Farzaneh; Mostafaei, Ali

    2017-04-01

    Semen cryopreservation produces significant amounts of reactive oxygen species (ROS), which may lead to impairment of sperm morphology, function, and ultimately, male fertility. Since Tribulus terrestris has antioxidant and free-radical-scavenging properties, this study aims to reveal the effect of the Tribulus terrestris extract on motility and vitality of human sperms after cryopreservation. Semen specimens from 80 healthy volunteers were divided into eight groups: fresh control (group I), freeze control (group II), groups III, IV, and V, which had 20, 40, and 50 μg/mL doses of Tribulus terrestris extract added before cryopreservation, and groups VI, VII, and VIII, which were supplemented by these extract doses after the freeze-thaw process. To evaluate the effects of the Tribulus terrestris extract, the semen samples were incubated with the extract and evaluated with a light microscope for motility and viability. After cryopreservation, a significant improvement in spermatozoa viability was observed in group VII. In groups VII and VIII, motility, according to World Health Organization (WHO) criteria, increased considerably (p Tribulus terrestris, which improves human sperm motility and viability, may be due to its antioxidant properties. On the basis of the results, the researchers concluded that Tribulus terrestris can be used as a safe therapeutic alternative to current modalities for the management of motility dysfunction in males. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Rooster sperm plasma membrane protein and phospholipid organization and reorganization attributed to cooling and cryopreservation

    Science.gov (United States)

    Cholesterol to phospholipid ratio is used as a representation for membrane fluidity, and predictor of cryopreservation success but results are not consistent across species and ignore the impact of membrane proteins. Therefore, this research explored the modulation of membrane fluidity and protein ...

  9. Cryopreserved Cadaveric Arterial Allograft for Arterial Reconstruction in Patients with Prosthetic Infection.

    Science.gov (United States)

    Lejay, Anne; Delay, Charline; Girsowicz, Elie; Chenesseau, Bettina; Bonnin, Emilie; Ghariani, Mohamed-Zied; Thaveau, Fabien; Georg, Yannick; Geny, Bernard; Chakfe, Nabil

    2017-11-01

    The aim of this study was to report outcomes of cryopreserved arterial allografts used as a vascular substitute in the setting of prosthetic material infection. A retrospective analysis of prospectively collected data was conducted including all consecutive interventions performed with cryopreserved arterial allografts used for vascular reconstruction in the setting of prosthetic material infection between January 2005 and December 2014. Five year outcomes included allograft related re-interventions, survival, primary patency, and limb salvage rates. Fifty-three procedures were performed using cryopreserved allografts for vascular prosthetic infection: 25 procedures (47%) were performed at aorto-iliac level (Group 1) and 28 procedures (53%) at peripheral level (Group 2). The mean follow-up was 52 months. Five year allograft related re-intervention was 55% in Group 1 (6 allograft ruptures and 5 allograft aneurysm degenerations) and 33% in Group 2 (2 allograft ruptures and 7 allograft aneurysm degenerations). Five year survival was 40% and 68%, primary patency was 89% and 59% and limb salvage was 100% and 89% for Group 1 and 2 respectively. Use of cryopreserved arterial allografts provides acceptable results but is tempered by suboptimal 5 year outcomes with high re-intervention rates. Copyright © 2017 European Society for Vascular Surgery. Published by Elsevier Ltd. All rights reserved.

  10. Identification of a highly successful cryopreservation method (droplet-vitrification) for petunia

    Science.gov (United States)

    Petunia (Petunia × hybrida Vilm.) is a very important crop conserved in the National Genebank of China. Petunia cultivar “Niu 2” was used to develop a droplet-vitrification protocol to cryopreserve shoot tips. Six variables (age of the in vitro plants, concentration of sucrose in the preculture solu...

  11. Potentials of short term and long term cryopreserved sperm of the ...

    African Journals Online (AJOL)

    To service the growing demand for male African giant catfish (Clarias gariepinus) broodstock for aquaculture in Nigeria, and to conserve valuable genetic resources, we improved both short-term (in deep freezer at -35°C) and long-term cryopreservation (in liquid nitrogen at -296°C) of catfish sperm. Catfish sperm ...

  12. In Vitro Propagarion and Cryopreservation of Important Grape Cultivars (Vitis Vinifera L. and Rootstocks

    Directory of Open Access Journals (Sweden)

    F. CELEBI TOPRAK

    2014-06-01

    Full Text Available Grape (Vitis vinifera L. is among the most important species that is cultivated almost all around the world. There are over one thousand varieties that are grown for raisin, fresh consumption and wine making purposes. The grape germplasm resources are generally maintained as whole plants under field conditions. The traditional way of germplasm preservation is very risky due to natural uncertainties. In vitro technologies can help producing healthy propagation materials free from viroids, viruses, bacteria, phytoplasmas, fungi, and nematodes. When combined with cryopreservation technologies in vitro preservation systems can allow safe protection and propagation of valuable Vitis genetic resources. In this study, 12 commercial cultivar and two rootstock materials were tested for the applicability of long term preservation by in vitro clonal propagation and cryopreservation techniques. Axillary shoot tips collected from newly emerging shoots were placed in Magenda boxes containing 30 g/l sucrose on MS medium and cultured in a growth chamber adjusted to 16 h ligth/25o C and 8 h dark/17o C. All grape genotypes tested responded well to this application and produced healthy root and shoots. Shoot explants from these in vitro stocks were subcultured in every three months for one year. Apical dome explants excised from in vitro grape plants were stored in liquid nitrogen for cryopreservation. Genotypes varied in their responses to cryopservation treatment. Five genotypes showed shoot or callus formation. Regenerated shoots continued to grow and produced normal shoots and roots, but no plants could be developed from calli. Flow cytometry analysis of regenerants from continuous subculture and cryopreservation did not show any chromosome number abnormalities. In vitro micropropagation is an excellent choice for a long-term conservation of grape germplasm, which allows access to actively growing plant materials without seasonal restriction. Such cultures are

  13. Improved quality of cryopreserved cheetah (Acinonyx jubatus) spermatozoa after centrifugation through Accudenz.

    Science.gov (United States)

    Crosier, Adrienne E; Henghali, Josephine N; Howard, Jogayle; Pukazhenthi, Budhan S; Terrell, Kimberly A; Marker, Laurie L; Wildt, David E

    2009-01-01

    Sperm cryopreservation, in combination with assisted reproductive techniques, is a valuable tool for the genetic management of endangered felids. However, the acrosome of the cheetah spermatozoon is especially sensitive to cryopreservation, with approximately 40% of spermatozoa experiencing acrosomal damage immediately after thawing and then another approximately 15% loss during the next 4 hours in vitro. Additionally, thawing causes a reduction in sperm motility by approximately 20% with another decrease of approximately 12% during subsequent incubation in vitro. We hypothesized that slow removal of glycerol from cryopreserved cheetah spermatozoa using an Accudenz gradient would improve acrosomal integrity, sperm motility longevity, and structural morphology. Accudenz was compared with traditional cheetah sperm processing methods for glycerol removal that involves washing, multistep resuspension, and swim-up processing. Electroejaculates (n = 21 total from 8 males) were washed in Ham F10 medium, and sperm pellets were resuspended in TEST-yolk buffer with 0% glycerol. Samples were cryopreserved in straws in 4% final glycerol, thawed, and assessed for percent intact acrosomes (% IA), percent motility (% M), and forward progressive status (FPS; scale, 0-5). Sperm motility index (SMI) was calculated as (% M + [FPS x 20]) / 2. In study 1, glycerol removal by centrifugation through an Accudenz gradient (4%, 10%) was compared with traditional sperm washing (control) and multistep resuspension protocols. At each time after centrifugation (hourly for 4 hours), % IA was improved (P cheetah sperm mitigates the significant loss in sperm quality that occurs after freeze-thawing. This alleviation of cellular damage resulting from cryopreservation contributes to a more than 10% improvement in overall sperm motility and, more importantly, allows retention of 40% or more of sperm with intact acrosomes.

  14. Immature dendritic cells generated from cryopreserved human monocytes show impaired ability to respond to LPS and to induce allogeneic lymphocyte proliferation.

    Directory of Open Access Journals (Sweden)

    Guilherme Ferreira Silveira

    Full Text Available Dendritic cells play a key role in the immune system, in the sensing of foreign antigens and triggering of an adaptive immune response. Cryopreservation of human monocytes was investigated to understand its effect on differentiation into immature monocyte-derived dendritic cells (imdDCs, the response to inflammatory stimuli and the ability to induce allogeneic lymphocyte proliferation. Cryopreserved (crp-monocytes were able to differentiate into imdDCs, albeit to a lesser extent than freshly (frh-obtained monocytes. Furthermore, crp-imdDCs had lower rates of maturation and cytokine/chemokine secretion in response to LPS than frh-imdDCs. Lower expression of Toll-like receptor 4 (at 24 and 48 h and higher susceptibility to apoptosis in crp-imdDCs than in fresh cells would account for the impaired maturation and cytokine/chemokine secretion observed. A mixed leukocyte reaction showed that lymphocyte proliferation was lower with crp-imdDCs than with frh-imdDCs. These findings suggested that the source of monocytes used to generate human imdDCs could influence the accuracy of results observed in studies of the immune response to pathogens, lymphocyte activation, vaccination and antigen sensing. It is not always possible to work with freshly isolated monocytes but the possible effects of freezing/thawing on the biology and responsiveness of imdDCs should be taken into account.

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

    Science.gov (United States)

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

    2011-12-01

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

  16. Improving the ablation efficiency of excimer laser systems with higher repetition rates through enhanced debris removal and optimized spot pattern.

    Science.gov (United States)

    Arba-Mosquera, Samuel; Klinner, Thomas

    2014-03-01

    To evaluate the reasons for the required increased radiant exposure for higher-repetition-rate excimer lasers and determine experimentally possible compensations to achieve equivalent ablation profiles maintaining the same single-pulse energies and radiant exposures for laser repetition rates ranging from 430 to 1000 Hz. Schwind eye-tech-solutions GmbH and Co. KG, Kleinostheim, Germany. Experimental study. Poly(methyl methacrylate) (PMMA) plates were photoablated. The pulse laser energy was maintained during all experiments; the effects of the flow of the debris removal, the shot pattern for the correction, and precooling the PMMA plates were evaluated in terms of achieved ablation versus repetition rate. The mean ablation performance ranged from 88% to 100%; the variability between the profile measurements ranged from 1.4% to 6.2%. Increasing the laser repetition rate from 430 Hz to 1000 Hz reduced the mean ablation performance from 98% to 91% and worsened the variability from 1.9% to 4.3%. Increasing the flow of the debris removal, precooling the PMMA plates to -18°C, and adapting the shot pattern for the thermal response of PMMA to excimer ablation helped stabilize the variability. Only adapting the shot pattern for the thermal response of PMMA to excimer ablation helped stabilize the mean ablation performance. The ablation performance of higher-repetition-rate excimer lasers on PMMA improved with improvements in the debris removal systems and shot pattern. More powerful debris removal systems and smart shot patterns in terms of thermal response improved the performance of these excimer lasers. Copyright © 2014 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.

  17. Developmental competence of oocytes isolated from surplus medulla tissue in connection with cryopreservation of ovarian tissue for fertility preservation

    DEFF Research Database (Denmark)

    Wilken-Jensen, Helle N; Kristensen, Stine G; Jeppesen, Janni V

    2014-01-01

    OBJECTIVE: Evaluating the developmental competence of immature oocytes collected from surplus medulla tissue in connection with ovarian tissue cryopreservation for fertility preservation. DESIGN: Cohort comparative study. SETTING: University laboratory in Denmark from 2011-2012. POPULATION: 69...

  18. Effects of transportation time after extraction on the magnetic cryopreservation of pulp cells of rat dental pulp

    Directory of Open Access Journals (Sweden)

    Mao-Suan Huang

    2011-03-01

    Conclusions: The freezing technique used in this animal study provided positive effects on pulp cell storage. In addition, the storage time before the freezing procedure is an important issue for cryopreserving pulp cells in intact teeth.

  19. Cryopreservation of testicular tissue before long-term testicular cell culture does not alter in vitro cell dynamics

    NARCIS (Netherlands)

    Baert, Yoni; Braye, Aude; Struijk, Robin B.; van Pelt, Ans M. M.; Goossens, Ellen

    2015-01-01

    To assess whether testicular cell dynamics are altered during long-term culture after testicular tissue cryopreservation. Experimental basic science study. Reproductive biology laboratory. Testicular tissue with normal spermatogenesis was obtained from six donors. None. Detection and comparison of

  20. Comparative proteome analysis of cryopreserved flagella and head plasma membrane proteins from sea bream spermatozoa: effect of antifreeze proteins.

    Science.gov (United States)

    Zilli, Loredana; Beirão, José; Schiavone, Roberta; Herraez, Maria Paz; Gnoni, Antonio; Vilella, Sebastiano

    2014-01-01

    Cryopreservation induces injuries to fish spermatozoa that in turn affect sperm quality in terms of fertilization ability, motility, DNA and protein integrity and larval survival. To reduce the loss of sperm quality due to freezing-thawing, it is necessary to improve these procedures. In the present study we investigated the ability of two antifreeze proteins (AFPI and AFPIII) to reduce the loss of quality of sea bream spermatozoa due to cryopreservation. To do so, we compared viability, motility, straight-line velocity and curvilinear velocity of fresh and (AFPs)-cryopreserved spermatozoa. AFPIII addition to cryopreservation medium improved viability, motility and straight-line velocity with respect to DMSO or DMSO plus AFPI. To clarify the molecular mechanism(s) underlying these findings, the protein profile of two different cryopreserved sperm domains, flagella and head plasma membranes, was analysed. The protein profiles differed between fresh and frozen-thawed semen and results of the image analysis demonstrated that, after cryopreservation, out of 270 proteins 12 were decreased and 7 were increased in isolated flagella, and out of 150 proteins 6 showed a significant decrease and 4 showed a significant increase in head membranes. Mass spectrometry analysis identified 6 proteins (4 from isolated flagella and 2 present both in flagella and head plasma membranes) within the protein spots affected by the freezing-thawing procedure. 3 out of 4 proteins from isolated flagella were involved in the sperm bioenergetic system. Our results indicate that the ability of AFPIII to protect sea bream sperm quality can be, at least in part, ascribed to reducing changes in the sperm protein profile occurring during the freezing-thawing procedure. Our results clearly demonstrated that AFPIII addition to cryopreservation medium improved the protection against freezing respect to DMSO or DMSO plus AFPI. In addition we propose specific proteins of spermatozoa as markers related to

  1. Sperm DNA fragmentation induced by cryopreservation: new insights and effect of a natural extract from Opuntia ficus-indica.

    Science.gov (United States)

    Meamar, Mehrdad; Zribi, Nassira; Cambi, Marta; Tamburrino, Lara; Marchiani, Sara; Filimberti, Erminio; Fino, Maria Grazia; Biggeri, Annibale; Menezo, Yves; Forti, Gianni; Baldi, Elisabetta; Muratori, Monica

    2012-08-01

    To analyze the effect of cryopreservation on sperm DNA fragmentation (SDF) in two cytometric sperm populations, PI(brighter) and PI(dimmer), and to test the effects of Opuntia ficus-indica (OFI) extracts, which contain antioxidants and flavanoids, and of resveratrol on cryopreservation of human semen. In vitro prospective study. Institutional study. Twenty-one normozoospermic men undergoing semen analysis for couple infertility. Cryopreservation using the routine method in the presence of OFI extracts or resveratrol. Measurement of SDF by TUNEL/PI flow cytometric method to evaluate sperm motility (by automated motion analysis, CASA system) and viability (by eosin/nigrosin staining) in the two populations of sperm PI(br) and PI(dim). Cryopreservation induced an increase of SDF only in the PI(br) sperm population. The increase was negatively dependent on the basal values of SDF in the same population. Addition of OFI extracts and resveratrol to the cryopreservation medium slightly but statistically significantly reduced SDF in the PI(br) population without affecting the deleterious effect of cryopreservation on sperm motion parameters or viability. The increase of SDF in the PI(br) population, which is unrelated to semen quality, suggests that caution must be taken in using cryopreserved semen, as morphologically normal and motile sperm may be damaged. The addition of substances with multifunctional properties such as OFI extracts to cryopreservation medium is only slightly effective in preventing the dramatic effects on SDF. Copyright © 2012 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  2. The Effect of a GnRH Agonist Injection or Progesterone Implant at Diestrus in Cryopreserved Embryo Transferred Cows

    OpenAIRE

    KIRBAŞ, Mesut; BÜLBÜL, Bülent; KÖSE, Mehmet; DURSUN, Şükrü; ÇOLAK, Mehmet

    2014-01-01

    In this study, the effect of a single dose of GnRH on d 13 or progesterone implant for 7 days between d 13 and 20 on plasma progesterone levels and pregnancy rates on cryopreserved embryo transferred cows were investigated. Synchronized 48 Brown Swiss recipient cows were used as animal material. Seven days after estrus detection, cryopreserved cattle embryos were transferred into recipients and cows were assigned randomly into three groups. In GnRH group (n=16), cows were intramuscularly inje...

  3. Cryopreservation of chayote (Sechium edule JACQ.SW.) zygotic embryos and shoot-tips from in vitroplantlets

    OpenAIRE

    Abdelnour-Esquivel, Ana; Engelmann, Florent

    2002-01-01

    This paper presents the development of cryopreservation protocols for zygotic embryos and apices of chayote (Sechium edule Jacq. Sw.), a tropical plant species with recalcitrant seeds. Zygotic embryos of two cultivars, Ccocro negro (CN) and Claudio (Cl) could withstand cryopreservation, with survival percentages of 10 and 30 %, after desiccation to 23 and 19 % moisture content (fresh weight basis), respectively. Apices sampled on in vitro plantlets of cultivars Cl, 13 and JM we...

  4. Cryopreservation of human oocytes, zygotes, embryos and blastocysts: A comparison study between slow freezing and ultra rapid (vitrification methods

    Directory of Open Access Journals (Sweden)

    Tahani Al-Azawi

    2013-12-01

    Full Text Available Preservation of female genetics is currently done primarily by means of oocyte and embryo cryopreservation. The field has seen much progress during its four-decade history, progress driven predominantly by research in humans. It can also be done by preservation of ovarian tissue or entire ovary for transplantation, followed by oocyte harvesting or natural fertilization. Two basic cryopreservation techniques rule the field, slow-rate freezing, the first to be developed and vitrification which in recent years, has gained a foothold. The slow-rate freezing method previously reported had low survival and pregnancy rates, along with the high cost of cryopreservation. Although there are some recent data indicating better survival rates, cryopreservation by the slow freezing method has started to discontinue. Vitrification of human embryos, especially at early stages, became a more popular alternative to the slow rate freezing method due to reported comparable clinical and laboratory outcomes. In addition, vitrification is relatively simple, requires no expensive programmable freezing equipment, and uses a small amount of liquid nitrogen for freezing. Moreover, oocyte cryopreservation using vitrification has been proposed as a solution to maintain women’s fertility by serving and freezing their oocytes at the optimal time. The aim of this research is to compare slow freezing and vitrification in cryopreservation of oocytes, zygotes, embryos and blastocysts during the last twelve years. Therefore, due to a lot of controversies in this regard, we tried to achieve an exact idea about the subject and the best technique used.

  5. INTERRELATION BETWEEN TYPE AND CULTIVAR OF BERRY-LIKE CROP AND CUTTINGS CRYOPRESERVATION RESULTS

    Directory of Open Access Journals (Sweden)

    L. V. Gorbunov

    2013-04-01

    Full Text Available Optimal cryopreservation parameters were determined for the berry-like crop cuttings: blackcurrant (Dachnica, Yuviley, Sofiivska, Kitayskaya; redcurrant (Kitaivska, Djoker, Svyatkova; gooseberries (Krasen, Malachite, Kolobok; raspberries (Novost’ Kuz’mina, Struyka, Skromnica. This procedure allowed increasing viability of deconservation samples in 2–5 times compared to the existing methods of cryopreservation. Maximal values of cutting viability were obtained after their temperature adaptation at –10 °C during 14–60 days, stepwise cooling of the samples with 0.1–0.5 °C/hr rate down to –30 °C (exposure 3–7 days, direct plunging into liquid nitrogen, storage from 1 to 30 days and thawing rate of 70 °C/min. It was found that the in the region of allowable values for maximum viability of investigated birch (control cuttings humidity values correspond 32±40%, for blackcurrant are 40±50%, and raspberries 37±40 % in affirmative temperature span and 14±28 %, 30±40 %, 20±23 % in below zero. Using dispersion analysis it has been established that the development probability of frozenthawed plant cuttings significantly depends on the selected cultivar and cryopreservation method. It is noted that the force of influence η2А, due to the individual properties of the studied cultivars of berries, is comparable to that of the force of influence η2В of cryopreservation procedures. The differences of average indexes of viability of the studied cultivars were showed within a single species. It was found that these rates are different for black currant by 92%, redcurrant — 54%, gooseberries — 48%, raspberries — 70%. Using paired Student’s t-test has reduced to 3 times the error of representativeness of the sample that enabled to improve the reliability of the significance of differences compared to the values of the P ≥0,99. Selection of the cuttings’ cultivars and cryopreservation method significantly affect the

  6. Alginate beads as a tool to handle, cryopreserve and culture isolated human primordial/primary follicles.

    Science.gov (United States)

    Camboni, A; Van Langendonckt, A; Donnez, J; Vanacker, J; Dolmans, M M; Amorim, C A

    2013-08-01

    One major concern of grafting cryopreserved ovarian tissue to restore fertility in cancer patients is the possibility of reintroducing tumor cells. Cryopreservation of isolated primordial/primary follicles (PFs) may circumvent this problem. The aim of our work was to compare dimethyl sulfoxide (ME2SO) and ethylene glycol (EG) as cryoprotectants (CPAs) for slow-freezing of isolated human PFs in alginate. Ovarian biopsies from four women were processed for follicle isolation. PFs were embedded in alginate (5-15 per group). Follicles were frozen-thawed using 1.4M ME2SO or 1.5M EG as CPAs. Fresh and cryopreserved isolated follicles were in vitro cultured (IVC) for 7 days. At different time periods (after isolation, cryopreservation and IVC), follicles were evaluated with live/dead assay (using fluorescent probes) and diameter measurement. Follicle viability was calculated according to the percentage of dead follicular cells and the presence of a live/dead oocyte. A total of 841 PFs were isolated, embedded in alginate and cryopreserved with ME2SO (n=424) or EG (n=259), or used as controls (n=158). After 7 days of IVC, a significant increase in follicle size was observed in the fresh and ME2SO groups, but not in the EG group. The percentage of totally viable PFs was not significantly different before or after seven days of culture in fresh (100% and 82%) or ME2SO (93.2% and 85.1%) tissue. The EG group showed significantly lower viability before (63.9%) and after IVC (66.2%) than the fresh and ME2SO groups. Our results show that 1.4M ME2SO yields better preservation of isolated PF viability after thawing and 7 days of IVC than 1.5M EG. Alginate constitutes an easy, safe hydrogel matrix to handle and cryopreserve isolated human follicles using ME2SO as a CPA. Copyright © 2013 Elsevier Inc. All rights reserved.

  7. Optimal Thawing of Cryopreserved Peripheral Blood Mononuclear Cells for Use in High-Throughput Human Immune Monitoring Studies

    Directory of Open Access Journals (Sweden)

    Ramu A. Subbramanian

    2012-07-01

    Full Text Available Cryopreserved peripheral blood mononuclear cells (PBMC constitute an important component of immune monitoring studies as they allow for efficient batch- testing of samples as well as for the validation and extension of original studies in the future. In this study, we systematically test the permutations of PBMC thawing practices commonly employed in the field and identify conditions that are high and low risk for the viability of PBMC and their functionality in downstream ELISPOT assays. The study identifies the addition of ice-chilled washing media to thawed cells at the same temperature as being a high risk practice, as it yields significantly lower viability and functionality of recovered PBMC when compared to warming the cryovials to 37 °C and adding a warm washing medium. We found thawed PBMC in cryovials could be kept up to 30 minutes at 37 °C in the presence of DMSO before commencement of washing, which surprisingly identifies exposure to DMSO as a low risk step during the thawing process. This latter finding is of considerable practical relevance since it permits batch-thawing of PBMC in high-throughput immune monitoring environments.

  8. Morphology and function of cryopreserved whole ovine ovaries after heterotopic autotransplantation

    Directory of Open Access Journals (Sweden)

    Sharma Rakesh K

    2008-04-01

    Full Text Available Abstract Background The objective of this study was to perform complex characterization of cryopreserved and then autotransplanted ovaries including determination of the ability to respond to in vivo follicle stimulating hormone (FSH-treatment, fertilizability of retrieved oocytes, and morphology, vascularization, cellular proliferation and apoptosis in sheep. Methods Mature crossbred ewes were divided into two groups; an intact (control group (n = 4, and autotransplanted group (n = 4 in which oophorectomy was performed laparoscopically and ovaries with intact vascular pedicles frozen, thawed and transplanted back into the same animal at a different site. Approximately five months after autotransplantation, estrus was synchronized, ewes were treated with FSH, and ovaries were collected. For all ovaries, number of visible follicles was determined, and collected cumulus oocyte complexes (COC were matured and fertilized in vitro. Remaining ovarian tissues were fixed for evaluation of morphology, expression of factor VIII (marker of endothelial cells, vascular endothelial growth factor (VEGF; expressed by pericytes and smooth muscle cells, and smooth muscle cell actin (SMCA; marker of pericytes and smooth muscle cells, and cellular proliferation and apoptosis. Two fully functional ovaries were collected from each control ewe (total 8 ovaries. Results Out of eight autotransplanted ovaries, a total of two ovaries with developing follicles were found. Control ewes had 10.6 +/- 2.7 follicles/ovary, oocytes were in vitro fertilized and developed to the blastocyst stage. One autotransplanted ewe had 4 visible follicles from which 3 COC were collected, but none of them was fertilized. The morphology of autotransplanted and control ovaries was similar. In control and autotransplanted ovaries, primordial, primary, secondary, antral and preovulatory follicles were found along with fully functional vascularization which was manifested by expression of factor

  9. Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development.

    Science.gov (United States)

    Torres, Leticia; Hu, E; Tiersch, Terrence R

    2016-01-07

    Cryopreservation in aquatic species in general has been constrained to research activities for more than 60 years. Although the need for application and commercialisation pathways has become clear, the lack of comprehensive quality assurance and quality control programs has impeded the progress of the field, delaying the establishment of germplasm repositories and commercial-scale applications. In this review we focus on the opportunities for standardisation in the practices involved in the four main stages of the cryopreservation process: (1) source, housing and conditioning of fish; (2) sample collection and preparation; (3) freezing and cryogenic storage of samples; and (4) egg collection and use of thawed sperm samples. In addition, we introduce some key factors that would assist the transition to commercial-scale, high-throughput application.

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

    Science.gov (United States)

    2012-08-01

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

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

    Science.gov (United States)

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

    2012-12-01

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

  12. Polyproline as a Minimal Antifreeze Protein Mimic That Enhances the Cryopreservation of Cell Monolayers.

    Science.gov (United States)

    Graham, Ben; Bailey, Trisha L; Healey, Joseph R J; Marcellini, Moreno; Deville, Sylvain; Gibson, Matthew I

    2017-12-11

    Tissue engineering, gene therapy, drug screening, and emerging regenerative medicine therapies are fundamentally reliant on high-quality adherent cell culture, but current methods to cryopreserve cells in this format can give low cell yields and require large volumes of solvent "antifreezes". Herein, we report polyproline as a minimum (bio)synthetic mimic of antifreeze proteins that is accessible by solution, solid-phase, and recombinant methods. We demonstrate that polyproline has ice recrystallisation inhibition activity linked to its amphipathic helix and that it enhances the DMSO cryopreservation of adherent cell lines. Polyproline may be a versatile additive in the emerging field of macromolecular cryoprotectants. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    Science.gov (United States)

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

    2010-02-01

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

  14. Effect of the addition of two superoxide dismutase analogues (Tempo and Tempol) to alpaca semen extender for cryopreservation.

    Science.gov (United States)

    Santiani, Alexei; Acosta, Alexei Santiani; Evangelista, Shirley; Vargas, Shirley Evangelista; Valdivia, Martha; Cuya, Martha Valdivia; Risopatrón, Jennie; González, Jennie Risopatrón; Sánchez, Raúl; Gutiérrez, Raúl Sánchez

    2013-03-15

    The main objective was to study the effects, on sperm function, of the addition of two superoxide dismutase (SOD) analogues (Tempo and Tempol) to alpaca semen extender for cryopreservation. Twelve alpaca semen samples were collected using an artificial vagina and then diluted at a 1:3 ratio in an extender based on skim milk, egg yolk, and fructose. Each semen sample was divided into three equal parts to form the following groups: control, Tempo (1 mM), and Tempol (1 mM). Groups were cooled to 5 °C in 90 minutes (-1 °C in 3 minutes); when samples reached approximately 10 °C, SOD analogues were added to the respective groups. At 5 °C, ethylene glycol (final concentration, 0.1 M) was added to each group. After 30 minutes at 5 °C, samples were loaded in 0.25 mL plastic straws, placed in liquid nitrogen vapor for 15 minutes, and then plunged. Percentages of sperm motility, functional sperm membrane integrity, and viable sperm with intact acrosomes were evaluated before and after freeze-thaw using visual analysis, the hypoosmotic swelling test, and the double-stain trypan blue/giemsa technique, respectively. The Terminal deoxymucleotidyl transferase dUTP Nick End Labeling assay was performed for evaluation of sperm DNA fragmentation of frozen-thawed sperm. Sperm motility was higher (P < 0.05) in the Tempol and Tempo groups than in the control group (mean, 22.1%, 19.7%, and 11.2%, respectively), with similar results for functional sperm membrane integrity. Additionally, DNA fragmentation was lower (P < 0.05) in the Tempol group (16.7%) than in the control group (38.8%). Viable sperm with intact acrosomes were not affected by the use of SOD analogues. There was a negative correlation (r = -0.58) between DNA fragmentation of alpaca sperm and sperm motility after freeze-thawing, but DNA damage was neither related to functional membrane integrity nor viable sperm with intact acrosomes. We concluded that DNA fragmentation and loss of motility during cryopreservation of

  15. Assisted Reproductive Technology and Newborn Size in Singletons Resulting from Fresh and Cryopreserved Embryos Transfer

    OpenAIRE

    Levi Dunietz, Galit; Holzman, Claudia; Zhang, Yujia; Talge, Nicole M.; Li, Chenxi; Todem, David; Boulet, Sheree L.; McKane, Patricia; Kissin, Dmitry M.; Copeland, Glenn; Bernson, Dana; Diamond, Michael P.

    2017-01-01

    Objectives and Study Design The aim of this study was two-fold: to investigate the association of Assisted Reproductive Technology (ART) and small newborn size, using standardized measures; and to examine within strata of fresh and cryopreserved embryos transfer, whether this association is influenced by parental infertility diagnoses. We used a population-based retrospective cohort from Michigan (2000?2009), Florida and Massachusetts (2000?2010). Our sample included 28,946 ART singletons con...

  16. ANALYSIS OF INDIVIDUAL BIOLOGICAL CHARACTERISTICS OF AMUR CARP (CYPRINUS CARPIO HAEMATOPTERUS REPRODUCED USING CRYOPRESERVED SPERM

    Directory of Open Access Journals (Sweden)

    N. Kolisnyk

    2014-12-01

    Full Text Available Purpose. To reproduce Amur carp population using cryopreserved sperm and analyze some biological and fish culture peculiarities of the reproduced fish stock. Methodology. Generally accepted methods for fish culture [1]. Experimental reproduction was carried out in pond conditions of «Carpathian vodogray» LTD (Lisnevychi village, Pustomytivsky district, Lviv region. Hydrochemical analysis was carried out classically by O. Alуokin (1970 [2], hydrobiological studies in the fatting ponds according to V. Zhadin (1956, 1960 [3, 4]. Haemoglobin concentration was determined by hemocyanin method of G. Dervis, A. Vorobiov [5]. Blood for this method was collected from fish heart with the use of Pasteur pipettes in Eppendorf tubes with heparin. Following exterior morphometric parameters were analysed: body weight (m, g, standard fish body length (l, cm, largest body height (H, cm and body circumference (O cm. Following exterior indices were calculated based on these parameters: body depth index (l/H, body circumference index (l/O and Fulton’s condition factor (Kv. The study was carried out using two groups of carp: control and experimental. The first group was reproduced from the native sperm, the second from the cryopreserved sperm. Findings. Carp reproduction and growing was carried out using native and cryopreserved sperm. This work contains the results of growing 1+ Amur carp of experimental and control groups. Hydrochemical and hydrobiological parameters of the fattening ponds were studied. Peculiarities of the exterior and some hematological parameters of the carp of different origin were characterized. Originality. For the first time we performed a comparison of some biological parameters of Amur carp reproduced using native and cryopreserved sperm. Practical Value. Considering the economic importance of Amur carp due to its use in hybridization, reproduction of its population plays an important role in the development of the stocks of the pure

  17. Thermo-mechanical stress analysis of cryopreservation in cryobags and the potential benefit of nanowarming.

    Science.gov (United States)

    Solanki, Prem K; Bischof, John C; Rabin, Yoed

    2017-06-01

    Cryopreservation by vitrification is the only promising solution for long-term organ preservation which can save tens of thousands of lives across the world every year. One of the challenges in cryopreservation of large-size tissues and organs is to prevent fracture formation due to the tendency of the material to contract with temperature. The current study focuses on a pillow-like shape of a cryobag, while exploring various strategies to reduce thermo-mechanical stress during the rewarming phase of the cryopreservation protocol, where maximum stresses are typically found. It is demonstrated in this study that while the level of stress may generally increase with the increasing amount of CPA filled in the cryobag, the ratio between width and length of the cryobag play a significant role. Counterintuitively, the overall maximum stress is not found when the bag is filled to its maximum capacity (when the filled cryobag resembles a sphere). Parametric investigation suggests that reducing the initial rewarming rate between the storage temperature and the glass transition temperature may dramatically decrease the thermo-mechanical stress. Adding a temperature hold during rewarming at the glass transition temperature may reduce the thermo-mechanical stress in some cases, but may have an adverse effect in other cases. Finally, it is demonstrated that careful incorporation of volumetric heating by means on nanoparticles in an alternating magnetic field, or nanowarming, can dramatically reduce the resulting thermo-mechanical stress. These observations display the potential benefit of a thermo-mechanical design of the cryopreservation protocols in order to prevent structural damage. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Optimized cryopreservation of mixed microbial communities for conserved functionality and diversity.

    Directory of Open Access Journals (Sweden)

    Frederiek-Maarten Kerckhof

    Full Text Available The use of mixed microbial communities (microbiomes for biotechnological applications has steadily increased over the past decades. However, these microbiomes are not readily available from public culture collections, hampering their potential for widespread use. The main reason for this lack of availability is the lack of an effective cryopreservation protocol. Due to this critical need, we evaluated the functionality as well as the community structure of three different types of microbiomes before and after cryopreservation with two cryoprotective agents (CPA. Microbiomes were selected based upon relevance towards applications: (1 a methanotrophic co-culture (MOB, with potential for mitigation of greenhouse gas emissions, environmental pollutants removal and bioplastics production; (2 an oxygen limited autotrophic nitrification/denitrification (OLAND biofilm, with enhanced economic and ecological benefits for wastewater treatment, and (3 fecal material from a human donor, with potential applications for fecal transplants and pre/probiotics research. After three months of cryopreservation at -80 °C, we found that metabolic activity, in terms of the specific activity recovery of MOB, aerobic ammonium oxidizing bacteria (AerAOB and anaerobic AOB (AnAOB, anammox in the OLAND mixed culture, resumes sooner when one of our selected CPA [dimethyl sulfoxide (DMSO and DMSO plus trehalose and tryptic soy broth (DMSO+TT] was added. However, the activity of the fecal community was not influenced by the CPA addition, although the preservation of the community structure (as determined by 16S rRNA gene sequencing was enhanced by addition of CPA. In summary, we have evaluated a cryopreservation protocol that succeeded in preserving both community structure and functionality of value-added microbiomes. This will allow individual laboratories and culture collections to boost the use of microbiomes in biotechnological applications.

  19. Cryopreserved Ultra-Thick Human Amniotic Membrane for Conjunctival Surface Reconstruction After Excision of Conjunctival Tumors.

    Science.gov (United States)

    Tanaka, Thais S; Demirci, Hakan

    2016-04-01

    Cryopreserved ultra-thick human amniotic membrane (AM) is used for glaucoma surgery. We evaluated the use of cryopreserved ultra-thick human AM for conjunctival surface reconstruction after excision of a conjunctival tumor. We retrospectively reviewed the medical records of 28 patients who underwent conjunctival surface reconstruction with cryopreserved ultra-thick human AM after excision of the tumor. The AM was secured to the surrounding conjunctiva and underlying sclera with interrupted 8-0 Vicryl sutures. Clinical data regarding demographics, diagnosis, size and location of conjunctival tumors, patient outcome, and complications were gathered. Of 28 patients, 6 (21.4%) had malignant melanoma, 4 (14.3%) had squamous cell carcinoma, 6 (21.4%) had conjunctival intraepithelial neoplasia, 1 (3.6%) had sebaceous carcinoma, 1 (3.6%) had mucoepidermoid carcinoma, 1 (3.6%) had conjunctival intraepithelial dysplasia, 5 (17.9%) had pterygium, 2 (7.1%) had compound nevus, 1 (3.6%) had a large epithelial inclusion cyst, and 1 (3.6%) patient had a granuloma. The mean area of graft size was 156 ± 120 mm2. Postoperatively, the graft was well tolerated with no failure, discomfort, or dehiscence. During the 17-month mean follow-up, symblepharon, which was clinically nonsignificant, developed in 3 (11%) patients and partial stem cell deficiency was noted in 5 (18%) patients. Cryopreserved ultra-thick human AM is a well-tolerated, effective graft material that is easy to handle. It is a viable alternative for conjunctival surface reconstruction after excision of a conjunctival tumor.

  20. Recovery from supercooling, freezing, and cryopreservation stress in larvae of the drosophilid fly, Chymomyza costata

    Czech Academy of Sciences Publication Activity Database

    Štětina, Tomáš; Hůla, Petr; Moos, Martin; Šimek, Petr; Šmilauer, P.; Košťál, Vladimír

    2018-01-01

    Roč. 8, č. 1 (2018), č. článku 4414. ISSN 2045-2322 R&D Projects: GA ČR(CZ) GA16-06374S Institutional support: RVO:60077344 Keywords : insect * cold tolerance * cryopreservation Subject RIV: ED - Physiology OBOR OECD: Biology (theoretical, mathematical, thermal, cryobiology, biological rhythm), Evolutionary biology Impact factor: 4.259, year: 2016 http://www.nature.com/articles/s41598-018-22757-0

  1. Recent advances in the field of ovarian tissue cryopreservation and opportunities for research.

    Science.gov (United States)

    Ladanyi, Camille; Mor, Amir; Christianson, Mindy S; Dhillon, Namisha; Segars, James H

    2017-06-01

    The purpose of this study was to summarize the latest advances and successes in the field of ovarian tissue cryopreservation while identifying gaps in current knowledge that suggest opportunities for future research. A systematic review was performed according to PRISMA guidelines for all relevant full-text articles in PubMed published in English that reviewed or studied historical or current advancements in ovarian tissue cryopreservation and auto-transplantation techniques. Ovarian tissue auto-transplantation in post-pubertal women is capable of restoring fertility with over 80 live births currently reported with a corresponding pregnancy rate of 23 to 37%. The recently reported successes of live births from transplants, both in orthotopic and heterotopic locations, as well as the emerging methods of in vitro maturation (IVM), in vitro culture of primordial follicles, and possibility of in vitro activation (IVA) suggest new fertility options for many women and girls. Vitrification, as an ovarian tissue cryopreservation technique, has also demonstrated successful live births and may be a more cost-effective method to freezing with less tissue injury. Further, transplantation via the artificial ovary with an extracellular tissue matrix (ECTM) scaffolding as well as the effects of sphingosine-1-phosphate (SIP) and fibrin modified with heparin-binding peptide (HBP), heparin, and a vascular endothelial growth factor (VEGF) have demonstrated important advancements in fertility preservation. As a fertility preservation method, ovarian tissue cryopreservation and auto-transplantation are currently considered experimental, but future research may pave the way for these modalities to become a standard of care for women facing the prospect of sterility from ovarian damage.

  2. In vitro reactivity to concanavalin A of bovine lymphocytes after cryopreservation

    NARCIS (Netherlands)

    Kuil, H.

    1984-01-01

    Cryopreservation of bovine peripheral lymphocytes and its effect on the in vitro response to concanavalin A tested in a microculture system is described. Using DMSO as cryoprotectant in the medium, the cells were cooled to −30°C at 1.3°C/minute and further to −80°C at 6°C/minute and then rapidly to

  3. Biomass boilers: towards a higher efficiency

    International Nuclear Information System (INIS)

    Petitot, Pauline; Signoret, Stephane; Mary, Olivier; Dejeu, Mathieu; Tachet, Jean-Pierre

    2014-01-01

    A set of articles proposes an overview of the situation and perspectives of biomass fuelled boilers in France. As outlined in an interview, professionals are supported by ADEME and the Heat Fund (Fonds Chaleur) for a continuous development of wood-energy in order to reach national objectives for renewable energies by 2020. The next article discusses issues related to wood supply, with some concerns regarding forest exploitation, and needs to find new management ways and to use other sources than forests. The technical status and perspectives of smoke condensation in wood-fuelled boilers are discussed. The example of a malt-house near Issoudun fuelled by biomass since 2013 is presented. Other examples concern a small town of Burgundy which developed and is still improving a heat network, a wood-fuelled heat network in Saint-Denis, and a biomass wood-fuelled heat production plant for the Toulouse University hospital. Graphs indicate evolutions of prices for different wood-based fuel products. The last article outlines the role of forests and the importance of their protection in the struggle against climate change, and discusses problems faced to support this preservation and its financing

  4. Sperm quality and cryopreservation of Brazilian freshwater fish species: a review.

    Science.gov (United States)

    Viveiros, A T M; Godinho, H P

    2009-03-01

    The Brazilian freshwater fish diversity is the richest in the world. Only 0.7% of all Brazilian species have had any aspect of their sperm biology addressed up to this date. The majority of the fish species described in this review migrate during the spawning season (a phenomenon known as piracema). Urbanization, pollution, hydroelectric dams and deforestation are some of the causes of stock depletion or even local extinction of some of these species. The knowledge concerning sperm quality and minimum sperm:egg ratio is important to maximize the use of males without reducing hatching rates. Furthermore, sperm cryopreservation and gene banking can guarantee the conservation of genetic diversity and development of adequate breeding programs of native fish species. In this review, we present and evaluate the existing information on Brazilian fish species that have been subject to sperm quality and cryopreservation studies. The following parameters were evaluated: volume of extractable sperm, sperm motility, sperm concentration, freezing media, freezing methods, and post-thaw sperm quality. Although the existing protocols yield relatively high post-thaw motility and fertilization rates, the use of cryopreserved sperm in routine hatchery production is still limited in Brazil.

  5. Noninvasive embryo assessment technique based on buoyancy and its association with embryo survival after cryopreservation.

    Science.gov (United States)

    Wessels, Cara; Penrose, Lindsay; Ahmad, Khaliq; Prien, Samuel

    2017-11-01

    Embryo cryopreservation offers many benefits by allowing genetic preservation, genetic screening, cost reduction, global embryo transport and single embryo transfer. However, freezing of embryos decreases embryo viability, as intracellular ice crystal formation often damages embryos. Success rates of frozen embryo transfer are expected to be 15-20% less than fresh embryo transfer. We have developed a noninvasive embryo assessment technique (NEAT) which enables us to predict embryo viability based on buoyancy. The purpose of this research was twofold. First was to determine if a NEAT, through a specific gravity device can detect embryo survival of cryopreservation. Second, it was to relate embryo buoyancy to embryo viability for establishing pregnancies in sheep. Blastocysts descent times were measured on one-hundred sixty-nine mice blastocysts before cryopreservation, according to standard protocol and post-thawing blastocysts descent times were measured again. There was a significant difference in blastocyst post-thaw descent times with NEAT in those blastocysts which demonstrated viability from those that did not (P embryos. Further studies on a larger scale commercial setting will evaluate the efficacy of NEAT. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Cryopreservation of artificial gut microbiota produced with in vitro fermentation technology.

    Science.gov (United States)

    Bircher, Lea; Schwab, Clarissa; Geirnaert, Annelies; Lacroix, Christophe

    2018-01-01

    Interest in faecal microbiota transplantation (FMT) has increased as therapy for intestinal diseases, but safety issues limit its widespread use. Intestinal fermentation technology (IFT) can produce controlled, diverse and metabolically active 'artificial' colonic microbiota as potential alternative to common FMT. However, suitable processing technology to store this artificial microbiota is lacking. In this study, we evaluated the impact of the two cryoprotectives, glycerol (15% v/v) and inulin (5% w/v) alone and in combination, in preserving short-chain fatty acid formation and recovery of major butyrate-producing bacteria in three artificial microbiota during cryopreservation for 3 months at -80°C. After 24 h anaerobic fermentation of the preserved microbiota, butyrate and propionate production were maintained when glycerol was used as cryoprotectant, while acetate and butyrate were formed more rapidly with glycerol in combination with inulin. Glycerol supported cryopreservation of the Roseburia spp./Eubacterium rectale group, while inulin improved the recovery of Faecalibacterium prausnitzii. Eubacterium hallii growth was affected minimally by cryopreservation. Our data indicate that butyrate producers, which are key organisms for gut health, can be well preserved with glycerol and inulin during frozen storage. This is of high importance if artificially produced colonic microbiota is considered for therapeutic purposes. © 2017 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  7. Seedling development and evaluation of genetic stability of cryopreserved Dendrobium hybrid mature seeds.

    Science.gov (United States)

    Galdiano, Renato Fernandes; de Macedo Lemos, Eliana Gertrudes; de Faria, Ricardo Tadeu; Vendrame, Wagner Aparecido

    2014-03-01

    Vitrification, a simple, fast, and recommended cryopreservation method for orchid germplasm conservation, was evaluated for Dendrobium hybrid "Dong Yai" mature seeds. The genetic stability of regenerated seedlings was also evaluated using flow cytometry. Mature seeds from this hybrid were submitted to plant vitrification solution (PVS2) for 0, 0.5, 1, 2, 3, 4, 5, or 6 h at 0 °C. Subsequently, they were plunged into liquid nitrogen (LN) at -196 °C for 1 h and recovered in half-strength Murashige and Skoog culture medium (1/2 MS), and seed germination was evaluated after 30 days. Seeds directly submitted to LN did not germinate after cryopreservation. Seeds treated with PVS2 between 1 and 3 h presented the best germination (between 51 and 58%), although longer exposure to PVS2 returned moderated germination (39%). Germinated seeds were further subcultured in P-723 culture medium and developed whole seedlings in vitro after 180 days, with no abnormal characteristics, diseases, or nutritional deficiencies. Seedlings were successfully acclimatized under greenhouse conditions with over 80% survival. Flow cytometry analysis revealed no chromosomal changes on vitrified seedlings, as well as seedlings germinated from the control treatment (direct exposure to LN). These findings indicate that vitrification is a feasible and safe germplasm cryopreservation method for commercial Dendrobium orchid hybrid conservation.

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

    Science.gov (United States)

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

    2013-04-01

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

  9. Behavior of lateral buds of Hancornia speciosa after cryopreservation by encapsulation-vitrification

    Directory of Open Access Journals (Sweden)

    Débora de Oliveira Prudente

    2017-05-01

    Full Text Available Hancornia speciosa is a fruitful species from Cerrado biome with high economic potential. However, the intense and disordered extractivism have caused a reduction of its population in its endemic area. In addition, seed recalcitrance negatively affects the conventional conservation of the species. Aiming to find alternatives that enable the long-term conservation of this species, the study’s objective was to assess the behavior of lateral bud’s regrowth after cryopreservation procedures by encapsulation-vitrification technique. Sodium alginate capsules containing lateral buds were pre-cultured in liquid WPM supplemented with 1.0 M glycerol, and subsequently exposed to different concentrations of sucrose (0.3; 0.75 and 1.0 M for 24 or 48 hours. The capsules were subjected to dehydration in silica gel or airflow hood for 0, 1, 2 and 3 hours before different incubation times in PVS2 (0, 15, 30, 60 and 120 minutes at 0°C. A high regeneration percentage of lateral buds was observed after cryopreservation of capsules treated with 0.75 M sucrose plus 1.0 M glycerol (24 hours, associated with dehydration in an airflow hood (1 hour and immersion in PVS2 (15 minutes. Encapsulation-vitrification allowed the long-term conservation, and provided high plant material survival rates after cryopreservation of Hancornia speciosa sensitive explants.

  10. Ice Recrystallization Inhibiting Polymers Enable Glycerol-Free Cryopreservation of Micro-organisms.

    Science.gov (United States)

    Hasan, Muhammad; Fayter, Alice E R; Gibson, Matthew I

    2018-06-22

    All modern molecular biology and microbiology is underpinned not only by the tools to handle and manipulate microorganisms, but also those to store, bank and transport them. Glycerol is the current gold-standard cryoprotectant but it is intrinsically toxic to most micro-organisms: only a fraction of cells survive freezing and the presence of glycerol can impact down-stream applications and assays. Extremophile organisms survive repeated freeze/thaw cycles by producing antifreeze proteins which are potent ice recrystallization inhibitors. Here we introduce a new concept for the storage/transport of micro-organisms by using ice recrystallization inhibiting poly(vinyl alcohol) in tandem with poly(ethylene glycol). This cryopreserving formulation is shown to result in a 4-fold increase in E. coli yield post-thaw, compared to glycerol, utilizing lower concentrations, with successful cryopreservation at just 1.1 weight percent of additive. The mechanism of protection is demonstrated to be linked to inhibiting ice recrystallization (by comparison to a recombinant antifreeze protein) but also to the significantly lower toxicity of the polymers compared to glycerol. Optimized formulations are presented and shown to be broadly applicable to the cryopreservation of a panel of Gram negative, Gram positive and Mycobacteria strains. This represents a step-change in how micro-organisms will be stored by the design of new macromolecular ice growth inhibitors; it should enable a transition from traditional solvent-based to macromolecular microbiology storage methods.

  11. Mechanical compliance and immunological compatibility of fixative-free decellularized/cryopreserved human pericardium.

    Directory of Open Access Journals (Sweden)

    Maria Cristina Vinci

    Full Text Available BACKGROUND: The pericardial tissue is commonly used to produce bio-prosthetic cardiac valves and patches in cardiac surgery. The procedures adopted to prepare this tissue consist in treatment with aldehydes, which do not prevent post-graft tissue calcification due to incomplete xeno-antigens removal. The adoption of fixative-free decellularization protocols has been therefore suggested to overcome this limitation. Although promising, the decellularized pericardium has not yet used in clinics, due to the absence of proofs indicating that the decellularization and cryopreservation procedures can effectively preserve the mechanical properties and the immunologic compatibility of the tissue. PRINCIPAL FINDINGS: The aim of the present work was to validate a procedure to prepare decellularized/cryopreserved human pericardium which may be implemented into cardiovascular homograft tissue Banks. The method employed to decellularize the tissue completely removed the cells without affecting ECM structure; furthermore, uniaxial tensile loading tests revealed an equivalent resistance of the decellularized tissue to strain, before and after the cryopreservation, in comparison with the fresh tissue. Finally, immunological compatibility, showed a minimized host immune cells invasion and low levels of systemic inflammation, as assessed by tissue transplantation into immune-competent mice. CONCLUSIONS: Our results indicate, for the first time, that fixative-free decellularized pericardium from cadaveric tissue donors can be banked according to Tissue Repository-approved procedures without compromising its mechanical properties and immunological tolerance. This tissue can be therefore treated as a safe homograft for cardiac surgery.

  12. Neural differentiation of novel multipotent progenitor cells from cryopreserved human umbilical cord blood

    International Nuclear Information System (INIS)

    Lee, Myoung Woo; Moon, Young Joon; Yang, Mal Sook; Kim, Sun Kyung; Jang, In Keun; Eom, Young-woo; Park, Joon Seong; Kim, Hugh C.; Song, Kye Yong; Park, Soon Cheol; Lim, Hwan Sub; Kim, Young Jin

    2007-01-01

    Umbilical cord blood (UCB) is a rich source of hematopoietic stem cells, with practical and ethical advantages. To date, the presence of other stem cells in UCB remains to be established. We investigated whether other stem cells are present in cryopreserved UCB. Seeded mononuclear cells formed adherent colonized cells in optimized culture conditions. Over a 4- to 6-week culture period, colonized cells gradually developed into adherent mono-layer cells, which exhibited homogeneous fibroblast-like morphology and immunophenotypes, and were highly proliferative. Isolated cells were designated 'multipotent progenitor cells (MPCs)'. Under appropriate conditions for 2 weeks, MPCs differentiated into neural tissue-specific cell types, including neuron, astrocyte, and oligodendrocyte. Differentiated cells presented their respective markers, specifically, NF-L and NSE for neurons, GFAP for astrocytes, and myelin/oligodendrocyte for oligodendrocytes. In this study, we successfully isolated MPCs from cryopreserved UCB, which differentiated into the neural tissue-specific cell types. These findings suggest that cryopreserved human UCB is a useful alternative source of neural progenitor cells, such as MPCs, for experimental and therapeutic applications

  13. Ovarian tissue cryopreservation in girls undergoing haematopoietic stem cell transplant: experience of a single centre.

    Science.gov (United States)

    Biasin, E; Salvagno, F; Berger, M; Nesi, F; Quarello, P; Vassallo, E; Evangelista, F; Marchino, G L; Revelli, A; Benedetto, C; Fagioli, F

    2015-09-01

    Fertility after childhood haemopoietic stem cell transplant (HSCT) is a major concern. Conditioning regimens before HSCT present a high risk (>80%) of ovarian failure. Since 2000, we have proposed cryopreservation of ovarian tissue to female patients undergoing HSCT at our centre, to preserve future fertility. After clinical and haematological evaluation, the patients underwent ovarian tissue collection by laparoscopy. The tissue was analysed by histologic examination to detect any tumour contamination and then frozen following the slow freezing procedure and cryopreserved in liquid nitrogen. From August 2000 to September 2013, 47 patients planned to receive HSCT, underwent ovarian tissue cryopreservation. The median age at diagnosis was 11.1 years and at the time of procedure it was 13 years, respectively. Twenty-four patients were not pubertal at the time of storage, whereas 23 patients had already experienced menarche. The median time between laparoscopy and HSCT was 25 days. Twenty-six out of 28 evaluable patients (93%) developed hypergonadotropic hypogonadism at a median time of 23.3 months after HSCT. One patient required autologous orthotopic transplantation that resulted in one live birth. Results show a very high rate of iatrogenic hypergonadotropic hypogonadism, highlighting the need for fertility preservation in these patients.

  14. Cryopreservation of adenovirus-transfected dendritic cells (DCs) for clinical use.

    Science.gov (United States)

    Gülen, D; Maas, S; Julius, H; Warkentin, P; Britton, H; Younos, I; Senesac, J; Pirruccello, Samuel M; Talmadge, J E

    2012-05-01

    In this study, we examined the effects of cryoprotectant, freezing and thawing, and adenovirus (Adv) transduction on the viability, transgene expression, phenotype, and function of human dendritic cells (DCs). DCs were differentiated from cultured peripheral blood (PB) monocytes following Elutra isolation using granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 6 days and then transduced using an Adv vector with an IL-12 transgene. Fresh, cryopreserved, and thawed transduced immature DCs were examined for their: 1) cellular concentration and viability; 2) antigenicity using an allogeneic mixed lymphocyte reaction (MLR); 3) phenotype (HLA-DR and CD11c) and activation (CD83); and 4) transgene expression based on IL-12 secretion. Stability studies revealed that transduced DCs could be held in cryoprotectant for as long as 75 min at 2-8°C prior to freezing with little effect on their viability and cellularity. Further, cryopreservation, storage, and thawing reduced the viability of the transduced DCs by an average of 7.7%; and had no significant impact on DC phenotype and activation. In summary, cryopreservation, storage, and thawing had no significant effect on DC viability, function, and transgene expression by Adv-transduced DCs. Copyright © 2012 Elsevier B.V. All rights reserved.

  15. Endocrine disruption screening by protein and gene expression of vitellogenin in freshly isolated and cryopreserved rainbow trout hepatocytes.

    Science.gov (United States)

    Markell, Lauren K; Mingoia, Robert T; Peterson, Heather M; Yao, Jianhong; Waters, Stephanie M; Finn, James P; Nabb, Diane L; Han, Xing

    2014-08-18

    Xenobiotics may activate the estrogen receptor, resulting in alteration of normal endocrine functions in animals and humans. Consequently, this necessitates development of assay end points capable of identifying estrogenic xenobiotics. In the present study, we screened the potential estrogenicity of chemicals via their ability to induce vitellogenin (VTG) expression in cultured primary hepatocytes from male trout. A routine method for VTG detection measures the secretion of the protein by enzyme-linked immunosorbent assay (ELISA) in freshly isolated trout hepatocytes. However, this lengthy (6 days) culturing procedure requires that hepatocyte isolation is performed each time the assay is run. We optimized this methodology by investigating the utility of cryopreserved hepatocytes, shortening the incubation time, performing a quantitative real-time PCR (qPCR) method for VTG quantification, and verifying the model system with reference chemicals 17β-estradiol, estrone, diethylstilbestrol, hexestrol, genistein, and a negative control, corticosterone. To test the performance of both freshly isolated and cryopreserved hepatocytes, mRNA was collected from hepatocytes following 24 h treatment for VTG gene expression analysis, whereas cell culture media was collected for a VTG ELISA 96 h post-treatment. EC50 values were obtained for each reference chemical except for corticosterone, which exhibited no induction of VTG gene or protein level. Our results show linear concordance between ELISA and qPCR detection methods. Although there was approximately 50% reduction in VTG inducibility following cryopreservation, linear concordance of EC50 values was found between freshly isolated and cryopreserved hepatocytes, indicating that cryopreservation does not alter the functional assessment of estrogen receptor activation and therefore VTG expression. These studies demonstrate that qPCR is a sensitive and specific method for detecting VTG gene expression that can be used together

  16. Meyer, H.-D., St. John, E.P., Chankseliani, M. & Uribe, L. (Eds.. (2013. Fairness in Access to Higher Education in a Global Perspective: Reconciling Excellence, Efficiency, and Justice. Rotterdam: Sense Publishers.

    Directory of Open Access Journals (Sweden)

    Brewis Elisa

    2017-12-01

    Full Text Available Fairness in Access to Higher Education in a Global Perspective: Reconciling Excellence, Efficiency, and Justice is an edited volume that aims to address the central question of how and why we can promote policies for fair access to higher education (HE. It takes a fairly common view of equity, exploring fair access in terms of racial, socio-economic and rural/urban background. The chapters on China and Georgia also consider fairness in the context of political favouritism and nepotism. The book’s primary focus is on access to HE, in other words, academic preparation, selection of students and affordability of HE. To a lesser extent, the book also explores the question of how to sustain participation in and completion of HE among disadvantaged groups.

  17. Bactericidal active ingredient in cryopreserved plasma-treated water with the reduced-pH method for plasma disinfection

    Science.gov (United States)

    Kitano, Katsuhisa; Ikawa, Satoshi; Nakashima, Yoichi; Tani, Atsushi; Yokoyama, Takashi; Ohshima, Tomoko

    2016-09-01

    For the plasma disinfection of human body, plasma sterilization in liquid is crucial. We found that the plasma-treated water (PTW) has strong bactericidal activity under low pH condition. Physicochemical properties of PTW is discussed based on chemical kinetics. Lower temperature brings longer half-life and the bactericidal activity of PTW can be kept by cryopreservation. High performance PTW, corresponding to the disinfection power of 22 log reduction (B. subtilis spore), can be obtained by special plasma system equipped with cooling device. This is equivalent to 65% H2O2, 14% sodium hypochlorite and 0.33% peracetic acid, which are deadly poison for human. But, it is deactivated soon at higher temperature (4 sec. at body temperature), and toxicity to human body seems low. For dental application, PTW was effective on infected models of human extracted tooth. Although PTW has many chemical components, respective chemical components in PTW were isolated by ion chromatography. In addition to peaks of H2O2, NO2- and NO3-, a specific peak was detected. and only this fraction had bactericidal activity. Purified active ingredient of PTW is the precursor of HOO, and further details will be discussed in the presentation. MEXT (15H03583, 23340176, 25108505). NCCE (23-A-15).

  18. Equine ovarian tissue viability after cryopreservation and in vitro culture

    Science.gov (United States)

    The efficiency of several cryoprotective agents were compared using both slow-freezing and vitrification methods. Results indicate that the viability of ovarian tissue cells increases when DMSO (slow-freezing) and ethylene glycol (vitrification) are used....

  19. Higher Education

    African Journals Online (AJOL)

    Kunle Amuwo: Higher Education Transformation: A Paradigm Shilt in South Africa? ... ty of such skills, especially at the middle management levels within the higher ... istics and virtues of differentiation and diversity. .... may be forced to close shop for lack of capacity to attract ..... necessarily lead to racial and gender equity,.

  20. Knowledge, attitudes, and intentions toward fertility awareness and oocyte cryopreservation among obstetrics and gynecology resident physicians.

    Science.gov (United States)

    Yu, L; Peterson, B; Inhorn, M C; Boehm, J K; Patrizio, P

    2016-02-01

    What knowledge, attitudes and intentions do US obstetrics and gynecology (OB/GYN) residents have toward discussing age-related fertility decline and oocyte cryopreservation with their patients? Most OB/GYN residents believe that age-related fertility decline, but not oocyte cryopreservation, should be discussed during well-woman annual exams; furthermore, nearly half of residents overestimated the age at which female fertility markedly declines. Oocyte cryopreservation can be utilized to preserve fertility potential. Currently, no studies of US OB/GYN residents exist that question their knowledge, attitudes, and intentions toward discussing age-related fertility decline and oocyte cryopreservation with patients. A cross-sectional online survey was conducted during the fall of 2014 among residents in American Council for Graduate (ACOG) Medical Education-approved OB/GYN residency programs. Program directors were emailed via the ACOG Council on Resident Education in Obstetrics and Gynecology server listing and asked to solicit resident participation. Participants included 238 residents evenly distributed between post-graduate years 1-4 with varied post-residency plans; 90% of residents were women and 75% were 26-30 years old. The survey was divided into three sections: demographics, fertility awareness, and attitudes toward discussing fertility preservation options with patients. Descriptive and inferential statistics were conducted. A strong majority of residents (83%) believed an OB/GYN should initiate discussions about age-related fertility decline with patients (mean patient age 31.8), and 73% percent believed these discussions should be part of an annual exam. One third of residents overestimated the age at which there is a slight decline in female fertility, while nearly half of residents overestimated the age at which female fertility markedly declines. Over three-quarters of residents (78.4%) also overestimated the likelihood of success using assisted

  1. Production of F1 interspecies hybrid offspring with cryopreserved sperm from a live-bearing fish, the swordtail Xiphophorus helleri.

    Science.gov (United States)

    Yang, Huiping; Hazlewood, Leona; Heater, Sheila J; Guerrero, Paula A; Walter, Ronald B; Tiersch, Terrence R

    2007-03-01

    Despite study of sperm cryopreservation in more than 200 fish species, production of broods from cryopreserved sperm in live-bearing fish has not been demonstrated. This has not been due to a lack of effort, but instead is a result of the unique morphology, biology, and biochemistry of reproduction in viviparous fishes. For example, sperm of Xiphophorus helleri have a cylindrical nucleus, can swim for days after being activated, have glycolytic capabilities, and can reside in the female reproduction tract for months before fertilization. These traits are not found in fishes with external fertilization. The long-standing research use of the genus Xiphophorus has led to development of over 60 pedigreed lines among the 26 species maintained around the world. These species and lines serve as contemporary models in medical research, although they must be maintained as live populations. Previous attempts at establishing sperm cryopreservation protocols for Xiphophorus have not produced live young. To address this we have been studying the parameters surrounding cryobiology of Xiphophorus sperm and applying this information to an improved understanding of internal fertilization and reproduction. Here we report the first successful fertilization and offspring production by cryopreserved sperm in any live-bearing fish. This claim is supported by our use of artificial insemination between two species that yield distinct hybrid offspring to verify paternity via cryopreserved sperm. We provide a practical approach for preservation of valuable genetic resources from live-bearing fish species, a group that is rapidly being lost due to destruction of native habitats.

  2. Development of cryopreservation for Loxocarya cinerea---an endemic Australian plant species important for post-mining restoration.

    Science.gov (United States)

    Kaczmarczyk, Anja; Funnekotter, Bryn; Turner, Shane R; Bunn, Eric; Bryant, Gary; Hunt, Taavi E; Mancera, Ricardo L

    2013-01-01

    We report the development of a cryopreservation protocol for the endemic Western Australian plant species Loxocarya cinerea (Restionaceae). Shoot tips from two genotypes, SXH404 and SXH804, were cryopreserved using the droplet-vitrification technique. Control explants, which were cryoprotected, but not cooled, showed regeneration for both genotypes (SXH404, 22.1 +/- 5.9%; SXH804, 67.7 +/- 9.6%). Extension of incubation in PVS2 from 30 to 60 min did not lead to survival after cryopreservation. Thermal analysis using differential scanning calorimetry confirmed the beneficial effect of a loading phase but also revealed no or very little ice formation after cryoprotection of shoot tips in other treatments. Regeneration following cryopreservation was obtained for genotype SXH804 (4.3 +/- 2.1%) but not for SXH404. Regenerated explants of L. cinerea SXH804 were morphologically identical to tissue-cultured plants. As an alternative to shoot tips, callus tissues of clone SXH404 were successfully cryopreserved (> 66.7% post LN survival) using the same protocol.

  3. Second-harmonic generation microscopy used to evaluate the effect of the dimethyl sulfoxide in the cryopreservation process in collagen fibers of differentiated chondrocytes

    Science.gov (United States)

    Andreoli-Risso, M. F.; Duarte, A. S. S.; Ribeiro, T. B.; Bordeaux-Rego, P.; Luzo, A.; Baratti, M. O.; Adur, J.; de Thomaz, A. A.; Pelegati, V. B.; Carvalho, H. F.; Cesar, C. L.; Kharmadayan, P.; Costa, F. F.; Olalla-Saad, S. T.

    2012-03-01

    Cartilaginous lesions are a significant public health problem and the use of adult stem cells represents a promising therapy for this condition. Cryopreservation confers many advantages for practitioners engaged in cell-based therapies. However, conventional slow freezing has always been associated with damage and mortality due to intracellular ice formation, cryoprotectant toxicity, and dehydration. The aim of this work is to observe the effect of the usual Dimethyl Sulfoxide (DMSO) cryopreservation process on the architecture of the collagen fiber network of chondrogenic cells from mesenchymal stem cells by Second Harmonic Generation (SHG) microscopy. To perform this study we used Mesenchymal Stem Cells (MSC) derived from adipose tissue which presents the capacity to differentiate into other lineages such as osteogenic, adipogenic and chondrogenic lineages. Mesenchymal stem cells obtained after liposuction were isolated digested by collagenase type I and characterization was carried out by differentiation of mesodermic lineages, and flow cytometry using specific markers. The isolated MSCs were cryopreserved by the DMSO technique and the chondrogenic differentiation was carried out using the micromass technique. We then compared the cryopreserved vs non-cryopreserved collagen fibers which are naturally formed during the differentiation process. We observed that noncryopreserved MSCs presented a directional trend in the collagen fibers formed which was absent in the cryopreserved MSCs. We confirmed this trend quantitatively by the aspect ratio obtained by Fast Fourier Transform which was 0.76 for cryopreserved and 0.52 for non-cryopreserved MSCs, a statistical significant difference.

  4. Evaluation of Glycerol Removal Techniques, Cryoprotectants, and Insemination Methods for Cryopreserving Rooster Sperm with Implications for Breed and/or Line Regeneration

    Science.gov (United States)

    A series of experiments was designed to evaluate the quality of cryopreserved rooster sperm and its fertility so that programs needing to bank germplasm and recreate animals can do so utilizing a minimal amount of cryopreserved semen. In Experiment 1, semen from roosters was collected and cryoprese...

  5. Ovarian function after removal of an entire ovary for cryopreservation of pieces of cortex prior to gonadotoxic treatment: a follow-up study

    DEFF Research Database (Denmark)

    Rosendahl, M.; Andersen, Claus Yding; Ernst, E.

    2008-01-01

    menstruation was shown to be a good indicator of ovarian function. The cryopreservation procedure rarely complicated cancer treatment (5%) and 84% felt comforted because they had potentially secured their fertility. CONCLUSIONS: Cryopreservation of ovarian tissue should be considered in young female patients...

  6. Cell banking for regulatory T cell-based therapy: strategies to overcome the impact of cryopreservation on the Treg viability and phenotype.

    Science.gov (United States)

    Gołąb, Karolina; Grose, Randall; Placencia, Veronica; Wickrema, Amittha; Solomina, Julia; Tibudan, Martin; Konsur, Evelyn; Ciepły, Kamil; Marek-Trzonkowska, Natalia; Trzonkowski, Piotr; Millis, J Michael; Fung, John; Witkowski, Piotr

    2018-02-09

    The first clinical trials with adoptive Treg therapy have shown safety and potential efficacy. Feasibility of such therapy could be improved if cells are cryopreserved and stored until optimal timing for infusion. Herein, we report the evaluation of two cell-banking strategies for Treg therapy: 1) cryopreservation of CD4 + cells for subsequent Treg isolation/expansion and 2) cryopreservation of ex-vivo expanded Tregs (CD4 + CD25 hi CD127 lo/- cells). First, we checked how cryopreservation affects cell viability and Treg markers expression. Then, we performed Treg isolation/expansion with the final products release testing. We observed substantial decrease in cell number recovery after thawing and overnight culture. This observation might be explained by the high percentage of necrotic and apoptotic cells found just after thawing. Furthermore, we noticed fluctuations in percentage of CD4 + CD25 hi CD127 - and CD4 + FoxP3 + cells obtained from cryopreserved CD4 + as well as Treg cells. However, after re-stimulation Tregs expanded well, presented a stable phenotype and fulfilled the release criteria at the end of expansions. Cryopreservation of CD4 + cells for subsequent Treg isolation/expansion and cryopreservation of expanded Tregs with re-stimulation and expansion after thawing, are promising solutions to overcome detrimental effects of cryopreservation. Both of these cell-banking strategies for Treg therapy can be applied when designing new clinical trials.

  7. Establishment and cryopreservation of a skin fibroblast cell line derived from Yunnan semi-fine wool sheep in the presence of synthetic ice blocker.

    Science.gov (United States)

    Wu, Shuai Shuai; Li, Dong Jiang; Lv, Chun Rong; Yang, Hong Yuan; Zhu, Lan; Li, Wei Juan; Quan, Guo Bo; Hong, Qiong Hua

    2013-01-01

    In this study, the fibroblasts cell line derived from ear marginal tissue of Yunnan semi-fine wool sheep was successfully established using the primary explants technique and cryopreservation technology. Additionally, the protective effect of synthetic ice blocker (SIB) including 1, 3-cyclohexanediol (1, 3-CHD) and 1, 4-cyclohexanediol (1, 4-CHD) on frozen fibroblast cells was also assessed and compared. Propidium iodide (PI) was used to stain the dead cells following cryopreservation and thawing. The results showed that compared with Medium 199 (M199) and Dulbecco's modified Eagle's medium : Nutrient Mixture F-12 (1 : 1) Mixture (DMEM/F12), Dulbecco's modified Eagle's medium (DMEM) may be more suitable for the primary culture of fibroblast cells of Yunnan semi-fine wool sheep. The growth curve of cells is a typical "S" type. After subculture for four days, the cells entered the plateau phase and began to degenerate. Biological analysis showed that the population doubling time (PDT) for subculturing fibroblast cells was approximately 26h. The Karyotyping data indicated that the percentage of fibroblast cells with normal chromosome number 2n = 54 was over 90% following subculture for 10 passages. Moreover, the tests for bacteria, fungi, viruses and mycoplasma were negative. After serial subculture for 5 generations, the fibroblast cells were cryopreserved in the presence or absence of 1, 3-CHD or 1, 4-CHD. The data indicated that with increase of the synthetic ice blocker concentrations, the viability of frozen-thawed fibroblast cells was firstly increased and then decreased. When the concentration of 1, 3-CHD or 1, 4-CHD was 50 mM, the viable percentage of frozen-thawed fibroblast cells was 91.93% +/- 2.24% and 94.13% +/- 0.55% respectively and significantly higher than that of the cells frozen in the absence of synthetic ice blockers (88.10% +/- 1.49%, P skin fibroblast cell line of Yunnan semi-fine wool sheep was firstly established in this study. Additionally

  8. Higher Education

    Science.gov (United States)

    & Development (LDRD) National Security Education Center (NSEC) Office of Science Programs Richard P Databases National Security Education Center (NSEC) Center for Nonlinear Studies Engineering Institute Scholarships STEM Education Programs Teachers (K-12) Students (K-12) Higher Education Regional Education

  9. Cryopreservation, semen use and the likelihood of fatherhood in male Hodgkin lymphoma survivors: an EORTC-GELA Lymphoma Group cohort study.

    Science.gov (United States)

    van der Kaaij, M A E; van Echten-Arends, J; Heutte, N; Meijnders, P; Abeilard-Lemoisson, E; Spina, M; Moser, E C; Allgeier, A; Meulemans, B; Lugtenburg, P J; Aleman, B M P; Noordijk, E M; Fermé, C; Thomas, J; Stamatoullas, A; Fruchart, C; Eghbali, H; Brice, P; Smit, W G J M; Sebban, C; Doorduijn, J K; Roesink, J M; Gaillard, I; Coiffier, B; Lybeert, M L M; Casasnovas, O; André, M; Raemaekers, J M M; Henry-Amar, M; Kluin-Nelemans, J C

    2014-03-01

    How does the successful cryopreservation of semen affect the odds of post-treatment fatherhood among Hodgkin lymphoma (HL) survivors? Among 334 survivors who wanted to have children, the availability of cryopreserved semen doubled the odds of post-treatment fatherhood. Cryopreservation of semen is the easiest, safest and most accessible way to safeguard fertility in male patients facing cancer treatment. Little is known about what proportion of patients achieve successful semen cryopreservation. To our knowledge, neither the factors which influence the occurrence of semen cryopreservation nor the rates of fatherhood after semen has been cryopreserved have been analysed before. This is a cohort study with nested case-control analyses of consecutive Hodgkin survivors treated between 1974 and 2004 in multi-centre randomized controlled trials. A written questionnaire was developed and sent to 1849 male survivors. Nine hundred and two survivors provided analysable answers. The median age at treatment was 31 years. The median follow-up after cryopreservation was 13 years (range 5-36). Three hundred and sixty-three out of 902 men (40%) cryopreserved semen before the start of potentially gonadotoxic treatment. The likelihood of semen cryopreservation was influenced by age, treatment period, disease stage, treatment modality and education level. Seventy eight of 363 men (21%) used their cryopreserved semen. Men treated between 1994 and 2004 had significantly lower odds of cryopreserved semen use compared with those treated earlier, whereas alkylating or second-line (chemo)therapy significantly increased the odds of use; no other influencing factors were identified. We found an adjusted odds ratio of 2.03 (95% confidence interval 1.11-3.73, P = 0.02) for post-treatment fatherhood if semen cryopreservation was performed. Forty-eight out of 258 men (19%) who had children after HL treatment became a father using cryopreserved semen. Data came from questionnaires and so this

  10. [Ultrastructural changes in the MP3 neuron of the mollusk Lymnaea stagnalis after cryopreservation of the isolated brain].

    Science.gov (United States)

    Dmitrieva, E V; Moshkov, D A; Gakhova, E N

    2006-01-01

    Investigation of a possibility of long-term storage of frozen (-196 degrees C) viable neurons and nervous tissue is one of the central present day problems. In this study ultrastructural changes in neurons of frozen-thawed snail brain were examined as a function of time. We studied the influence of cryopreservation, cryoprotectant (Me2SO), cooling to 4-6 degrees C, and a prolonged incubation in physiological solution at 4-6 degrees C on dictyosomes of Golgi apparatus, endoplasmic reticulum (ER) cisternae and mitochondria. It has been found that responses of these intracellular structures of cryopreserved neurons to the above influences are similar: dissociation of Golgi dictyosomes, swelling of endoplasmic reticulum cisternae and mitochondrial cristae. Both freezing-thawing and cryoprotectant were seen to cause an increase in the number of lysosomes, liposomes, myelin-like structures, and to form large vacuoles. The structural changes in molluscan neurons caused by cryopreservation with Me2SO (2 M) were reversible.

  11. Higher Education.

    Science.gov (United States)

    Hendrickson, Robert M.

    This chapter reports 1982 cases involving aspects of higher education. Interesting cases noted dealt with the federal government's authority to regulate state employees' retirement and raised the questions of whether Title IX covers employment, whether financial aid makes a college a program under Title IX, and whether sex segregated mortality…

  12. Effect of cryopreservation on the pre-hatching behavior in the Mexican fruit fly Anastrepha ludens Loew (Diptera, Tephritidae).

    Science.gov (United States)

    Rajamohan, Arun; Rinehart, Joseph P; Leopold, Roger A

    2018-02-01

    In a sampling of untreated embryos of the economically important fruit pest species, Anastrepha ludens, the cumulative hatch percentage in the lab was noted to be ∼85%. Approximately 70% of the larvae had eclosed through the posterior pole of the egg. This process is effected by the act of Pole Reversal (PR) of the fully developed pre-hatch larva from the wider anterior to the narrower posterior pole of the egg. Investigation of the effects of cryopreservation and various pretreatments prior to cryostorage on the PR behavior was prompted by the observation of significantly lower proportion of cryopreserved embryos exhibiting the PR behavior. Pretreatments (dechorionation and permeabilization) followed by vitrification resulted in delayed hatching, reflecting a slower embryonic development rate of ∼10 h. A smaller proportion of the treated embryos either eclosed from the anterior end of the egg or did not eclose at all despite complete development and prehatch gnawing activity. In the untreated controls, 24.0% of the embryos eclosed from the anterior pole. After permeabilization and cryopreservation, 83% and 55% (adjusted hatch) of the embryos were noted to hatch this way, respectively. An analysis of the hatch count after the treatments shows that factors contributing to the embryos' inability to properly invert polarity is not solely due to cryopreservation but also due to the pretreatment procedures including dechorionation and permeabilization. In fact, the permeabilization pre-treatment contributed the highest to this phenomenon lending support to the view that chemical toxicity rather than physical effects of cryopreservation play a major role in post-cryopreservation effects. Published by Elsevier Inc.

  13. Development of soya milk extender for semen cryopreservation of Karan Fries (crossbreed cattle).

    Science.gov (United States)

    Singh, V K; Singh, A K; Kumar, R; Atreja, S K

    2013-01-01

    Egg yolk based semen extenders are used widely, with the potential risk of xenobiotic contamination. This study was designed to develop a soya milk based extender to substitute egg yolk based extender for bovine semen cryopreservation. In the first experiment soya milk was prepared from fresh soya bean (Glycine max). Concentration of soya milk in tris based extender was standardized based on quality parameters of spermatozoa during liquid preservation at 5°C up to 72 h and compared with egg yolk tris (EYT) extender. Sperm in soya milk tris (SMT) extender with 25 percent soya milk showed no significant (P > 0.05) differences in all the quality parameters like motility, viability, membrane integrity and acrosome integrity, as compared to sperm in EYT extender up to 72h in liquid dilution. In the second experiment the Karan Fries semen was cryopreserved in SMT extender with 25 percent soya milk (selected from the first experiment) using different concentration of glycerol, as cryoprotectant, ranging from 6-7 percent with a difference of 0.2 percent to standardize optimum concentration based on post thaw motility of spermatozoa. Glycerol at a final concentration of 6.4 percent was found to be the best among all. Further, semen samples were split and cryopreserved in newly developed SMT extender containing 6.4 percent glycerol and compared with conventional EYT extender for post thaw sperm quality parameters and degree of cryocapacitation. There were no significant (P > 0.05) differences between sperm in EYT extender and SMT extender for post thaw motility, viability, membrane integrity, acrosome integrity and cryocapacitation. In conclusion, the newly developed SMT extender maintained comparable semen quality as compared to EYT extender hence it can.

  14. Antioxidant activity of aqueous extract of noni in dilutent for ram semen cryopreservation

    Directory of Open Access Journals (Sweden)

    Ana Lauren Costa Nascimento

    2016-03-01

    Full Text Available Noni (Morinda citrifolia L. is a fruit consumed worldwide because of its nutritional and therapeutic properties resulting from the large amount of phenolic compounds, which has aroused interest of the scientific community. In order to identify new natural sources of antioxidants, the objective of this study was to evaluate the performance of noni in diluent for ram semen cryopreservation. A completely randomized design consisting of four treatments and three repetitions per treatment was used. The treatments differed in terms of the concentration of the aqueous extract of noni added to the diluent: control, no addition of the extract, and three concentrations (24, 72, and 120 µg/mL. The physical and chemical variables of the mature fruit were evaluated: total acidity (8.78, pH (4.12, and soluble solids (8.18%. The vitamin C content was 309.42 mg per 100 g fresh matter. The aqueous extract of noni was also evaluated regarding the quantity of total phenolic compounds, antioxidant activity, and lipid peroxidation inhibition capacity. The aqueous extract contained a moderate amount of phenolic compounds (47.96 ± 1.95 mg gallic acid equivalent/100 g extract. The concentrations of the aqueous extract of 72 and 120 µg/mL in diluent used for semen cryopreservation inhibited lipid peroxidation by 21.75% and 51.32%, respectively. There was no positive effect of the lowest concentration (24 µg/mL. The antioxidant activity index of noni was 33.33, corresponding to very strong antioxidant activity. The aqueous extract of noni exhibits very strong antioxidant activity and its addition to the diluent for semen cryopreservation at a concentration of 72 µg/mL is able to inhibit lipid peroxidation.

  15. Prevent the degradation of algicidal ability in Scenedesmus-lysing bacteria using optimized cryopreservation.

    Science.gov (United States)

    Liao, Chunli; Liu, Xiaobo

    2016-03-01

    With the anthropogenic nutrient loading increasing, the frequency and impacts of harmful algal blooms (HABs) have intensified in recent years. To biocontrol HABs, many corresponding algal-lysing bacteria have been exploited successively. However, there are few studies on an effective algal-lysing culture collection to prevent cells from death and particularly the degradation of algicidal ability to their hosts. An optimized cryopreservation was developed and experiments on the validation of this method on preventing algicidal degradation and effects of this optimized cryopreservation on the survival rate of Scenedesmus-lysing bacterium, Enterobacter NP23, isolated from Scenedesmus sp. community, China, on the algicidal dynamic of Scenedesmus wuhanensis was investigated. The optimized cryoprotectant composition consists of 30.0 g/L gelatin, 48.5 g/L sucrose, and 28.4 g/L glycerol, respectively. Using this approach, the survival rate of NP23 cells can still maintain above 90 % and the algal-lysing rate only decline 4 % after the 18-month cryoprotection. Moreover, the 16 generations' passage experiment showed a significant (p < 0.05) genetic stability of algicidal capacity after 18 months. The growth dynamic of S. wuhanensis was investigated in a 5-L bioreactor during 132 h in the absence or presence of NP23. As a result, NP23 has a significant (p < 0.05) inhibition to S. wuhanensis growth when injected into algal culture in the exponential phase at 60th hour. In addition, S. wuhanensis culture initially with NP23 exhibited a slow growth, performing a prolonged lag phase without a clear stationary phase and then rapidly decreased. Our findings, combined with the capacity of preventing the degradation of algicidal ability collectively suggest that the use of this opitimized cryopreservation may be a promising strategy for maintaining algicidal cells.

  16. Optimization of a protocol for cryopreservation of mouse spermatozoa using cryotubes.

    Science.gov (United States)

    Hasegawa, Ayumi; Yonezawa, Kazuya; Ohta, Akihiko; Mochida, Keiji; Ogura, Atsuo

    2012-01-01

    The rapid increase in the number of genetically modified mouse strains has produced a high demand for their frozen spermatozoa from laboratories and mouse banking facilities. Historically, plastic straws have been used preferentially as containers for frozen mammalian spermatozoa because spermatozoa frozen in plastic straws have a high survival rate after thawing. However, plastic straws are more fragile and are used less often than the cryotubes used for conventional cell freezing. In this study, we sought to develop a new protocol for sperm freezing using cryotubes as the container to increase the accessibility of mouse sperm cryopreservation. Epididymal spermatozoa were collected from mature ICR or C57BL/6J (B6) males and were suspended in 18% raffinose and 3% skim milk solution. We then optimized the following conditions using the sperm survival rate as an index: 1) distance of cryotubes from the surface of the liquid nitrogen at freezing, 2) volume of the sperm suspension in the cryotube and 3) temperature of warming sperm during thawing. The best result was obtained when cryotubes containing 10 µl of sperm suspension were immersed 1 cm below the surface of the liquid nitrogen and then thawed at 50 C. The fertilization rates using spermatozoa frozen and thawed using this method were 63.1% in ICR mice and 28.2% in B6 mice. The latter rate was increased to 62.3% by adding reduced glutathione to the fertilization medium. After embryo transfer, 68% and 62% of the fertilized oocytes developed into normal offspring in the ICR and B6 strains, respectively. These results show that cryotubes can be used for cryopreservation of mouse spermatozoa under optimized conditions. This protocol is easy and reproducible, and it may be used in laboratories that do not specialize in sperm cryopreservation.

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

    Science.gov (United States)

    Pojprasath, T; Lohachit, C; Techakumphu, M; Stout, T; Tharasanit, T

    2011-06-01

    Cryopreservation of stallion semen is often associated with poor post-thaw sperm quality. Sugars are among the important components of a freezing extender and act as non-permeating cryoprotectants. This study aimed to compare the quality of stallion sperm frozen with glucose, fructose or sorbitol-containing freezing extenders. Semen was collected from six stallions of proven fertility and cryopreserved using a freezing extender containing different types of monosaccharide sugars (glucose, fructose or sorbitol). After thawing, the semen was examined for sperm motility, viability, acrosome integrity, plasma membrane functionality and sperm longevity. The fertility of semen frozen in the presence of sorbitol was also tested by artificial insemination. Sperm quality was significantly decreased following freezing and thawing (P sorbitol and glucose (P sorbitol-based extender when examined at 2 and 4 h post-thaw, all of these parameters plus plasma membrane functionality were improved for sperm frozen in the sorbitol extender than in the glucose extender when examined 10 min post-thaw. Two of four mares (50%) inseminated with semen frozen with a sorbitol-containing freezing extender became pregnant. It is concluded that different sugars have different abilities to protect against cryoinjury during freezing and thawing of stallion sperm. This study demonstrated that an extender containing sorbitol as primary sugar can be used to successfully cryopreserve equine sperm; moreover, the quality of frozen-thawed sperm appeared to be better than when glucose or fructose was the principle sugar in the freezing extender. Copyright © 2011 Elsevier Inc. All rights reserved.

  18. Ejaculate traits and sperm cryopreservation in the endangered Baird's tapir (Tapirus bairdii).

    Science.gov (United States)

    Pukazhenthi, Budhan S; Togna, Gina Della; Padilla, Luis; Smith, Diorene; Sanchez, Carlos; Pelican, Katey; Sanjur, Oris I

    2011-01-01

    There is little information on the reproductive biology of the male Baird's tapir (Tapirus bairdii). In this study, we characterized the ejaculate traits and evaluated the efficacy of 2 cryodiluents on sperm cryosurvival. Ejaculates were assessed for volume, pH, sperm motility, forward progression, osmolality, sperm concentration, sperm morphology, and acrosomal integrity. For cryopreservation, ejaculates with >50% total sperm motility were washed, and sperm pellets were resuspended in either Botu-Crio (CryoVital, Grandau, Germany) or INRA 96 containing 2% egg yolk and 2.5% each of methyl- and dimethylformamide (INRA 96), and they were cryopreserved over liquid nitrogen vapor. Thawed samples were incubated in vitro (25 °C) and evaluated for percent total sperm motility, forward progression, and acrosomal integrity at hourly intervals for 4 hours. Spermic ejaculates were obtained from all males, and the mean seminal volume, sperm concentration per milliliter, percent sperm motility, progressive status, and percent morphologically normal cells were 20.4 ± 4.3 mL, 101.2 ± 24.0 × 10(6)/mL, 46.1% ± 5.0%, 2.9 ± 0.1, and 6.9% ± 1.4%, respectively. There was a positive significant correlation between percent normal sperm and animal age (r = 0.66; P tapir; demonstrate that tapir spermatozoa can be cryopreserved in diluents containing amides alone or in combination with glycerol; and provide fundamental information critical for development of assisted reproductive technologies for the Baird's tapir.

  19. Consequences of metaphase II oocyte cryopreservation on mRNA content.

    Science.gov (United States)

    Chamayou, S; Bonaventura, G; Alecci, C; Tibullo, D; Di Raimondo, F; Guglielmino, A; Barcellona, M L

    2011-04-01

    We studied the consequences of freezing/thawing processes on mRNA contents in MII oocytes after slow-freezing/rapid thawing (SF/RT) and vitrification/warming (V/W) protocols, and compared the results to fresh MII oocytes. We quantified the nuclear transcript mRNA responsible for the translation of proteins belonging either to trans-regulatory protein family or to functional structural proteins such as proteins involved in DNA structural organization (NAP1L1, TOP1, H1F0H1), chromosomal structure maintenance (SMC, SCC3, RAD21, SMC1A, SMC1B, STAG3, REC8), mitochondrial energetic pathways (ATP5GJ, SDHC), cell cycle regulation and processes (CLTA, MAPK6, CKS2) and staminal cell potency-development competence stage (DPPA3, OCT4, FOXJ2). Surplus MII oocytes were donated from patients in IVF cycles and divided in three groups of 15 oocytes. Group 1 was comprised of non-cryopreserved oocytes and Groups 2 and 3 underwent SF/RT and V/W procedures, respectively. There was an overall decrease of mRNA extracted from cryopreserved oocytes compared to control group. Only 39.4% of mRNA content were preserved after SF/RT while 63.3% of mRNA content were maintained after V/W. Oocyte cryopreservation is associated with molecular injury associated with the decrease of stored mRNA. However the V/W protocol is more conservative than SF/RT resulting in a level of mRNA sufficient to maintain biologic functions in the subsequent fertilized oocyte. Copyright © 2011 Elsevier Inc. All rights reserved.

  20. The Effect of Cryopreserved Human Placental Tissues on Biofilm Formation of Wound-Associated Pathogens.

    Science.gov (United States)

    Mao, Yong; Singh-Varma, Anya; Hoffman, Tyler; Dhall, Sandeep; Danilkovitch, Alla; Kohn, Joachim

    2018-01-08

    Biofilm, a community of bacteria, is tolerant to antimicrobial agents and ubiquitous in chronic wounds. In a chronic DFU (Diabetic Foot Ulcers) clinical trial, the use of a human cryopreserved viable amniotic membrane (CVAM) resulted in a high rate of wound closure and reduction of wound-related infections. Our previous study demonstrated that CVAM possesses intrinsic antimicrobial activity against a spectrum of wound-associated bacteria under planktonic culture conditions. In this study, we evaluated the effect of CVAM and cryopreserved viable umbilical tissue (CVUT) on biofilm formation of S. aureus and P. aeruginosa , the two most prominent pathogens associated with chronic wounds. Firstly, we showed that, like CVAM, CVUT released antibacterial activity against multiple bacterial pathogens and the devitalization of CVUT reduced its antibacterial activity. The biofilm formation was then measured using a high throughput method and an ex vivo porcine dermal tissue model. We demonstrate that the formation of biofilm was significantly reduced in the presence of CVAM- or CVUT-derived conditioned media compared to control assay medium. The formation of P. aeruginosa biofilm on CVAM-conditioned medium saturated porcine dermal tissues was reduced 97% compared with the biofilm formation on the control medium saturated dermal tissues. The formation of S. auerus biofilm on CVUT-conditioned medium saturated dermal tissues was reduced 72% compared with the biofilm formation on the control tissues. This study is the first to show that human cryopreserved viable placental tissues release factors that inhibit biofilm formation. Our results provide an explanation for the in vivo observation of their ability to support wound healing.

  1. The Effect of Cryopreserved Human Placental Tissues on Biofilm Formation of Wound-Associated Pathogens

    Directory of Open Access Journals (Sweden)

    Yong Mao

    2018-01-01

    Full Text Available Biofilm, a community of bacteria, is tolerant to antimicrobial agents and ubiquitous in chronic wounds. In a chronic DFU (Diabetic Foot Ulcers clinical trial, the use of a human cryopreserved viable amniotic membrane (CVAM resulted in a high rate of wound closure and reduction of wound-related infections. Our previous study demonstrated that CVAM possesses intrinsic antimicrobial activity against a spectrum of wound-associated bacteria under planktonic culture conditions. In this study, we evaluated the effect of CVAM and cryopreserved viable umbilical tissue (CVUT on biofilm formation of S. aureus and P. aeruginosa, the two most prominent pathogens associated with chronic wounds. Firstly, we showed that, like CVAM, CVUT released antibacterial activity against multiple bacterial pathogens and the devitalization of CVUT reduced its antibacterial activity. The biofilm formation was then measured using a high throughput method and an ex vivo porcine dermal tissue model. We demonstrate that the formation of biofilm was significantly reduced in the presence of CVAM- or CVUT-derived conditioned media compared to control assay medium. The formation of P. aeruginosa biofilm on CVAM-conditioned medium saturated porcine dermal tissues was reduced 97% compared with the biofilm formation on the control medium saturated dermal tissues. The formation of S. auerus biofilm on CVUT-conditioned medium saturated dermal tissues was reduced 72% compared with the biofilm formation on the control tissues. This study is the first to show that human cryopreserved viable placental tissues release factors that inhibit biofilm formation. Our results provide an explanation for the in vivo observation of their ability to support wound healing.

  2. Effect of green tea (Camellia sinensis) extract and pre-freezing equilibration time on the post-thawing quality of ram semen cryopreserved in a soybean lecithin-based extender.

    Science.gov (United States)

    Mehdipour, Mahdieh; Daghigh Kia, Hossein; Najafi, Abouzar; Vaseghi Dodaran, Hossein; García-Álvarez, Olga

    2016-12-01

    The aim of this study was to determine the effect of Camellia sinensis extract as antioxidant supplement and pre-freezing equilibration times in a soybean lecithin extender for freezing ram semen. In this study, a total of 20 ejaculates were collected from four Ghezel rams and diluted with extenders (1.5% soybean lecithin, 7% glycerol) containing no supplements (control) and Camellia sinensis extract (5, 10, and 15 mg/L) and cryopreserved, immediately after thermal equilibrium was reached at 5 °C (0 h), or 4 h after equilibration. Sperm motility characteristics, membrane integrity, abnormal morphology, mitochondria activity, apoptotic status, MDA and antioxidant activities (GPx, SOD and total antioxidant capacity (TAC)) were evaluated following freeze-thawing. Camellia sinensis extract at level 10 mg/L led to the highest total and progressive motilities percentages, in comparison to other treatments (P extract at level of 5 and 10 mg/L led to higher plasma membrane integrity, mitochondria activity and Total antioxidant capacity (TAC) in comparison to the level of 15 mg/L and control group (P extract at 10 mg/L level produced the highest percentage of live spermatozoa and the lowest apoptotic spermatozoa in comparison to all treatments (P concentration, 10 mg/L, compared to all treatments (P  0.05) were observed between equilibration times (0 h vs. 4 h) for sperm samples incubated with or without different concentrations of Camellia sinensis extract. In conclusion, addition of Camellia sinensis extract at level of 10 mg/L can improve post-thawing quality of ram semen cryopreserved in a soybean lecithin extender. However, further research is needed to standardize the process of Camellia sinensis extraction and specially for identifying which compounds are responsible of its beneficial effect on ram sperm cryopreservation. Copyright © 2016 Elsevier Inc. All rights reserved.

  3. The Effect of Breed on Cryopreservation of Cock Semen

    African Journals Online (AJOL)

    USER

    sperm motility and viability rates were significantly higher in sperm from indigenous roosters than from. Ross males. At 30 min post-thaw the sperm survival rate was lower in all breeds without significant differences between them. This observation indicates a decreased survival rate with extended storage period of semen ...

  4. A new portable device for automatic controlled-gradient cryopreservation of blood mononuclear cells

    DEFF Research Database (Denmark)

    Hviid, L; Albeck, G; Hansen, B

    1993-01-01

    Protection of the functional integrity of mononuclear cells stored in liquid N2 requires careful control of the freezing procedure. Consequently, optimal quality of cryopreserved cells is usually assured by freezing according to a specified time-temperature gradient generated by computer......-controlled freezing devices. While such equipment offers large capacity and secures maximum survival and functional integrity of the lymphocytes upon thawing, it is quite costly and strictly stationary. We have previously developed and tested an alternative, manual device for controlled-gradient lymphocyte freezing...

  5. Cryopreservation of ovarian tissue for fertility preservation in young female oncological patients

    DEFF Research Database (Denmark)

    Andersen, Claus Yding; Kristensen, Stine Gry; Greve, Tine

    2012-01-01

    Girls and women suffering from a cancer that requires treatment with gonadotoxic drugs may experience cessation of reproductive function as a side effect due to obliteration of the ovarian pool of follicles. Techniques are now available for fertility preservation, such as cryopreservation of mature...... and growth of follicles, giving rise to menstrual cycles and hormone production for several years. Worldwide, the procedure has resulted in the birth of 15 healthy children. Many cancer patients including girls and young women want fertility preservation, and the techniques are now being further developed...

  6. Cholesterol addition aids the cryopreservation of dromedary camel (Camelus dromedarius) spermatozoa.

    Science.gov (United States)

    Crichton, Elizabeth G; Pukazhenthi, Budhan S; Billah, M; Skidmore, Julian A

    2015-01-15

    The cryopreservation of dromedary camel (Camelus dromedarius) sperm has proved challenging with little success reported. The routine application of artificial insemination with frozen semen would assist the flow of valuable genetic material nationally and internationally. The current study sought to examine the effects of cholesterol (cholesterol-loaded cyclodextrin [CLC]) preloading on camel sperm cryosurvival. Ejaculates (n = 3 males; 3 ejaculates per male) were collected using an artificial vagina during the breeding season and extended in HEPES-buffered Tyrode's albumin lactate pyruvate (TALP) and allowed to liquefy in the presence of papain (0.1 mg/mL) before removal of the seminal plasma by centrifugation. Sperm pellets were resuspended (120 million/mL) in fresh TALP and incubated (15 minutes; 37 °C) with 0, 1.5, or 4.5 mg CLC/mL. Sperm suspensions were then centrifuged and reconstituted in INRA-96 containing 20% (v:v) egg yolk and 2.5% (v:v) methylformamide, loaded in 0.5-mL plastic straws, sealed, and cooled for 20 minutes at 4 °C. Straws were frozen over liquid nitrogen (4 cm above liquid; 15 minutes), plunged, and stored. Sperm motility, forward progressive status, and acrosomal integrity were recorded at 0 and 3 hours after thawing and compared with these same parameters before freezing. Aliquots also were stained with chlortetracycline hydrochloride to assess spontaneous sperm capacitation status before freezing and post-thaw. Pretreatment with CLC (1.5 and 4.5 mg/mL) enhanced cryosurvival. Post-thaw sperm motility was highest (P < 0.05) in 1.5 mg CLC/mL immediately after thawing (44%) and after 3 hours incubation at room temperature (34%). Highest post-thaw sperm progressive status was also achieved in the presence of 1.5 CLC. Greater proportions of spermatozoa retained acrosomal membrane integrity when cryopreserved in the presence of CLC, but there was no difference between 1.5 and 4.5 CLC. Although thawed spermatozoa underwent spontaneous

  7. Reconstruction of Chest Wall by Cryopreserved Sternal Allograft after Resection of Aneurysmal Bone Cyst of Sternum

    Directory of Open Access Journals (Sweden)

    Kambiz Sheikhy

    2017-01-01

    Full Text Available A 20-year-old female was referred to our hospital due to deformity and bulging in anterior aspect of chest wall in sternal area. Chest X-ray and CT scan confirmed a large mass with destruction of sternum. Pathologic diagnosis after incisional biopsy was compatible with aneurysmal bone cyst. We resected sternum completely and reconstructed large anterior defect by a cryopreserved sternal allograft. In follow-up of patient there was no unstability of chest wall with good cosmetic result.

  8. Effect of the cryopreservation method used, the embryonic stage and the use of conjugated linoleic acid isomers on the cryotolerance of in vitro-produced bovine embryos

    Directory of Open Access Journals (Sweden)

    Luciana Simões Rafagnin Marinho

    2015-12-01

    Full Text Available Conjugated linoleic acid (CLA might be able to improve the cryotolerance of in vitro-produced (IVP embryos. The effect of two CLA isomers on the cryotolerance of bovine IVP embryos, as well as that of the stage of embryonic development and the method used for cryopreservation was evaluated by three experiments. In Experiment 1, oocytes (n = 3,917 were fertilized in vitro and cultured with 0, 50, 100, or 200 ?M trans-10, cis-12 (t10, c12 CLA. In Experiment 2, fertilized oocytes (n = 2,131 were cultured with 100 ?M t10, c12 or cis-9, trans-11 (c9, t11 CLA, or a combination of both isomers. The embryos were vitrified at the blastocyst (BL or the expanded blastocyst (EB stage. In Experiment 3, oocytes (n = 1,720 were fertilized and cultured with or without 100 ?M t10, c12 CLA, and the blastocysts were vitrified or frozen. Blastocyst development rate as well as the rates of re-expansion and hatching after thawing was recorded. Moreover, the mean cell number and mRNA expression of acetyl-CoA carboxylase (ACC1 and stearoyl-CoA desaturase (SCD1 as well as fatty acid synthase (FASN multienzyme complex were determined. In Experiment 1, the highest concentration of t10, c12 CLA that did not reduce blastocyst development rate was 100 ?M. In Experiment 2, the rates of re-expansion and hatching among the EBs obtained through IVP after supplementation with t10, c12 CLA (73.1% and 57.7%, with c9, t11 CLA (80.0% and 68.6%, with the combination (78.3% and 52.2%, and with the control group (85.4% and 58.3% were similar. At the BL stage, the rates of re-expansion and hatching were lower than those at the EB stage, and CLA combination allowed a hatching rate (8.0% lower than that observed in the control group (40.0%. In Experiment 3, the hatching rates for vitrified EBs (vitrified control; 67.4% and vitrified CLA EBs (65.8% were higher than those obtained for frozen EBs, exposed (13.3% or not exposed (28.6% to CLA. In addition, in Experiment 3, the hatching rate was

  9. Cryopreservation of adult unrelated donor products in hematopoietic cell transplantation: the OneMatch experience and systematic review of the literature.

    Science.gov (United States)

    Aziz, Joseph; Morris, Gail; Rizk, Mina; Shorr, Risa; Mercer, Dena; Young, Kimberly; Allan, David

    2017-11-01

    The frequency of cryopreserving blood stem or progenitor products from unrelated donors is not known and the underlying reasons are poorly documented. Greater insight is needed to develop policies on cryopreservation that balance donor safety with patient needs. Cryopreservation requests between January 1, 2014, and May 31, 2016, at the OneMatch Stem Cell and Marrow Network at Canadian Blood Services were reviewed and a systematic review of the literature was performed. Thirty products of 719 (4.2%) unrelated donor collections facilitated by OneMatch were cryopreserved. Patient-related reasons were most common and included the need to delay transplant for continued antimicrobial treatment (six patients), patient too deconditioned to proceed with scheduled transplant (five patients), and/or need for more treatment for relapsed disease (three patients). Donor-related issues leading to cryopreservation requests were less common (five cases), mainly due to lack of donor availability after attempting to reschedule. Cryopreservation of a product that was never infused occurred infrequently (two cases, 7%). In our systematic review of the literature, 993 cases were identified in 32 published reports. Both patient-related and donor-related reasons were cited but not specifically reported, precluding quantitative insight regarding the relative frequency of causes. The impact of cryopreservation on hematopoietic engraftment appears negligible when compared to controls in a subset of studies; however, reporting of outcomes was inconsistent. Future studies with standard outcome measures are needed to clarify the impact of cryopreservation on engraftment and other transplant outcomes. International guidelines that consider the ethical framework surrounding requests for donor product cryopreservation are needed. © 2017 AABB.

  10. Elective single embryo transfer with cryopreservation improves the outcome and diminishes the costs of IVF/ICSI.

    Science.gov (United States)

    Veleva, Zdravka; Karinen, Petri; Tomás, Candido; Tapanainen, Juha S; Martikainen, Hannu

    2009-07-01

    Although elective single embryo transfer (eSET) minimizes the multiple birth rate after in vitro fertilization (IVF)/intra cytoplasmic sperm injection (ICSI), there remain concerns in many countries that it is less effective and more expensive than conventional double embryo transfer (DET). We compared the clinical outcome achieved in the years 1995-1999, in which eSET was rarely used (4.2% of women, DET period) with that of the years 2000-2004, in which eSET was more widely used (46.2%, eSET period). In the DET period, 826 women had 1359 fresh embryo cycles followed by 589 frozen-thawed embryo transfer (FET) cycles. In the eSET period, 684 women had 1027 fresh and 683 FET cycles. The cumulative term live birth rate/woman was the primary clinical outcome measure. An incremental cost-effectiveness ratio of a term live birth was also calculated based on hospital charges and medication prices of IVF/ICSI treatment. The cumulative pregnancy rate/oocytes pickup (38.2 versus 33.1%, P = 0.01), cumulative live birth rate/oocytes pickup (28.0 versus 22.5%, P = 0.002) and cumulative live birth rate/woman (41.7 versus 36.6%, P = 0.04) were all higher in the eSET period than in the DET period. The cumulative multiple birth rate was significantly lower in the eSET period than in the DET period (8.9 versus 19.6%, P < 0.0001). A term live birth in the eSET period was 19 889 euros less expensive than in the DET period. This study shows that eSET with cryopreservation is more effective and less expensive than DET and should be adopted as a treatment of choice.

  11. Analysis of the impact of cryopreservation and theophylline on motility of sperm

    Directory of Open Access Journals (Sweden)

    Elaheh Gorji

    2018-06-01

    Full Text Available Objective: Sperm parameters, particularly motility, decrease during cryopreservation. Theophylline generally enhances sperm motility. We analyzed effects of theophylline and freezing on sperm motility.Design: Experimental study.Setting: Private IVF lab.Setting: IVF lab of Mehrgan Hospital. Method: 22–55 year-old men participated in this study (30 fresh ejaculation and 8 TESE samples. After sperm analysis, we added theophylline (40 mM to half of our samples as case group to compare motility with the remaining samples as control group. Cryopreservation was performed in two groups. After thawing, motility of both groups was recorded. Furthermore, theophylline (40 mM was applied to both groups after thawing again. Result: After adding theophylline, sperm motility improved significantly in all samples. Sperm motility reduced in control group more than the study group after freeze-thaw procedure (P < 0.002, normal morphology <5%. Sperm motility was not enhanced significantly by re-adding of theophylline to the two groups. Interactions between stages and groups were statistically significant in semen and biopsy samples (p < 0.001. Conclusion: Adding theophylline before freezing can preserve motility of sperms in samples with different parameters and even sperms extracted in testicular biopsy. Theophylline may have protective impact on sperms in freezing procedure. Keywords: Sperm motility, Theophylline, Freezing, Morphology, Biopsy

  12. Initial 10-year Experience of Sperm Cryopreservation Services for Cancer Patients

    Directory of Open Access Journals (Sweden)

    Hong-Chiang Chang

    2006-01-01

    Full Text Available Offering sperm cryopreservation to preserve the fertility of male cancer patients is a relatively recent service in Asia. This study analyzed the types of cancer, timing of collection, sperm quality, and utilization for reproductive services by patients during a 10-year period at a medical center in Taiwan. A total of 75 oncology patients elected to freeze sperm for fertility preservation at our medical center during the initial 10 years of the availability of this service. The mean age of the patients was 25.7 years. Storage was discontinued in 13 (17% patients and their survival duration was 13.1 ±11.1 months. The utilization rate of sperm cryo-preservation was 2.8% (75/2642. The types of cancer varied, with leukemia (35%, lymphoma (25%, and testicular cancer (13% comprising the largest groups. A significantly lower sperm count was found in patients with chronic myelogenous leukemia, suggesting the need for earlier sperm collection after initiation of cancer treatment. Only three (4% patients utilized their specimens for reproductive purposes. There was no clinical pregnancy during the study period, although one biochemical pregnancy was achieved. The low rates of sperm cryostorage for fertility preservation in cancer patients in this study suggest that there is a need for greater emphasis of this option for male oncology patients whose fertility is likely to be affected by chemotherapeutic treatment.

  13. Biomechanical and ultrastructural comparison of cryopreservation and a novel cellular extraction of porcine aortic valve leaflets.

    Science.gov (United States)

    Courtman, D W; Pereira, C A; Omar, S; Langdon, S E; Lee, J M; Wilson, G J

    1995-12-01

    Heart valve substitutes of biological origin often fail by degenerative mechanisms. Many authors have hypothesized that mechanical fatigue and structural degradation are instrumental to in vivo failure. Since the properties of the structural matrix at implantation may predetermine failure, we have examined the ultrastructure, fracture, mechanics, and uniaxial high-strain-rate viscoelastic properties of: (1) fresh, (2) cryopreserved, and (3) cellular extracted porcine aortic valve leaflets. The cellular extraction process is being developed in order to reduce immunological attack and calcification. Cryopreservation causes cellular disruption and necrotic changes throughout the tissue, whereas extraction removes all cells and lipid membranes. Both processes leave an intact collagen and elastin structural matrix and preserve the high-strain-rate viscoelastic characteristics of the fresh leaflets. Extraction does cause a 20% reduction in the fracture tension and increases tissue extensibility, with the percent strain at fracture rising to 45.3 +/- 4 (mean +/- SEM) from 31.5 +/- 3 for fresh leaflets. However, extraction does preserve matrix structure and mechanics over the physiological loading range. Glutaraldehyde fixation produces increased extensibility, increased elastic behavior, and, when applied to extracted leaflets, it causes a marked drop in fracture tension, to 50% of that for fresh leaflets. The combination of extraction and fixation may lead to early degenerative failure. The cellular extraction technique alone may be a useful alternative to glutaraldehyde fixation in preparing bioprosthetic heart valves.

  14. EFFECT OF DIFFERENT CRYOPROTECTIVE AGENTS ON SKIM MILK AND DIMITROPOULUS EXTENDER FOR STALLION SEMEN CRYOPRESERVATION

    Directory of Open Access Journals (Sweden)

    R.I. Arifiantini

    2014-10-01

    Full Text Available s to assess different CPAs on stallion semen cryopreservation. Skim milk (SM and Dimitropoulos(DV were the extenders used in this study; each was added by glycerol (Gly, combination of ethyleneglycol-glycerol (EG+Gly or dimethilformamide (DMF. Each semen sample was evaluated and dividedequally into six tubes; semen in the three tubes was diluted 1:1 with (SM, while in the remaining tubesthe semen was diluted 1:1 by DV. After being diluted, all tubes were centrifuged at 1006xg for 10minutes. The supernatan discarded, the pellet was rediluted by SM trehalosa or DV trehalose, and addedby G, EG+Gly, or DMF to reach the final sperm concentration of 200x106/ml. The extended semen wasindividually packed in 0.3 ml minitube, equilibrated at 4oC for 2 hours, frozen in liquid nitrogen vaporfor 10 minutes, and then was stored in liquid nitrogen container at -196 oC. After 24 hours, the semenwas thawed at 37 oC for 30 second. There were no significantly different (p>0.05 on the percentages ofmotile and viable sperm in SMT (21.7% and 43.4%, respectively compared with those extended withDV T extender (26.9% and 50.8%, respectively. DMF demonstrated better results as CPA compared tothe others; and DVTDMF combination had the best protection during cryopreservation in this study.

  15. Establishment and cryopreservation of a giant panda skeletal muscle-derived cell line.

    Science.gov (United States)

    Yu, Fang-Jian; Zeng, Chang-Jun; Zhang, Yan; Wang, Cheng-Dong; Xiong, Tie-Yi; Fang, Sheng-Guo; Zhang, He-Min

    2015-06-01

    The giant panda Ailuropoda melanoleuca is an endangered species and is a symbol for wildlife conservation. Although efforts have been made to protect this rare and endangered species through breeding and conservative biology, the long-term preservation of giant panda genome resources (gametes, tissues, organs, genomic libraries, etc.) is still a practical option. In this study, the giant panda skeletal muscle-derived cell line was successfully established via primary explants culture and cryopreservation techniques. The population doubling time of giant panda skeletal cells was approximately 33.8 h, and this population maintained a high cell viability before and after cryopreservation (95.6% and 90.7%, respectively). The two skeletal muscle-specific genes SMYD1 and MYF6 were expressed and detected by RT-PCR in the giant panda skeletal muscle-derived cell line. Karyotyping analysis revealed that the frequencies of giant panda skeletal muscle cells showing a chromosome number of 2n=42 ranged from 90.6∼94.2%. Thus, the giant panda skeletal muscle-derived cell line provides a vital resource and material platform for further studies and is likely to be useful for the protection of this rare and endangered species.

  16. Random-start ovarian stimulation in women desiring elective cryopreservation of oocytes.

    Science.gov (United States)

    Pereira, Nigel; Voskuilen-Gonzalez, Anna; Hancock, Kolbe; Lekovich, Jovana P; Schattman, Glenn L; Rosenwaks, Zev

    2017-10-01

    The current study investigates the utility of random-start ovarian stimulation in women desiring elective oocyte cryopreservation. Women in the study cohort underwent random-start ovarian stimulation, and were subdivided based on the phase of the menstrual cycle that ovarian stimulation began, i.e. early follicular, late follicular or luteal phase. Women undergoing conventional cycle day (CD) 2/3 ovarian stimulation start were controls. A total of 1302 women were included - 859 (66.0%) conventional CD 2/3, 342 (26.3%) early follicular, 42 (3.2%) late follicular and 59 (4.5%) luteal ovarian stimulation starts. There was no difference in the demographics or baseline ovarian stimulation characteristics. The duration of ovarian stimulation (11 versus 9 days; P start group. The number of total and MII oocytes in the control and random-start groups was similar. A non-significant trend towards increased cycle cancellation was noted in the late follicular start group (7.1%). Study findings indicate the number of total and MII oocytes derived from random-start protocols initiated during any phase of the menstrual cycle is similar to conventional CD 2/3 ovarian stimulation start protocols in women desiring elective oocyte cryopreservation. Copyright © 2017 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  17. Effects of some cryopreservation procedures on recalcitrant zygotic embryos of Ammocharis coranica.

    Science.gov (United States)

    Nomali, Z; Ngobese; Sershen; Berjak, P; Pammenter, N W

    2014-01-01

    Cryopreservation, the most promising method for the long-term conservation of recalcitrant (desiccation-sensitive) seed germplasm, is often associated with high viability losses. Cryo-procedures involve a sequence of steps which must be optimised to reduce the impact of the stresses. This study reports on the effects of some of the steps of cryopreservation on the recalcitrant zygotic embryos of the amaryllid, Ammocharis coranica. Embryos were subjected to cryoprotection with glycerol and/or DMSO, rapid (flash) drying, and rapid (>100 degree C s(-1)) or slow (1 degree C s(-1)) cooling. Rapid dehydration (from c. 2.7 to 0.9 g g(-1) over 60 min) and cooling had a detrimental effect on the viability of the embryos, which was exacerbated when these steps were applied sequentially. After cooling, seedling production (30%) was obtained only from embryos that had been cryoprotected with glycerol prior to drying and rapid cooling, while 30% of non-treated embryos and 70% of those that had undergone cathodic protection during flash drying produced callus. Noting that no post-cryo survival of A. coranica embryos had previously been obtained, this study identified cryoprotection with glycerol and the incorporation of cathodic protection during flash drying as promising intervention points for future studies.

  18. Effect of cryopreservation on mitochondrial activity in buffalo sperm Bubalus bubalis

    Directory of Open Access Journals (Sweden)

    O. Kandil

    2010-02-01

    Full Text Available Sperm mitochondrial activity is investigated and used as “in vitro” spermatozoa vitality indicator and about quality effectiveness of different sperm diluents. It was studied the cytochemically activity of NADPH-diaphorase and LDH-C4 in cryopreserved buffalo sperm. Low intensity of the enzyme reaction was established in all examined sperm samples in both enzymes, regardless from the used cryoprotectors. The main part of the enzyme reaction was localized in mitochondrial sheath and in a very small degree in the head base of spermatozoa. No increase of the enzymes activities or the spermatozoa motility has been found after the incubating with Sp-TALP medium although the established caffeine stimulating effect on the glycolysis and fresh spermatozoa motility. Established by us low sperm motility after cryopreservation may be due to low LDH and NADPH-diaphorase activity due to glycolisis disturbances and ATP synthesis. This method allows to estimate quality of buffalo semen and to find some different disturbances in mitochondrial sheath, which could not be found by routine morphological studies and could be used in practice ejaculates with high number of metabolic active sperm cells.

  19. Value of lymphocyte cryo-preservation after a radiological or nuclear accident

    International Nuclear Information System (INIS)

    Laroche, P.; Lataillade, J.J.; Chambrette, V.; Voisin, Ph.

    1997-01-01

    The conventional cytogenetic method in biological dosimetry is most useful for the estimation of the received radiation dose. It shows resulting unstable chromosomal aberrations (dicentrics, centric rings and fragments) in peripheral blood lymphocytes. This method has been used over the past 30 years and is used in forensic medicine. Nevertheless, it is long and fastidious. Accordingly, the number of simultaneous analyses of blood samples is limited and depends on the capacity of specialized laboratories. This capacity may be insufficient in the case of large scale radiological or nuclear accidents. Cryo-preservation is the usual method to store cells before analysis or use, for instance for biological dosimetry purposes. Some investigations have shown that thawing following freezing may induce cell injury but few studies have been made on the effect of cryo-preservation on cells containing radiation-induced unstable chromosomal aberrations. In this work, lymphocytes were irradiated with 1 to 4 Gy gamma rays and stored in liquid nitrogen. The dicentric and centric ring yields were analysed after storage periods of 1 week, 1 month, 3 months, and 1 year. No difference in aberration frequency from control, unfrozen samples was observed over this period. Lymphocytes stored at -196 deg C for up to least 1 year may therefore be used for chromosome aberration scoring when overexposure to ionizing radiation is suspected. (author)

  20. B Cells and B Cell Blasts Withstand Cryopreservation While Retaining Their Functionality for Producing Antibody

    Directory of Open Access Journals (Sweden)

    Philipp Fecher

    2018-05-01

    Full Text Available In individuals who have once developed humoral immunity to an infectious/foreign antigen, the antibodies present in their body can mediate instant protection when the antigen re-enters. Such antigen-specific antibodies can be readily detected in the serum. Long term humoral immunity is, however, also critically dependent on the ability of memory B cells to engage in a secondary antibody response upon re-exposure to the antigen. Antibody molecules in the body are short lived, having a half-life of weeks, while memory B cells have a life span of decades. Therefore, the presence of serum antibodies is not always a reliable indicator of B cell memory and comprehensive monitoring of humoral immunity requires that both serum antibodies and memory B cells be assessed. The prevailing view is that resting memory B cells and B cell blasts in peripheral blood mononuclear cells (PBMC cannot be cryopreserved without losing their antibody secreting function, and regulated high throughput immune monitoring of B cell immunity is therefore confined to—and largely limited by—the need to test freshly isolated PBMC. Using optimized protocols for freezing and thawing of PBMC, and four color ImmunoSpot® analysis for the simultaneous detection of all immunoglobulin classes/subclasses we show here that both resting memory B cells and B cell blasts retain their ability to secrete antibody after thawing, and thus demonstrate the feasibility of B cell immune monitoring using cryopreserved PBMC.

  1. Use of commercial extenders and alternatives to prevent sperm agglutination for cryopreservation of brown bear semen.

    Science.gov (United States)

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

    2014-08-01

    The objective of this study was to evaluate different bovine and canine commercial semen extenders for cryopreservation of brown bear ejaculates and the effect of semen collection directly into extender on sperm agglutination. Semen samples were obtained by electroejaculation from 13 adult males. In experiment 1, eleven ejaculates from eight bears were used to evaluate Bioxcell and Andromed as extenders, whereas in experiment 2, nine ejaculates from six bears were used to evaluate Triladyl canine, CaniPro, and Extender 2 as extenders. An extender specifically developed for brown bears (Test-Tris-fructose-egg yolk-glycerol, TTF-ULE/bear) served as a control extender in both experiments. After thawing, total and progressive sperm motility and sperm viability were greater (P bear and Andromed extenders than for Bioxcell in experiment 1 and greater (P bear extender than for Triladyl Canine, CaniPro, and Extender 2 in experiment 2. In experiment 3, addition of handling extender (TTF-H) to the semen collection tube for eight ejaculates from seven bears resulted in less (P bear is the most suitable extender for brown bear semen cryopreservation, but comparable results can be obtained with the commercial extender Andromed. In addition, collection of ejaculates directly in TTF-H extender decreases sperm agglutination in fresh samples. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Optimization of conditions for long-term prefreezing storage of brown bear sperm before cryopreservation.

    Science.gov (United States)

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

    2015-10-15

    Brown bear ejaculates are usually collected in field conditions and may need to be shipped to a laboratory for the application of reproductive biotechnologies before cryopreservation. The aim of this study was to extend the prefreezing step to 48 hours (1 hour vs. long-term storage [LS] to 24 and 48 hours) to enable the sample to be transported. The effects of storage temperature (experiment 1), glycerol concentration (experiment 2), and dilution rate (experiment 3) on sperm were evaluated. Electroejaculates from brown bears were stored under different experimental conditions and cryopreserved. The sperm motility and viability, apoptotic status, and acrosomal status of sperm were assessed before freezing (prefreezing), after thawing, and after 2-hour incubation at 37 °C (thermal stress test). In all experiments, one control sample was frozen using a standard protocol (control). In experiment 1, three temperatures during LS with 6% glycerol were tested: 5 °C (T5), 15 °C (T15), and room temperature (RT). The LS-T5 sample yielded the highest postthawing results for viability (42.4%), progressive motility (15.6%), and intact acrosome (83.1%) after 24 hours in comparison with the other temperatures (P bear electroejaculates are at 5 °C, at a concentration of 100 × 10(6) sperm/mL, and with 6% glycerol. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Development of a vitrification-based cryopreservation protocol for the storage of saltcedar (Tamarix boveana Bunge).

    Science.gov (United States)

    Cano-Castillo, M; Casas, J L

    2012-01-01

    We cryopreserved in vitro shoot tips of saltcedar (Tamarix boveana Bunge) using the vitrification technique. The success of the cryopreservation protocol was strongly affected by preculture, loading duration, dehydration duration in plant vitrification solution 2 (PVS2), and medium composition during post-warming regrowth. The highest explant regrowth (50 percent) occurred when the following conditions were employed: preculture in 0.4 M glycerol; treatment with a loading solution (LS) consisting of 2 M glycerol + 0.4 M sucrose in culture medium for 40 min at room temperature; and dehydration in PVS2 at 0 degree C for 45 min before rapid immersion in liquid nitrogen (LN). Rewarming was performed in a water-bath at 40 degree C for 2 min. Explants were then immersed in unloading solution for 10 min before plating on recovery medium supplemented with 0.01 mg per liter thidiazuron (TDZ). TDZ was progressively eliminated from the medium over a period of 6 weeks. Plantlets were transferred to a double-layer medium to enhance rooting. This protocol was successfully applied to three individuals of T. boveana harvested from the wild.

  4. B Cells and B Cell Blasts Withstand Cryopreservation While Retaining Their Functionality for Producing Antibody.

    Science.gov (United States)

    Fecher, Philipp; Caspell, Richard; Naeem, Villian; Karulin, Alexey Y; Kuerten, Stefanie; Lehmann, Paul V

    2018-05-31

    In individuals who have once developed humoral immunity to an infectious/foreign antigen, the antibodies present in their body can mediate instant protection when the antigen re-enters. Such antigen-specific antibodies can be readily detected in the serum. Long term humoral immunity is, however, also critically dependent on the ability of memory B cells to engage in a secondary antibody response upon re-exposure to the antigen. Antibody molecules in the body are short lived, having a half-life of weeks, while memory B cells have a life span of decades. Therefore, the presence of serum antibodies is not always a reliable indicator of B cell memory and comprehensive monitoring of humoral immunity requires that both serum antibodies and memory B cells be assessed. The prevailing view is that resting memory B cells and B cell blasts in peripheral blood mononuclear cells (PBMC) cannot be cryopreserved without losing their antibody secreting function, and regulated high throughput immune monitoring of B cell immunity is therefore confined to-and largely limited by-the need to test freshly isolated PBMC. Using optimized protocols for freezing and thawing of PBMC, and four color ImmunoSpot ® analysis for the simultaneous detection of all immunoglobulin classes/subclasses we show here that both resting memory B cells and B cell blasts retain their ability to secrete antibody after thawing, and thus demonstrate the feasibility of B cell immune monitoring using cryopreserved PBMC.

  5. Cryopreservation of human insulin expressing cells macro-encapsulated in a durable therapeutic immunoisolating device theracyte.

    Science.gov (United States)

    Yakhnenko, Ilya; Wong, Wallace K; Katkov, Igor I; Itkin-Ansari, Pamela

    2012-01-01

    Encapsulating insulin producing cells (INPCs) in an immunoisolation device have been shown to cure diabetes in rodents without the need for immunosuppression. However, micro-encapsulation in semi-solid gels raises longevity and safety concerns for future use of stem cell derived INPCs. We have focused on a durable and retrievable macro-encapsulation (> 10(6) cells) device (TheraCyte). Cryopreservation (CP) of cells preloaded into the device is highly desirable but may require prolonged exposure to cryoprotectants during loading and post-thaw manipulations. Here, we are reporting survival and function of a human islet cell line frozen as single cells or as islet-like cell clusters. The non-clusterized cells exhibited high cryosurvival after prolonged pre-freeze or post-thaw exposure to 10 percent DMSO. However, both clusterization and especially loading INPCs into the device reduced viable yield even without CP. The survived cryopreserved macro-encapsulated INPCs remained fully functional suggesting that CP of macro-encapsulated cells is a promising tool for cell based therapies.

  6. Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) by vitrification.

    Science.gov (United States)

    Sakai, A; Kobayashi, S; Oiyama, I

    1990-06-01

    The nucellar cells of navel orange(Citrus sinensis Osb. var. brasiliensis Tanaka) were successfully cryopreserved by vitrification. In this method, cells were sufficiently dehydrated with highly concentrated cryoprotective solution(PVS2) prior to direct plunge in liquid nitrogen. The PVS2 contains(w/v) 30% glycerol, 15% ethylene glycol and 15% DMSO in Murashige-Tucker medium(MT) containing 0.15 M sucrose. Cells were treated with 60% PVS2 at 25°C for 5 min and then chilled PVS2 at 0°C for 3 min. The cell suspension of about 0.1 ml was loaded in a 0.5 ml transparent plastic straw and directly plunged in liquid nitrogen for 30 min. After rapid warming, the cell suspension was expelled in 2 ml of MT medium containing 1.2 M sucrose. The average rate of survival was about 80%. The vitrified cells regenerated plantlets. This method is very simple and the time required for cryopreservation is only about 10 min.

  7. The Development of Optimal Cryopreservation Media For Longspine Scraper (Capoeta trutta Sperm

    Directory of Open Access Journals (Sweden)

    Erdinç Şahinöz

    2018-03-01

    Full Text Available This study is performed to determine some of sperm quality after applying freezing / thawing process. Thus, the aim of this study is to examine different cryprotective agents with additives in terms of their effects at different pH on the cryopreservation process of longspine scraper (Capoeta trutta. The present study, twelve media were prepared by mixing three different cryoprotectants (dimethyl sulfoxide (DMSO, (CH32SO; methanol (CH3OH; methyl glycol (MG, CH3O (CH22OH with an extenders (glucose at four different pH (7.2, 7.6, 8.0 and 8.4 for longspine scraper semen. Considering the findings from the examination (The motility rate after thawing process and duration of motility obtained in DMSO as 81% and 20 min, in methanol as 73% and 12 min, in methyl glycol as 60% and 15 min., we can conclude that the DMSO is the best freezing media in order to create new essays in cryopreservation for sperm of Capoeta trutta in the future.

  8. Schistosoma mansoni: vaccination of mice with 10-krad-irradiated, cryopreserved schistosomules

    International Nuclear Information System (INIS)

    Lewis, F.A.; Stirewalt, M.A.; Leef, J.L.

    1984-01-01

    Protection against a Schistosoma mansoni cercarial challenge was evaluated in mice immunized with a vaccine composed of 10-krad-irradiated, cryopreserved schistosomules. The level of resistance induced in C57B1/6 or NMRI (CV) mice increased with the number of schistosomules injected. Up to 83% reduction in challenge worm burden was achieved when 5000 schistosomules were injected per mouse. Intramuscular injection of the vaccine was superior to subcutaneous. Multiple immunizations, up to 3 at 4-week intervals, did not increase the resistance induced by a single immunization. A high level of protection developed in as little as 2 weeks and was maintained through at least 12 weeks postimmunization. The vaccine irradiated with 10 krad from either a 60-cobalt or 137-cesium source induced equivalent levels of resistance, and no differences were found in the immunogenicity of vaccines comprised of organisms irradiated as cercariae or as 1- to 3-hr-old schistosomules. These findings are basic to the development of a cryopreserved, live vaccine against schistosomiasis of humans or domestic animals

  9. Boar spermatozoa cryopreservation in low glycerol/trehalose enriched freezing media improves cellular integrity.

    Science.gov (United States)

    Gutiérrez-Pérez, Oscar; Juárez-Mosqueda, María de Lourdes; Carvajal, Salvador Uribe; Ortega, María Elena Trujillo

    2009-06-01

    The use of glycerol for boar semen cryopreservation results in low fertility, possibly due to toxicity. This has led to recommend the use of solutions with less than 4% glycerol. Trehalose is a disaccharide known to stabilize proteins and biologic membranes during processes such as cryopreservation. Thus, it was decided to evaluate the cryoprotective effect of glycerol/trehalose mixtures. Effects on motility (M), viability (Vb) and acrosomal integrity (nA) were evaluated. Sperm samples were frozen in three different extenders: G4 contained 4% glycerol; T1 contained 1% glycerol plus 250 mM trehalose and T0.5 was constituted by 0.5% glycerol plus 250 mM trehalose. All extenders yielded similar post-freezing/thawing motility rates. Viability was diminished in T0.5 as compared to the others. In regard to acrosome integrity, it was twice as high (Pextender. Thus, T1 twice as many spermatozoa were alive, motile and intact, than in either T0.5 or G4, i.e. during freeze/thawing the use of T1 resulted in twice as many fertile cells as when using the other extenders. During our study, we noted that there were wide individual variations both in sperm viability and in motility.

  10. Dimethylformamide is not better than glycerol for cryopreservation of boar semen.

    Science.gov (United States)

    Malo, C; Gil, L; Cano, R; Martínez, F; García, A; Jerez, R A

    2012-05-01

    To improve the boar sperm cryopreservation process, the influence of the sugar (lactose, trehalose) source and the cryoprotectant [glycerol, dimethylformamide (DMF)] on the success of freezing was investigated. Sperm samples were frozen in one of six extenders: lactose plus 3% glycerol (LG); lactose plus 1.5% glycerol and 1.5% DMF (LGD); lactose plus 3% DMF (LD); trehalose plus 3% glycerol (TG); trehalose plus 1.5% glycerol and 1.5% DMF (TGD); trehalose plus 3% DMF (TD). Effects on motility, viability, acrosome integrity and hypoosmotic test (HOST) were measured. The results showed that extender containing 3% glycerol retained the highest motility percentages. In regard to viability and acrosome integrity, all extenders yielded similar rates except for the decreasing values of TD. Endosmosis was diminished in TD and LD at 2 h (P = 0.0018), as compared with the others. The results of the study demonstrated that the use of DMF as a cryoprotectant adversely affected boar sperm quality after cryopreservation. © 2011 Blackwell Verlag GmbH.

  11. On the Effects of Thermal History on the Development and Relaxation of Thermo-Mechanical Stress in Cryopreservation.

    Science.gov (United States)

    Eisenberg, David P; Steif, Paul S; Rabin, Yoed

    2014-01-01

    This study investigates the effects of the thermal protocol on the development and relaxation of thermo-mechanical stress in cryopreservation by means of glass formation, also known as vitrification. The cryopreserved medium is modeled as a homogeneous viscoelastic domain, constrained within either a stiff cylindrical container or a highly compliant bag. Annealing effects during the cooling phase of the cryopreservation protocol are analyzed. Results demonstrate that an intermediate temperature-hold period can significantly reduce the maximum tensile stress, thereby decreasing the potential for structural damage. It is also demonstrated that annealing at temperatures close to glass transition significantly weakens the dependency of thermo-mechanical stress on the cooling rate. Furthermore, a slower initial rewarming rate after cryogenic storage may drastically reduce the maximum tensile stress in the material, which supports previous experimental observations on the likelihood of fracture at this stage. This study discusses the dependency of the various stress components on the storage temperature. Finally, it is demonstrated that the stiffness of the container wall can affect the location of maximum stress, with implications on the development of cryopreservation protocols.

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

    Science.gov (United States)

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

    2013-01-01

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

  13. Long term storage of Pleurotus ostreatus and Trametes versicolor isolates using different cryopreservation techniques and its impact on laccase activity

    Czech Academy of Sciences Publication Activity Database

    Eichlerová, Ivana; Homolka, Ladislav; Tomšovský, M.; Lisá, Ludmila

    2015-01-01

    Roč. 119, č. 12 (2015), s. 1345-1353 ISSN 1878-6146 R&D Projects: GA ČR GAP504/12/0709 Institutional support: RVO:61388971 Keywords : Basidiomycetes * Cryopreservant * Liquid nitrogen Subject RIV: EE - Microbiology, Virology Impact factor: 2.244, year: 2015

  14. DMSO-free cryopreservation of adipose-derived mesenchymal stromal cells: expansion medium affects post-thaw survival

    Czech Academy of Sciences Publication Activity Database

    Rogulska, O.; Petrenko, Yuriy; Petrenko, A.

    2017-01-01

    Roč. 69, č. 2 (2017), s. 265-276 ISSN 0920-9069 Institutional support: RVO:68378041 Keywords : human adiposederived mesenchymalstromal cells * DMSO-free cryopreservation * plateletlysate Subject RIV: FP - Other Medical Disciplines OBOR OECD: Cell biology Impact factor: 1.857, year: 2016

  15. Ovarian tissue cryopreserved for fertility preservation from patients with Ewing or other sarcomas appear to have no tumour cell contamination

    DEFF Research Database (Denmark)

    Greve, Tine; Wielenga, Vera Timmermans; Grauslund, Morten

    2013-01-01

    The chemotherapy required to treat patients with sarcoma may as a side-effect induce infertility in girls and