Sample records for vitrification permitting program

  1. World first in high level waste vitrification - A review of French vitrification industrial achievements

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

    Brueziere, J.; Chauvin, E.; Piroux, J.C.

    2013-07-01

    AREVA has more than 30 years experience in operating industrial HLW (High Level radioactive Waste) vitrification facilities (AVM - Marcoule Vitrification Facility, R7 and T7 facilities). This vitrification technology was based on borosilicate glasses and induction-heating. AVM was the world's first industrial HLW vitrification facility to operate in-line with a reprocessing plant. The glass formulation was adapted to commercial Light Water Reactor fission products solutions, including alkaline liquid waste concentrates as well as platinoid-rich clarification fines. The R7 and T7 facilities were designed on the basis of the industrial experience acquired in the AVM facility. The AVM vitrification process wasmore » implemented at a larger scale in order to operate the R7 and T7 facilities in-line with the UP2 and UP3 reprocessing plants. After more than 30 years of operation, outstanding record of operation has been established by the R7 and T7 facilities. The industrial startup of the CCIM (Cold Crucible Induction Melter) technology with enhanced glass formulation was possible thanks to the close cooperation between CEA and AREVA. CCIM is a water-cooled induction melter in which the glass frit and the waste are melted by direct high frequency induction. This technology allows the handling of highly corrosive solutions and high operating temperatures which permits new glass compositions and a higher glass production capacity. The CCIM technology has been implemented successfully at La Hague plant.« less

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

    PubMed

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

    2016-10-15

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

  3. Safeguarding Fertility With Whole Ovary Cryopreservation and Microvascular Transplantation: Higher Follicular Survival With Vitrification Than With Slow Freezing in a Ewe Model.

    PubMed

    Torre, Antoine; Vertu-Ciolino, Delphine; Mazoyer, Claire; Selva, Jacqueline; Lornage, Jacqueline; Salle, Bruno

    2016-09-01

    In young women, ovarian cortex cryopreservation before gonadotoxic chemotherapy and its avascular grafting after cancer healing permitted fertility restoration. However, ischemia reduced the grafts' lifespan. Microvascular transplantation of cryopreserved whole ovary may allow immediate revascularization, ensuring better fertility preservation, but the best cryopreservation method is unknown. We aimed to compare slow freezing and vitrification of whole ovary for fertility preservation purposes, in an ewe model. Twelve ewes were allocated at random to slow freezing (n = 6) or vitrification group (n = 6). Ewes' left ovary was removed and cryopreserved. Dimethyl sulfoxide 2 M was used as cryoprotector for slow freezing. Vitrification was obtained using increasing concentrations of a vitrification solution of the latest generation (VM3) and gradual temperature lowering to minimize toxicity. After a month, the right ovary was removed, the left ovary was thawed/warmed, and its vessels were anastomosed to the right pedicle. Fertility and ovarian function were assessed for 3 years. Ovarian follicles in native and transplanted ovaries were counted and compared at study completion. Hormonal secretion resumed in all ewes of both groups. One ewe of the slow-freezing group delivered healthy twins 1 year 9 months and 12 days after transplantation. Estimated whole follicle survival was very low in both groups but significantly higher after vitrification than after slow freezing (0.3% ± 0.5% vs 0.017% ± 0.019%, respectively; p < 0.05). Further progress is needed before whole-ovary cryopreservation can be considered an option for safeguarding fertility. Whole ovary vitrification provides better follicular survival compared to slow freezing and may be a valuable cryopreservation option.

  4. GEOSAFE CORPORATION IN SITU VITRIFICATION: INNOVATIVE TECHNOLOGY EVALUATION REPORT

    EPA Science Inventory

    This report summarizes the findings associated with a Demonstration of the Geosafe Corporation (Geosafe) In Situ Vitrification (ISV) Process. The Geosafe ISV Technology was evaluated under the EPA Superfund Innovative Technology Evaluation (SITE) Program in conjuction with remedi...

  5. Hanford High-Level Waste Vitrification Program at the Pacific Northwest National Laboratory: technology development - annotated bibliography

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

    Larson, D.E.

    1996-09-01

    This report provides a collection of annotated bibliographies for documents prepared under the Hanford High-Level Waste Vitrification (Plant) Program. The bibliographies are for documents from Fiscal Year 1983 through Fiscal Year 1995, and include work conducted at or under the direction of the Pacific Northwest National Laboratory. The bibliographies included focus on the technology developed over the specified time period for vitrifying Hanford pretreated high-level waste. The following subject areas are included: General Documentation; Program Documentation; High-Level Waste Characterization; Glass Formulation and Characterization; Feed Preparation; Radioactive Feed Preparation and Glass Properties Testing; Full-Scale Feed Preparation Testing; Equipment Materials Testing; Meltermore » Performance Assessment and Evaluations; Liquid-Fed Ceramic Melter; Cold Crucible Melter; Stirred Melter; High-Temperature Melter; Melter Off-Gas Treatment; Vitrification Waste Treatment; Process, Product Control and Modeling; Analytical; and Canister Closure, Decontamination, and Handling« less

  6. Comparison of two closed carriers for vitrification of human blastocysts in a donor program.

    PubMed

    Guerrero, Jaime; Gallardo, Miguel; Rodríguez-Arnedo, Adoración; Ten, Jorgen; Bernabeu, Rafael

    2018-04-01

    The survival of human blastocysts to vitrification with two different carriers is compared. Both vitrification carriers used in this study are in the category of closed carriers, as they completely isolate the samples from direct contact with liquid nitrogen or its vapours during cooling and storage, until warming. This characteristic is appealing because it reduces or eliminates the theoretical risk of cross-contamination during that period of time. The two closed vitrification systems used present very different design and features: in the High Security Vitrification device, the carrier straw containing the embryos is encapsulated inside an external straw before plunging in liquid nitrogen, resulting in thermal insulation during cooling. On the other hand, in the SafeSpeed carrier embryos are loaded in a thin-walled, narrow capillary designed to maximize the thermal transference. Both closed carriers achieved comparable outcomes in terms of survival of blastocysts to the vitrification process, with 97.5% vs. 96.1% survival with HSV and SafeSpeed, respectively. In conclusion, the cooling and warming rates at which these carriers operate, in combination with the cytosolic solute concentration in the cells of the cryopreserved blastocysts attained after a cryoprotectant-loading protocol, result in successful vitrification of human blastocysts for human assisted reproduction. Copyright © 2018. Published by Elsevier Inc.

  7. Property evolution during vitrification of dimethacrylate photopolymer networks

    PubMed Central

    Abu-Elenain, Dalia; Lewis, Steven H.; Stansbury, Jeffrey W.

    2013-01-01

    Objectives This study seeks to correlate the interrelated properties of conversion, shrinkage, modulus and stress as dimethacrylate networks transition from rubbery to glassy states during photopolymerization. Methods An unfilled BisGMA/TEGDMA resin was photocured for various irradiation intervals (7–600 s) to provide controlled levels of immediate conversion, which was monitored continuously for 10 min. Fiber optic near-infrared spectroscopy permitted coupling of real-time conversion measurement with dynamic polymerization shrinkage (linometer), modulus (dynamic mechanical analyzer) and stress (tensometer) development profiles. Results The varied irradiation conditions produced final conversion ranging from 6 % to more than 60 %. Post-irradiation conversion (dark cure) was quite limited when photopolymerization was interrupted either at very low or very high levels of conversion while significant dark cure contributions were possible for photocuring reactions suspended within the post-gel, rubbery regime. Analysis of conversion-based property evolution during and subsequent to photocuring demonstrated that the shrinkage rate increased significantly at about 40 % conversion followed by late-stage suppression in the conversion-dependent shrinkage rate that begins at about 45–50 % conversion. The gradual vitrification process over this conversion range is evident based on the broad but well-defined inflection in the modulus versus conversion data. As limiting conversion is approached, modulus and, to a somewhat lesser extent, stress rise precipitously as a result of vitrification with the stress profile showing little if any late-stage suppression as seen with shrinkage. Significance Near the limiting conversion for this model resin, the volumetric polymerization shrinkage rate slows while an exponential rise in modulus promotes the vitrification process that appears to largely dictate stress development. PMID:24080378

  8. [Successful pregnancies after oocyte and embryo vitrification].

    PubMed

    Salazar, Francisco Hernández; Loza, Erik Omar Okhuysen; Lucas, Maria Teresa Huerta J; Gutiérrez, Gustavo Romero

    2008-02-01

    Cryopreservation of human oocytes represents a solution for ethic conflict about frozen embryo storage for patients with risk to develop ovarian hyperstimulation syndrome; also is an available technique to preserve fertility in women with cancer under treatment, in poor response patients, in case of premature ovarian failure or aging and for other medical or social conditions that require to delay pregnancies, as well as to make easier oocyte donation programs. This paper reports two cases of successful pregnancies after embryo and oocyte vitrification, as well as their results. The technique of vitrification with the cryotop method is an excellent alternative, efficient, fast and cheap for oocyte and embryo cryopreservation with high ranges of fertilization, cleavage and pregnancies with a normal evolution.

  9. Industrial scale-plant for HLW partitioning in Russia

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

    Dzekun, E.G.; Glagolenko, Y.V.; Drojko, E.G.

    1996-12-31

    Radiochemical plant of PA <> at Ozersk, which was come on line in December 1948 originally for weapon plutonium production and reoriented on the reprocessing of spent fuel, till now keeps on storage HLW of the military program. Application of the vitrification method since 1986 has not essentially reduced HLW volumes. So, as of September 1, 1995 vitrification installations had been processed 9590 m{sup 3} HLW and 235 MCi of radionuclides was included in glass. However only 1100 m{sup 3} and 20.5 MCi is part of waste of the military program. The reason is the fact, that the technology andmore » equipment of vitrification were developed for current waste of Purex-process, for which low contents of corrosion-dangerous impurity to materials of vitrification installation is characteristic of. With reference to HLW, which are growing at PA <> in the course of weapon plutonium production, the program of Science-Research Works includes the following main directions of work. Development of technology and equipment of installations for immobilising HLW with high contents of impurity into a solid form at induction melter. Application of High-temperature Adsorption Method for sorption of radionuclides from HLW on silica gel. Application of Partitioning Method of radionuclides from HLW, based on extraction cesium and strontium into cobalt dicarbollyde or crown-ethers, but also on recovery of cesium radionuclides by sorption on inorganic sorbents. In this paper the results of work on creation of first industrial scale-plant for partitioning HLW by the extraction and sorption methods are reported.« less

  10. Property evolution during vitrification of dimethacrylate photopolymer networks.

    PubMed

    Abu-elenain, Dalia A; Lewis, Steven H; Stansbury, Jeffrey W

    2013-11-01

    This study seeks to correlate the interrelated properties of conversion, shrinkage, modulus and stress as dimethacrylate networks transition from rubbery to glassy states during photopolymerization. An unfilled BisGMA/TEGDMA resin was photocured for various irradiation intervals (7-600 s) to provide controlled levels of immediate conversion, which was monitored continuously for 10 min. Fiber optic near-infrared spectroscopy permitted coupling of real-time conversion measurement with dynamic polymerization shrinkage (linometer), modulus (dynamic mechanical analyzer) and stress (tensometer) development profiles. The varied irradiation conditions produced final conversion ranging from 6% to more than 60%. Post-irradiation conversion (dark cure) was quite limited when photopolymerization was interrupted either at very low or very high levels of conversion while significant dark cure contributions were possible for photocuring reactions suspended within the post-gel, rubbery regime. Analysis of conversion-based property evolution during and subsequent to photocuring demonstrated that the shrinkage rate increased significantly at about 40% conversion followed by late-stage suppression in the conversion-dependent shrinkage rate that begins at about 45-50% conversion. The gradual vitrification process over this conversion range is evident based on the broad but well-defined inflection in the modulus versus conversion data. As limiting conversion is approached, modulus and, to a somewhat lesser extent, stress rise precipitously as a result of vitrification with the stress profile showing little if any late-stage suppression as seen with shrinkage. Near the limiting conversion for this model resin, the volumetric polymerization shrinkage rate slows while an exponential rise in modulus promotes the vitrification process that appears to largely dictate stress development. Copyright © 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

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

    PubMed

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

    2017-03-01

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

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

    PubMed Central

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

    2017-01-01

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

  13. INNOVATIVE FOSSIL FUEL FIRED VITRIFICATION TECHNOLOGY FOR SOIL REMEDIATION

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

    J. Hnat; L.M. Bartone; M. Pineda

    2001-07-13

    This Summary Report summarizes the progress of Phases 3, 3A and 4 of a waste technology Demonstration Project sponsored under a DOE Environmental Management Research and Development Program and administered by the U.S. Department of Energy National Energy Technology Laboratory-Morgantown (DOE-NETL) for an ''Innovative Fossil Fuel Fired Vitrification Technology for Soil Remediation''. The Summary Reports for Phases 1 and 2 of the Program were previously submitted to DOE. The total scope of Phase 3 was to have included the design, construction and demonstration of Vortec's integrated waste pretreatment and vitrification process for the treatment of low level waste (LLW), TSCA/LLWmore » and mixed low-level waste (MLLW). Due to funding limitations and delays in the project resulting from a law suit filed by an environmental activist and the extended time for DOE to complete an Environmental Assessment for the project, the scope of the project was reduced to completing the design, construction and testing of the front end of the process which consists of the Material Handling and Waste Conditioning (MH/C) Subsystem of the vitrification plant. Activities completed under Phases 3A and 4 addressed completion of the engineering, design and documentation of the Material Handling and Conditioning System such that final procurement of the remaining process assemblies can be completed and construction of a Limited Demonstration Project be initiated in the event DOE elects to proceed with the construction and demonstration testing of the MH/C Subsystem.« less

  14. Equine ovarian tissue viability after cryopreservation and in vitro culture.

    PubMed

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

    2017-07-15

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

  15. Vitrification, encapsulation-vitrification and droplet-vitrification: a review.

    PubMed

    Sakai, Akira; Engelmann, Florent

    2007-01-01

    This paper discusses the importance of the successive steps of the vitrification technique and reviews the current development and use of vitrification and of the two derived protocols, encapsulation-vitrification and droplet-vitrification. Vitrification refers to the physical process by which a highly concentrated cryoprotective solution supercools to very low temperatures and finally solidifies into a metastable glass, without undergoing crystallization at a practical cooling rate. Samples are thus cryopreserved without detrimental intracellular ice formation. In a standard vitrification protocol, excised explants are precultured on medium enriched with sucrose, treated (loaded) with a loading solution composed of 2 M glycerol + 0.4 M sucrose, dehydrated with a highly concentrated vitrification solution [e.g. the PVS2 vitrification solution, which contains 30 percent (w/v) glycerol, 15 percent (w/v) ethylene glycol and 15 percent (w/v) DMSO and 0.4 M sucrose], frozen and rewarmed rapidly, unloaded with basal culture medium supplemented with 1.2 M sucrose, and then transferred to standard culture conditions. In the encapsulation-vitrification technique, the explants are encapsulated in alginate beads, loaded and dehydrated with a vitrification solution before rapid immersion in liquid nitrogen. In the droplet-freezing technique, excised explants are loaded, treated with the vitrification solution and frozen in individual microdroplets of vitrification solution placed on aluminium foils, which are immersed rapidly in liquid nitrogen. These three techniques have been applied to different tissues of over 100 plant species from temperate and tropical origins and the number of cases where they are being tested on a large scale or applied routinely is increasing.

  16. Vitrification as an alternative means of cryopreserving ovarian tissue.

    PubMed

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

    2011-08-01

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

  17. In vitro conservation of Dendrobium germplasm.

    PubMed

    Teixeira da Silva, Jaime A; Zeng, Songjun; Galdiano, Renato Fernandes; Dobránszki, Judit; Cardoso, Jean Carlos; Vendrame, Wagner A

    2014-09-01

    Dendrobium is a large genus in the family Orchidaceae that exhibits vast diversity in floral characteristics, which is of considerable importance to orchid breeders, biotechnologists and collectors. Native species have high value as a result of their medicinal properties, while their hybrids are important as ornamental commodities, either as cut flowers or potted plants and are thus veritable industrial crops. Thus, preservation of Dendrobium germplasm is valuable for species conservation, breeding programs and the floriculture industry. Cryopreservation represents the only safe, efficient and cost-effective long-term storage option to facilitate the conservation of genetic resources of plant species. This review highlights 16 years of literature related to the preservation of Dendrobium germplasm and comprises the most comprehensive assessment of thorough studies performed to date, which shows reliable and reproducible results. Air-drying, encapsulation-dehydration, encapsulation-vitrification, vitrification and droplet-vitrification are the current cryopreservation methodologies that have been used to cryopreserve Dendrobium germplasm. Mature seeds, pollen, protoplasts, shoot primordia, protocorms and somatic embryos or protocorm-like bodies (PLBs) have been cryopreserved with different levels of success. Encapsulation-vitrification and encapsulation-dehydration are the most used protocol, while PLBs represent the main explant explored.

  18. TECHNICAL ASSESSMENT OF BULK VITRIFICATION PROCESS & PRODUCT FOR TANK WASTE TREATMENT AT THE DEPARTMENT OF ENERGY HANFORD SITE

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

    SCHAUS, P.S.

    At the U.S. Department of Energy (DOE) Hanford Site, the Waste Treatment Plant (WTP) is being constructed to immobilize both high-level waste (IUW) for disposal in a national repository and low-activity waste (LAW) for onsite, near-surface disposal. The schedule-controlling step for the WTP Project is vitrification of the large volume of LAW, current capacity of the WTP (as planned) would require 50 years to treat the Hanford tank waste, if the entire LAW volume were to be processed through the WTP. To reduce the time and cost for treatment of Hanford Tank Waste, and as required by the Tank Wastemore » Remediation System Environmental Impact Statement Record of Decision and the Hanford Federal Facility Consent Agreement (Tn-Party Agreement), DOE plans to supplement the LAW treatment capacity of the WTP. Since 2002, DOE, in cooperation with the Environmental Protection Agency and State of Washington Department of Ecology has been evaluating technologies that could provide safe and effective supplemental treatment of LAW. Current efforts at Hanford are intended to provide additional information to aid a joint agency decision on which technology will be used to supplement the WTP. A Research, Development and Demonstration permit has been issued by the State of Washington to build and (for a limited time) operate a Demonstration Bulk Vitrification System (DBVS) facility to provide information for the decision on a supplemental treatment technology for up to 50% of the LAW. In the Bulk Vitrification (BV) process, LAW, soil, and glass-forming chemicals are mixed, dried, and placed in a refractory-lined box, Electric current, supplied through two graphite electrodes in the box, melts the waste feed, producing a durable glass waste-form. Although recent modifications to the process have resulted in significant improvements, there are continuing technical concerns.« less

  19. THE DOE OFFICE OF ENVIRONMENTAL MANAGEMENT INTERNATIONAL COOPERATIVE PROGRAM: OVERVIEW OF TECHNICAL TASKS AND RESULTS

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

    Marra, J.; Fox, K.; Farfan, E.

    2009-12-08

    The DOE Office of Environmental Management (DOE-EM) Office of Engineering and Technology is responsible for implementing EM's International Cooperative Program. Over the past 15 years, collaborative work has been conducted through this program with researchers in Russia, Ukraine, France, United Kingdom and Republic of Korea. Currently, work is being conducted with researchers in Russia and Ukraine. Efforts aimed at evaluating and advancing technologies to support U.S. high-level waste (HLW) vitrification initiatives are being conducted in collaboration with Russian researchers. Work at Khlopin Radium Institute (KRI) is targeted at improving the throughput of current vitrification processes by increasing melting rate. Thesemore » efforts are specifically targeted at challenging waste types identified at the Savannah River Site (SRS) and Hanford Site. The objectives of current efforts at SIA Radon are to gain insight into vitrification process limits for the cold crucible induction melter (CCIM) technology. Previous demonstration testing has shown that the CCIM offers the potential for dramatic increases in waste loading and waste throughput. However, little information is known regarding operational limits that could affect long-term, efficient CCIM operations. Collaborative work with the Russian Electrotechnical University (ETU) 'LETI' is aimed at advancing CCIM process monitoring, process control and design. The goal is to further mature the CCIM technology and to establish it as a viable HLW vitrification technology. The greater than two year effort conducted with the International Radioecology Laboratory in the Ukraine recently completed. The objectives of this study were: to assess the long-term impacts to the environment from radiation exposure in the Chernobyl Exclusion Zone (ChEZ); and to provide information on remediation guidelines and ecological risk assessment within radioactively contaminated territories around the Chernobyl Nuclear Power Plant (ChNPP) based on the results of long-term field monitoring, analytical measurements, and numerical modeling of soils and groundwater radioactive contamination.« less

  20. Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species.

    PubMed

    Tiersch, Nolan J; Childress, William M; Tiersch, Terrence R

    2018-05-16

    Vitrification is a method of cryopreservation that freezes samples rapidly, while forming an amorphous solid ("glass"), typically in small (μL) volumes. The goal of this project was to create, by three-dimensional (3D) printing, open vitrification devices based on an elliptical loop that could be efficiently used and stored. Vitrification efforts can benefit from the application of 3D printing, and to begin integration of this technology, we addressed four main variables: thermoplastic filament type, loop length, loop height, and method of loading. Our objectives were to: (1) design vitrification loops with varied dimensions; (2) print prototype loops for testing; (3) evaluate loading methods for the devices; and (4) classify vitrification responses to multiple device configurations. The various configurations were designed digitally using 3D CAD (Computer Aided Design) software, and prototype devices were produced with MakerBot ® 3D printers. The thermoplastic filaments used to produce devices were acrylonitrile butadiene styrene (ABS) and polylactic acid (PLA). Vitrification devices were characterized by the film volumes formed with different methods of loading (pipetting or submersion). Frozen films were classified to determine vitrification quality: zero (opaque, or abundant crystalline ice formation); one (translucent, or partial vitrification), or two (transparent, or substantial vitrification, glass). A published vitrification solution was used to conduct experiments. Loading by pipetting formed frozen films more reliably than by submersion, but submersion yielded fewer filling problems and was more rapid. The loop designs that yielded the highest levels of vitrification enabled rapid transfer of heat, and most often were characterized as being longer and consisting of fewer layers (height). 3D printing can assist standardization of vitrification methods and research, yet can also provide the ability to quickly design and fabricate custom devices when needed.

  1. Cryopreservation of umbilical cord blood-derived mesenchymal stem cells without dimethyl sulfoxide.

    PubMed

    Wang, Hai-Yan; Lun, Zhao-Rong; Lu, Shu-Shen

    2011-01-01

    Cryopreservation of umbilical cord blood-derived mesenchymal stem cells (UCB-derived MSCs) is crucial step for its clinical applications in cell transplantation therapy. In the cryopreservation of MSCs, dimethyl sulfoxide has been widely used as a cryoprotectant (CPA). However, it has been proved that DMSO has toxic side effects to human body. In this study, DMSO-free CPA solutions which contained ethylene glycol (EG), 1, 2-propylene glycol (PG) and sucrose as basic CPAs, supplemented with polyvinyl alcohol (PVA) as an additive, were developed for the cryopreservation of UCB-derived MSCs. The cryopreservation of UCB-derived MSCs was achieved by vitrification via plunging into liquid nitrogen and by programmed freezing via an optical-DSC system respectively. The viability of thawed UCB-derived MSCs was tested by trypan blue exclusion assay. Results showed that the viability of thawed UCB-derived MSCs was enhanced from 71.2% to 95.4% in the presence of PVA for vitrification, but only < 10% to 45% of viability was found for programmed freezing. These results indicate that PVA exerts a beneficial effect on the cryopreservation of UCB-derived MSCs and suggest the vitrification in combination with the dimethyl sulfoxide free CPA solutions supplemented with PVA would be an efficient protocol for the cryopreservation of UCB-derived MSCs.

  2. Technical information report: Plasma melter operation, reliability, and maintenance analysis

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

    Hendrickson, D.W.

    1995-03-14

    This document provides a technical report of operability, reliability, and maintenance of a plasma melter for low-level waste vitrification, in support of the Hanford Tank Waste Remediation System (TWRS) Low-Level Waste (LLW) Vitrification Program. A process description is provided that minimizes maintenance and downtime and includes material and energy balances, equipment sizes and arrangement, startup/operation/maintence/shutdown cycle descriptions, and basis for scale-up to a 200 metric ton/day production facility. Operational requirements are provided including utilities, feeds, labor, and maintenance. Equipment reliability estimates and maintenance requirements are provided which includes a list of failure modes, responses, and consequences.

  3. CRYOPRESERVATION STRATEGY FOR TISSUE ENGINEERING CONSTRUCTS CONSISTING OF HUMAN MESENHYMAL STEM CELLS AND HYDROGEL BIOMATERIALS.

    PubMed

    Wu, Y; Wen, F; Gouk, S S; Lee, E H; Kuleshova, L

    2015-01-01

    The development of vitrification strategy for cell-biomaterial constructs, particularly biologically inspired nanoscale materials and hydrogels mimicking the in vivo environment is an active area. A cryopreservation strategy mimicking the in vivo environment for cell-hydrogel constructs may enhance cell proliferation and biological function. To demonstrate the efficacy of vitrification as a platform technology involving tissue engineering and human mesenchymal stem cells (hMSCs). Microcarriers made from alginate coated with chitosan and collagen are used. Conventional freezing and vitrification were compared. The vitrification strategy includes 10 min step-wise exposure to a vitrification solution (40% v/v EG, 0.6M sucrose) and immersion into liquid nitrogen. Confocal imaging of live/dead staining of hMSCs cultured on the surface of microcarriers demonstrated that vitrified cells had excellent appearance and prolonged spindle shape morphology. The proliferation ability of post-vitrified cells arbitrated to protein Ki-67 gene expression was not significantly different in comparison to untreated control, while that of post-freezing cells was almost lost. The ability of hMSCs cultured on the surface of microcarriers to proliferate has been not affected by vitrification and it was significantly better after vitrification than after conventional freezing during continuous culture. Collagen II related mRNA expression by 4 weeks post-vitrification and post-freezing showed that ability to differentiate into cartilage was sustained during vitrification and reduced during conventional freezing. No significant difference was found between control and vitrification groups only. Vitrification strategy coupled with advances in hMSC-expansion platform that completely preserves the ability of stem cells to proliferate and subsequently differentiate allows not only to reach a critical cell number, but also demonstrate prospects for effective utilization and transportation of cells with their support system, creating demand for novel biodegradable materials.

  4. Pretreatment of Hanford medium-curie wastes by fractional crystallization.

    PubMed

    Nassif, Laurent; Dumont, George; Alysouri, Hatem; Rousseau, Ronald W

    2008-07-01

    Acceleration of the schedule for decontamination of the Hanford site using bulk vitrification requires implementation of a pretreatment operation. Medium-curie waste must be separated into two fractions: one is to go to a waste treatment and immobilization plant and a second, which is low-activity waste, is to be processed by bulk vitrification. The work described here reports research on using fractional crystallization for that pretreatment. Sodium salts are crystallized by evaporation of water from solutions simulating those removed from single-shell tanks, while leaving cesium in solution. The crystalline products are then recovered and qualified as low-activity waste, which is suitable upon redissolution for processing by bulk vitrification. The experimental program used semibatch operation in which a feed solution was continuously added to maintain a constant level in the crystallizer while evaporating water. The slurry recovered at the end of a run was filtered to recover product crystals, which were then analyzed to determine their composition. The results demonstrated that targets on cesium separation from the solids, fractional recovery of sodium salts, and sulfate content of the recovered salts can be achieved by the process tested.

  5. Improved low-CPA vitrification of mouse oocytes using quartz microcapillary.

    PubMed

    Choi, Jung Kyu; Huang, Haishui; He, Xiaoming

    2015-06-01

    Cryopreservation by low-cryoprotectant (CPA) vitrification has the potential to combine all the advantages of the conventional high-CPA vitrification and slow-freezing approaches while avoiding their drawbacks. However, current low-CPA vitrification protocol for cryopreservation of oocytes requires a lengthy and multi-step procedure for unloading CPAs. In this study, we report a much-simplified procedure of using quartz microcapillary (QMC) for low-CPA vitrification of mouse oocytes with only one step for unloading CPAs. The immediate viability of oocytes after the improved low-CPA vitrification was determined to be more than 90%. Moreover, no significant difference was observed in terms of embryonic development from the two-cell to blastocyst stages between the fresh and vitrified oocytes after in vitro fertilization (IVF). This improved low-CPA vitrification technology has the potential for efficient cryopreservation of oocytes to preserve the fertility of mammals including humans for assisted reproductive medicine, maintenance of animal resource and endangered species, and livestock management. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Vitrification-based cryopreservation of shoot-tips of Pinus kesiya Royle ex. Gord.

    PubMed

    Kalita, V; Choudhury, H; Kumaria, S; Tandon, P

    2012-01-01

    The present investigation was aimed at developing a protocol for long-term preservation of germplasm of Pinus kesiya Royle ex. Gord. through vitrification. Some of the critical components affecting explant tolerance to cryopreservation, such as effects of preculture, vitrification solutions, exposure time to vitrification solutions, volume of vitrification solution and its toxicity, washing of vitrified tissues after thawing, were analysed. The results showed that shoot regrowth of P. kesiya shoot-tips was considerably affected when exposed to cryoprotectants for longer periods of time (longer than 10 min). Among different vitrification solutions studied, maximum survival (76 percent) of shoot-tips was achieved with mVSL (using 0.6 ml of the solution) in MS basal medium containing 4.0 mg l-1 N6-benzyladenine (BA).

  7. Tank 241-AZ-101 Mixer Pump Test Vapor Sampling and Analysis Plan

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

    TEMPLETON, A.M.

    2000-03-06

    This sampling and analysis plan (SAP) identifies characterization objectives pertaining to sample collection, laboratory analytical evaluation, and reporting requirements for vapor samples obtained during the operation of mixer pumps in tank 241-AZ-101. The primary purpose of the mixer pump test (MPT) is to demonstrate that the two 300 horsepower mixer pumps installed in tank 241-AZ-101 can mobilize the settled sludge so that it can be retrieved for treatment and vitrification. Sampling will be performed in accordance with Tank 241-AZ-101 Mixer Pump Test Data Quality Objective (Banning 1999) and Data Quality Objectives for Regulatory Requirements for Hazardous and Radioactive Air Emissionsmore » Sampling and Analysis (Mulkey 1999). The sampling will verify if current air emission estimates used in the permit application are correct and provide information for future air permit applications.« less

  8. Tank 241-AZ-101 Mixer Pump Test Vapor Sampling and Analysis Plan

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

    TEMPLETON, A.M.

    2000-01-31

    This sampling and analysis plan (SAP) identifies characterization objectives pertaining to sample collection, laboratory analytical evaluation, and reporting requirements for vapor samples obtained during the operation of mixer pumps in tank 241-AZ-101. The primary purpose of the mixer pump test (MPT) is to demonstrate that the two 300 horsepower mixer pumps installed in tank 241-AZ-101 can mobilize the settled sludge so that it can be retrieved for treatment and vitrification Sampling will be performed in accordance with Tank 241-AZ-101 Mixer Pump Test Data Quality Objective (Banning 1999) and Data Quality Objectives for Regulatory Requirements for Hazardous and Radioactive Air Emissionsmore » Sampling and Analysis (Mulkey 1999). The sampling will verify if current air emission estimates used in the permit application are correct and provide information for future air permit applications.« less

  9. Tank 241-AZ-101 Mixer Pump Test Vapor Sampling and Analysis Plan

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

    TEMPLETON, A.M.

    2000-04-10

    This sampling and analysis plan (SAP) identifies characterization objectives pertaining to sample collection, laboratory analytical evaluation, and reporting requirements for vapor samples obtained during the operation of mixer pumps in tank 241-AZ-101. The primary purpose of the mixer pump test (MPT) is to demonstrate that the two 300 horsepower mixer pumps installed in tank 241-AZ-101 can mobilize the settled sludge so that it can be retrieved for treatment and vitrification. Sampling will be performed in accordance with Tank 241-AZ-101 Mixer Pump Test Data Quality Objective (Banning 1999) and Data Quality Objectives for Regulatory Requirements for Hazardous and Radioactive Air Emissionsmore » Sampling and Analysis (Mulkey 1999). The sampling will verify if current air emission estimates used in the permit application are correct and provide information for future air permit applications.« less

  10. Comparison of apoptosis pathway following the use of two protocols for vitrification of immature mouse testicular tissue.

    PubMed

    Hajiaghalou, Samira; Ebrahimi, Bita; Shahverdi, Abdolhossein; Sharbatoghli, Mina; Beigi Boroujeni, Nasim

    2016-11-01

    Our objective was to evaluate the apoptosis incidence in immature mouse testicular tissue after two different protocols of vitrification and short-term culture. Testes of 7-day-old Naval Medical Research Institute mice were isolated and distributed into control and vitrification groups. In vitrification 1 group, testes were vitrified using a combination of ethylene glycol and DMSO in three steps, and in vitrification 2 group, testes were vitrified using a combination of ethylene glycol and sucrose in five steps. Then, fresh and vitrified-warmed testis fragments were cultured for 20 hours. Morphology, cell viability, apoptosis incidence, and apoptosis gene expression (BAX, BCL2, Caspase 3, Fas, Fas ligand, p53) were evaluated at 0, 3, and 20 hours of culture by light microscopy, flow cytometry, and real-time polymerase chain reaction, respectively. Significant decrease of early apoptosis (annexin V+/PI- cells in vitrification 1 and 2 groups at 0 hours of culture, 37.34 ± 0.91 and 30.72 ± 2.2, and at 20 hours of culture, 1.46 ± 0.28 and 0.76 ± 0.11, respectively), increase of late apoptosis (annexin V+/PI+ cells in vitrification 1 group at 0 hours of culture, 14.46 ± 0.86, and at 20 hours of culture, 37.18 ± 2.34), and BAX/BCL-2 ratio (in vitrification 1 and 2 groups at 0 hours of culture, 7.31 ± 0.31 and 6.83 ± 1.38, and at 20 hours of culture, 24.08 ± 4.32 and 9.35 ± 1.91, respectively) were observed in vitrification groups during culture period. Caspase 3 expression was significantly decreased in all groups after 3 hours of culture (in control, vitrification 1, and vitrification 2 groups at 0 hours of culture, 1.00 ± 0.0, 1.56 ± 0.09, and 0.79 ± 0.06, and at 20 hours of culture, 0.37 ± 0.0, 0.96 ± 0.10, and 0.12 ± 0.03, respectively). Expression of p53 was significantly lower in vitrification 1 (0.32 ± 0.02) and control (0.50 ± 0.03) groups in 20 hours of culture as compared with vitrification 2 (0.88 ± 0.14) group. Fas (in vitrification 1 and 2 groups at 0 hours of culture, 2.29 ± 0.23 and 1.14 ± 0.15, and at 20 hours of culture, 12.43 ± 0.46 and 6.7 ± 0.48, respectively) and Fas Ligand (in vitrification 1 and 2 groups at 0 hours of culture, 1.2 ± 0.28 and 5.24 ± 0.32, and at 20 hours of culture, 21.75 ± 2.00 and 25.82 ± 2.15, respectively) expressions significantly increased in vitrification groups after 20 hours of culture. Although both vitrification protocols cause cell death via apoptotic and necrotic pathway, it seems that vitrification 1 protocol induces cell death more via apoptotic pathway than via necrosis. The apoptosis incidence after vitrification may have occurred independent of p53. Copyright © 2016 Elsevier Inc. All rights reserved.

  11. Appendix C: Automated Vitrification of Mammalian Embryos on a Digital Microfluidic Device.

    PubMed

    Liu, Jun; Pyne, Derek G; Abdelgawad, Mohamed; Sun, Yu

    2017-01-01

    This chapter introduces a digital microfluidic device that automates sample preparation for mammalian embryo vitrification. Individual microdroplets manipulated on the microfluidic device were used as microvessels to transport a single mouse embryo through a complete vitrification procedure. Advantages of this approach, compared to manual operation and channel-based microfluidic vitrification, include automated operation, cryoprotectant concentration gradient generation, and feasibility of loading and retrieval of embryos.

  12. Niv versus dropping vitrification in cryopreservation of human ovarian tissue.

    PubMed

    Xiao, Z; Li, S W; Zhang, Y Y; Wang, Y; Li, L L; Fan, W

    2014-01-01

    The containers for vitrification of tissues include cryovials, copper grids, Pasteur pipettes, the solid-surface method and etc. Recently the acupuncture needle was used to achieve better result in vitrification of human ovarian tissue. To determine if the needle immersed vitrification method (NIV) is a promising approach to vitrify the human ovarian tissue. Human ovarian biopsies from five patients were vitrified using NIV and Dropping vitrification. After 14 days of in vitro culture, the incidence of apoptotic primordial follicles from fresh and vitrified groups was assessed by TUNEL assay. 17β-estradiol (E2) and progesterone (P4) were detected in the media after culturing of vitrified and fresh ovarian tissues. The incidence of apoptotic primordial follicles was significantly higher in the dropping vitrification group than in the NIV group (P < 0.05). E2 and P4 concentrations were significantly higher in NIV groups than in Dropping vitrification group (P < 0.05). NIV was an appropriate method to vitrify ovarian tissue by improving the growth potential of frozen-warmed ovarian tissue in vitro culture.

  13. Vitrification and levitation of a liquid droplet on liquid nitrogen.

    PubMed

    Song, Young S; Adler, Douglas; Xu, Feng; Kayaalp, Emre; Nureddin, Aida; Anchan, Raymond M; Maas, Richard L; Demirci, Utkan

    2010-03-09

    The vitrification of a liquid occurs when ice crystal formation is prevented in the cryogenic environment through ultrarapid cooling. In general, vitrification entails a large temperature difference between the liquid and its surrounding medium. In our droplet vitrification experiments, we observed that such vitrification events are accompanied by a Leidenfrost phenomenon, which impedes the heat transfer to cool the liquid, when the liquid droplet comes into direct contact with liquid nitrogen. This is distinct from the more generally observed Leidenfrost phenomenon that occurs when a liquid droplet is self-vaporized on a hot plate. In the case of rapid cooling, the phase transition from liquid to vitrified solid (i.e., vitrification) and the levitation of droplets on liquid nitrogen (i.e., Leidenfrost phenomenon) take place simultaneously. Here, we investigate these two simultaneous physical events by using a theoretical model containing three dimensionless parameters (i.e., Stefan, Biot, and Fourier numbers). We explain theoretically and observe experimentally a threshold droplet radius during the vitrification of a cryoprotectant droplet in the presence of the Leidenfrost effect.

  14. Vitrification and levitation of a liquid droplet on liquid nitrogen

    PubMed Central

    Song, Young S.; Adler, Douglas; Xu, Feng; Kayaalp, Emre; Nureddin, Aida; Anchan, Raymond M.; Maas, Richard L.; Demirci, Utkan

    2010-01-01

    The vitrification of a liquid occurs when ice crystal formation is prevented in the cryogenic environment through ultrarapid cooling. In general, vitrification entails a large temperature difference between the liquid and its surrounding medium. In our droplet vitrification experiments, we observed that such vitrification events are accompanied by a Leidenfrost phenomenon, which impedes the heat transfer to cool the liquid, when the liquid droplet comes into direct contact with liquid nitrogen. This is distinct from the more generally observed Leidenfrost phenomenon that occurs when a liquid droplet is self-vaporized on a hot plate. In the case of rapid cooling, the phase transition from liquid to vitrified solid (i.e., vitrification) and the levitation of droplets on liquid nitrogen (i.e., Leidenfrost phenomenon) take place simultaneously. Here, we investigate these two simultaneous physical events by using a theoretical model containing three dimensionless parameters (i.e., Stefan, Biot, and Fourier numbers). We explain theoretically and observe experimentally a threshold droplet radius during the vitrification of a cryoprotectant droplet in the presence of the Leidenfrost effect. PMID:20176969

  15. Tc removal from the waste treatment and immobilization plant low-activity waste vitrification off-gas recycle

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

    Taylor-Pashow, Kathryn M. L.; McCabe, Daniel J.; Nash, Charles A.

    Vitrification of Low Activity Waste in the Hanford Waste Treatment and Immobilization Plant generates a condensate stream from the off-gas processes. Components in this stream are partially volatile and accumulate to high concentrations through recycling, which impacts the waste glass loading and facility throughput. The primary radionuclide that vaporizes and accumulates in the stream is 99Tc. This program is investigating Tc removal via reductive precipitation with stannous chloride to examine the potential for diverting this stream to an alternate disposition path. As a result, research has shown stannous chloride to be effective, and this paper describes results of recent experimentsmore » performed to further mature the technology.« less

  16. Tc removal from the waste treatment and immobilization plant low-activity waste vitrification off-gas recycle

    DOE PAGES

    Taylor-Pashow, Kathryn M. L.; McCabe, Daniel J.; Nash, Charles A.

    2017-03-16

    Vitrification of Low Activity Waste in the Hanford Waste Treatment and Immobilization Plant generates a condensate stream from the off-gas processes. Components in this stream are partially volatile and accumulate to high concentrations through recycling, which impacts the waste glass loading and facility throughput. The primary radionuclide that vaporizes and accumulates in the stream is 99Tc. This program is investigating Tc removal via reductive precipitation with stannous chloride to examine the potential for diverting this stream to an alternate disposition path. As a result, research has shown stannous chloride to be effective, and this paper describes results of recent experimentsmore » performed to further mature the technology.« less

  17. The effect of minimal concentration of ethylene glycol (EG) combined with polyvinylpyrrolidone (PVP) on mouse oocyte survival and subsequent embryonic development following vitrification.

    PubMed

    Wang, Yao; Okitsu, Osamu; Zhao, Xiao-Ming; Sun, Yun; Di, Wen; Chian, Ri-Cheng

    2014-01-01

    Vitrification techniques employ a relatively high concentration of cryoprotectant in vitrification solutions. Exposure of oocytes to high concentrations of cryoprotectant is known to damage the oocytes via both cytotoxic and osmotic effects. Therefore, the key to successful vitrification of oocytes is to strike a balance between the usage of minimal concentration of cryoprotectant without compromising their cryoprotective actions. The minimal concentration of ethylene glycol (EG) on mouse oocyte survival and subsequent embryonic development was evaluated following vitrification-warming and parthenogenetic activation. Polyvinylpyrrolidone (PVP) combined with EG on mouse oocyte survival and subsequent embryonic development as well as morphology of the spindle and chromosome alignment were also evaluated. Vitrification system was adapted with JY Straw and the cooling rate was approximately 442-500 °C/min. In contrast, the warming rate was approximately 2,210-2,652 °C/min. Survival rate of oocytes increased significantly when 15 % EG was combined with 2 % PVP in vitrification solution (VS). The effect of combination of EG and PVP was not significant when the concentration of EG was 20 % and higher. Although there were no significant differences in embryonic development, the percentage of abnormal spindle and chromosome alignment was significantly higher in the oocytes without 2 % PVP in VS. Our data provide a proof of principle for oocyte vitrification that may not require a high concentration of cryoprotectant. There are synergic effects of EG combined with PVP for oocyte vitrification, which may provide important information to the field in developing less cytotoxic VS.

  18. Cellular damage suffered by equine embryos after exposure to cryoprotectants or cryopreservation by slow-freezing or vitrification.

    PubMed

    Hendriks, W K; Roelen, B A J; Colenbrander, B; Stout, T A E

    2015-11-01

    Equine embryos are cryopreserved by slow-freezing or vitrification. While small embryos (<300 μm) survive cryopreservation reasonably well, larger embryos do not. It is not clear if slow-freezing or vitrification is less damaging to horse embryos. To compare the type and extent of cellular damage suffered by small and large embryos during cryopreservation by slow-freezing vs. vitrification. Sixty-three Day 6.5-7 embryos were subdivided by size and assigned to one of 5 treatments: control, exposure to slow-freezing or vitrification cryoprotectants (CPs), and cryopreservation by either technique. After thawing/CP removal, embryos were stained with fluorescent stains for various parameters of cellular integrity, and assessed by multiphoton microscopy. Exposing large embryos to vitrification CPs resulted in more dead cells (6.8 ± 1.3%: 95% confidence interval [CI], 3.1-10.4%) than exposure to slow-freezing media (0.3 ± 0.1%; 95% CI 0.0-0.6%: P = 0.001). Cryopreservation by either technique induced cell death and cytoskeleton disruption. Vitrification of small embryos resulted in a higher proportion of cells with fragmented or condensed (apoptotic) nuclei (P = 0.002) than slow-freezing (6.7 ± 1.5%, 95% CI 3.0-10.4% vs. 5.0 ± 2.1%, 95% CI 4.0-14.0%). Slow-freezing resulted in a higher incidence of disintegrated embryos (P = 0.01) than vitrification. Mitochondrial activity was low in control embryos, and was not differentially affected by cryopreservation technique, whereas vitrification changed mitochondrial distribution from a homogenous crystalline pattern in control embryos to a heterogeneous granulated distribution in vitrified embryos (P = 0.05). Cryopreservation caused more cellular damage to large embryos than smaller ones. While vitrification is more practical, it is not advisable for large embryos due to a higher incidence of dead cells. The choice is less obvious for small embryos, as vitrification led to occasionally very high percentages of dead or damaged cells, but a lower incidence of embryo disintegration. Modifications that reduce the level of cellular damage induced by vitrification are required before it can be considered the method of choice for cryopreserving equine embryos. © 2014 EVJ Ltd.

  19. Label-free characterization of vitrification-induced morphology changes in single-cell embryos with full-field optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Zarnescu, Livia; Leung, Michael C.; Abeyta, Michael; Sudkamp, Helge; Baer, Thomas; Behr, Barry; Ellerbee, Audrey K.

    2015-09-01

    Vitrification is an increasingly popular method of embryo cryopreservation that is used in assisted reproductive technology. Although vitrification has high post-thaw survival rates compared to other freezing techniques, its long-term effects on embryo development are still poorly understood. We demonstrate an application of full-field optical coherence tomography (FF-OCT) to visualize the effects of vitrification on live single-cell (2 pronuclear) mouse embryos without harmful labels. Using FF-OCT, we observed that vitrification causes a significant increase in the aggregation of structures within the embryo cytoplasm, consistent with reports in literature based on fluorescence techniques. We quantify the degree of aggregation with an objective metric, the cytoplasmic aggregation (CA) score, and observe a high degree of correlation between the CA scores of FF-OCT images of embryos and of fluorescence images of their mitochondria. Our results indicate that FF-OCT shows promise as a label-free assessment of the effects of vitrification on embryo mitochondria distribution. The CA score provides a quantitative metric to describe the degree to which embryos have been affected by vitrification and could aid clinicians in selecting embryos for transfer.

  20. Cryotop vitrification as compared to conventional slow freezing for human embryos at the cleavage stage: survival and outcomes.

    PubMed

    Lin, Tseng-Kai; Su, Jin-Tsung; Lee, Fa-Kung; Lin, Yu-Ru; Lo, Hsiao-Ching

    2010-09-01

    This study was conducted to compare the efficacy of cryotop vitrification of human cleavage-stage embryos to that of conventional slow freezing of these embryos with respect to survival. A second objective was to compare the two cryopreservation techniques with respect to outcomes for a cohort of women. Cleavage-stage embryos from 102 patients were cryopreserved either by vitrification (57 patients) or by traditional slow freezing (45 patients). After thawing, rates of embryo survival, implantation, and clinical pregnancy were determined. Survival of embryos was significantly higher with the vitrification procedure as compared to traditional slow freezing [287/298 (96.3%) vs. 294/446 (65.9%); p < 0.05). Rates of implantation and clinical pregnancy were also significantly higher using vitrification procedure as compared to the slow freezing procedure (24.3% vs. 7.1% and 35.6% vs. 15.6% respectively, p < 0.05). As compared to conventional slow freezing, cryopreservation of human cleavage-stage embryo using vitrification results in higher rates of embryo survival, implantation, and clinical pregnancy. Vitrification therefore represents the superior cryopreservation technique for cleavage-stage embryos. Copyright © 2010 Taiwan Association of Obstetric & Gynecology. Published by Elsevier B.V. All rights reserved.

  1. RNA-Seq transcriptome profiling of mouse oocytes after in vitro maturation and/or vitrification.

    PubMed

    Gao, Lei; Jia, Gongxue; Li, Ai; Ma, Haojia; Huang, Zhengyuan; Zhu, Shien; Hou, Yunpeng; Fu, Xiangwei

    2017-10-16

    In vitro maturation (IVM) and vitrification have been widely used to prepare oocytes before fertilization; however, potential effects of these procedures, such as expression profile changes, are poorly understood. In this study, mouse oocytes were divided into four groups and subjected to combinations of in vitro maturation and/or vitrification treatments. RNA-seq and in silico pathway analysis were used to identify differentially expressed genes (DEGs) that may be involved in oocyte viability after in vitro maturation and/or vitrification. Our results showed that 1) 69 genes were differentially expressed after IVM, 66 of which were up-regulated. Atp5e and Atp5o were enriched in the most significant gene ontology term "mitochondrial membrane part"; thus, these genes may be promising candidate biomarkers for oocyte viability after IVM. 2) The influence of vitrification on the transcriptome of oocytes was negligible, as no DEGs were found between vitrified and fresh oocytes. 3) The MII stage is more suitable for oocyte vitrification with respect to the transcriptome. This study provides a valuable new theoretical basis to further improve the efficiency of in vitro maturation and/or oocyte vitrification.

  2. Cryopreservation of in vitro grown shoot tips of Diospyros kaki thunb. using different methods.

    PubMed

    Niu, Y L; Luo, Z R; Zhang, Y F; Zhang, Q L

    2012-01-01

    The objective of this study was to compare the potential of different cryopreservation strategies for in vitro shoot tips of Diospyros kaki Thunb. The treatments consisted of three different cryopreservation methods: vitrification, droplet-vitrification and modified droplet-vitrification. The following variables were assessed: cold acclimation, sucrose concentration in the preculture medium and PVS2 treatment time. A higher average survival level was obtained using the modified droplet-vitrification method compared to the other two methods.

  3. Vitrification of zona-free rabbit expanded or hatching blastocysts: a possible model for human blastocysts.

    PubMed

    Cervera, R P; Garcia-Ximénez, F

    2003-10-01

    The purpose of this study was to test the effectiveness of one two-step (A) and two one-step (B1 and B2) vitrification procedures on denuded expanded or hatching rabbit blastocysts held in standard sealed plastic straws as a possible model for human blastocysts. The effect of blastocyst size was also studied on the basis of three size categories (I: diameter <200 micro m; II: diameter 200-299 micro m; III: diameter >/==" BORDER="0">300 micro m). Rabbit expanded or hatching blastocysts were vitrified at day 4 or 5. Before vitrification, the zona pellucida was removed using acidic phosphate buffered saline. For the two-step procedure, prior to vitrification, blastocysts were pre- equilibrated in a solution containing 10% dimethyl sulphoxide (DMSO) and 10% ethylene glycol (EG) for 1 min. Different final vitrification solutions were compared: 20% DMSO and 20% EG with (A and B1) or without (B2) 0.5 mol/l sucrose. Of 198 vitrified blastocysts, 181 (91%) survived, regardless of the vitrification procedure applied. Vitrification procedure A showed significantly higher re-expansion (88%), attachment (86%) and trophectoderm outgrowth (80%) rates than the two one-step vitrification procedures, B1 and B2 (46 and 21%, 20 and 33%, and 18 and 23%, respectively). After warming, blastocysts of greater size (II and III) showed significantly higher attachment (54 and 64%) and trophectoderm outgrowth (44 and 58%) rates than smaller blastocysts (I, attachment: 29%; trophectoderm outgrowth: 25%). These result demonstrate that denuded expanded or hatching rabbit blastocysts of greater size can be satisfactorily vitrified by use of a two-step procedure. The similarity of vitrification solutions used in humans could make it feasible to test such a procedure on human denuded blastocysts of different sizes.

  4. Method for initiating in-situ vitrification using an impregnated cord

    DOEpatents

    Carter, John G.

    1991-01-01

    In-situ vitrification of soil is initiated by placing a cord of dielectric material impregnated with conductive material in thermally-conductive contact with the soil, and energizing the cord with an electric current for heating the cord and starting the vitrification process.

  5. Cryoprotectant-free vitrification of human spermatozoa in new artificial seminal fluid.

    PubMed

    Agha-Rahimi, A; Khalili, M A; Nottola, S A; Miglietta, S; Moradi, A

    2016-11-01

    Vitrification is a new method that has been recently introduced in Assisted Reproduction Technique programs. The aim of this study was to design a new medium similar to normal human seminal fluid (SF), formulation artificial seminal fluid (ASF), and to compare the cryoprotective potency of this medium with SF and human tubal fluid (HTF) medium. Thirty normal ejaculates were processed with the swim-up technique and sperm suspensions were divided into four aliquots: (i) fresh sample (control); (ii) vitrification in HTF medium supplemented with 5 mg/mL human serum albumin and 0.25 mol sucrose (Vit HTF); (iii) vitrification with patients' SF (Vit SF); and (iv) vitrification in ASF (Vit ASF). After warming, sperm parameters of motility, viability, and morphology were analyzed using WHO criteria. Also, sperm pellets were fixed in 2.5% glutaraldehyde and processed for scanning electron microscopy and transmission electron microscopy observations. The results showed that progressive motility (46.09 ± 10.33 vs. 36.80 ± 13.75), grade A motility (36.59 ± 11.40 vs. 16.41 ± 11.24), and normal morphology (18.74 ± 8.35 vs. 11.85 ± 5.84) and viability (68.22 ± 10.83 vs. 60.86 ± 11.72) of spermatozoa were significantly higher in Vit ASF than in Vit HTF. All parameters were better in Vit ASF than in Vit SF, but only viability was significantly different (p = 0.006). After cryopreservation, deep invagination in cytoplasm and mechanically weak point sites and folded tail were commonly observed. But, this phenomenon was more significant in Vit HTF and Vit SF than in ASF (p < 0.05). In transmission electron microscopy evaluation, acrosome damage, plasma membrane loss, chromatin vacuolation, and disruption of mitochondria arrangement and structures were observed in all vitrified groups. Adherence of several tail sections together was also seen in all cryo groups. But this was seen more in Vit HTF and Vit SF than in ASF (p < 0.05). In conclusion, vitrification of human spermatozoa with ASF can effectively preserve the quality of sperm motility in comparison with Vit HTF. © 2016 American Society of Andrology and European Academy of Andrology.

  6. Nanoliter droplet vitrification for oocyte cryopreservation.

    PubMed

    Zhang, Xiaohui; Khimji, Imran; Shao, Lei; Safaee, Hooman; Desai, Khanjan; Keles, Hasan Onur; Gurkan, Umut Atakan; Kayaalp, Emre; Nureddin, Aida; Anchan, Raymond M; Maas, Richard L; Demirci, Utkan

    2012-04-01

    Oocyte cryopreservation remains largely experimental, with live birth rates of only 2-4% per thawed oocyte. In this study, we present a nanoliter droplet technology for oocyte vitrification. An ejector-based droplet vitrification system was designed to continuously cryopreserve oocytes in nanoliter droplets. Oocyte survival rates, morphologies and parthenogenetic development after each vitrification step were assessed in comparison with fresh oocytes. Oocytes were retrieved after cryoprotectant agent loading/unloading, and nanoliter droplet encapsulation showed comparable survival rates to fresh oocytes after 24 h in culture. Also, oocytes recovered after vitrification/thawing showed similar morphologies to those of fresh oocytes. Additionally, the rate of oocyte parthenogenetic activation after nanoliter droplet encapsulation was comparable with that observed for fresh oocytes. This nanoliter droplet technology enables the vitrification of oocytes at higher cooling and warming rates using lower cryoprotectant agent levels (i.e., 1.4 M ethylene glycol, 1.1 M dimethyl sulfoxide and 1 M sucrose), thus making it a potential technology to improve oocyte cryopreservation outcomes.

  7. Nanoliter droplet vitrification for oocyte cryopreservation

    PubMed Central

    Zhang, Xiaohui; Khimji, Imran; Shao, Lei; Safaee, Hooman; Desai, Khanjan; Keles, Hasan Onur; Gurkan, Umut Atakan; Kayaalp, Emre; Nureddin, Aida; Anchan, Raymond M; Maas, Richard L; Demirci, Utkan

    2011-01-01

    Aim Oocyte cryopreservation remains largely experimental, with live birth rates of only 2–4% per thawed oocyte. In this study, we present a nanoliter droplet technology for oocyte vitrification. Materials & methods An ejector-based droplet vitrification system was designed to continuously cryopreserve oocytes in nanoliter droplets. Oocyte survival rates, morphologies and parthenogenetic development after each vitrification step were assessed in comparison with fresh oocytes. Results Oocytes were retrieved after cryoprotectant agent loading/unloading, and nanoliter droplet encapsulation showed comparable survival rates to fresh oocytes after 24 h in culture. Also, oocytes recovered after vitrification/thawing showed similar morphologies to those of fresh oocytes. Additionally, the rate of oocyte parthenogenetic activation after nanoliter droplet encapsulation was comparable with that observed for fresh oocytes. This nanoliter droplet technology enables the vitrification of oocytes at higher cooling and warming rates using lower cryoprotectant agent levels (i.e., 1.4 M ethylene glycol, 1.1 M dimethyl sulfoxide and 1 M sucrose), thus making it a potential technology to improve oocyte cryopreservation outcomes. PMID:22188180

  8. Cryopreservation of redwood (Sequoia sempervirens) in vitro buds using vitrification-based techniques.

    PubMed

    Ozudogru, E A; Kirdok, E; Kaya, E; Capuana, M; Benelli, C; Engelmann, E

    2011-01-01

    In this study, the efficiency of three vitrification-based cryopreservation techniques, i.e. vitrification, encapsulation-vitrification and droplet-vitrification were compared for cryopreserving Sequoia sempervirens apical and basal buds sampled from in vitro shoot cultures. The effect of cold-hardening of mother-plants and of bud culture medium and sucrose preculture was also investigated. Culture of apical and basal buds sampled from cold-hardened mother-plants on Quoirin and Lepoivre medium with activated charcoal had a positive effect on regrowth. Only droplet-vitrification ensured survival and regrowth after cryopreservation. After cryopreservation, regeneration of apical buds was possible for PVS2 exposure durations between 90 and 180 min but it remained low, with a maximum of 18 percent after 135 min treatment. With basal buds, regeneration after cryopreservation was possible over a larger range of PVS2 treatment durations, between 30 and 180 min. The highest regeneration percentage was slightly higher (22 percent) than that measured with apical buds, and was also achieved after 135 min PVS2 exposure.

  9. ANNUAL RADIOACTIVE WASTE TANK INSPECTION PROGRAM 2008

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

    West, B.; Waltz, R.

    2009-06-11

    Aqueous radioactive wastes from Savannah River Site (SRS) separations and vitrification processes are contained in large underground carbon steel tanks. Inspections made during 2008 to evaluate these vessels and other waste handling facilities along with evaluations based on data from previous inspections are the subject of this report.

  10. Method for initiating in-situ vitrification using an impregnated cord

    DOEpatents

    Carter, J.G.

    1991-04-02

    In-situ vitrification of soil is initiated by placing a cord of dielectric material impregnated with conductive material in thermally-conductive contact with the soil, and energizing the cord with an electric current for heating the cord and starting the vitrification process. 1 figure.

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

    USDA-ARS?s Scientific Manuscript database

    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...

  12. Comparison of survival rate of cleavage stage embryos produced from in vitro maturation cycles after slow freezing and after vitrification.

    PubMed

    Son, Weon-Young; Chung, Jin-Tae; Gidoni, Yariv; Holzer, Hananel; Levin, Dan; Chian, Ri-Cheng; Tan, Seang Lin

    2009-09-01

    Significantly more embryos survived the vitrification procedure compared to slow freezing (85.5% vs. 61.8%) in cleavage-stage human embryos produced from in vitro maturation cycles, suggesting that vitrification is more efficient than slow freezing for cryopreservation.

  13. Vitrification of plutonium at Rocky Flats the argument for a pilot plant

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

    Moore, L.

    1996-05-01

    Current plans for stabilizing and storing the plutonium at Rocky Flats Plant fail to put the material in a form suitable for disposition and resistant to proliferation. Vitrification should be considered as an alternate technology. The vitrification should begin with a small-scale pilot plant.

  14. High-Throughput Non-Contact Vitrification of Cell-Laden Droplets Based on Cell Printing

    NASA Astrophysics Data System (ADS)

    Shi, Meng; Ling, Kai; Yong, Kar Wey; Li, Yuhui; Feng, Shangsheng; Zhang, Xiaohui; Pingguan-Murphy, Belinda; Lu, Tian Jian; Xu, Feng

    2015-12-01

    Cryopreservation is the most promising way for long-term storage of biological samples e.g., single cells and cellular structures. Among various cryopreservation methods, vitrification is advantageous by employing high cooling rate to avoid the formation of harmful ice crystals in cells. Most existing vitrification methods adopt direct contact of cells with liquid nitrogen to obtain high cooling rates, which however causes the potential contamination and difficult cell collection. To address these limitations, we developed a non-contact vitrification device based on an ultra-thin freezing film to achieve high cooling/warming rate and avoid direct contact between cells and liquid nitrogen. A high-throughput cell printer was employed to rapidly generate uniform cell-laden microdroplets into the device, where the microdroplets were hung on one side of the film and then vitrified by pouring the liquid nitrogen onto the other side via boiling heat transfer. Through theoretical and experimental studies on vitrification processes, we demonstrated that our device offers a high cooling/warming rate for vitrification of the NIH 3T3 cells and human adipose-derived stem cells (hASCs) with maintained cell viability and differentiation potential. This non-contact vitrification device provides a novel and effective way to cryopreserve cells at high throughput and avoid the contamination and collection problems.

  15. High-Throughput Non-Contact Vitrification of Cell-Laden Droplets Based on Cell Printing

    PubMed Central

    Shi, Meng; Ling, Kai; Yong, Kar Wey; Li, Yuhui; Feng, Shangsheng; Zhang, Xiaohui; Pingguan-Murphy, Belinda; Lu, Tian Jian; Xu, Feng

    2015-01-01

    Cryopreservation is the most promising way for long-term storage of biological samples e.g., single cells and cellular structures. Among various cryopreservation methods, vitrification is advantageous by employing high cooling rate to avoid the formation of harmful ice crystals in cells. Most existing vitrification methods adopt direct contact of cells with liquid nitrogen to obtain high cooling rates, which however causes the potential contamination and difficult cell collection. To address these limitations, we developed a non-contact vitrification device based on an ultra-thin freezing film to achieve high cooling/warming rate and avoid direct contact between cells and liquid nitrogen. A high-throughput cell printer was employed to rapidly generate uniform cell-laden microdroplets into the device, where the microdroplets were hung on one side of the film and then vitrified by pouring the liquid nitrogen onto the other side via boiling heat transfer. Through theoretical and experimental studies on vitrification processes, we demonstrated that our device offers a high cooling/warming rate for vitrification of the NIH 3T3 cells and human adipose-derived stem cells (hASCs) with maintained cell viability and differentiation potential. This non-contact vitrification device provides a novel and effective way to cryopreserve cells at high throughput and avoid the contamination and collection problems. PMID:26655688

  16. Preparation of glass-forming materials from granulated blast furnace slag

    NASA Astrophysics Data System (ADS)

    Alonso, M.; Sáinz, E.; Lopez, F. A.

    1996-10-01

    Glass precursor materials, to be used for the vitrification of hazardous wastes, have been prepared from blast furnace slag powder through a sol-gel route. The slag is initially reacted with a mixture of alcohol (ethanol or methanol) and mineral acid (HNO3 or H2SO4) to give a sol principally consisting of Si, Ca, Al, and Mg alkoxides. Gelation is carried out with variable amounts of either ammonia or water. The gelation rate can be made as fast as desired by adding excess hydrolizing agent or else by distilling the excess alcohol out of the alkoxide solution. The resulting gel is first dried at low temperature and ground. The powder thus obtained is then heat treated at several temperatures. The intermediate and final materials are characterized by thermal analysis, infrared (IR) spectroscopy, X-ray diffraction, scanning electron microscopy (SEM), and chemical analysis. From the results, the operating conditions yielding a variety of glass precursors differing in their composition are established. The method, in comparison with direct vitrification of slag, presents a number of advantages: (1) the glass precursor obtained devitrifies at higher temperatures; (2) it enables the adjustment, to a certain extent, of the chemical composition of the glass precursor; and (3) it permits recovering marketable materials at different stages of the process.

  17. Cryotolerance of Day 2 or Day 6 in vitro produced ovine embryos after vitrification by Cryotop or Spatula methods.

    PubMed

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

    2015-02-01

    This study was conducted to evaluate the cryotolerance of in vitro produced ovine embryos submitted to vitrification at different developmental stages using two methods of minimum volume and rapid cooling rate. Embryos were vitrified at early stage (2 to 8-cells) on Day 2 or at advanced stage (morulae and blastocysts) on Day 6 after in vitro fertilization. Vitrification procedure consisted of the Cryotop (Day 2, n=165; Day 6, n=174) or the Spatula method (Day 2, n=165; Day 6, n=175). Non vitrified embryos were maintained in in vitro culture as a control group (n=408). Embryo survival was determined at 3h and 24h after warming, development and hatching rates were evaluated on Day 6 and Day 8 after fertilization, and total cell number was determined on expanded blastocysts. Embryo survival at 24h after warming increased as the developmental stage progressed (P<0.05) and was not affected by the vitrification method. The ability for hatching of survived embryos was not affected by the stage of the embryos at vitrification or by the vitrification method. Thus, the proportion of hatching from vitrified embryos was determined by the survival rate and was lower for Day 2 than Day 6 vitrified embryos. The percentage of blastocysts on Day 8 was lower for the embryos vitrified on Day 2 than Day 6 (P<0.05), and was lower for both days of vitrification than for non-vitrified embryos (P<0.05). No interaction of embryo stage by vitrification method was found (P=NS) and no significant difference was found in the blastocyst cell number among vitrified and non-vitrified embryos. In conclusion, both methods using minimum volume and ultra-rapid cooling rate allow acceptable survival and development rates in Day 2 and Day 6 in vitro produced embryos in sheep. Even though early stage embryos showed lower cryotolerance, those embryos that survive the vitrification-warming process show high development and hatching rates, similar to vitrification of morulae or blastocysts. Copyright © 2014 Elsevier Inc. All rights reserved.

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

    PubMed

    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 MIH-enhanced rewarming. Therefore, the novel MIH-enhanced cell vitrification is valuable to facilitate the long-term storage of hUCM-MSCs and possibly many other important cells to meet their ever-increasing demand by the burgeoning cell-based medicine. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  19. Successful pregnancies from vitrified embryos in the dromedary camel: Avoidance of a possible toxic effect of sucrose on embryos.

    PubMed

    Herrid, M; Billah, M; Skidmore, J A

    2017-12-01

    Successful embryo cryopreservation facilitates the wider application of assisted reproduction technologies and also provides a useful method for gene banking of valuable genetics. Unfortunately attempts to establish an effective cryopreservation protocol for camelid embryos have been unsuccessful. In the current study, a modified vitrification protocol with three steps was investigated, whereby embryos were exposed to solutions containing increasing amounts of glycerol and ethylene glycol for fixed time periods. Embryos were then loaded into an Open Pull Straw (OPS) and plunged directly into liquid nitrogen for storage. Three experiments were designed to investigate the effect of 1) artificial shrinkage (AS) of embryos, 2) the addition of sucrose to the vitrification solutions, and 3) the replacement of sucrose by galactose in the warming solution, on the outcome of vitrification. The results showed that neither AS of hatched embryos prior to vitrification, nor the addition of sucrose into vitrification solutions improves the outcome of vitrification, while replacement of sucrose with galactose in warming solution increases the survival and developmental rates of vitrified embryos in culture. Transfer of vitrified embryos that were warmed in galactose resulted in a pregnancy rate of 42.8% per embryo or 46.1% per recipient. Collectively, these results suggest a possible species-specific toxic effect of sucrose on camel embryos, and that avoiding its use either in vitrification or warming solution is critical for establishing an effective protocol. This study may also be applicable to the vitrification of embryos of other camelid species including alpaca and llamas. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. The evaluation of xenotransplantation of feline ovarian tissue vitrified by needle immersed vitrification technique into male immunodeficient mice.

    PubMed

    Demirel, Mürşide Ayşe; Acar, Duygu Baki; Ekim, Burcu; Çelikkan, Ferda Topal; Alkan, Kübra Karakaş; Salar, Seçkin; Erdemli, Esra Atabenli; Özkavukçu, Sinan; Yar, Seda Sağlam; Kanca, Halit; Baştan, Ayhan

    2018-03-01

    In this study, the efficiency of the "Needle Immersed Vitrification" technique was tested on cryopreserved feline ovarian tissue. For vitrification, ovarian fragments (0.5-1.5 mm 2 ) from each ovary were collected; the grafts were exposed to 7.5-15% ethylene glycol and 7.5-15% dimethyl sulfoxide at room temperature and stored in liquid nitrogen at least 1 week. Morphologic examinations, expression of genes such as B cell lymphoma 2, B-cell lymphoma-2-associated X protein, Bone morphogenetic protein 15, zone of polarizing activity, zona pellucida C protein and DNA (cytosine-5)-methyltransferase 1, ultrastructural analysis and viability tests were carried out from collected grafts. Light microscopy examinations revealed the percentage of morphologically normal primordial follicles in a fresh group which was significantly higher than the treatment groups (p < 0.001). Terminal deoxynucleotidyl transferase dUTP nick end labeling and anti-caspase-3 staining observed in oocytes, follicle cells, interstitial tissue showed higher rates of apoptosis for post-vitrification and -transplantation groups than freshly grafted ovarian tissues. Furthermore, we observed significant downregulation of zone of polarizing activity and zona pellucida C protein gene expression in vitrified ovarian tissue grafts than in the fresh grafts (p < 0.05). In conclusion, we suggest that the needle immersed vitrification method is a convenient, cheap, and feasible vitrification method for cat ovarian tissues. However, further studies need to be performed to determine more optimal vitrification solutions and equilibration times for the needle immersed vitrification method in order to adapt it for cat ovaries.

  1. In Vitro Cryopreservation of Date Palm Caulogenic Meristems.

    PubMed

    Fki, Lotfi; Chkir, Olfa; Kriaa, Walid; Nasri, Ameni; Baklouti, Emna; Masmoudi, Raja B; Rival, Alain; Drira, Noureddine; Panis, Bart

    2017-01-01

    Cryopreservation is the technology of choice not only for plant genetic resource preservation but also for virus eradication and for the efficient management of large-scale micropropagation. In this chapter, we describe three cryopreservation protocols (standard vitrification, droplet vitrification, and encapsulation vitrification) for date palm highly proliferating meristems that are initiated from vitro-cultures using plant growth regulator-free MS medium. The positive impact of sucrose preculture and cold hardening treatments on survival rates is significant. Regeneration rates obtained with standard vitrification, encapsulation-vitrification, and droplet-vitrification protocols can reach 30, 40, and 70%, respectively. All regenerated plants from non-cryopreserved or cryopreserved explants don't show morphological variation by maintaining genetic integrity without adverse effect of cryogenic treatment. Cryopreservation of date palm vitro-cultures enables commercial tissue culture laboratories to move to large-scale propagation from cryopreserved cell lines producing true-to-type plants after clonal field-testing trials. When comparing the cost of cryostorage and in-field conservation of date palm cultivars, tissue cryopreservation is the most cost-effective. Moreover, many of the risks linked to field conservation like erosion due to climatic, edaphic, and phytopathologic constraints are circumvented.

  2. Numerical investigations of transient heat transfer characteristics and vitrification tendencies in ultra-fast cell cooling processes.

    PubMed

    Jiao, Anjun; Han, Xu; Critser, John K; Ma, Hongbin

    2006-06-01

    During freezing, cells are often damaged directly or indirectly by ice formation. Vitrification is an alternative approach to cryopreservation that avoids ice formation. The common method to achieve vitrification is to use relatively high concentrations of cryoprotectant agents (CPA) in combination with a relatively slow cooling rate. However, high concentrations of CPAs have potentially damaging toxic and/or osmotic effects on cells. Therefore, establishing methods to achieve vitrification with lower concentrations of CPAs through ultra-fast cooling rates would be advantageous in these aspects. These ultra-fast cooling rates can be realized by a cooling system with an ultra-high heat transfer coefficient (h) between the sample and coolant. The oscillating motion heat pipe (OHP), a novel cooling device utilizing the pressure change to excite the oscillation motion of the liquid plugs and vapor bubbles, can significantly increase h and may fulfill this aim. The current investigation was designed to numerically study the effects of different values of h on the transient heat transfer characteristics and vitrification tendencies of the cell suspension during the cooling processes in an ultra-thin straw (100 microm in diameter). The transient temperature distribution, the cooling rate and the volume ratio (x) of the ice quantity to the maximum crystallizable ice of the suspension were calculated. From these numerical results, it is concluded that the ultra-high h (>10(4) W/m2 K) obtained by OHPs could facilitate vitrification by efficiently decreasing x as well as the time to pass through the dangerous temperature region where the maximum ice formation happens. For comparison, OHPs can decrease both of the parameters to less than 20% of those from the widely used open pulled straw methods. Therefore, the OHP method will be a promising approach to improving vitrification tendencies of CPA solutions and could also decrease the required concentration of CPAs for vitrification, both of which are of great importance for the successful cryopreservation of cells by vitrification.

  3. Comparison of sucrose and trehalose media modification as an update of oocyte vitrification: A study of apoptotic level

    NASA Astrophysics Data System (ADS)

    Lestari, Silvia W.; Fitriyah, Nurin N.; Pangestu, Mulyoto; Pratama, Gita; Margiana, Ria

    2018-02-01

    Number of women who are not being able to have offspring in their reproductive life is increasing which might be influenced by several factors. As a consequence, oocyte cryopreservation could be an ensuring solution for women fertility preservation. A good vitrification could be conducted by combining an appropriate of type and concentration of cryoprotectants. One of the marks of successful vitrification is the vitrified oocytes could avoid apoptosis. This study aimed to evaluate the modification of cryoprotectant media as un update of oocyte vitrification as follow: the combination and the concentration of cryoprotectant media of oocytes vitrification, based on their effects on the apoptosis or DNA damage of oocytes. A total of 84 MII stage oocytes from adult female Deutschland, Denken and Yoken (DDY) mice (7-8 weeks old) were used in this study. Vitrification procedure was performed by using VS1 contained 15% EG, 15% DMSO, 0.5 mol/l sucrose (Merck, Darmstadt, Germany) and VS2 contained 15% EG, 15% DMSO, 0.5 mol/l trehalose (Merck, Darmstadt, Germany) in HM. Furthermore, warming solution (WS) was divided into four groups. There were: WS1a contained 0.3 mol/l sucrose, WS1b contained 0.15 mol/l sucrose, WS2a contained 0.3 mol/l trehalose, and WS2b contained 0.15 mol/l trehalose. Apoptotic level was performed by staining the oocytes with TUNEL and propidium iodide (PI) based on Brison and Schultz method then examined under confocal microscope. The rate of apoptosis in oocytes after vitrification and warming was higher compared to the fresh control oocytes. Furthermore, the rate of apoptosis in the vitrified oocytes by sucrose media (28%) was higher compared to the vitrified oocytes by trehalose media (16%). The results of this study indicated that vitrification increased apoptosis in the vitrified oocytes related to the oocyte injury after vitrification. Moreover, the vitrification increased apoptosis more in the vitrified oocytes by sucrose media than the vitrified oocytes by trehalose media. The exposure to the 16.5% EG, 16.5% DMSO and 0.5 mol/l trehalose as cryoprotectant media decreased their viability and increased the number of DNA-fragmented nuclei in the oocytes, lesser than sucrose. Trehalose was proved to be the more suitable extracellular cryoprotectant media in oocyte vitrification based on the apoptotic level, compared to that of sucrose. A modification of cryoprotectant media as an update of oocyte vitrification consisted 0.5 mol/l trehalose concentration as extracellular cryoprotectant and combined with 16.5% EG and 16.5% DMSO as intracellular cryoprotectant has produced.

  4. Vitrification and xenografting of human ovarian tissue.

    PubMed

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

    2012-11-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2014-03-01

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

  6. Vitrification of neat semen alters sperm parameters and DNA integrity.

    PubMed

    Khalili, Mohammad Ali; Adib, Maryam; Halvaei, Iman; Nabi, Ali

    2014-05-06

    Our aim was to evaluate the effect of neat semen vitrification on human sperm vital parameters and DNA integrity in men with normal and abnormal sperm parameters. Semen samples were 17 normozoospermic samples and 17 specimens with abnormal sperm parameters. Semen analysis was performed according to World Health Organization (WHO) criteria. Then, the smear was provided from each sample and fixed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Vitrification of neat semen was done by plunging cryoloops directly into liquid nitrogen and preserved for 7 days. The samples were warmed and re-evaluated for sperm parameters as well as DNA integrity. Besides, the correlation between sperm parameters and DNA fragmentation was assessed pre- and post vitrification. Cryopreserved spermatozoa showed significant decrease in sperm motility, viability and normal morphology after thawing in both normal and abnormal semen. Also, the rate of sperm DNA fragmentation was significantly higher after vitrification compared to fresh samples in normal (24.76 ± 5.03 and 16.41 ± 4.53, P = .002) and abnormal (34.29 ± 10.02 and 23.5 ± 8.31, P < .0001), respectively. There was negative correlation between sperm motility and sperm DNA integrity in both groups after vitrification. Vitrification of neat ejaculates has negative impact on sperm parameters as well as DNA integrity, particularly among abnormal semen subjects. It is, therefore, recommend to process semen samples and vitrify the sperm pellets.

  7. Process for vitrification of contaminated sodium oxide

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

    Blair, H.T.; Mellinger, G.B.

    1983-03-01

    A glass composition was developed to accommodate 30 wt % sodium oxide and resist devitrification and leaching. An in-can melting process that is compatible with a comtaminated sodium calciner developed by Argonne National Laboratory was tested both on a laboratory and on an engineering scale and found to be viable. The Liquid Metal Fast Breeder Reactor experimental program continues to produce elemental sodium contaminated with radionuclides. This material is presently in temporary storage facilities because the current criterion will not permit alkali metals to be disposed of in shallow land burials. As a first step in treatment, Argonne National Laboratorymore » (ANL) has developed a calciner that will convert the sodium metal to an oxide. In work supported by the U.S. Department of Energy, Pacific Northwest Laboratory (PNL) is developing and demonstrating a process that is compatible with the calciner and facilities at ANL-West for incorporating sodium oxide into a glass. Glass, which normally contains sodium oxide, was chosen as the waste form because it is chemically durable and nondispersible. It is simple to produce, and the technology for incorporating nuclear wastes into glass is well developed.« less

  8. Vitrification of mouse embryo-derived ICM cells: a tool for preserving embryonic stem cell potential?

    PubMed

    Desai, Nina; Xu, Jing; Tsulaia, Tamara; Szeptycki-Lawson, Julia; AbdelHafez, Faten; Goldfarb, James; Falcone, Tommaso

    2011-02-01

    Vitrification technology presents new opportunities for preservation of embryo derived stem cells without first establishing a viable ESC line. This study tests the feasibility of cryopreserving ICM cells using vitrification. ICMs from mouse embryos were isolated and vitrified in HSV straws or on cryoloops. Upon warming, the vitrified ICMs were cultured and observed for attachment and morphology. Colonies were passaged every 3-6 days. ICMs and ICM-derived ESC colonies were tested for expression of stem cell specific markers. ICMs vitrified on both the cryoloop and the HSV straw had high survival rates. ICM derived ESCs remained undifferentiated for several passages and demonstrated expression of typical stem cell markers; SSEA-1, Sox-2, Oct 4 and alkaline phosphatase. This is the first report on successful vitrification of isolated ICMs and the subsequent derivation of ESC colonies. Vitrification of isolated ICMs is a novel approach for preservation of the "stem cell source" material.

  9. Aseptic minimum volume vitrification technique for porcine parthenogenetically activated blastocyst.

    PubMed

    Lin, Lin; Yu, Yutao; Zhang, Xiuqing; Yang, Huanming; Bolund, Lars; Callesen, Henrik; Vajta, Gábor

    2011-01-01

    Minimum volume vitrification may provide extremely high cooling and warming rates if the sample and the surrounding medium contacts directly with the respective liquid nitrogen and warming medium. However, this direct contact may result in microbial contamination. In this work, an earlier aseptic technique was applied for minimum volume vitrification. After equilibration, samples were loaded on a plastic film, immersed rapidly into factory derived, filter-sterilized liquid nitrogen, and sealed into sterile, pre-cooled straws. At warming, the straw was cut, the filmstrip was immersed into a 39 degree C warming medium, and the sample was stepwise rehydrated. Cryosurvival rates of porcine blastocysts produced by parthenogenetical activation did not differ from control, vitrified blastocysts with Cryotop. This approach can be used for minimum volume vitrification methods and may be suitable to overcome the biological dangers and legal restrictions that hamper the application of open vitrification techniques.

  10. Droplet vitrification technique for cryopreservation of different pineapple (Ananas comosus L. Merrill) accessions

    USDA-ARS?s Scientific Manuscript database

    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 ...

  11. Final Report - Crystal Settling, Redox, and High Temperature Properties of ORP HLW and LAW Glasses, VSL-09R1510-1, Rev. 0, dated 6/18/09

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

    Kruger, Albert A.; Wang, C.; Gan, H.

    2013-11-13

    The radioactive tank waste treatment programs at the U. S. Department of Energy (DOE) have featured joule heated ceramic melter technology for the vitrification of high level waste (HLW). The Hanford Tank Waste Treatment and Immobilization Plant (WTP) employs this same basic technology not only for the vitrification of HLW streams but also for the vitrification of Low Activity Waste (LAW) streams. Because of the much greater throughput rates required of the WTP as compared to the vitrification facilities at the West Valley Demonstration Project (WVDP) or the Defense Waste Processing Facility (DWPF), the WTP employs advanced joule heated meltersmore » with forced mixing of the glass pool (bubblers) to improve heat and mass transport and increase melting rates. However, for both HLW and LAW treatment, the ability to increase waste loadings offers the potential to significantly reduce the amount of glass that must be produced and disposed and, therefore, the overall project costs. This report presents the results from a study to investigate several glass property issues related to WTP HLW and LAW vitrification: crystal formation and settling in selected HLW glasses; redox behavior of vanadium and chromium in selected LAW glasses; and key high temperature thermal properties of representative HLW and LAW glasses. The work was conducted according to Test Plans that were prepared for the HLW and LAW scope, respectively. One part of this work thus addresses some of the possible detrimental effects due to considerably higher crystal content in waste glass melts and, in particular, the impact of high crystal contents on the flow property of the glass melt and the settling rate of representative crystalline phases in an environment similar to that of an idling glass melter. Characterization of vanadium redox shifts in representative WTP LAW glasses is the second focal point of this work. The third part of this work focused on key high temperature thermal properties of representative WTP HLW and LAW glasses over a wide range of temperatures, from the melter operating temperature to the glass transition.« less

  12. Cryopreservation by vitrification: a promising approach for transplant organ banking.

    PubMed

    Finger, Erik B; Bischof, John C

    2018-06-01

    The objective of this review is to describe the physical and biological barriers to organ cryopreservation, historic approaches for conventional cryopreservation and evolving techniques for ice-free cryopreservation by vitrification. Vitrification is a process whereby a biologic substance is cooled to cryogenic temperatures without the destructive phase transition of liquid to solid ice. Recent advances in cryoprotective solutions, organ perfusion techniques and novel heating technologies have demonstrated the potential for vitrification and rewarming organs on a scale applicable for human transplantation. Successful strategies for organ cryopreservation could enable organ banking, which would recast the entire process in which organs are recovered, allocated, stored and prepared for transplant.

  13. Effect of open pulled straw (OPS) vitrification on the fertilisation rate and developmental competence of porcine oocytes.

    PubMed

    Varga, Erika; Gardón, J C; Papp, Agnes Bali

    2006-03-01

    Freezing technologies are very important to preserve gametes and embryos of animals with a good pedigree or those having high genetic value. The aim of this work was to compare immature and in vitro matured porcine oocytes regarding their morphology and ability to be fertilised after vitrification by the open pulled straw (OPS) method. In four experiments 830 oocytes were examined. To investigate the effect of cumulus cells on oocyte survival after OPS vitrification, both denuded and cumulus-enclosed oocytes were vitrified at the germinal vesicle (GV) stage, then after vitrification they were matured in vitro. Besides, in vitro matured oocytes surrounded with a cumulus and those without a cumulus were also vitrified. The survival of oocytes was evaluated by their morphology. After in vitro fertilisation the rates of oocytes penetrated by spermatozoa were compared. Our results suggest that the vitrification/warming procedure is the most effective in cumulus-enclosed oocytes (22.35 +/- 1.75%). There was no difference between the order of maturation and vitrification in cumulus-enclosed oocytes, which suggests the importance of cumulus cells in protecting the viability of oocytes during cryopreservation.

  14. Cryopreservation of human embryos by vitrification or slow freezing: which one is better?

    PubMed

    Kolibianakis, Efstratios M; Venetis, Christos A; Tarlatzis, Basil C

    2009-06-01

    To summarize the available evidence from randomized controlled trials comparing vitrification versus slow freezing for cryopreservation of human embryos. Vitrification, as compared with slow freezing, appears to be better in terms of postthawing survival rates both for cleavage-stage embryos [odds ratio (OR): 6.35, 95% confidence interval (CI): 1.14-35.26, random effects model] and for blastocysts (OR: 4.09, 95% CI: 2.45-6.84, random effects model). Furthermore, postthawing blastocyst development of embryos cryopreserved in the cleavage stage is significantly higher with vitrification as compared with slow freezing (OR: 1.56, 95% CI: 1.07-2.27, fixed effects model). No significant difference in clinical pregnancy rates per transfer could be detected between the two cryopreservation methods (OR: 1.66, 95% CI: 0.98-2.79). Currently, vitrification does not appear to be associated with an increased probability of pregnancy. However, a significant advantage of vitrification over slow freezing in terms of postthawing survival rates is present for embryos cryopreserved both at the cleavage and at the blastocyst stages. The above conclusions are based on limited data, and thus further properly designed randomized controlled trials are needed.

  15. Thermal and clinical performance of a closed device designed for human oocyte vitrification based on the optimization of the warming rate.

    PubMed

    Gallardo, Miguel; Hebles, María; Migueles, Beatriz; Dorado, Mónica; Aguilera, Laura; González, Mercedes; Piqueras, Paloma; Montero, Lorena; Sánchez-Martín, Pascual; Sánchez-Martín, Fernando; Risco, Ramón

    2016-08-01

    Although it was qualitatively pointed out by Fahy et al. (1984), the key role of the warming rates in non-equillibrium vitrification has only recently been quantitatively established for murine oocytes by Mazur and Seki (2011). In this work we study the performance of a closed vitrification device designed under the new paradigm, for the vitrification of human oocytes. The vitrification carrier consists of a main straw in which a specifically designed capillary is mounted and where the oocytes are loaded by aspiration. It can be hermetically sealed before immersion in liquid nitrogen for vitrification, and it is warmed in a sterile water bath at 37 °C. Measured warming rates achieved with this design were of 600.000 ºC/min for a standard DMEM solution and 200.000 ºC/min with the vitrification solution for human oocytes. A cohort of 143 donor MII sibling human oocytes was split into two groups: control (fresh) and vitrified with SafeSpeed device. Similar results were found in both groups: survival (97.1%), fertilization after ICSI (74.7% in control vs. 77.3% in vitrified) and good quality embryos at day three (54.3% in control vs. 58.1% in vitrified) were settled as performance indicators. The pregnancy rate was 3/6 (50%) for the control, 2/3 (66%) for vitrified and 4/5 (80%) for mixed transfers. Copyright © 2016. Published by Elsevier Inc.

  16. Review of FY 2001 Development Work for Vitrification of Sodium Bearing Waste

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

    Taylor, Dean Dalton; Barnes, Charles Marshall

    2002-09-01

    Treatment of sodium-bearing waste (SBW) at the Idaho Nuclear Technology and Engineering Center (INTEC) within the Idaho National Engineering and Environmental Laboratory is mandated by the Settlement Agreement between the Department of Energy and the State of Idaho. This report discusses significant findings from vitrification technology development during 2001 and their impacts on the design basis for SBW vitrification.

  17. Innovative vitrification for soil remediation

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

    Jetta, N.W.; Patten, J.S.; Hart, J.G.

    1995-12-01

    The objective of this DOE demonstration program is to validate the performance and operation of the Vortec Cyclone Melting System (CMS{trademark}) for the processing of LLW contaminated soils found at DOE sites. This DOE vitrification demonstration project has successfully progressed through the first two phases. Phase 1 consisted of pilot scale testing with surrogate wastes and the conceptual design of a process plant operating at a generic DOE site. The objective of Phase 2, which is scheduled to be completed the end of FY 95, is to develop a definitive process plant design for the treatment of wastes at amore » specific DOE facility. During Phase 2, a site specific design was developed for the processing of LLW soils and muds containing TSCA organics and RCRA metal contaminants. Phase 3 will consist of a full scale demonstration at the DOE gaseous diffusion plant located in Paducah, KY. Several DOE sites were evaluated for potential application of the technology. Paducah was selected for the demonstration program because of their urgent waste remediation needs as well as their strong management and cost sharing financial support for the project. During Phase 2, the basic nitrification process design was modified to meet the specific needs of the new waste streams available at Paducah. The system design developed for Paducah has significantly enhanced the processing capabilities of the Vortec vitrification process. The overall system design now includes the capability to shred entire drums and drum packs containing mud, concrete, plastics and PCB`s as well as bulk waste materials. This enhanced processing capability will substantially expand the total DOE waste remediation applications of the technology.« less

  18. Phase Stability Determinations of DWPF Waste Glasses

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

    Marra, S.L.

    1999-10-22

    Liquid high-level nuclear waste will be immobilized at the Savannah River Site (SRS) by vitrification in borosilicate glass. To fulfill this requirement, glass samples were heat treated at various times and temperatures. These results will provide guidance to the repository program about conditions to be avoided during shipping, handling and storage of DWPF canistered waste forms.

  19. Cryopreservation of day 2-3 embryos by vitrification yields better outcome than slow freezing.

    PubMed

    Levron, Jacob; Leibovitz, Oshrit; Brengauz, Masha; Gitman, Hila; Yerushalmi, Gil M; Katorza, Eldad; Gat, Itai; Elizur, Shai E

    2014-03-01

    To compare the outcome of vitrification versus slow freezing cryopreservation for cleavage stage day 2-3 embryos. A retrospective observational study. All thawed embryos assisted reproduction cycles between January 2010 and December 2012 at a single IVF laboratory of a Tertiary Medical Center. Five hundred and thirty-nine cycles of day 2-3 thawed embryos. In 327 of the thawed cycles, the embryos were vitrified and in 212 of the cycles the embryos were derived from slow freezing embryos. Embryo survival rate, blastomere surviving rate and pregnancy rate. Embryo survival rate was significantly higher after vitrification compared with slow freezing (81.6%, 647/793 versus 70.0%, 393/562 embryos, p < 0.0001). The clinical pregnancy rate per ET was significantly higher following vitrification compared to slow freezing, 20.0%, 63/314 versus 11.9%, 23/193, respectively (p = 0.02). Vitrification of day 2-3 cleavage stage embryos yields better cycle outcome in all the parameters compared to slow freezing.

  20. Glass Property Models, Constraints, and Formulation Approaches for Vitrification of High-Level Nuclear Wastes at the US Hanford Site

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

    Kim, Dong-Sang

    2015-03-02

    The legacy nuclear wastes stored in underground tanks at the US Department of Energy’s Hanford site is planned to be separated into high-level waste and low-activity waste fractions and vitrified separately. Formulating optimized glass compositions that maximize the waste loading in glass is critical for successful and economical treatment and immobilization of nuclear wastes. Glass property-composition models have been developed and applied to formulate glass compositions for various objectives for the past several decades. The property models with associated uncertainties and combined with composition and property constraints have been used to develop preliminary glass formulation algorithms designed for vitrification processmore » control and waste form qualification at the planned waste vitrification plant. This paper provides an overview of current status of glass property-composition models, constraints applicable to Hanford waste vitrification, and glass formulation approaches that have been developed for vitrification of hazardous and highly radioactive wastes stored at the Hanford site.« less

  1. Generation of live offspring from vitrified embryos with synthetic polymers SuperCool X-1000 and SuperCool Z-1000.

    PubMed

    Marco-Jimenez, F; Jimenez-Trigos, E; Lavara, R; Vicente, J S

    2014-01-01

    Ice growth and recrystallisation are considered important factors in determining vitrification outcomes. Synthetic polymers inhibit ice formation during cooling or warming of the vitrification process. The aim of this study was to assess the effect of adding commercially available synthetic polymers SuperCool X-1000 and SuperCool Z-1000 to vitrification media on in vivo development competence of rabbit embryos. Four hundred and thirty morphologically normal embryos recovered at 72 h of gestation were used. The vitrification media contained 20% dimethyl sulphoxide and 20% ethylene glycol, either alone or in combination with 1% of SuperCool X-1000 and 1% SuperCool. Our results show that embryos can be successfully vitrified using SuperCool X-1000 and SuperCool Z-1000 and when embryos are transferred, live offspring can be successfully produced. In conclusion, our results demonstrated that we succeeded for the first time in obtaining live offspring after vitrification of embryos using SuperCool X-1000 and SuperCool Z-1000 polymers.

  2. Alginate Hydrogel Microencapsulation Inhibits Devitrification and Enables Large-Volume Low-CPA Cell Vitrification

    PubMed Central

    Huang, Haishui; Choi, Jung Kyu; Rao, Wei; Zhao, Shuting; Agarwal, Pranay; Zhao, Gang

    2015-01-01

    Cryopreservation of stem cells is important to meet their ever-increasing demand by the burgeoning cell-based medicine. The conventional slow freezing for stem cell cryopreservation suffers from inevitable cell injury associated with ice formation and the vitrification (i.e., no visible ice formation) approach is emerging as a new strategy for cell cryopreservation. A major challenge to cell vitrification is intracellular ice formation (IIF, a lethal event to cells) induced by devitrification (i.e., formation of visible ice in previously vitrified solution) during warming the vitrified cells at cryogenic temperature back to super-zero temperatures. Consequently, high and toxic concentrations of penetrating cryoprotectants (i.e., high CPAs, up to ~8 M) and/or limited sample volumes (up to ~2.5 μl) have been used to minimize IIF during vitrification. We reveal that alginate hydrogel microencapsulation can effectively inhibit devitrification during warming. Our data show that if ice formation were minimized during cooling, IIF is negligible in alginate hydrogel-microencapsulated cells during the entire cooling and warming procedure of vitrification. This enables vitrification of pluripotent and multipotent stem cells with up to ~4 times lower concentration of penetrating CPAs (up to 2 M, low CPA) in up to ~100 times larger sample volume (up to ~250 μl, large volume). PMID:26640426

  3. Alginate Hydrogel Microencapsulation Inhibits Devitrification and Enables Large-Volume Low-CPA Cell Vitrification.

    PubMed

    Huang, Haishui; Choi, Jung Kyu; Rao, Wei; Zhao, Shuting; Agarwal, Pranay; Zhao, Gang; He, Xiaoming

    2015-11-25

    Cryopreservation of stem cells is important to meet their ever-increasing demand by the burgeoning cell-based medicine. The conventional slow freezing for stem cell cryopreservation suffers from inevitable cell injury associated with ice formation and the vitrification ( i.e. , no visible ice formation) approach is emerging as a new strategy for cell cryopreservation. A major challenge to cell vitrification is intracellular ice formation (IIF, a lethal event to cells) induced by devitrification ( i.e. , formation of visible ice in previously vitrified solution) during warming the vitrified cells at cryogenic temperature back to super-zero temperatures. Consequently, high and toxic concentrations of penetrating cryoprotectants ( i.e. , high CPAs, up to ~8 M) and/or limited sample volumes (up to ~2.5 μl) have been used to minimize IIF during vitrification. We reveal that alginate hydrogel microencapsulation can effectively inhibit devitrification during warming. Our data show that if ice formation were minimized during cooling, IIF is negligible in alginate hydrogel-microencapsulated cells during the entire cooling and warming procedure of vitrification. This enables vitrification of pluripotent and multipotent stem cells with up to ~4 times lower concentration of penetrating CPAs (up to 2 M, low CPA) in up to ~100 times larger sample volume (up to ~250 μl, large volume).

  4. Ultrastructural changes of sheep cumulus-oocyte complexes following different methods of vitrification.

    PubMed

    Ebrahimi, Bita; Valojerdi, Mojtaba Rezazadeh; Eftekhari-Yazdi, Poopak; Baharvand, Hossein

    2012-05-01

    To determine the ultrastructural changes of sheep cumulus-oocyte complexes (COCs) following different methods of vitrification, good quality isolated COCs (GV stage) were randomly divided into the non-vitrified control, conventional straw, cryotop and solid surface vitrification groups. In both conventional and cryotop methods, vitrified COCs were respectively loaded by conventional straws and cryotops, and then plunged directly into liquid nitrogen (LN2); whereas in the solid surface group, vitrified COCs were first loaded by cryotops and then cooled before plunging into LN2. Post-warming survivability and ultrastructural changes of healthy COCs in the cryotop group especially in comparison with the conventional group revealed better viability rate and good preservation of the ooplasm organization. However in all vitrification groups except the cryotop group, mitochondria were clumped. Solely in the conventional straw group, the mitochondria showed different densities and were extremely distended. Moreover in the latter group, plenty of large irregular connected vesicles in the ooplasm were observed and in some parts their membrane ruptured. Also, in the conventional and solid surface vitrification groups, cumulus cells projections became retracted from the zona pellucida in some parts. In conclusion, the cryotop vitrification method as compared with other methods seems to have a good post-warming survivability and shows less deleterious effects on the ultrastructure of healthy vitrified-warmed sheep COCs.

  5. Structural and microstructural aspects of asbestos-cement waste vitrification

    NASA Astrophysics Data System (ADS)

    Iwaszko, Józef; Zawada, Anna; Przerada, Iwona; Lubas, Małgorzata

    2018-04-01

    The main goal of the work was to evaluate the vitrification process of asbestos-cement waste (ACW). A mixture of 50 wt% ACW and 50 wt% glass cullet was melted in an electric furnace at 1400 °C for 90 min and then cast into a steel mold. The vitrified product was subjected to annealing. Optical microscopy, scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) were used to evaluate the effects of the vitrification. The chemical constitution of the material before and after the vitrification process was also analyzed. It was found that the vitrified product has an amorphous structure in which the components of asbestos-cement waste are incorporated. MIR spectroscopy showed that the absorption bands of chrysotile completely disappeared after the vitrification process. The results of the spectroscopic studies were confirmed by X-ray studies - no diffraction reflections from the chrysotile crystallographic planes were observed. As a result of the treatment, the fibrous asbestos construction, the main cause of its pathogenic properties, completely disappeared. The vitrified material was characterized by higher resistance to ion leaching in an aquatic environment than ACW and a smaller volume of nearly 72% in relation to the apparent volume of the substrates. The research has confirmed the high effectiveness of vitrification in neutralizing hazardous waste containing asbestos and the FT-IR spectroscopy was found to be useful to identify asbestos varieties and visualizing changes caused by the vitrification process. The work also presents the current situation regarding the utilization of asbestos-containing products.

  6. System for enhanced destruction of hazardous wastes by in situ vitrification of soil

    DOEpatents

    Timmerman, Craig L.

    1991-01-01

    The present invention comprises a system for promoting the destruction of volatile and/or hazardous contaminants present in waste materials during in situ vitrification processes. In accordance with the present invention, a cold cap (46) comprising a cohesive layer of resolidified material is formed over the mass of liquefied soil and waste (40) present between and adjacent to the electrodes (10, 12, 14, 16) during the vitrification process. This layer acts as a barrier to the upward migration of any volatile type materials thereby increasing their residence time in proximity to the heated material. The degree of destruction of volatile and/or hazardous contaminants by pyrolysis is thereby improved during the course of the vitrification procedure.

  7. Destruction and Vitrification of Asbestos Using Plasma Arc Technology (Construction Productivity Advancement Research (CPAR) Program)

    DTIC Science & Technology

    1993-09-01

    DOCUMENTATION PAGE M N Public mpsMIs b~ b•r tOr coleton of ia nlormmn ias edsimad to aevue 1 hour pw response , indudng the Snm ta raing e. taeu"tig aining...Asbestos Emergency Response Act of 1986 (AHERA), which mandates inspection in school grades Kindergarten through 12. Congress is currently...CRC), Georgia Institute of Technology, Atlanta, GA. The CRC was responsible for the technical portion of the research program; i.e., developing the test

  8. Vitrification as an alternative to landfilling of tannery sewage sludge

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

    Celary, Piotr, E-mail: pcelary@is.pcz.czest.pl; Sobik-Szołtysek, Jolanta, E-mail: jszoltysek@is.pcz.czest.pl

    Highlights: • The possibility of vitrification of tannery sewage sludge was investigated. • Glass cullet was substituted with different wastes of mineral character. • Component ratio in the processed mixtures was optimized. • Environmental safety of the acquired vitrificates was verified. • An alternative management approach of usually landfilled waste was presented. - Abstract: Due to high content of heavy metals such as chromium, tannery sewage sludge is a material which is difficult to be biologically treated as it is in the case of organic waste. Consequently, a common practice in managing tannery sewage sludge is landfilling. This poses amore » potential threat to both soil and water environments and it additionally generates costs of construction of landfills that meet specific environment protection requirements. Vitrification of this kind of sewage sludge with the addition of mineral wastes can represent an alternative to landfilling. The aim of this study was to investigate the possibility of obtaining an environmentally safe product by means of vitrification of tannery sewage sludge from a flotation wastewater treatment process and chemical precipitation in order to address the upcoming issue of dealing with sewage sludge from the tannery industry which will be prohibited to be landfilled in Poland after 2016. The focus was set on determining mixtures of tannery sewage sludge with additives which would result in the lowest possible heavy metal leaching levels and highest hardness rating of the products obtained from their vitrification. The plasma vitrification process was carried out for mixtures with various amounts of additives depending on the type of sewage sludge used. Only the materials of waste character were used as additives. One finding of the study was an optimum content of mineral additives in vitrified mixture of 30% v/v waste molding sands with 20% v/v carbonate flotation waste from the zinc and lead industry for the formulations with flotation sewage sludge, and 45% v/v and 5% v/v, respectively, for precipitation sewage sludge. These combinations allowed for obtaining products with negligible heavy metal leaching levels and hardness similar to commercial glass, which suggests they could be potentially used as construction aggregate substitutes. Incineration of sewage sludge before the vitrification process lead to increased hardness of the vitrificates and reduced leaching of some heavy metals.« less

  9. Competition between crystallization and vitrification of the rigid amorphous fraction in poly(3-hydroxybutyrate)

    NASA Astrophysics Data System (ADS)

    Di Lorenzo, Maria Laura; Righetti, Maria Cristina; Gazzano, Massimo

    2012-07-01

    Semicrystalline polymers have a metastable nanophase structure, where the various nanophases can be crystal, liquid, glass, or mesophase. This multi-level structure is determined by a competition among self-organization, crystallization, and vitrification of the amorphous segments and is established during material processing. The kinetics of such competition is here determined for poly(3-hydroxybutyrate) (PHB), as vitrification/devitrification of the rigid amorphous fraction strongly affects crystallization kinetics of PHB.

  10. Vitrification versus slow freezing gives excellent survival, post warming embryo morphology and pregnancy outcomes for human cleaved embryos.

    PubMed

    Rezazadeh Valojerdi, Mojtaba; Eftekhari-Yazdi, Poopak; Karimian, Leila; Hassani, Fatemeh; Movaghar, Bahar

    2009-06-01

    The objective of this retrospective study was to evaluate the efficacy of vitrification and slow freezing for the cryopreservation of human cleavage stage embryos in terms of post-warming survival rate, post-warming embryo morphology and clinical outcomes. The embryos of 305 patients at cleavage stages were cryopreserved either with vitrification (153 patients) or slow-freezing (152 patients) methods. After warming; the survival rate, post-warmed embryo morphology, clinical pregnancy and implantation rates were evaluated and compared between the two groups. In the vitrification group versus slow freezing group, the survival rate (96.9% vs. 82.8%) and the post-warmed excellent morphology with all blastomeres intact (91.8% vs. 56.2%) were higher with an odds ratio of 6.607 (95% confidence interval; 4.184-10.434) and 8.769 (95% confidence interval; 6.460-11.904), respectively. In this group, the clinical pregnancy rate (40.5% vs. 21.4%) and the implantation rate (16.6% vs. 6.8%) were also higher with an odds ratio of 2.427 (95%confidence interval; 1.461-4.033) and 2.726 (95% confidence interval; 1.837-4.046), respectively. Vitrification in contrast to slow freezing is an efficient method for cryopreservation of human cleavage stage embryos. Vitrification provides a higher survival rate, minimal deleterious effects on post-warming embryo morphology and it can improve clinical outcomes.

  11. Prospective randomized comparison of human oocyte cryopreservation with slow-rate freezing or vitrification.

    PubMed

    Smith, Gary D; Serafini, Paulo C; Fioravanti, Joyce; Yadid, Isaac; Coslovsky, Marcio; Hassun, Pericles; Alegretti, José Roberto; Motta, Eduardo L

    2010-11-01

    To compare cryopreservation of mature human oocytes with slow-rate freezing and vitrification and determine which is most efficient at establishing a pregnancy. Prospective randomized. Academically affiliated, private fertility center. Consenting patients with concerns about embryo cryopreservation and more than nine mature oocytes at retrieval were randomized to slow-rate freezing or vitrification of supernumerary (more than nine) oocytes. Oocytes were frozen or vitrified, and upon request oocytes were thawed or warmed, respectively. Oocyte survival, fertilization, embryo development, and clinical pregnancy. Patient use has resulted in 30 thaws and 48 warmings. Women's age at time of cryopreservation was similar. Oocyte survival was significantly higher following vitrification/warming (81%) compared with freezing/thawing (67%). Fertilization was more successful in oocytes vitrified/warmed compared with frozen/thawed. Fertilized oocytes from vitrification/warming had significantly better cleavage rates (84%) compared with freezing/thawing (71%) and resulted in embryos with significantly better morphology. Although similar numbers of embryos were transferred, embryos resulting from vitrified oocytes had significantly enhanced clinical (38%) pregnancy rates compared with embryos resulting from frozen oocyte (13%). Miscarriage and/or spontaneous abortion rates were similar. Our results suggest that vitrification/warming is currently the most efficient means of oocyte cryopreservation in relation to subsequent success in establishing pregnancy. Copyright © 2010 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  12. The Increased Expression of Connexin and VEGF in Mouse Ovarian Tissue Vitrification by Follicle Stimulating Hormone

    PubMed Central

    Yang, Yanzhou; Chen, Jie; Wu, Hao; Pei, Xiuying; Chang, Qing; Ma, Wenzhi; Ma, Huiming; Hei, Changchun; Zheng, Xiaomin; Cai, Yufang; Zhao, Chengjun; Yu, Jia; Wang, Yanrong

    2015-01-01

    Ovarian follicular damages were caused by cryoinjury during the process of ovarian vitrification and ischemia/reperfusion during the process of ovarian transplantation. And appropriate FSH plays an important role in antiapoptosis during ovarian follicle development. Therefore, in this study, 0.3 IU/mL FSH was administered into medium during mouse ovarian cryopreservation by vitrification to ascertain the function of FSH on ovarian vitrification and avascular transplantation. The results suggested that the expressions of Cx37, Cx43, apoptotic molecular caspase-3, and angiogenesis molecular VEGF were confirmed using immunohistochemistry, western blotting, and real-time PCR, and the results suggested that the treatment with FSH remarkably increased the number of morphologically normal follicles in vitrified/warmed ovaries by upregulating the expression of Cx37, Cx43, VEGF, and VEGF receptor 2, but downregulating the expression of caspase-3. In addition, the vitrified/warmed ovaries were transplanted, and the related fertility was analyzed, and the results suggested that the fertility, neoangiogenesis, and follicle reserve were remarkably increased in the FSH administrated group. Taken together, administration of 0.3 IU/mL FSH during ovarian cryopreservation by vitrification can maintain ovarian survival during ovarian vitrification and increases the blood supply with avascular transplantation via upregulation of Cx43, Cx37, and VEGF/VEGFR2, as well as through its antiapoptotic effects. PMID:26539488

  13. Super cool X-1000 and Super cool Z-1000, two ice blockers, and their effect on vitrification/warming of mouse embryos.

    PubMed

    Badrzadeh, H; Najmabadi, S; Paymani, R; Macaso, T; Azadbadi, Z; Ahmady, A

    2010-07-01

    To evaluate the survival and blastocyst formation rates of mouse embryos after vitrification/thaw process with different ice blocker media. We used X-1000 and Z-1000 separately and mixed using V-Kim, a closed vitrification system. Mouse embryos were vitrified using ethylene glycol based medium supplemented with Super cool X-1000 and/or Super cool Z-1000. Survival rates for the control, Super cool X-1000, Super cool Z-1000, and Super cool X-1000/Z-1000 groups were 74%, 72%, 68%, and 85% respectively, with no significant difference among experimental and control groups; however, a significantly higher survival rate was noticed in the Super cool X-1000/Z-1000 group when compared with the Super cool Z-1000 group. Blastocyst formation rates for the control, Super cool X-1000, Super cool Z-1000, and Super cool X-1000/Z-1000 groups were 71%, 66%, 65%, and 72% respectively. There was no significant difference in this rate among control and experimental groups. In a closed vitrification system, addition of ice blocker Super cool X-1000 to the vitrification solution containing Super cool Z-1000 may improve the embryo survival rate. We recommend combined ice blocker usage to optimize the vitrification outcome. Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.

  14. Hydroxypropyl cellulose as an option for supplementation of cryoprotectant solutions for embryo vitrification in human assisted reproductive technologies.

    PubMed

    Mori, Chiemi; Yabuuchi, Akiko; Ezoe, Kenji; Murata, Nana; Takayama, Yuko; Okimura, Tadashi; Uchiyama, Kazuo; Takakura, Kei; Abe, Hiroyuki; Wada, Keiko; Okuno, Takashi; Kobayashi, Tamotsu; Kato, Keiichi

    2015-06-01

    Hydroxypropyl cellulose (HPC) was investigated as a replacement for serum substitute supplement (SSS) for use in cryoprotectant solutions for embryo vitrification. Mouse blastocysts from inbred (n = 1056), hybrid (n = 128) strains, and 121 vitrified blastocysts donated by infertile patients (n = 102) were used. Mouse and human blastocysts, with or without zona pellucida, were vitrified and warmed in either 1% or 5% HPC or in 5% or 20% SSS-supplemented media using the Cryotop (Kitazato BioPharma Co. Ltd, Fuji, Japan) method, and the survival and oxygen consumption rates were assessed. Viscosity of each vitrification solution was compared. Survival rates of mouse hybrid blastocysts and human zona pellucida-intact blastocysts were comparable among the groups. Mouse and human zona pellucida-free blastocysts, which normally exhibit poor cryoresistance, showed significantly higher survival rates in 5% HPC than 5% SSS (P < 0.05). The 5% HPC-supplemented vitrification solution showed a significantly higher viscosity (P < 0.05). The blastocysts were easily detached from the Cryotop strip during warming when HPC-supplemented vitrification solution was used. The oxygen consumption rates were similar between non-vitrified and 5% HPC groups. The results suggest possible use of HPC for supplementation of cryoprotectant solutions and provide useful information to improve vitrification protocols. Copyright © 2015 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  15. Leaching characteristics of copper flotation waste before and after vitrification.

    PubMed

    Coruh, Semra; Ergun, Osman Nuri

    2006-12-01

    Copper flotation waste from copper production using a pyrometallurgical process contains toxic metals such as Cu, Zn, Co and Pb. Because of the presence of trace amounts of these highly toxic metals, copper flotation waste contributes to environmental pollution. In this study, the leaching characteristics of copper flotation waste from the Black Sea Copper Works in Samsun, Turkey have been investigated before and after vitrification. Samples obtained from the factory were subjected to toxicity tests such as the extraction procedure toxicity test (EP Tox), the toxicity characteristic leaching procedure (TCLP) and the "method A" extraction procedure of the American Society of Testing and Materials. The leaching tests showed that the content of some elements in the waste before vitrification exceed the regulatory limits and cannot be disposed of in the present form. Therefore, a stabilization or inertization treatment is necessary prior to disposal. Vitrification was found to stabilize heavy metals in the copper flotation waste successfully and leaching of these metals was largely reduced. Therefore, vitrification can be an acceptable method for disposal of copper flotation waste.

  16. Measurement of Thermal Conductivities of Two Cryoprotective Agent Solutions for Vitreous Cryopreservation of Organs at the Temperature Range of 77 K-300 K Using a Thermal Sensor Made of Microscale Enamel Copper Wire.

    PubMed

    Li, Yufang; Zhao, Gang; Hossain, S M Chapal; Panhwar, Fazil; Sun, Wenyu; Kong, Fei; Zang, Chuanbao; Jiang, Zhendong

    2017-06-01

    Biobanking of organs by cryopreservation is an enabling technology for organ transplantation. Compared with the conventional slow freezing method, vitreous cryopreservation has been regarded to be a more promising approach for long-term storage of organs. The major challenges to vitrification are devitrification and recrystallization during the warming process, and high concentrations of cryoprotective agents (CPAs) induced metabolic and osmotic injuries. For a theoretical model based optimization of vitrification, thermal properties of CPA solutions are indispensable. In this study, the thermal conductivities of M22 and vitrification solution containing ethylene glycol and dimethyl sulfoxide (two commonly used vitrification solutions) were measured using a self-made microscaled hot probe with enameled copper wire at the temperature range of 77 K-300 K. The data obtained by this study will further enrich knowledge of the thermal properties for CPA solutions at low temperatures, as is of primary importance for optimization of vitrification.

  17. Stability of the cytoskeleton of matured buffalo oocytes pretreated with cytochalasin B prior to vitrification.

    PubMed

    Wang, C L; Xu, H Y; Xie, L; Lu, Y Q; Yang, X G; Lu, S S; Lu, K H

    2016-06-01

    Stabilizing the cytoskeleton system during vitrification can improve the post-thaw survival and development of vitrified oocytes. The cytoskeleton stabilizer cytochalasin B (CB) has been used in cryopreservation to improve the developmental competence of vitrified oocytes. To assess the effect of pretreating matured buffalo oocytes with CB before vitrification, we applied 0, 4, 8, or 12 μg/mL CB for 30 min. The optimum concentration of CB treatment (8 μg/mL for 30 min) was then used to evaluate the distribution of microtubules and microfilaments, the expression of the cytoskeleton proteins actin and tubulin, and the developmental potential of matured oocytes that were vitrified-warmed by the Cryotop method. Western blotting demonstrated that vitrification significantly decreased tubulin expression, but that the decrease was attenuated for oocytes pretreated with 8 μg/mL CB before vitrification. After warming and intracytoplasmic sperm injection, oocytes that were pretreated with 8 μg/mL CB before vitrification yielded significantly higher 8-cell and blastocyst rates than those that were vitrified without CB pretreatment. The values for the vitrified groups in all experiments were significantly lower (P < 0.01) than those of the control groups. In conclusion, pretreatment with 8 μg/mL CB for 30 min significantly improves the cytoskeletal structure, expression of tubulin, and development capacity of vitrified matured buffalo oocytes. Copyright © 2016 Elsevier Inc. All rights reserved.

  18. [Testicular tissue vitrification: evolution or revolution?].

    PubMed

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

    2013-09-01

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

  19. Vitrification in human and domestic animal embryology: work in progress.

    PubMed

    Vajta, Gábor

    2013-01-01

    According to the analysis of papers published in major international journals, rapidly increasing application of vitrification is one of the greatest achievements in domestic animal and especially human embryology during the first decade of our century. This review highlights factors supporting or hampering this progress, summarises results achieved with vitrification and outlines future tasks to fully exploit the benefits of this amazing approach that has changed or will change many aspects of laboratory (and also clinical) embryology. Supporting factors include the simplicity, cost efficiency and convincing success of vitrification compared with other approaches in all species and developmental stages in mammalian embryology, while causes that slow down the progress are mostly of human origin: inadequate tools and solutions, superficial teaching, improper application and unjustified concerns resulting in legal restrictions. Elimination of these hindrances seems to be a slower process and more demanding task than meeting the biological challenge. A key element of future progress will be to pass the pioneer age, establish a consensus regarding biosafety requirements, outline the indispensable features of a standard approach and design fully-automated vitrification machines executing all phases of the procedure, including equilibration, cooling, warming and dilution steps.

  20. Selecting a plutonium vitrification process

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

    Jouan, A.

    1996-05-01

    Vitrification of plutonium is one means of mitigating its potential danger. This option is technically feasible, even if it is not the solution advocated in France. Two situations are possible, depending on whether or not the glass matrix also contains fission products; concentrations of up to 15% should be achievable for plutonium alone, whereas the upper limit is 3% in the presence of fission products. The French continuous vitrification process appears to be particularly suitable for plutonium vitrification: its capacity is compatible with the required throughout, and the compact dimensions of the process equipment prevent a criticality hazard. Preprocessing ofmore » plutonium metal, to convert it to PuO{sub 2} or to a nitric acid solution, may prove advantageous or even necessary depending on whether a dry or wet process is adopted. The process may involve a single step (vitrification of Pu or PuO{sub 2} mixed with glass frit) or may include a prior calcination step - notably if the plutonium is to be incorporated into a fission product glass. It is important to weigh the advantages and drawbacks of all the possible options in terms of feasibility, safety and cost-effectiveness.« less

  1. Improved cryotolerance and developmental potential of in vitro and in vivo matured mouse oocytes by supplementing with a glutathione donor prior to vitrification.

    PubMed

    Trapphoff, Tom; Heiligentag, Martyna; Simon, Jenny; Staubach, Nora; Seidel, Thorsten; Otte, Kathrin; Fröhlich, Thomas; Arnold, Georg J; Eichenlaub-Ritter, Ursula

    2016-12-01

    Can supplementation of media with a glutathione (GSH) donor, glutathione ethyl ester (GEE), prior to vitrification protect the mouse oocyte from oxidative damage and critical changes in redox homeostasis, and thereby improve cryotolerance? GEE supplementation supported redox regulation, rapid recovery of spindle and chromosome alignment after vitrification/warming and improved preimplantation development of mouse metaphase II (MII) oocytes. Cryopreservation may affect mitochondrial functionality, induce oxidative stress, and thereby affect spindle integrity, chromosome segregation and the quality of mammalian oocytes. GEE is a membrane permeable GSH donor that promoted fertilization and early embryonic development of macaque and bovine oocytes after IVM. Two experimental groups consisted of (i) denuded mouse germinal vesicle (GV) oocytes that were matured in vitro in the presence or absence of 1 mM GEE (IVM group 1) and (ii) in vivo ovulated (IVO) MII oocytes that were isolated from the ampullae and exposed to 1 mM GEE for 1 h prior to vitrification (IVO group 2). Recovery of oocytes from both groups was followed after CryoTop vitrification/warming for up to 2 h and parthenogenetic activation. Reactive oxygen species (ROS), spindle morphology and chromosome alignment were analyzed by confocal laser scanning microscopy (CLSM) and polarization microscopy in control and GEE-supplemented MII oocytes. The relative overall intra-oocyte GSH content was assessed by analysis of monochlorobimane (MBC)-GSH adduct fluorescence in IVM MII oocytes. The GSH-dependent intra-mitochondrial redox potential (E m GSH ) of IVM MII oocytes was determined after microinjection with specific mRNA at the GV stage to express a redox-sensitive probe within mitochondria (mito-Grx1-roGFP2). The absolute negative redox capacity (in millivolts) was determined by analysis of fluorescence of the oxidized versus the reduced form of sensor by CLSM and quantification according to Nernst equation. Proteome analysis was performed by quantitative 2D saturation gel electrophoresis (2D DIGE). Since microinjection and expression of redox sensor mRNA required removal of cumulus cells, and IVM of denuded mouse oocytes in group 1 induces zona hardening, the development to blastocysts was not assessed after IVF but instead after parthenogenetic activation of vitrified/warmed MII oocytes from both experimental groups. IVM of denuded mouse oocytes in the presence of 1 mM GEE significantly increased intra-oocyte GSH content. ROS was not increased by CryoTop vitrification but was significantly lower in the IVM GEE group compared to IVM without GEE before vitrification and after recovery from vitrification/warming (P < 0.001). Vitrification alone significantly increased the GSH-dependent intra-mitochondrial redox capacity after warming (E m GSH , P < 0.001) in IVM oocytes, presumably by diffusion/uptake of cytoplasmic GSH into mitochondria. The presence of 1 mM GEE during IVM increased the redox capacity before vitrification and there was no further increase after vitrification/warming. None of the reproducibly detected 1492 spots of 2D DIGE separated proteins were significantly altered by vitrification or GEE supplementation. However, IVM of denuded oocytes significantly affected spindle integrity and chromosome alignment right after warming from vitrification (0 h) in group 1 and spindle integrity in group 2 (P < 0.05). GEE improved recovery in IVM group as numbers of oocytes with unaligned chromosomes and aberrant spindles was not significantly increased compared to unvitrified controls. The supplementation with GEE for 1 h before vitrification also supported more rapid recovery of spindle birefringence. GEE improved significantly development to the 2-cell stage for MII oocytes that were activated directly after vitrification/warming in both experimental groups, and also the blastocyst rate in the IVO GEE-supplemented group compared to the controls (P < 0.05). None LIMITATIONS, REASONS FOR CAUTION: The studies were carried out in a mouse model, in IVM denuded rather than cumulus-enclosed oocytes, and in activated rather than IVF MII oocytes. Whether the increased GSH-dependent intra-mitochondrial redox capacity also improves male pronuclear formation needs to be studied further experimentally. The influence of GEE supplementation requires also further examination and optimization in human oocytes before it can be considered for clinical ART. Although GEE supplementation did not alter the proteome at MII, the GSH donor may support cellular homeostasis and redox regulation and, thus, increase developmental competence. While human MII oocyte vitrification is an established procedure, GEE might be particularly beneficial for oocytes that suffer from oxidative stress and reduced redox capacity (e.g. aged oocytes) or possess low GSH due to a reduced supply of GSH from cumulus. It might also be of relevance for immature human oocytes that develop without cumulus to MII in vitro (e.g. in ICSI cycles) for ART. The study has been supported by the German Research Foundation (DFG FOR 1041; EI 199/3-2). There are no conflict of interests. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  2. Successful vitrification and autografting of baboon (Papio anubis) ovarian tissue.

    PubMed

    Amorim, Christiani A; Jacobs, Sophie; Devireddy, Ram V; Van Langendonckt, Anne; Vanacker, Julie; Jaeger, Jonathan; Luyckx, Valérie; Donnez, Jacques; Dolmans, Marie-Madeleine

    2013-08-01

    Can a vitrification protocol using an ethylene glycol/dimethyl sulphoxide-based solution and a cryopin successfully cryopreserve baboon ovarian tissue? Our results show that baboon ovarian tissue can be successfully cryopreserved with our vitrification protocol. Non-human primates have already been used as an animal model to test vitrification protocols for human ovarian tissue cryopreservation. Ovarian biopsies from five adult baboons were vitrified, warmed and autografted for 5 months. After grafting, follicle survival, growth and function and also the quality of stromal tissue were assessed histologically and by immunohistochemistry. The influence of the vitrification procedure on the cooling rate was evaluated by a computer model. After vitrification, warming and long-term grafting, follicles were able to grow and maintain their function, as illustrated by Ki67, anti-Müllerian hormone (AMH) and growth differentiation factor-9 (GDF-9) immunostaining. Corpora lutea were also observed, evidencing successful ovulation in all the animals. Stromal tissue quality did not appear to be negatively affected by our cryopreservation procedure, as demonstrated by vascularization and proportions of fibrotic areas, which were similar to those found in fresh ungrafted ovarian tissue. Despite our promising findings, before applying this technique in a clinical setting, we need to validate it by achieving pregnancies. In addition to encouraging results obtained with our vitrification procedure for non-human ovarian tissue, this study also showed, for the first time, expression of AMH and GDF-9 in ovarian follicles. This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (grant Télévie No. 7.4507.10, grant 3.4.590.08 awarded to Marie-Madeleine Dolmans), Fonds Spéciaux de Recherche, Fondation St Luc, Foundation Against Cancer, and Department of Mechanical Engineering at Louisiana State University (support to Ram Devireddy), and donations from Mr Pietro Ferrero, Baron Frère and Viscount Philippe de Spoelberch. None of the authors has any competing interests to declare.

  3. Vitrification as an alternative to landfilling of tannery sewage sludge.

    PubMed

    Celary, Piotr; Sobik-Szołtysek, Jolanta

    2014-12-01

    Due to high content of heavy metals such as chromium, tannery sewage sludge is a material which is difficult to be biologically treated as it is in the case of organic waste. Consequently, a common practice in managing tannery sewage sludge is landfilling. This poses a potential threat to both soil and water environments and it additionally generates costs of construction of landfills that meet specific environment protection requirements. Vitrification of this kind of sewage sludge with the addition of mineral wastes can represent an alternative to landfilling. The aim of this study was to investigate the possibility of obtaining an environmentally safe product by means of vitrification of tannery sewage sludge from a flotation wastewater treatment process and chemical precipitation in order to address the upcoming issue of dealing with sewage sludge from the tannery industry which will be prohibited to be landfilled in Poland after 2016. The focus was set on determining mixtures of tannery sewage sludge with additives which would result in the lowest possible heavy metal leaching levels and highest hardness rating of the products obtained from their vitrification. The plasma vitrification process was carried out for mixtures with various amounts of additives depending on the type of sewage sludge used. Only the materials of waste character were used as additives. One finding of the study was an optimum content of mineral additives in vitrified mixture of 30% v/v waste molding sands with 20% v/v carbonate flotation waste from the zinc and lead industry for the formulations with flotation sewage sludge, and 45% v/v and 5% v/v, respectively, for precipitation sewage sludge. These combinations allowed for obtaining products with negligible heavy metal leaching levels and hardness similar to commercial glass, which suggests they could be potentially used as construction aggregate substitutes. Incineration of sewage sludge before the vitrification process lead to increased hardness of the vitrificates and reduced leaching of some heavy metals. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Equine ovarian tissue viability after cryopreservation and in vitro culture

    USDA-ARS?s Scientific Manuscript database

    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....

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

    PubMed

    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.

  6. Glass transition behavior of the vitrification solutions containing propanediol, dimethyl sulfoxide and polyvinyl alcohol.

    PubMed

    Wang, Hai-Yan; Lu, Shu-Shen; Lun, Zhao-Rong

    2009-02-01

    Knowledge of the glass transition behavior of vitrification solutions is important for research and planning of the cryopreservation of biological materials by vitrification. This brief communication shows the analysis for the glass transition and glass stability of the multi-component vitrification solutions containing propanediol (PE), dimethyl sulfoxide (Me2SO) and polyvinyl alcohol (PVA) by using differential scanning calorimetry (DSC) during the cooling and subsequent warming between 25 and -150 degrees C. The glass formation of the solutions was enhanced by introduction of PVA. Partial glass formed during cooling and the fractions of free water in the partial glass matrix increased with the increasing of PVA concentration, which caused slight decline of glass transition temperature, T(g). Exothermic peaks of devitrification were delayed and broadened, which may result from the inhibition of ice nucleation or recrystallization of PVA.

  7. Thermal Analyses of a Human Kidney and a Rabbit Kidney During Cryopreservation by Vitrification.

    PubMed

    Ehrlich, Lili E; Fahy, Gregory M; Wowk, Brian G; Malen, Jonathan A; Rabin, Yoed

    2018-01-01

    This study focuses on thermal analysis of the problem of scaling up from the vitrification of rabbit kidneys to the vitrification of human kidneys, where vitrification is the preservation of biological material in the glassy state. The basis for this study is a successful cryopreservation protocol for a rabbit kidney model, based on using a proprietary vitrification solution known as M22. Using the finite element analysis (FEA) commercial code ANSYS, heat transfer simulations suggest that indeed the rabbit kidney unquestionably cools rapidly enough to be vitrified based on known intrarenal concentrations of M22. Scaling up 21-fold, computer simulations suggest less favorable conditions for human kidney vitrification. In this case, cooling rates below -100 °C are sometimes slower than 1 °C/min, a rate that provides a clear-cut margin of safety at all temperatures based on the stability of rabbit kidneys in past studies. Nevertheless, it is concluded in this study that vitrifying human kidneys is possible without significant ice damage, assuming that human kidneys can be perfused with M22 as effectively as rabbit kidneys. The thermal analysis suggests that cooling rates can be further increased by a careful design of the cryogenic protocol and by tailoring the container to the shape of the kidney, in contrast to the present cylindrical container. This study demonstrates the critical need for the thermal analysis of experimental cryopreservation and highlights the unmet need for measuring the thermophysical properties of cryoprotective solutions under conditions relevant to realistic thermal histories.

  8. Efficient vitrification of mouse embryos using the Kitasato Vitrification System as a novel vitrification device.

    PubMed

    Momozawa, Kenji; Matsuzawa, Atsushi; Tokunaga, Yukio; Abe, Shiori; Koyanagi, Yumi; Kurita, Miho; Nakano, Marina; Miyake, Takao

    2017-04-24

    Currently, the cryopreservation of embryos and oocytes is essential for assisted reproductive technology (ART) laboratories worldwide. This study aimed to evaluate the efficacy of the Kitasato Vitrification System (KVS) as a vitrification device for the cryopreservation of mouse embryos to determine whether this novel device can be adapted to the field of ART. In Experiment 1, blastocysts were vitrified using the KVS. Vitrified blastocysts were warmed and subsequently cultured for 72 h. In Experiment 2, 2-cell-stage embryos were vitrified using the KVS, and vitrified embryos were warmed and subsequently cultured for 96 h. In Experiment 3, we evaluated the in vivo developmental potential of vitrified 2-cell-stage embryos using the KVS, and in Experiment 4, we evaluated the cooling and warming rates for these devices using a numerical simulation. In Experiment 1, there were no significant differences between the survival rates of the KVS and a control device. However, re-expanded (100%) and hatching (91.8%) rates were significantly higher for blastocysts vitrified using the KVS. In Experiment 2, there were no significant differences between the survival rates, or rates of development to the blastocyst stage, of vitrified and fresh embryos. In Experiment 3, after embryo transfer, 41% of the embryos developed into live offspring. In Experiment 4, the cooling and warming rates of the KVS were 683,000 and 612,000 °C/min, respectively, exceeding those of the control device. Our study clearly demonstrates that the KVS is a novel vitrification device for the cryopreservation of mouse embryos at the blastocyst and 2-cell stage.

  9. ENGINEERING BULLETIN: IN SITU VITRIFICATION TREATMENT

    EPA Science Inventory

    In situ vitrification (ISV) uses electrical power to heat and melt soil, sludge, mine tailings, buried wastes, and sediments contaminated with organic, inorganic, and metal-bearing hazardous wastes. The molten material cools to form a hard, monolithic, chemically inert, stable...

  10. DEMONSTRATION BULLETIN: IN SITU VITRIFICATION - GEOSAFE CORPORATION

    EPA Science Inventory

    in Situ Vitrification (ISV) is designed to treat soils, sludges, sediments, and mine tailings contaminated with organic and inorganic compounds. The process uses electrical current to heat (mett) and vitrify the soil in place. Organic contaminants are decomposed by the extreme h...

  11. GEOTECH, INC., COLD TOP EX-SITU VITRIFICATION SYSTEM; INNOVATIVE TECHNOLOGY EVALUATION REPORT

    EPA Science Inventory

    A Superfund Innovative Technology Evaluation (SITE) technology demonstration was conducted in February and March 1997 to evaluate the Geotech Development Corporation (Geotech) Cold Top ex-situ vitrification technology in chromium-contaminated soils. The demonstration was conduct...

  12. SITE TECHNOLOGY CAPSULE: GEOSAFE CORPORATION IN SITU VITRIFICATION TECHNOLOGY

    EPA Science Inventory

    The Geosafe In Situ Vitrification (ISV) Technology is designed to treat soils, sludges, sediments, and mine tallings contaminated with organic, inorganic, and radioactive compounds. The organic compounds are pyrolyzed and reduced to simple gases which are collected under a treatm...

  13. Vitrification and devitrification of micro-droplets

    NASA Astrophysics Data System (ADS)

    Ryoun Youn, Jae; Song, Young Seok

    2012-11-01

    Vitrification can be achieved by flash freezing and thawing (i.e. quenching) when ice crystal formation is inhibited in a cryogenic environment. Such ultra-rapid cooling and rewarming occurs due to the large temperature difference between the liquid and its surrounding medium. Here, we analyze the crystallization behavior of a droplet (i.e. vitrification and devitrification) by using a numerical model. The numerical results were found to explain the experimental observations successfully. The findings showed that for successful cryopreservation not only sufficiently fast cooling, but also rewarming processes should be designed and controlled to avoid devitrification of a droplet.

  14. Vitrification

    NASA Astrophysics Data System (ADS)

    A. Takahashi, Tsuneo

    Vitrification is an alternative to customary approaches to cryopreserve cell, tissue and organ. In this method, ice formation can be prevented by a combination of high solute concentration and rapid cooling, a solution become glassy without ice crystalline formation at temperatures below-115°C. The cell and tissue damage associated with ice formation is avoided, but thawing should be rapid enough to prevent ice growth during warming and they should be equilibrated with the vitrification medium without injury. This approach has been extensively studied in the past few years, and has the potential to be an alternative approach to the cryopreservation of a wide range of biological systems.

  15. In-situ vitrification of waste materials

    DOEpatents

    Powell, J.R.; Reich, M.; Barletta, R.

    1997-10-14

    A method for the in-situ vitrification of waste materials in a disposable can that includes an inner container and an outer container is disclosed. The method includes the steps of adding frit and waste materials to the inner container, removing any excess water, heating the inner container such that the frit and waste materials melt and vitrify after cooling, while maintaining the outer container at a significantly lower temperature than the inner container. The disposable can is then cooled to ambient temperatures and stored. A device for the in-situ vitrification of waste material in a disposable can is also disclosed. 7 figs.

  16. In-situ vitrification of waste materials

    DOEpatents

    Powell, James R.; Reich, Morris; Barletta, Robert

    1997-11-14

    A method for the in-situ vitrification of waste materials in a disposable can that includes an inner container and an outer container is disclosed. The method includes the steps of adding frit and waste materials to the inner container, removing any excess water, heating the inner container such that the frit and waste materials melt and vitrify after cooling, while maintaining the outer container at a significantly lower temperature than the inner container. The disposable can is then cooled to ambient temperatures and stored. A device for the in-situ vitrification of waste material in a disposable can is also disclosed.

  17. Unraveling protein stabilization mechanisms: vitrification and water replacement in a glass transition temperature controlled system.

    PubMed

    Grasmeijer, N; Stankovic, M; de Waard, H; Frijlink, H W; Hinrichs, W L J

    2013-04-01

    The aim of this study was to elucidate the role of the two main mechanisms used to explain the stabilization of proteins by sugar glasses during drying and subsequent storage: the vitrification and the water replacement theory. Although in literature protein stability is often attributed to either vitrification or water replacement, both mechanisms could play a role and they should be considered simultaneously. A model protein, alkaline phosphatase, was incorporated in either inulin or trehalose by spray drying. To study the storage stability at different glass transition temperatures, a buffer which acts as a plasticizer, ammediol, was incorporated in the sugar glasses. At low glass transition temperatures (<50°C), the enzymatic activity of the protein strongly decreased during storage at 60°C. Protein stability increased when the glass transition temperature was raised considerably above the storage temperature. This increased stability could be attributed to vitrification. A further increase of the glass transition temperature did not further improve stability. In conclusion, vitrification plays a dominant role in stabilization at glass transition temperatures up to 10 to 20°C above storage temperature, depending on whether trehalose or inulin is used. On the other hand, the water replacement mechanism predominantly determines stability at higher glass transition temperatures. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. DEMONSTRATION BULLETIN: COLD TOP EX-SITU VITRIFICATION PROCESS - GEOTECH DEVELOPMENT CORPORATION

    EPA Science Inventory

    The Cold Top Vitrification process, developed by Geotech Development Corporation, is an ex-situ, submerged-electrode, resistance-melting technology. The technology is designed to transform heavy metal contaminated soil into a glassy, amorphous, non-leachable mass composed of inte...

  19. Impact of prolonged oocyte incubation time before vitrification on oocyte survival, embryo formation, and embryo quality in mice.

    PubMed

    Karami, Azade; Bakhtiari, Mitra; Azadbakht, Mehri; Ghorbani, Rostam; Khazaei, Mozafar; Rezaei, Mansour

    2017-06-01

    Oocyte incubation time before freezing is one of the factors affecting oocyte vitrification. In the assisted reproductive technology (ART) clinics, it is sometimes decided to perform oocyte vitrification after a long period of incubation time due to various conditions, such as inability to collect semen samples, unsuccessful urological interventions (PESA, TESE, etc.), or unexpected conditions. A time factor of up to 6 h has been studied in the available reports. Therefore, this study was designed to evaluate oocyte incubation time before freezing at 0, 6, 12, 18, and 24 h after retrieval. Metaphase II (MII) oocytes were obtained from NMRI female mice after being randomly divided into the five groups of 0, 6, 12, 18, and 24 h of freezing via hormonal stimulation following retrieval and entered into the vitrification-warming process. The thawed oocytes were evaluated according to the survival criteria and then inseminated with the sperms of male mice for in vitro fertilization. The next day, the embryo formation rate and embryo quality were assessed. Our results demonstrated that even after 24 h of incubation, the survival rate of oocytes was 51.35% with the embryo formation rate of 73.21%. However, the survival and embryo formation rates significantly decreased within 12, 18, and 24 h after retrieval compared to the groups vitrified at 0 h. The embryo quality was significantly reduced by vitrification at 0 to 24 h after retrieval. According to our data, although a prolonged incubation time before freezing reduced the survival rate, there was still a chance for oocytes to stay alive with acceptable embryo formation and quality rates after vitrification warming of oocytes.

  20. Effect of MEM vitamins and forskolin on embryo development and vitrification tolerance of in vitro-produced pig embryos.

    PubMed

    Cuello, C; Gomis, J; Almiñana, C; Maside, C; Sanchez-Osorio, J; Gil, M A; Sánchez, A; Parrilla, I; Vazquez, J M; Roca, J; Martinez, E A

    2013-01-30

    The aims of this study were (1) to determine the effect of in vitro maturation (IVM) medium supplementation with MEM vitamins on in vitro embryo development and sensitivity to vitrification of Day 6 blastocysts and (2) to evaluate whether the addition of forskolin to in vitro culture (IVC) medium enhances blastocyst survival following Super Open Pulled Straw (SOPS) vitrification. Cumulus-oocyte complexes (COCs; n=4000) were matured with 0.0% or 0.05% (v/v) MEM vitamins. After 44h of IVM, the oocytes were in vitro fertilized, and presumptive zygotes were cultured. At Day 5 of IVC, embryos from both experimental groups were cultured for 24h with 0 or 10μM forskolin, achieving a 2×2 factorial design. The blastocyst formation rate was assessed on Day 6, and subsets of samples from the four experimental groups were vitrified (n=469) or kept fresh (n=546). Fresh and vitrified-warmed blastocysts were cultured for 24h prior to embryo survival and total blastocyst cell number assessment. The MEM vitamins increased (P<0.001) the blastocyst formation rate at Day 6, but they did not affect embryo survival after vitrification. In contrast, the addition of forskolin to the culture medium enhanced (P<0.05) the blastocyst vitrification tolerance. The total blastocyst cell number was similar among the groups. In conclusion, supplementation with 0.05% MEM vitamins improved the blastocyst formation rate, and the addition of 10μM forskolin to the culture medium increased survival in Day 6 in vitro-produced blastocysts after SOPS vitrification. Copyright © 2012 Elsevier B.V. All rights reserved.

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

    PubMed

    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.

  2. Alternation of apoptotic and implanting genes expression of mouse embryos after re-vitrification

    PubMed Central

    Majidi Gharenaz, Nasrin; Movahedin, Mansoureh; Mazaheri, Zohreh; Pour beiranvand, Shahram

    2016-01-01

    Background: Nowadays, oocytes and embryos vitrification has become a routine technique. Based on clinical judgment, re-vitrification maybe required. But little is known about re-vitrification impact on genes expression. Objective: The impact of re-vitrification on apoptotic and implanting genes, Bax, Bcl-2 and ErbB4, at compaction stage embryos were evaluated in this study. Materials and Methods: In this experimental study, 8 cell embryos (n=240) were collected from female mature mice, 60-62 hr post HCG injection. The embryos were divided randomly to 3 groups included: fresh (n=80), vitrified at 8 cell stage (n=80), vitrified at 8 cell stage thawed and re-vitrified at compaction stage (n=80). Embryos were vitrified by using cryolock, (open system) described by Kuwayama. Q-PCR was used to examine the expression of Bax, Bcl2 ErbB4 genes in derived blastocysts. Results: Our result showed that expanded blastocyst rate was similar between vitrified and re-vitrified groups, while re-vitrified embryos showed significant decrease in expanded blastocyst rate comparing with fresh embryos (p=0.03). In addition, significant difference was observed on apoptotic gene expression when comparing re-vitrified and fresh embryos (p=0.004), however expression of Bax and Bcl-2 (apoptotic) genes didn't demonstrate a significant difference between re-vitrified and vitrified groups. The expression rate of ErbB4, an implantation gene was decreased in re-vitrified embryos comparing with fresh embryos (p=0.003), but it was similar between re-vitrified and vitrified embryos. Conclusion: Re-vitrification can alter the expression of Bax, Bcl-2 and ErbB4 genes and developmental rate of mouse embryos in compaction stage. PMID:27679826

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

    PubMed

    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.

  4. Cryopreservation of mango (Mangifera indica L.) embryogenic cultures.

    PubMed

    Wu, Yong-Jie; Huang, Xue-Ling; Xiao, Jie-Ning; Li, Xiao-Ju; Zhou, Ming-De; Engelmann, Florent

    2003-01-01

    Three techniques were compared for cryopreserving embryogenic masses (EMs) sampled from mango (Mangifera indica L.) cv. Zihua embryogenic cultures: (i) encapsulation/dehydration; (ii) pregrowth/dehydration; and (iii) vitrification. In all experiments, EMs were sampled from embryogenic cultures during their exponential growth phase and pretreated for 24 h on solid medium containing 0.5 M sucrose before freezing. No recovery was achieved after cryopreservation using the encapsulation/dehydration technique, whatever the moisture content (fresh weight basis) of EMs, which ranged from 78.3% without dehydration to 40.8% after 6 h dehydration. With the pregrowth plus dehydration technique, limited recovery (8.3%) was achieved after desiccation of EMs for 1 h, to 58.5% MC. Using the vitrification technique, recovery ranged from 94.3% after treatment of EMs with the PVS3 vitrification solution for 20 min (EM moisture content of 34.7%) to 10.9% after a 120 min treatment with the vitrification solution (EM moisture content of 26.0%).

  5. Advances in the cryopreservation of mammalian oocytes and embryos: Development of ultrarapid vitrification

    PubMed Central

    2002-01-01

    The cryopreservation of embryos has become a powerful tool in assisted reproduction in several mammalian species. Embryos are cryopreserved by slow freezing or by vitrification. However, consistently high survival has not been obtained in most oocytes and in some embryos. The main reasons for the low survival would be sensitivity to low temperatures, which leads to chilling injury, and low permeability of the cell membrane, which leads to the formation of intracellular ice. As a strategy aiming to overcome these injuries, modified vitrification methods have been devised in which the cooling and warming rate is markedly increased by minimizing the volume of the solution and the container. The modified methods use electron microscope grids, open‐pulled straws, cryoloops, or container‐less microdrops. In this article, recent developments in the ultrarapid vitrification of mammalian oocytes and embryos are reviewed based on the understanding of the mechanisms of cell injury in cryopreservation. (Reprod Med Biol 2002; 1: 1–9) PMID:29699066

  6. Current status of human oocyte and embryo cryopreservation.

    PubMed

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

    2011-08-01

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

  7. Chapter 17 Sterile Plate-Based Vitrification of Adherent Human Pluripotent Stem Cells and Their Derivatives Using the TWIST Method.

    PubMed

    Neubauer, Julia C; Stracke, Frank; Zimmermann, Heiko

    2017-01-01

    Due to their high biological complexity, e.g., their close cell-to-cell contacts, cryopreservation of human pluripotent stem cells with standard slow-rate protocols often is inefficient and can hardly be standardized. Vitrification that means ultrafast freezing already showed very good viability and recovery rates for this sensitive cell system, but is only applicable for low cell numbers, bears a high risk of contamination, and can hardly be implemented under GxP regulations. In this chapter, a sterile plate-based vitrification method for adherent pluripotent stem cells and their derivatives is presented based on a procedure and device for human embryonic stem cells developed by Beier et al. (Cryobiology 66:8-16, 2013). This protocol overcomes the limitations of conventional vitrification procedures resulting in the highly efficient preservation of ready-to-use adherent pluripotent stem cells with the possibility of vitrifying cells in multi-well formats for direct application in high-throughput screenings.

  8. Evolution of human oocyte cryopreservation: slow freezing versus vitrification.

    PubMed

    Levi-Setti, Paolo Emanuele; Patrizio, Pasquale; Scaravelli, Giulia

    2016-12-01

    The purpose is to determine the efficiency and efficacy of oocyte cryopreservation by slow freezing versus vitrification, recent data collected from the Italian National Assisted Reproductive Technology Register during the period 2009-2014 will be presented and reviewed. The data on oocyte cryopreservation were also compared with the results obtained with embryo cryopreservation and relative IVF with fresh oocytes. During the period 2009-2014 preservation of oocytes by vitrification had a significantly higher survival rate, implantation, and pregnancy rate than slow freezing; however, there are still large variations in success rates among centers in relation to the number of procedures performed. Vitrification has now become the method of choice for oocyte cryopreservation because of better results than slow freezing, but still requires a more standardized utilization. The transfer of fresh or cryopreserved embryo still shows a statistically significant better performance than transfers with embryos obtained with cryopreserved oocytes. Only in a few centers with much experience in cryopreservation are the results between transfers of frozen embryos or embryos obtained from oocyte cryopreservation comparable.

  9. Vitrification solution containing DMSO and EG can induce parthenogenetic activation of in vitro matured ovine oocytes and decrease sperm penetration.

    PubMed

    Tian, Shu-Jun; Yan, Chang-Liang; Yang, Hui-Xin; Zhou, Guang-Bin; Yang, Zhong-Qiang; Zhu, Shi-En

    2007-10-01

    This study was designed to examine the reduced incidence of normal fertilization in vitrified ovine oocytes. After in vitro maturation for 24 h, the oocytes were randomly allocated into three groups: (1) untreated (control), (2) exposed to vitrification solution (VS) without being plunged into liquid nitrogen (toxicity), or (3) vitrified by open-pulled straw method (vitrification). In experiment 1, the treated and control oocytes were matured for another 2 h, and the oocytes were then in vitro fertilized for 12 h to examine sperm penetration. The percentage of monospermy in toxicity group (29.3%) and vitrification group (28.2%) dramatically decreased compared to the control group (45.0%) (P<0.05). To find the mechanism that the VS decreased the monospermy, some treated and control oocytes were used to test the distribution of CG and the resistance of zona pellucida (ZP) to 0.1% pronase E immediately (IVM 24 h), after another 2 h of maturation (IVM 26 h), and after 12 h of in vitro fertilization (IVF 12 h) respectively. Others were used to examine female pronucleus formation after 12 h of culture in fertilization medium with the absence of sperm. The results showed that the percentage of CG completely release in the oocytes (IVM 24 and 26 h) of toxicity group (41.2% and 39.9%) and vitrification group (41.7% and 51.7%) was significantly higher than that of control group (7.1% and 18.4%) (P<0.05). The ZP digestion duration in the oocytes (IVM 26 h) of the toxicity group (435.6 s) and vitrification group (422.3 s) was longer than that of control group (381.6 s) (P<0.05). The percentage of female pronucleus formation in toxicity group (58.7%) and vitrification group (63.9%) was higher than that (8.2%) of control group (P<0.05). The data above demonstrated that the VS containing DMSO and EG could parthenogenetically activate in vitro matured ovine oocytes, resulting in ZP hardening and decreased sperm penetration.

  10. A Rinsing Effluent Evaporator for Dismantling Operations - 13271

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

    Rives, Rachel; Asou-Pothet, Marielle; Chambon, Frederic

    2013-07-01

    Between 1958 and 1997, the UP1 plant at Marcoule - located in the south of France - reprocessed and recycled nearly 20,000 MT of used fuel from special defense applications reactors, as well as fuel from the first generation of electricity generating reactors in France (natural uranium fuel, CO{sub 2}-cooled, graphite-moderated). Decommissioning and Dismantling of the UP1 plant and its associated units started in 1998. Since 2005, the UP1 facility has been operated by AREVA as the Marcoule Management and Operation contractor for French Atomic Energy Commission (CEA). An important part of this decommissioning program deals with the vitrification facilitymore » of Marcoule. This facility includes 20 tanks devoted to interim storage of highly active solutions, prior to vitrification. In 2006, a rinsing program was defined as part of the tank cleanup strategy. The main objective of the rinsing phases was to decrease activity in order to limit the volume of 'long-life active' waste produced during the decommissioning operations, so the tanks can be dismantled without the need of remote operations. To enable this rinsing program, and anticipating large volumes of generated effluent, the construction of an evaporation unit proved to be essential. The main objective of this unit was to concentrate the effluent produced during tank rinsing operations by a factor of approximately 10, prior to it being treated by vitrification. The evaporator design phase was launched in September 2006. The main challenge for the Project team was the installation of this new unit within a nuclear facility still in operation and in existing compartments not initially designed for this purpose. Cold operating tests were completed in 2008, and in May 2009, the final connections to the process were activated to start the hot test phase. During the first hot test operations performed on the first batches of clean-up effluent, the evaporator had a major operating problem. Extremely large quantities of foam were produced, affecting the evaporator operation, and creating the risk of a reduction in its capacity and throughput performance. A task force of AREVA process, operations, and safety experts from Marcoule and the La Hague reprocessing complex was assembled. New operating parameters were defined and tested to improve the process. Since then, the evaporator has performed very satisfactorily. The foam buildup phenomenon has been brought under complete control. All the different types of effluents produced during cleanup operations have been concentrated, and the results obtained in terms of quality and throughput, have ensured a consistent supply to the vitrification unit. The evaporator was operated until the end of April 2012, and enabled the production of 500 cubic meters of very high activity effluent, concentrating the fission products rinsed from the storage tanks. The evaporator will now be deactivated and decommissioned, with the first rinsing and cleanup operations scheduled to begin in 2014. (authors)« less

  11. Prediction of the glass transition in aqueous solutions of simple amides by molecular dynamics simulations

    NASA Astrophysics Data System (ADS)

    Kreck, Cara A.; Mandumpal, Jestin B.; Mancera, Ricardo L.

    2011-01-01

    Some simple amides in aqueous solution are used in the cryopreservation of biological tissues as they are believed to promote the vitrification of water, inhibiting its crystallisation and the ensuing damage from ice formation. Molecular dynamics annealing simulations reveal a broadening in the glass transition of aqueous acetamide and N-methylacetamide solutions, suggesting a thermodynamic stabilisation of the glassy state, which may be responsible for their increased tendency of vitrification and their cryoprotective ability. By contrast, aqueous formamide solutions do not exhibit broadening of the glass transition; instead, it is shifted to lower temperatures, which explains their lack of vitrification properties.

  12. TECHNOLOGY DEMONSTRATION SUMMARY. BABCOCK AND WILCOX CYCLONE FURNACE VITRIFICATION TECHNOLOGY (EPA/540/SR-92/017)

    EPA Science Inventory

    A Superfund Innovative Technology Evaluation (SITE) Demonstration of the Babcock & Wilcox Cyclone Furnace Vitrification Technology was conducted in November 1991. This Demonstration occurred at the Babcock & Wilcox (B&W) Alliance Research Center (ARC) in Alliance, OH. The B&W cyc...

  13. A simple vitrification method for cryobanking avian testicular tissue

    USDA-ARS?s Scientific Manuscript database

    Cryopreservation of testicular tissue is a promising method of preserving male reproductive potential for avian species. This study was conducted to assess whether a vitrification method can be used to preserve avian testicular tissue, using the Japanese quail (Coturnix japonica) as a model. A sim...

  14. SITE TECHNOLOGY CAPSULE: GEOTECH DEVELOPMENT CORPORATION COLD TOP EX-SITU VITRIFICATION TECHNOLOGY

    EPA Science Inventory

    A SITE technology demonstration was conducted in 1997 to evaluate the potential applicability and effectiveness of the Geotech Cold Top ex-situ vitrification technology on chromium-contaminated soils. The primary objective was to develop test data to evaluate whether the waste a...

  15. Cleanup Verification Package for the 300 VTS Waste Site

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

    S. W. Clark and T. H. Mitchell

    2006-03-13

    This cleanup verification package documents completion of remedial action for the 300 Area Vitrification Test Site, also known as the 300 VTS site. The site was used by Pacific Northwest National Laboratory as a field demonstration site for in situ vitrification of soils containing simulated waste.

  16. EMERGING TECHNOLOGY SUMMARY: VITRIFICATION OF SOILS CONTAMINATED BY HAZARDOUS AND/OR RADIOACTIVE WASTES

    EPA Science Inventory

    A performance summary of an advanced multifuel-capable combustion and melting system (CMS) for the vitrification of hazardous wastes is presented. Vortex Corporation has evaluated its patented CMS for use in the remediation of soils contaminated with heavy metals and radionuclid...

  17. Successful vitrification of bovine immature oocyte using liquid helium instead of liquid nitrogen as cryogenic liquid.

    PubMed

    Yu, Xue-Li; Xu, Ya-Kun; Wu, Hua; Guo, Xian-Fei; Li, Xiao-Xia; Han, Wen-Xia; Li, Ying-Hua

    2016-04-01

    The objectives of this study were to compare the effectiveness of liquid helium (LHe) and liquid nitrogen (LN2) as cryogenic liquid for vitrification of bovine immature oocytes with open-pulled straw (OPS) system and determine the optimal cryoprotectant concentration of LHe vitrification. Cumulus oocyte complexes were divided into three groups, namely, untreated group (control), LN2 vitrified with OPS group, and LHe vitrified with OPS group. Oocyte survival was assessed by morphology, nuclear maturation, and developmental capability. Results indicated that the rates of normal morphology, maturation, cleavage, and blastocyst (89.3%, 52.8%, 42.7%, and 10.1%, respectively) in the LHe-vitrified group were all higher than those (79.3%, 43.4%, 34.1%, and 4.7%) in the LN2-vitrified group (P < 0.05) although the corresponding rates in both treated groups decreased compared with the control group (100%, 75.0%, 64.9%, and 40.8%; P < 0.05). Normal calves were obtained after the transfer of blastocysts derived from LHe- and LN2-vitrified oocytes. The effects of the different vitrification solutions (EDS30, EDS35, EDS40, EDS45, and EDS50) in LHe vitrification for bovine immature oocytes vitrification were examined. No difference was found in the rates of morphologically normal oocytes among the EDS30 (87.9%), EDS35 (90.1%), EDS40 (89.4%), and EDS45 (87.2%) groups (P > 0.05). The maturation rate of the EDS35 group (65.0%) was higher than those of the EDS30 (51.3%), EDS40 (50.1%), EDS45 (52.1%), and EDS50 groups (36.9%; P < 0.05). No significant differences were observed in the cleavage and blastocyst rates between the EDS35 (49.0% and 12.1%) and EDS40 (41.7% and 10.2%) groups. However, the cleavage and blastocyst rates in the EDS35 group were higher (P < 0.05) than those of the EDS30 (36.2% and 6.8%), EDS45 (35.9% and 5.8%), and EDS50 (16.6% and 2.2%) groups. In conclusion, LHe can be used as a cryogenic liquid for vitrification of bovine immature oocytes, and it is more efficient than LN2-vitrified oocytes in terms of blastocyst production. EDS35 was the optimal cryoprotectant agent combination for LHe vitrification in this study. Copyright © 2016 Elsevier Inc. All rights reserved.

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

    PubMed

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

    2018-04-01

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

  19. Thermal treatment and vitrification of boiler ash from a municipal solid waste incinerator.

    PubMed

    Yang, Y; Xiao, Y; Voncken, J H L; Wilson, N

    2008-06-15

    Boiler ash generated from municipal solid waste (MSW) incinerators is usually classified as hazardous materials and requires special disposal. In the present study, the boiler ash was characterized for the chemical compositions, morphology and microstructure. The thermal chemical behavior during ash heating was investigated with thermal balance. Vitrification of the ash was conducted at a temperature of 1400 degrees C in order to generate a stable silicate slag, and the formed slag was examined with chemical and mineralogical analyses. The effect of vitrification on the leaching characteristics of various elements in the ash was evaluated with acid leaching. The study shows that the boiler ash as a heterogeneous fine powder contains mainly silicate, carbonate, sulfates, chlorides, and residues of organic materials and heavy metal compounds. At elevated temperatures, the boiler ash goes through the initial moisture removal, volatilization, decomposition, sintering, melting, and slag formation. At 1400 degrees C a thin layer of salt melt and a homogeneous glassy slag was formed. The experimental results indicate that leaching values of the vitrified slag are significantly reduced compared to the original boiler ash, and the vitrification could be an interesting alternative for a safer disposal of the boiler ash. Ash compacting, e.g., pelletizing can reduce volatilization and weight loss by about 50%, and would be a good option for the feed preparation before vitrification.

  20. LONG-TERM CONSERVATION OF PROTOCORMS OF Brassavola nodosa (L) LIND. (ORCHIDACEAE): EFFECT OF ABA AND A RANGE OF CRYOCONSERVATION TECHNIQUES.

    PubMed

    Mata-Rosas, M; Lastre-Puertos, E

    2015-01-01

    Populations of Brassavola nodosa have been severely affected by habitat destruction and illegal collecting, and as with the majority of orchid species, it is critical to take action to guarantee their continued survival. The present study aimed to establish protocols for the long-term conservation of protocorms of species. Four different cryogenic techniques were compared: encapsulation-dehydration (ED), encapsulation-vitrification (EV), encapsulation-dehydration-vitrification (EDV) and vitrification. Preculture of protocorms with ABA was a critical factor in obtaining high percentages of regrowth. With vitrification, 100% regrowth was achieved in five treatments, mainly when protocorms were dehydrated with PVS2 for 120 min. 100% regrowth was also obtained with EDV, where the protocorms were precultured with ABA 5 mg/l for 3 days and incubated with PVS2 for 60 min. With the ED, regrowth of 72% was achieved with the preculture of protocorms with ABA 5 mg/l for the three times of incubation used (3, 6 and 9 days). In the case of EV, 92% regrowth, was recorded when protocorms were precultured for 9 days with ABA 3 mg/l and incubated with PVS2 for 90 min. Although regrowth of protocorms was obtained with all the techniques used, the vitrification technique is preferred since it requires less labour and is less costly.

  1. New controlled environment vitrification system for preparing wet samples for cryo-SEM.

    PubMed

    Ge, H; Suszynski, W J; Davis, H T; Scriven, L E

    2008-01-01

    A new controlled environment vitrification system (CEVS) has been designed and constructed to facilitate examination by cryogenic scanning electron microscopy (Cryo-SEM) of initial suspension state and of microstructure development in latex, latex-composite and other coatings while they still contain solvent. The new system has a main chamber with provisions for coating as well as drying, and for well-controlled plunging into cryogen. An added subsidiary chamber holds samples for drying or annealing over minutes to days before they are returned to the main chamber and plunged from it. In the main chamber, samples are blade-coated on 5 x 7 mm pieces of silicon wafer and held at selected temperature and humidity for successively longer times, either there or after transfer along a rail into the subsidiary chamber. They are then placed in the sample holder mounted on the plunge rod, so as to permit adjustment of the sample's attitude when it plunges, at controlled speed, into liquid ethane at its freezing point, to a chosen depth, in order to solidify the sample without significant shear or freezing artifacts. The entries of plunging samples and related sample holders into liquid ethane were recorded with a high-speed, high-resolution Photron digital camera. The data were interpreted with a new hypothesis about the width of the band of extremely rapid cooling by deeply subcooled nucleate boiling below the line of entry. Complementary cryo-SEM images revealed that the freezing rate and surface shearing of a sample need to be balanced by adjusting the plunging attitude.

  2. Shoot regeneration and embryogenesis in lily shoot tips cryopreserved by droplet vitrification

    USDA-ARS?s Scientific Manuscript database

    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...

  3. 40 CFR 72.72 - Criteria for State operating permit program.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.72 Criteria for State operating permit program. A State operating permit program (including a State Acid Rain program) shall meet... withdrawal of all or part of the Acid Rain portion of an approved State operating permit program by the...

  4. 40 CFR 72.72 - Criteria for State operating permit program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.72 Criteria for State operating permit program. A State operating permit program (including a State Acid Rain program) shall meet... withdrawal of all or part of the Acid Rain portion of an approved State operating permit program by the...

  5. 40 CFR 72.72 - Criteria for State operating permit program.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.72 Criteria for State operating permit program. A State operating permit program (including a State Acid Rain program) shall meet... withdrawal of all or part of the Acid Rain portion of an approved State operating permit program by the...

  6. 40 CFR 72.72 - Criteria for State operating permit program.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.72 Criteria for State operating permit program. A State operating permit program (including a State Acid Rain program) shall meet... withdrawal of all or part of the Acid Rain portion of an approved State operating permit program by the...

  7. 40 CFR 72.72 - Criteria for State operating permit program.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.72 Criteria for State operating permit program. A State operating permit program (including a State Acid Rain program) shall meet... withdrawal of all or part of the Acid Rain portion of an approved State operating permit program by the...

  8. Different routes to the glass transition: A comparison between chemical and physical vitrification

    NASA Astrophysics Data System (ADS)

    Caponi, Silvia; Corezzi, Silvia

    2012-07-01

    Despite the differences in the molecular processes involved in chemical and physical vitrification, surprising similarities are observed in the dynamics and in the thermodynamical properties of the resulting glasses. We report on a systematic study of reactive glass-formers undergoing a process of progressive polymerization of the constituent molecules via the formation of irreversible chemical bonds. The formation of most of the materials used in engineering plastics and the hardening of natural and synthetic resins, including epoxy resins, are based on chemical vitrification. The clear analogies characterizing the dynamic evolution of physical and chemical glass-formers, on the time scale of the structural and the low-frequency vibrational dynamics, are briefly reviewed.

  9. Identification of a highly successful cryopreservation method (droplet-vitrification) for petunia

    USDA-ARS?s Scientific Manuscript database

    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...

  10. Cryopreservation of in vitro-grown apical meristems of wasabi (Wasabia japonica) by vitrification and subsequent high plant regeneration.

    PubMed

    Matsumoto, T; Sakai, A; Yamada, K

    1994-05-01

    In vitro-grown apical meristems of wasabi (Wasabia japonica Matsumura) were successfully cryopreserved by vitrification. Excised apical meristems precultured on solidified M S medium containing 0.3M sucrose at 20°C for 1 day were loaded with a mixture of 2M glycerol and 0.4M sucrose for 20 min at 25°C. Cryoprotected meristems were then sufficiently dehydrated with a highly concentrated vitrification solution (designated PVS2) for 10 min at 25°C prior to a plunge into liquid nitrogen. After rapid warming, the meristems were expelled into 2 ml of 1.2M sucrose for 20 min and then plated on solidified culture medium. Successfully vitrified and warmed meristems remained green after plating, resumed growth within 3 days, and directly developed shoots within two weeks. The average rate of normal shoot formation amounted to about 80 to 90% in the cryopreserved meristems. This method was successfully applied to three other cultivars of wasabi. This vitrification procedure promises to become a routine method for cryopreserving meristems of wasabi.

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

    PubMed Central

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

    2015-01-01

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

  12. Roles of intracellular ice formation, vitrification of cell water, and recrystallisation of intracellular ice on the survival of mouse embryos and oocytes.

    PubMed

    Mazur, Peter; Paredes, Estefania

    2016-03-01

    Mazur and collaborators began examining the validity of initial views regarding mouse oocyte and embryo vitrification and found that most are partially or fully wrong. First, the relative effects of warming and cooling rates on the survival of mouse oocytes subjected to a vitrification procedure were determined. The high sensitivity to warming rate strongly suggests that the lethality of slow warming is a consequence of either the crystallisation of intracellular glassy water during warming or the recrystallisation during slow warming of small intracellular crystals that had formed during cooling. Warming rates of 107°C min-1 were achieved in 0.1-µL drops of ethylene glycol-acetamide-Ficoll-sucrose (EAFS) solution plus a small amount of India ink on Cryotops warmed using an infrared laser pulse. Under these conditions, survival rates of 90% were obtained even when mouse oocytes were suspended in 0.3× EAFS, a concentration that falls in the range that many cells can tolerate. A second important finding was that the survival of oocytes is more dependent on the osmotic withdrawal of much of the intracellular water before vitrification than it is on the penetration of cryoprotective solutes into the cells. Herein we review the roles of internal ice formation, vitrification and recrystallisation. It remains to be seen how widely these findings will be applicable to other types of cells and tissues from other species.

  13. Simple, efficient and successful vitrification of bovine blastocysts using electron microscope grids.

    PubMed

    Park, S P; Kim, E Y; Kim, D I; Park, N H; Won, Y S; Yoon, S H; Chung, K S; Lim, J H

    1999-11-01

    This study demonstrates that higher survival of vitrified-thawed bovine blastocysts can be obtained using electron microscope (EM) grids as embryo containers at freezing, rather than plastic straws. In-vitro produced day 7 bovine blastocysts after in-vitro fertilization (IVF) were vitrified on grids or in straws with EFS40 freezing solution and their survival after thawing was compared. Embryo survival was assessed as re-expanded and hatched rates at 24 and 48 h after thawing respectively. When the effects of exposure to vitrification solution and chilling injury from the freezing procedure were examined, embryo survival in the exposure group (24 h: 100, 48 h: 73.3%) was not different compared with that in the control group (100, 84.4%). After vitrification, the hatched rate of the EM grid group 48 h after thawing (67.8%) was significantly higher than that of the straw group (53.3%) (P < 0.05). Fast developing embryos (expanded blastocyst and early hatching blastocyst stage) showed better resistance to freezing than delayed ones (early blastocyst stage), irrespective of embryo containers (early: 24 h, 57.1 and 48 h, 24.4%; expanded: 84.7 and 60.6%; early hatching: 91.7 and 80.0%) (P < 0.001). When using expanded and early hatching blastocysts, embryo survival rates in the vitrification-EM grid group (67.8, 95.0% respectively) were significantly higher than that of the vitrification-straw group (53.0, 65.0%) at 48 h.

  14. A Low Temperature Limit for Life on Earth

    PubMed Central

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

    2013-01-01

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

  15. Impact of delipidated estrous sheep serum supplementation on in vitro maturation, cryotolerance and endoplasmic reticulum stress gene expression of sheep oocytes

    PubMed Central

    dos Santos Neto, Pedro C.; Cuadro, Federico; Bosolasco, Diego; Mulet, Ana P.; Crispo, Martina

    2018-01-01

    High lipid content of oocytes and embryos in domestic animals is one of the well-known factors associated with poor cryosurvival. Herein, we wanted to determine whether the use of delipidated estrous sheep serum during in vitro maturation (IVM) of ovine oocytes reduces the cytoplasmic lipid droplets content and improves embryo development and cryotolerance after vitrification. Cumulus oocytes complexes (COCs) were matured in vitro for 24 h in medium supplemented with whole or delipidated estrous sheep serum prior to vitrification. Neutral lipid present in lipid droplets of COCs, cleavage rate, embryo development rate on Day 6 and Day 8, and hatching rate on Day 8, were compared among experimental groups. Endoplasmic reticulum stress genes were evaluated in in vitro matured COCs under different lipid conditions prior to vitrification. The lipid droplets’ content (mean fluorescence intensity) of oocytes cultured with IVM media supplemented with delipidated serum was lower than COCs matured with whole serum (7.6 ± 1.7 vs. 22.8 ± 5.0 arbitrary units, respectively; P< 0.05). Despite IVM treatment, oocytes subjected to vitrification showed impaired competence compared with the non-vitrified groups (P<0.05). No significant differences in embryo production were observed in non-vitrified COCs after maturation in delipidated or whole serum (33.4±4.9 vs 31.9 ±4.2). COCs matured in delipidated serum and subjected to vitrification showed increased expression of ATF4, ATF6, GRP78, and CHOP10 genes (ER stress markers). Collectively, our results demonstrate that although supplementation of IVM medium with delipidated estrous sheep serum reduces the presence of cytoplasmic lipid droplets in oocytes after maturation, oocyte cryotolerance is not improved. Notably, the expression of genes associated with the unfolded protein response (UPR) was increased in COCs, with fewer lipid droplets subjected to vitrification, suggesting that oocyte cryopreservation is associated with ER stress and activation of adaptive responses. PMID:29912910

  16. A Low Temperature Limit for Life on Earth.

    PubMed

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

    2013-01-01

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

  17. Timing of The First Zygotic Cleavage Affects Post-Vitrification Viability of Murine Embryos Produced In Vivo

    PubMed Central

    Jusof, Wan-Hafizah Wan; Khan, Nor-Ashikin Mohamed Noor; Rajikin, Mohd Hamim; Satar, Nuraliza Abdul; Mustafa, Mohd-Fazirul; Jusoh, Norhazlin; Dasiman, Razif

    2015-01-01

    Background Timing of the first zygotic cleavage is an accurate predictor of embryo quality. Embryos that cleaved early (EC) have been shown to exhibit higher develop- mental viability compared to those that cleaved at a later period (LC). However, the vi- ability of EC embryos in comparison to LC embryos after vitrification is unknown. The present study aims to investigate the post-vitrification developmental viability of murine EC versus LC embryos. Materials and Methods In this experimental study, female ICR mice (6-8 weeks old) were superovulated and cohabited with fertile males for 24 hours. Afterwards, their ovi- ducts were excised and embryos harvested. Embryos at the 2-cell stage were catego- rized as EC embryos, while zygotes with two pronuclei were categorized as LC embryos. Embryos were cultured in M16 medium supplemented with 3% bovine serum albumin (BSA) in a humidified 5% CO2atmosphere. Control embryos were cultured until the blastocyst stage without vitrification. Experimental embryos at the 2-cell stage were vitri- fied for one hour using 40% v/v ethylene glycol, 18% w/v Ficoll-70 and 0.5 M sucrose as the cryoprotectant. We recorded the numbers of surviving embryos from the control and experimental groups and their development until the blastocyst stage. Results were analyzed using the chi-square test. Results A significantly higher proportion of EC embryos (96.7%) from the control group developed to the blastocyst stage compared with LC embryos (57.5%, P<0.0001). Similarly, in the experimental group, a significantly higher percentage of vitrified EC embryos (69.4%) reached the blastocyst stage compared to vitrified LC embryos (27.1%, P<0.0001). Conclusion Vitrified EC embryos are more vitrification tolerant than LC embryos. Prese- lection of EC embryos may be used as a tool for selection of embryos that exhibit higher developmental competence after vitrification. PMID:26246881

  18. A COMPREHENSIVE TECHNICAL REVIEW OF THE DEMONSTRATION BULK VITRIFICATION SYSTEM

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

    SCHAUS, P.S.

    2006-09-29

    In May 2006, CH2M Hill Hanford Group, Inc. chartered an Expert Review Panel (ERP) to review the current status of the Demonstration Bulk Vitrification System (DBVS). It is the consensus of the ERP that bulk vitrification is a technology that requires further development and evaluation to determine its potential for meeting the Hanford waste stabilization mission. No fatal flaws (issues that would jeopardize the overall DBVS mission that cannot be mitigated) were found, given the current state of the project. However, a number of technical issues were found that could significantly affect the project's ability to meet its overall missionmore » as stated in the project ''Justification of Mission Need'' document, if not satisfactorily resolved. The ERP recognizes that the project has changed from an accelerated schedule demonstration project to a formally chartered project that must be in full compliance with DOE 413.3 requirements. The perspective of the ERP presented herein, is measured against the formally chartered project as stated in the approved Justification of Mission Need document. A justification of Mission Need document was approved in July 2006 which defined the objectives for the DBVS Project. In this document, DOE concluded that bulk vitrification is a viable technology that requires additional development to determine its potential applicability to treatment of a portion of the Hanford low activity waste. The DBVS mission need statement now includes the following primary objectives: (1) process approximately 190,000 gallons of Tank S-109 waste into fifty 100 metric ton boxes of vitrified product; (2) store and dispose of these boxes at Hanford's Integrated Disposal Facility (IDF); (3) evaluate the waste form characteristics; (4) gather pilot plant operability data, and (5) develop the overall life cycle system performance of bulk vitrification and produce a comparison of the bulk vitrification process to building a second LAW Immobilization facility or other supplemental treatment alternatives as provided in M-62-08.« less

  19. Supplemental Immobilization of Hanford Low-Activity Waste: Cast Stone Screening Tests

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

    Westsik, Joseph H.; Piepel, Gregory F.; Lindberg, Michael J.

    2013-09-30

    More than 56 million gallons of radioactive and hazardous waste are stored in 177 underground storage tanks at the U.S. Department of Energy’s (DOE’s) Hanford Site in southeastern Washington State. The Hanford Tank Waste Treatment and Immobilization Plant (WTP) is being constructed to treat the wastes and immobilize them in a glass waste form. The WTP includes a pretreatment facility to separate the wastes into a small volume of high-level waste (HLW) containing most of the radioactivity and a larger volume of low-activity waste (LAW) containing most of the nonradioactive chemicals. The HLW will be converted to glass in themore » HLW vitrification facility for ultimate disposal at an offsite federal repository. At least a portion (~35%) of the LAW will be converted to glass in the LAW vitrification facility and will be disposed of onsite at the Integrated Disposal Facility (IDF). The pretreatment and HLW vitrification facilities will have the capacity to treat and immobilize the wastes destined for each facility. However, a second LAW immobilization facility will be needed for the expected volume of LAW requiring immobilization. A cementitious waste form known as Cast Stone is being considered to provide the required additional LAW immobilization capacity. The Cast Stone waste form must be acceptable for disposal in the IDF. The Cast Stone waste form and immobilization process must be tested to demonstrate that the final Cast Stone waste form can comply with the waste acceptance criteria for the disposal facility and that the immobilization processes can be controlled to consistently provide an acceptable waste form product. Further, the waste form must be tested to provide the technical basis for understanding the long-term performance of the waste form in the disposal environment. These waste form performance data are needed to support risk assessment and performance assessment (PA) analyses of the long-term environmental impact of the waste disposal in the IDF. The PA is needed to satisfy both Washington State IDF Permit and DOE Order requirements. Cast Stone has been selected for solidification of radioactive wastes including WTP aqueous secondary wastes treated at the Effluent Treatment Facility (ETF) at Hanford. A similar waste form called Saltstone is used at the Savannah River Site (SRS) to solidify its LAW tank wastes.« less

  20. Enhanced LAW Glass Correlation - Phase 1

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

    Muller, Isabelle S.; Matlack, Keith S.; Pegg, Ian L.

    About 50 million gallons of high-level mixed waste is currently stored in underground tanks at the United States Department of Energy’s (DOE’s) Hanford site in the State of Washington. The Hanford Tank Waste Treatment and Immobilization Plant (WTP) will provide DOE’s Office of River Protection (ORP) with a means of treating this waste by vitrification for subsequent disposal. The tank waste will be separated into low- and high-activity waste fractions, which will then be vitrified respectively into Immobilized Low Activity Waste (ILAW) and Immobilized High Level Waste (IHLW) products. The ILAW product will be disposed in an engineered facility onmore » the Hanford site while the IHLW product is designed for acceptance into a national deep geological disposal facility for high-level nuclear waste. The ILAW and IHLW products must meet a variety of requirements with respect to protection of the environment before they can be accepted for disposal. Acceptable glass formulations for vitrification of Hanford low activity waste (LAW) must meet a variety of product quality, processability, and waste loading requirements. To this end, The Vitreous State Laboratory (VSL) at The Catholic University of America (CUA) developed and tested a number of glass formulations during Part A, Part B1 and Part B2 of the WTP development program. The testing resulted in the selection of target glass compositions for the processing of eight of the Phase I LAW tanks. The selected glass compositions were tested at the crucible scale to confirm their compliance with ILAW performance requirements. Duramelter 100 (DM100) and LAW Pilot Melter tests were then conducted to demonstrate the viability of these glass compositions for LAW vitrification at high processing rates.« less

  1. Vitrification of oocytes from endangered Mexican gray wolves (Canis lupus baileyi).

    PubMed

    Boutelle, S; Lenahan, K; Krisher, R; Bauman, K L; Asa, C S; Silber, S

    2011-03-01

    Careful genetic management, including cryopreservation of genetic material, is central to conservation of the endangered Mexican gray wolf. We tested a technique, previously used to vitrify human and domestic animal oocytes, on oocytes from domestic dogs as a model and from the endangered Mexican wolf. This method provided a way to conserve oocytes from genetically valuable older female Mexican wolves as an alternative to embryos for preserving female genes. Oocytes were aspirated from ovaries of 36 female dogs in December and March (0 to 65 oocytes per female) and from six female wolves (4 to 73 per female) during their physiologic breeding season, or following stimulation with the GnRH agonist deslorelin. Oocytes from dogs were pooled; half were immediately tested for viability and the remainder vitrified, then warmed and tested for viability. All oocytes were vitrified by being moved through media of increasing cryoprotectant concentration, placed on Cryotops, and plunged into liquid nitrogen. There was no difference in viability (propidium iodide staining) between fresh and vitrified, warmed dog oocytes (65.7 and 61.0%, respectively, P = 0.27). Oocyte viability after warming was similarly assessed in a subset of wolves (4 to 15 oocytes from each of three females; total 29 oocytes). Of these, 57.1% of the post-thaw intact oocytes were viable, which was 41.4% of all oocytes warmed. These were the first oocytes from a canid or an endangered species demonstrated to have maintained viability after vitrification and warming. Furthermore, our results demonstrated that vitrification of oocytes with the Cryotop technique was an option for preserving female gametes from Mexican wolves for future use in captive breeding programs, although in vitro embryo production techniques must first be developed in canids for this technique to be used. Copyright © 2011 Elsevier Inc. All rights reserved.

  2. Effects of vitrification cryopreservation on follicular morphology and stress relaxation behaviors of human ovarian tissues: sucrose versus trehalose as the non-permeable protective agent.

    PubMed

    Tian, Ting; Zhao, Gang; Han, Dan; Zhu, Kaixuan; Chen, Dawei; Zhang, Zhiguo; Wei, Zhaolian; Cao, Yunxia; Zhou, Ping

    2015-04-01

    Is sucrose more effective than trehalose in human ovarian tissue cryopreservation? The effect of sucrose as a cryoprotective agent (CPA) was not significantly different from that of trehalose in human ovarian tissue cryopreservation. Sugars have the ability to keep the cell membrane intact and can decrease the toxicity of CPAs. Sucrose is the most commonly used non-permeable CPA, while trehalose is rarely used in human ovarian tissue cryopreservation. Although various methods are utilized to evaluate the efficiency of human ovarian tissue cryopreservation, few studies have evaluated the effect of cryopreservation from the viewpoint of biomechanics. A total of 15 ovarian tissue samples were collected from 15 patients (20-41 years old) with benign ovarian tumors or malignancies, and each was dissected into six slices. Two slices were taken as the fresh control group. The remaining four slices were vitrified using different vitrification protocols. After warming, samples in each group were either fixed for histological evaluation or destined for stress relaxation test. The CPA solutions for the control and vitrified groups were composed of EDS and EDT (E, ethylene glycol; D, dimethylsulphoxide; S, sucrose; T, trehalose). The stress relaxation experiments were carried out at room temperature using a dynamic mechanical analyzer. Ovarian tissue samples were assessed for both their morphology and viscoelasticity. Stress relaxation data (SRD) were calculated as a percentage, representing the ability to maintain the initial stress after stretching. The percentage of morphologically normal follicles was compared between groups, which was represented by morphologic preservation ratio. The morphologic preservation ratio of the primordial follicles in the fresh control group (87.58%) was higher than that in group S (72.33%) (P = 0.000) and group T (79.56%) (P = 0.002). Although not statistically significant, compared with the S group, vitrification with T suggested a trend toward a higher morphologic preservation ratio of the primordial follicles. The SRD in the fresh control group (0.6433 ± 0.7233) was significantly different from that in group S (0.5200 ± 0.8331, P = 0.000) or in group T (0.5667 ± 0.6415, P = 0.000). However, no significant difference was found between groups S and T. Experimental samples were directly exposed to the air, which will result in a discrepancy in the viscoelastic properties between experimental tissues and in vivo tissues. Our study suggested a trend toward a higher morphologic preservation ratio of the primordial follicles after vitrification in trehalose compared with sucrose, which may provide a basis for further optimizing human ovarian tissue vitrification. In addition, it was possible to evaluate the effect of ovarian tissue cryopreservation from a biomechanics perspective. This study was supported by the grants from the Medical Scientific Research Subject, Health Ministry of Anhui Province (2010B014) and National Basic Research Program of China (973 Program) (2012CB944704), and the National Natural Science Foundation of China (Nos. 51276179 and 51476160). The authors declare that there is no conflict of interests regarding the publication of this original paper. © The Author 2015. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  3. Proceedings of the 21st DOE/NRC Nuclear Air Cleaning Conference; Sessions 1--8

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

    First, M.W.

    1991-02-01

    Separate abstracts have been prepared for the papers presented at the meeting on nuclear facility air cleaning technology in the following specific areas of interest: air cleaning technologies for the management and disposal of radioactive wastes; Canadian waste management program; radiological health effects models for nuclear power plant accident consequence analysis; filter testing; US standard codes on nuclear air and gas treatment; European community nuclear codes and standards; chemical processing off-gas cleaning; incineration and vitrification; adsorbents; nuclear codes and standards; mathematical modeling techniques; filter technology; safety; containment system venting; and nuclear air cleaning programs around the world. (MB)

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

    PubMed

    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.

  5. 40 CFR 122.46 - Duration of permits (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Permit Conditions § 122.46 Duration of permits (applicable to State programs, see § 123.25...

  6. Improvement of vitrification of in vitro produced buffalo embryos with special reference to sex ratio following vitrification

    PubMed Central

    Mahmoud, K. Gh. M; Scholkamy, T. H; Darwish, S. F

    2015-01-01

    Cryopreservation and sexing of embryos are integrated into commercial embryo transfer technologies. To improve the effectiveness of vitrification of in vitro produced buffalo embryos, two experiments were conducted. The first evaluated the effect of exposure time (2 and 3 min) and developmental stage (morula and blastocysts) on the viability and development of vitrified buffalo embryos. Morphologically normal embryos and survival rates (re-expansion) significantly increased when vitrified morulae were exposed for 2 min compared to 3 min (P<0.001). On the other hand, morphologically normal and survival rates of blastocysts significantly increased when exposed for 3 min compared to 2 min (P<0.001). However, there were no significant differences between the two developmental stages (morulae and blastocystes) in the percentages of morphologically normal embryos and re-expansion rates after a 24 h culture. The second experiment aimed to evaluate the effect of viability on the sex ratio of buffalo embryos after vitrification and whether male and female embryos survived vitrification differently. A total number of 61 blastocysts were vitrified for 3 min with the same cryoprotectant as experiment 1. Higher percentages of males were recorded for live as compared to dead embryos; however, this difference was not significant. In conclusion, the post-thaw survival and development of in vitro produced morulae and blastocysts were found to be affected by exposure time rather than developmental stage. Survivability had no significant effect on the sex ratio of vitrified blastocysts; nevertheless, the number of surviving males was higher than dead male embryos. PMID:27175197

  7. Thermal Flammable Gas Production from Bulk Vitrification Feed

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

    Scheele, Randall D.; McNamara, Bruce K.; Bagaasen, Larry M.

    2008-05-21

    The baseline bulk-vitrification (BV) process (also known as in-container vitrification ICV™) includes a mixer/dryer to convert liquid low-activity waste (LAW) into a dried, blended feed for vitrification. Feed preparation includes blending LAW with glass-forming minerals (GFMs) and cellulose and drying the mixture to a suitable dryness, consistency, and particle size for transport to the ICVTM container. The cellulose is to be added to the BV feed at a rate sufficient to destroy 75% of the nitrogen present as nitrate or nitrite. Concern exists that flammable gases may be produced during drying operations at levels that could pose a risk. Themore » drying process is conducted under vacuum in the temperature range of 60 to 80°C. These flammable gases could be produced either through thermal decomposition of cellulose or waste organics or as a by-product of the reaction of cellulose and/or waste organics with nitrate or the postulated small amount of nitrite present in the waste. To help address the concern about flammable gas production during drying, the Pacific Northwest National Laboratory (PNNL) performed studies to identify the gases produced at dryer temperatures and at possible process upset conditions. Studies used a thermogravimetric analyzer (TGA) up to 525°C and isothermal testing up to 120°C to determine flammable gas production resulting from the cellulose and organic constituents in bulk vitrification feed. This report provides the results of those studies to determine the effects of cellulose and waste organics on flammable gas evolution« less

  8. Toxicity Minimized Cryoprotectant Addition and Removal Procedures for Adherent Endothelial Cells

    PubMed Central

    Davidson, Allyson Fry; Glasscock, Cameron; McClanahan, Danielle R.; Benson, James D.; Higgins, Adam Z.

    2015-01-01

    Ice-free cryopreservation, known as vitrification, is an appealing approach for banking of adherent cells and tissues because it prevents dissociation and morphological damage that may result from ice crystal formation. However, current vitrification methods are often limited by the cytotoxicity of the concentrated cryoprotective agent (CPA) solutions that are required to suppress ice formation. Recently, we described a mathematical strategy for identifying minimally toxic CPA equilibration procedures based on the minimization of a toxicity cost function. Here we provide direct experimental support for the feasibility of these methods when applied to adherent endothelial cells. We first developed a concentration- and temperature-dependent toxicity cost function by exposing the cells to a range of glycerol concentrations at 21°C and 37°C, and fitting the resulting viability data to a first order cell death model. This cost function was then numerically minimized in our state constrained optimization routine to determine addition and removal procedures for 17 molal (mol/kg water) glycerol solutions. Using these predicted optimal procedures, we obtained 81% recovery after exposure to vitrification solutions, as well as successful vitrification with the relatively slow cooling and warming rates of 50°C/min and 130°C/min. In comparison, conventional multistep CPA equilibration procedures resulted in much lower cell yields of about 10%. Our results demonstrate the potential for rational design of minimally toxic vitrification procedures and pave the way for extension of our optimization approach to other adherent cell types as well as more complex systems such as tissues and organs. PMID:26605546

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

    PubMed Central

    Zheng, Yuanyuan; Panhwar, Fazil

    2016-01-01

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

  10. Local geology controlled the feasibility of vitrifying Iron Age buildings.

    PubMed

    Wadsworth, Fabian B; Heap, Michael J; Damby, David E; Hess, Kai-Uwe; Najorka, Jens; Vasseur, Jérémie; Fahrner, Dominik; Dingwell, Donald B

    2017-01-12

    During European prehistory, hilltop enclosures made from polydisperse particle-and-block stone walling were exposed to temperatures sufficient to partially melt the constituent stonework, leading to the preservation of glassy walls called 'vitrified forts'. During vitrification, the granular wall rocks partially melt, sinter viscously and densify, reducing inter-particle porosity. This process is strongly dependent on the solidus temperature, the particle sizes, the temperature-dependence of the viscosity of the evolving liquid phase, as well as the distribution and longevity of heat. Examination of the sintering behaviour of 45 European examples reveals that it is the raw building material that governs the vitrification efficiency. As Iron Age forts were commonly constructed from local stone, we conclude that local geology directly influenced the degree to which buildings were vitrified in the Iron Age. Additionally, we find that vitrification is accompanied by a bulk material strengthening of the aggregates of small sizes, and a partial weakening of larger blocks. We discuss these findings in the context of the debate surrounding the motive of the wall-builders. We conclude that if wall stability by bulk strengthening was the desired effect, then vitrification represents an Iron Age technology that failed to be effective in regions of refractory local geology.

  11. Successful ongoing pregnancies after vitrification of oocytes.

    PubMed

    Lucena, Elkin; Bernal, Diana Patricia; Lucena, Carolina; Rojas, Alejandro; Moran, Abby; Lucena, Andrés

    2006-01-01

    To demonstrate the efficiency of vitrifying mature human oocytes for different clinical indications. Descriptive case series. Cryobiology laboratory, Centro Colombiano de Fertilidad y Esterilidad-CECOLFES LTDA. (Bogotá, Colombia). Oocyte vitrification was offered as an alternative management for patients undergoing infertility treatment because of ovarian hyperstimulation syndrome, premature ovarian failure, natural ovarian failure, male factor, poor response, or oocyte donation. Mature oocytes were obtained from 33 donor women and 40 patients undergoing infertility treatment. Oocytes were retrieved by ultrasound-guided transvaginal aspiration and vitrified with the Cryotops method, with 30% ethylene glycol, 30% dimethyl sulfoxide, and 0.5 mol/L sucrose. Viability was assessed 3 hours after thawing. The surviving oocytes were inseminated by intracytoplasmic sperm injection. Fertilization was evaluated after 24 hours. The zygotes were further cultured in vitro for up to 72 hours until time of embryo transfer. Recovery, viability, fertilization, and pregnancy rates. Oocyte vitrification with the Cryotop method resulted in high rates of recovery, viability, fertilization, cleavage, and ongoing pregnancy. Vitrification with the Cryotop method is an efficient, fast, and economical method for oocyte cryopreservation that offers high rates of survival, fertilization, embryo development, and ongoing normal pregnancies, providing a new alternative for the management of female infertility.

  12. Dismantling of Highly Contaminated Process Installations of the German Reprocessing Facility (WAK) - Status of New Remote Handling Technology - 13287

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

    Dux, Joachim; Friedrich, Daniel; Lutz, Werner

    2013-07-01

    Decommissioning and dismantling of the former German Pilot Reprocessing Plant Karlsruhe (WAK) including the Vitrification Facility (VEK) is being executed in different Project steps related to the reprocessing, HLLW storage and vitrification complexes /1/. While inside the reprocessing building the total inventory of process equipment has already been dismantled and disposed of, the HLLW storage and vitrification complex has been placed out of operation since vitrification and tank rinsing procedures where finalized in year 2010. This paper describes the progress made in dismantling of the shielded boxes of the highly contaminated laboratory as a precondition to get access to themore » hot cells of the HLLW storage. The major challenges of the dismantling of this laboratory were the high dose rates up to 700 mSv/h and the locking technology for the removal of the hot cell installations. In parallel extensive prototype testing of different carrier systems and power manipulators to be applied to dismantle the HLLW-tanks and other hot cell equipment is ongoing. First experiences with the new manipulator carrier system and a new master slave manipulator with force reflection will be reported. (authors)« less

  13. Influence of Meiotic Stages on Developmental Competence of Goat’ Oocyte After Vitrification

    NASA Astrophysics Data System (ADS)

    Wahyuningsih, S.; Ihsan, M. N.

    2018-02-01

    This objective of this research was to investigate effect of goat oocyte meiotic stages on developmental competence after cryopreservation. Ovaries were collected from slaugterhouse and oocytes was aspirated from2-6 mm of follicles. Oocyte with compacted cumulus cells and evenly granulated ooplasm were selected for this experiment. The lenght of in vitro maturation before vitrification was 8 or 22 h in IVM media TCM 199 + FCS 10 % + PMSG 10 IU + hCG 10 IU at 38.5 °C in a humidified atmosphere of 5 % CO2 in air and were vitrified. After vitrification process, GVBD and MII oocyte were matured for 18 or 4 h to fullfill 26 h maturation requirement and then oocytes were subjected to IVF and culture. Cleavage and blastocyst formation rate were to asses their developmental competence. Cleavage rates were obtained for both GVBD ( 56.78 %) and MII (69.64 % ) oocytes (P<0.05). Proportion of cleaved embryos from vitrified MII oocytes develop into blastocysts higher (P<0.05) than those from vitrified GVBD oocytes (10.25% vs 3.54%) repectively. Goat oocytes in different maturation stages response to vitrification differently and MII stages have better developmental competence than GVBD.

  14. 40 CFR 72.70 - Relationship to title V operating permit program.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...) AIR PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.70 Relationship to... operating permit programs and acceptance of State Acid Rain programs, the procedure for including State Acid... of an accepted State program, to issue Phase II Acid Rain permits. (b) Relationship to operating...

  15. 40 CFR 72.70 - Relationship to title V operating permit program.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ...) AIR PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.70 Relationship to... operating permit programs and acceptance of State Acid Rain programs, the procedure for including State Acid... of an accepted State program, to issue Phase II Acid Rain permits. (b) Relationship to operating...

  16. 40 CFR 72.70 - Relationship to title V operating permit program.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ...) AIR PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.70 Relationship to... operating permit programs and acceptance of State Acid Rain programs, the procedure for including State Acid... of an accepted State program, to issue Phase II Acid Rain permits. (b) Relationship to operating...

  17. 40 CFR 72.70 - Relationship to title V operating permit program.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...) AIR PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.70 Relationship to... operating permit programs and acceptance of State Acid Rain programs, the procedure for including State Acid... of an accepted State program, to issue Phase II Acid Rain permits. (b) Relationship to operating...

  18. 40 CFR 72.70 - Relationship to title V operating permit program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ...) AIR PROGRAMS (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.70 Relationship to... operating permit programs and acceptance of State Acid Rain programs, the procedure for including State Acid... of an accepted State program, to issue Phase II Acid Rain permits. (b) Relationship to operating...

  19. 40 CFR 60.4120 - General Hg budget trading program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 6 2010-07-01 2010-07-01 false General Hg budget trading program... budget trading program permit requirements. (a) For each Hg Budget source required to have a title V operating permit, such permit shall include a Hg Budget permit administered by the permitting authority for...

  20. Commercial Ion Exchange Resin Vitrification in Borosilicate Glass

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

    Cicero-Herman, C.A.; Workman, P.; Poole, K.

    1998-05-01

    Bench-scale studies were performed to determine the feasibility of vitrification treatment of six resins representative of those used in the commercial nuclear industry. Each resin was successfully immobilized using the same proprietary borosilicate glass formulation. Waste loadings varied from 38 to 70 g of resin/100 g of glass produced depending on the particular resin, with volume reductions of 28 percent to 68 percent. The bench-scale results were used to perform a melter demonstration with one of the resins at the Clemson Environmental Technologies Laboratory (CETL). The resin used was a weakly acidic meth acrylic cation exchange resin. The vitrification processmore » utilized represented a approximately 64 percent volume reduction. Glass characterization, radionuclide retention, offgas analyses, and system compatibility results will be discussed in this paper.« less

  1. Melter Technologies Assessment

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

    Perez, J.M. Jr.; Schumacher, R.F.; Forsberg, C.W.

    1996-05-01

    The problem of controlling and disposing of surplus fissile material, in particular plutonium, is being addressed by the US Department of Energy (DOE). Immobilization of plutonium by vitrification has been identified as a promising solution. The Melter Evaluation Activity of DOE`s Plutonium Immobilization Task is responsible for evaluating and selecting the preferred melter technologies for vitrification for each of three immobilization options: Greenfield Facility, Adjunct Melter Facility, and Can-In-Canister. A significant number of melter technologies are available for evaluation as a result of vitrification research and development throughout the international communities for over 20 years. This paper describes an evaluationmore » process which will establish the specific requirements of performance against which candidate melter technologies can be carefully evaluated. Melter technologies that have been identified are also described.« less

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

    Dahl, Suzanne; Biyani, Rabindra; Holmes, Erika

    The United States Department of Energy's (US DOE's) Hanford Nuclear Site has 177 underground waste storage tanks located 19 to 24 km (12 to 15 miles) from the Columbia River in south-central Washington State. Hanford's tanks now hold about 212,000 cu m (56 million gallons) of highly radioactive and chemically hazardous waste. Sixty-seven tanks have leaked an estimated 3,785 cu m (1 million gallons) of this waste into the surrounding soil. Further releases to soil, groundwater, and the Columbia River are the inevitable result of the tanks continuing to age. The risk from this waste is recognized as a threatmore » to the Northwest by both State and Federal governments. US DOE and Bechtel National, Inc., are building the Waste Treatment and Immobilization Plant (WTP) to treat and vitrify (immobilize in glass) the waste from Hanford's tanks. As is usual for any groundbreaking project, problems have arisen that must be resolved as they occur if treatment is to take place as specified in the court-enforceable Hanford Federal Facility Agreement and Consent Order (Tri-Party Agreement) and the Consent Decree, entered into by US DOE, the U.S. Environmental Protection Agency, and the Washington State Department of Ecology (Ecology). At times, US DOE's approach to solving these critical issues seems to have caused undue wastes of time, energy, and, ultimately, public funds. Upon reviewing the history of Hanford's tank waste treatment project, Ecology hopes that constructive criticism of past failures and praise of successes will inspire US DOE to consider changing practices, be more transparent with regulatory agencies and the public, and take a 'lean production' approach to successfully completing this project. All three Tri-Party Agreement agencies share the goal of completing WTP on time, ensuring it is operational and in compliance with safety standards. To do this, Ecology believes US DOE should: - Maintain focus on the primary goal of completing the five major facilities of WTP. - Construct a supplemental low-activity waste (LAW) vitrification facility for the two-thirds balance of LAW that will not be treated by the vitrification facility under construction. - Prepare infrastructure for waste feed from the tanks and facilities to handle the WTP waste streams. To support this project track, Ecology expedites dangerous waste permitting by using a design-build approach to integrate WTP into the Resource Conservation and Recovery Act permit for Hanford. (authors)« less

  3. 47 CFR 5.305 - Program license not permitted.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 1 2013-10-01 2013-10-01 false Program license not permitted. 5.305 Section 5.305 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL EXPERIMENTAL RADIO SERVICE Program Experimental Radio Licenses § 5.305 Program license not permitted. Experiments are not permitted under this...

  4. 47 CFR 5.305 - Program license not permitted.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 1 2014-10-01 2014-10-01 false Program license not permitted. 5.305 Section 5.305 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL EXPERIMENTAL RADIO SERVICE Program Experimental Radio Licenses § 5.305 Program license not permitted. Experiments are not permitted under this...

  5. 40 CFR 122.43 - Establishing permit conditions (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Permit Conditions § 122.43 Establishing permit conditions (applicable to State...

  6. 40 CFR 122.43 - Establishing permit conditions (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Permit Conditions § 122.43 Establishing permit conditions (applicable to State...

  7. Vitrification of ion exchange resins

    DOEpatents

    Cicero-Herman, Connie A.; Workman, Rhonda Jackson

    2001-01-01

    The present invention relates to vitrification of ion exchange resins that have become loaded with hazardous or radioactive wastes, in a way that produces a homogenous and durable waste form and reduces the disposal volume of the resin. The methods of the present invention involve directly adding borosilicate glass formers and an oxidizer to the ion exchange resin and heating the mixture at sufficient temperature to produce homogeneous glass.

  8. Bio-inspired Cryo-ink Preserves Red Blood Cell Phenotype and Function during Nanoliter Vitrification

    PubMed Central

    Assal, Rami El; Guven, Sinan; Gurkan, Umut Atakan; Gozen, Irep; Shafiee, Hadi; Dalbeyber, Sedef; Abdalla, Noor; Thomas, Gawain; Fuld, Wendy; Illigens, Ben M.W.; Estanislau, Jessica; Khoory, Joseph; Kaufman, Richard; Zylberberg, Claudia; Lindeman, Neal; Wen, Qi; Ghiran, Ionita; Demirci, Utkan

    2014-01-01

    Current red blood cell cryopreservation methods utilize bulk volumes, causing cryo-injury of cells, which results in irreversible disruption of cell morphology, mechanics, and function. An innovative approach to preserve human red blood cell morphology, mechanics, and function following vitrification in nanoliter volumes is developed using a novel cryo-ink integrated with a bio-printing approach. PMID:25047246

  9. Testing Report: Littleford-Day Dryer Operation: Dryer Operation Impacts of Proposed MIS Mitigation Changes

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

    Shimskey, Rick W.; Buchmiller, William C.; Elmore, Monte R.

    2007-06-01

    Pacific Northwest National Laboratory performed a series of tests using the Littleford Day 22-liter dryer during investigations that evaluated changes in the melter-feed composition for the Demonstration Bulk Vitrification System. During testing, a new melter-feed formulation was developed that improved dryer performance while improving the retention of waste salts in the melter feed during vitrification.

  10. Effective Oocyte Vitrification and Survival Techniques for Bovine Somatic Cell Nuclear Transfer

    PubMed Central

    Park, Min Jee; Lee, Seung Eun; Lee, Jun Beom; Jeong, Chang Jin

    2015-01-01

    Abstract Bovine somatic cell nuclear transfer (SCNT) using vitrified–thawed (VT) oocytes has been studied; however, the cloning efficiency of these oocytes is not comparable with that of nonvitrified (non-V) fresh oocytes. This study sought to optimize the survival and cryopreservation of VT oocytes for SCNT. Co-culture with feeder cells that had been preincubated for 15 h significantly improved the survival of VT oocytes and their in vitro developmental potential following SCNT in comparison to co-culture with feeder cells that had been preincubated for 2, 5, or 24 h (p<0.05). Spindle assessment via the Oosight Microscopy Imaging System and microtubule staining revealed that vitrified metaphase II oocytes (VT group) were not suitable for SCNT. However, enucleating and/or activating oocytes prior to freezing enhanced their developmental potential and suitability for SCNT. The cloning efficiency of the enucleated–activated–vitrified–thawed (EAVT) group (21.6%) was better than that of the other vitrification groups [enucleated–vitrified–thawed (EVT) group, 13.7%; VT group, 15.0%; p<0.05] and was comparable with that of the non-V group (25.9%). The reactive oxygen species level was significantly lower in the EAVT group than in the other vitrification groups (p<0.05). mRNA levels of maternal genes (ZAR1, BMP15, and NLRP5) and a stress gene (HSF1) were lower in the vitrification groups than in the non-V group (p<0.05), whereas the level of phospho-p44/42 mitogen-activated protein kinase did not differ among the groups. Among the vitrification groups, blastocysts in the EAVT group had the best developmental potential, as judged by their high mRNA expression of developmental potential–related genes (POU5f1, Interferon-tau, and SLC2A5) and their low expression of proapoptotic (CASP3) and stress (Hsp70) genes. This study demonstrates that SCNT using bovine frozen–thawed oocytes can be successfully achieved using optimized vitrification and co-culture techniques. PMID:25984830

  11. Effects of Vitrification on Outcomes of In Vivo-Mature, In Vitro-Mature and Immature Human Oocytes.

    PubMed

    Song, Wen-Yan; Peng, Zhao-Feng; Chen, Xue-Mei; Jin, Hai-Xia; Yao, Gui-Dong; Shi, Sen-Lin; Yang, Hong-Yi; Zhang, Xiang-Yang; Sun, Ying-Pu

    2016-01-01

    To observe the effects of vitrification on spindle, zona pellucida, embryonic aneuploidy and DNA injury in in vivo-maruted, in vitro-mature and immature human oocytes. Between January 2009 and February 2015, 223 immature oocytes from 450 infertile patients, and 31 in vivo-mature oocytes from 3 infertile couples were collected. Of the 223 immature oocytes, 113 were used for in vitro culture before vitrification. Some oocytes were randomly divided into in vivo-mature group (group A, n = 15), in vitro-mature group (group B, n = 88) and immature group (group C, n = 85), and then the oocytes with spindle in these three groups after freezing-thawing were selected to use for Polscope imaging, embryonic aneuploidy screening and embryo development evaluation. Other oocytes were randomly divided into group A (n = 16), group B (n = 25) and group C (n = 25) for detecting DNA injury. After thawing, spindle occurrence rate, spindle Retardance value, and cleavage rate were significantly higher in groups A and B than in group C (all P < 0.05), but there were no statistical differences in fertility rate, high-quality embryo rate, blastulation rate and aneuploidy rate amongst the three groups (all P > 0.05). Zona pellucida density (ZPD) was significantly lower in group A than in groups B and C both before and after vitrification (all P < 0.05). ZPD was significantly higher after thawing than before vitrification (all P < 0.05), but zona pellucida thickness (ZPT) was not significantly changed in all the three groups (all P > 0.05). Rate of comet cells was significantly lower in group A than in groups B and C (all P < 0.01). Comet tail was significantly longer in group C than in groups B and A (all P < 0.05). In vivo- and in vitro-mature human oocytes are more suitable to vitrification than immature human oocytes. Spindle Retardance value has more predictive value for embryonic development potential than ZPD and ZPT. © 2016 The Author(s) Published by S. Karger AG, Basel.

  12. [A prospective study to compare the efficiency of oocyte vitrification using closed or open devices].

    PubMed

    Sarandi, S; Herbemont, C; Sermondade, N; Benoit, A; Sonigo, C; Poncelet, C; Benard, J; Gronier, H; Boujenah, J; Grynberg, M; Sifer, C

    2016-05-01

    Oocyte vitrification using an open device is thought to be a source of microbiological and chemical contaminations that can be avoided using a closed device. The principal purpose of this study was to compare the two vitrification protocols: closed and open system. The secondary aim was to study the effects of the storage in the vapor phase of nitrogen (VPN) on oocytes vitrified using an open system and to compare it to those of a storage in liquid nitrogen (LN). Forty-four patients have been included in our study between November 2014 and May 2015. Two hundred and fourteen oocytes have been vitrified at germinal vesicle (GV), metaphase I (0PB) and metaphase II (1PB) stages. We vitrified 96 oocytes (59 GV/37 0PB) using a closed vitrification device and 118 oocytes (57 GV/31 0PB/30 1PB) using an open device. The vitrified oocytes were then stored either in LN or in VPN. The main outcome measures were the survival rate after warming (SR), meiosis resumption rate (MRR) and maturation rate (MR). The global post-thaw SR was significantly higher for oocytes vitrified using an open system (93.2%) compared to those vitrified using a closed one (64.5%; P<0.001). On the contrary, there was no significant difference in terms of global MRR and MR (82.1% vs. 87.5% and 60.7% vs. 61.2% using closed and open system respectively). The SR, MRR and the MR were not significantly different when vitrified oocytes were stored in VPN or LN (91.6, 83.8, 64.5% vs. 93.9, 89.8, 59.1% respectively). Taking into account the limits of our protocol, the open vitrification system remains the more efficient system. The use of sterile liquid nitrogen for oocyte vitrification and the subsequent storage in vapor phase of nitrogen could minimize the hypothetical risks of biological and chemical contaminations. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  13. 40 CFR 97.22 - Information requirements for NOX Budget permit applications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Budget permit applications. 97.22 Section 97.22 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.22 Information requirements for NOX Budget permit applications. A complete NOX Budget permit...

  14. 76 FR 12926 - Clean Air Act Proposed Interim Approval of Title V Operating Permits Program; Southern Ute Indian...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-09

    ... Proposed Interim Approval of Title V Operating Permits Program; Southern Ute Indian Tribe AGENCY... Title V Operating Permits Program submitted by the Southern Ute Indian Tribe (Tribe). The Tribe's Title V Operating Permit Program (Title V Program) was submitted for the purpose of administering a tribal...

  15. Office of River Protection Advanced Low-Activity Waste Glass Research and Development Plan

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

    Peeler, David K.; Kim, Dong-Sang; Vienna, John D.

    2015-11-01

    The U.S. Department of Energy Office of River Protection (ORP) has initiated and leads an integrated Advanced Waste Glass (AWG) program to increase the loading of Hanford tank wastes in glass while meeting melter lifetime expectancies and process, regulatory, and product performance requirements. The integrated ORP program is focused on providing a technical, science-based foundation for making key decisions regarding the successful operation of the Hanford Tank Waste Treatment and Immobilization Plant (WTP) facilities in the context of an optimized River Protection Project (RPP) flowsheet. The fundamental data stemming from this program will support development of advanced glass formulations, keymore » product performance and process control models, and tactical processing strategies to ensure safe and successful operations for both the low-activity waste (LAW) and high-level waste vitrification facilities. These activities will be conducted with the objective of improving the overall RPP mission by enhancing flexibility and reducing cost and schedule. The purpose of this advanced LAW glass research and development plan is to identify the near-term, mid-term, and longer-term research and development activities required to develop and validate advanced LAW glasses, property-composition models and their uncertainties, and an advanced glass algorithm to support WTP facility operations, including both Direct Feed LAW and full pretreatment flowsheets. Data are needed to develop, validate, and implement 1) new glass property-composition models and 2) a new glass formulation algorithm. Hence, this plan integrates specific studies associated with increasing the Na2O and SO3/halide concentrations in glass, because these components will ultimately dictate waste loadings for LAW vitrification. Of equal importance is the development of an efficient and economic strategy for 99Tc management. Specific and detailed studies are being implemented to understand the fate of Tc throughout the WTP flowsheet and the underlying mechanisms that dictate its partitioning between streams within the LAW vitrification facility. These studies are aimed at increasing the single-pass Tc retention in glass and the potential use of high-temperature mineral phases to capture Tc. The Tc-bearing mineral phases would be thermally stable and resistant to Tc release during feed melting reactions or they could serve as alternative waste forms. The LAW glass research and development is focused on reducing the total volume of LAW glass produced and minimizing the impact of (or potentially eliminating) the need for recycle.« less

  16. Strain preservation of experimental animals: vitrification of two-cell stage embryos for multiple mouse strains.

    PubMed

    Eto, Tomoo; Takahashi, Riichi; Kamisako, Tsutomu

    2015-04-01

    Strain preservation of experimental animals is crucial for experimental reproducibility. Maintaining complete animal strains, however, is costly and there is a risk for genetic mutations as well as complete loss due to disasters or illness. Therefore, the development of effective vitrification techniques for cryopreservation of multiple experimental animal strains is important. We examined whether a vitrification method using cryoprotectant solutions, P10 and PEPeS, is suitable for preservation of multiple inbred and outbred mouse strains. First, we investigated whether our vitrification method using cryoprotectant solutions was suitable for two-cell stage mouse embryos. In vitro development of embryos exposed to the cryoprotectant solutions was similar to that of fresh controls. Further, the survival rate of the vitrified embryos was extremely high (98.1%). Next, we collected and vitrified two-cell stage embryos of 14 mouse strains. The average number of embryos obtained from one female was 7.3-33.3. The survival rate of vitrified embryos ranged from 92.8% to 99.1%, with no significant differences among mouse strains. In vivo development did not differ significantly between fresh controls and vitrified embryos of each strain. For strain preservation using cryopreserved embryos, two offspring for inbred lines and one offspring for outbred lines must be produced from two-cell stage embryos collected from one female. The expected number of surviving fetuses obtained from embryos collected from one female of either the inbred or outbred strains ranged from 2.9 to 19.5. The findings of the present study indicated that this vitrification method is suitable for strain preservation of multiple mouse strains. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

  17. Vitrification: an effective new approach to oocyte banking and preserving fertility in cancer patients.

    PubMed

    Cobo, A; Domingo, J; Pérez, S; Crespo, J; Remohí, J; Pellicer, A

    2008-05-01

    Oocyte cryopreservation is a useful tool for preserving the fertility of cancer patients at risk of losing ovarian function due to undergoing potentially sterilising therapies. Results obtained with different cryopreservation protocols have been disappointing, particularly those obtained with slow cooling procedures. The efficacy of vitrification as an application in clinical practice has recently been demonstrated. The aim of this study is to report results obtained with the Cryotop method of oocyte vitrification in a population of healthy women and to point out its potential usefulness for fertility preservation in oncological patients. The study population consisting of non-oncological patients included 47 oocyte donors and 57 recipients undergoing an oocyte donation cycle of assisted reproductive technology (ART). A total of 693 mature metaphase II oocytes were collected following ovarian stimulation using long protocol down-regulation plus gonadotropin administration. Vitrification was carried out by means of the Cryotop method. Oocytes were donated to a compatible recipient after endometrial preparation. Of the 693 oocytes, 666 (96.1%) survived. A total of 487 (73.1%) were fertilised successfully. One hundred and seventeen embryos were transferred to 57 recipients. Pregnancy rate per transfer and implantation rates were 63.2% and 38.5% respectively. Twenty-eight healthy babies were later born. Oocyte cryo-banking by means of the Cryotop vitrification method represents a viable option for healthy women, producing excellent survival rates and a clinical outcome similar to that obtained with fresh oocytes. This approach could potentially be used in cancer patients who want to safeguard their fertility. Cancer patients could potentially benefit from this approach by storing their oocytes before the onset of the oncological therapy.

  18. Forskolin effect on the cryosurvival of in vitro-produced bovine embryos in the presence or absence of fetal calf serum.

    PubMed

    Paschoal, Daniela Martins; Sudano, Mateus José; Guastali, Midyan Daroz; Dias Maziero, Rosiára Rosária; Crocomo, Letícia Ferrari; Oña Magalhães, Luis Carlos; da Silva Rascado, Tatiana; Martins, Alicio; da Cruz Landim-Alvarenga, Fernanda

    2014-05-01

    The objective of this study was to assess the viability and cryotolerance of zebu embryos produced in vitro with or without the addition of fetal calf serum (FCS) and forskolin (F). Embryos produced in vivo were used as a control. Presumptive zygotes were cultured in modified synthetic oviductal fluid supplemented with amino acids (SOFaa), bovine serum albumin (BSA) and with (2.5%) or without (0%) FCS. On day 6 of growth, the embryos from each group were divided into treatments with or without 10 μM F to induce embryonic lipolysis, comprising a total of four experimental groups: 2.5% FCS, 0% FCS, 2.5% + F and 0% + F. For vitrification, embryos were exposed to vitrification solution 1 (5 M EG (ethylene glycol)) for 3 min and then transferred to vitrification solution 2 (7 M EG, 0.5 M galactose solution and 18% (w/v) Ficoll 70) before being introduced to liquid nitrogen. The presence of FCS in the culture medium resulted in the production of embryos with a similar rate of damaged cells compared with in vivo-produced embryos. After vitrification, the 2.5% FCS group had a significantly higher rate of damaged cells when compared with the other groups (P < 0.05). The results of this experiment indicated that the omission of FCS and the addition of forskolin do not have deleterious effect on embryo production rates. In addition, embryos produced in the presence of FCS had greater sensitivity to cryopreservation, but this effect was reversed when forskolin was added to the medium, which improved embryo survival without affecting embryo development and quality after vitrification.

  19. Decreased pregnancy and live birth rates after vitrification of in vitro matured oocytes.

    PubMed

    Cohen, Yoni; St-Onge-St-Hilaire, Alexandra; Tannus, Samer; Younes, Grace; Dahan, Michael H; Buckett, William; Son, Weon-Young

    2018-06-04

    To assess effects on fertilization rate, embryo quality, pregnancy, and live birth rates of vitrification and warming of oocytes that matured in vitro (vIVM) compared to fresh in vitro maturation (fIVM) cycles. A retrospective cohort study conducted at a university hospital-affiliated IVF unit. Fifty-six cycles of vIVM cycles and 263 fIVM in women diagnosed with polycystic ovarian syndrome (PCOS) ovaries were included in the analysis. The study group included PCOS patients who failed ovulation induction with intrauterine insemination and were offered IVM cycle followed by oocyte vitrification and warming. The embryological aspects and clinical outcomes were compared to those of controls undergoing fresh IVM cycles during the same period. The main outcome measure was live birth rate. One thousand seventy oocytes were collected from 56 patients and underwent vitrification and warming. In the control group, 4781 oocytes were collected from 219 patients who had undergone a fresh IVM cycle. Oocyte maturation rates were similar between the groups (mean ± SD: 0.7 ± 0.2 vs. 0.6 ± 0.2, for vIVM and fIVM, respectively). Survival rate after warming was 59.8%. Fertilization and embryo cleavage rates per oocyte were significantly lower in the vIVM group. Clinical pregnancy (10.7 vs. 36.1%) and live birth rates (8.9 vs. 25.9%) per cycle were significantly lower in the vIVM group than those in the fIVM group (P = 0.005 and P < 0.001, respectively). Five healthy babies were born in the vIVM group. The reproductive potential of vitrified IVM oocytes is impaired. This injury likely occurs through vitrification and warming.

  20. Survival of mouse embryos after vitrification depending on the cooling rate of the cryoprotectant solution.

    PubMed

    Hredzák, R; Ostró, A; Zdilová, Viera; Maracek, I; Kacmárik, J

    2006-03-01

    The aim of the study was to determine the relationship between the rate of cooling of eight-cell mouse embryos to the temperature of liquid nitrogen (-196 degrees C) and their developmental capacity after thawing on the basis of their ability to leave the zona pellucida ('hatching') during in vitro culturing. Eight-cell embryos were obtained from superovulated female mice and divided into three experimental and one control group. Embryos from the experimental groups were cryopreserved by the vitrification method using ethylene glycol as cryoprotectant. The vitrification protocols used in the study differed in the rate of cooling of the cryoprotectant solution. Embryos from the first group were frozen in conventional 0.25-ml plastic straws, those from the second group in pipetting 'tips', and embryos from the third group, placed in vitrification solution, were introduced dropwise directly into liquid nitrogen. The control group of embryos was cultured in vitro without freezing in a culturing medium in an environment consisting of 95% air and 5% CO2. The developmental capacity of thawed embryos was assessed on the basis of their ability to leave the zona pellucida ('hatching') after three days of in vitro culturing. In the control group 95.1% of embryos 'hatched'. A significantly higher number of embryos that 'hatched' after thawing was observed in the group introduced dropwise directly into liquid nitrogen (60.0%) compared to the group frozen in pipetting 'tips' (37.9%). The group frozen in straws yielded significantly the lowest proportion of 'hatching' embryos (8.1%). These results showed that increasing cooling rates during vitrification of embryos improved their survival.

  1. Effects of vitrification on nuclear maturation, ultrastructural changes and gene expression of canine oocytes

    PubMed Central

    2010-01-01

    Background Cryopreservation of oocytes, which is an interesting procedure to conserve female gametes, is an essential part of reproductive biotechnology. The objective of the present study was to investigate the effects of vitrification on nuclear maturation, ultrastructural changes and gene expression of canine oocytes. Methods Immature oocytes (germinal vesicles) isolated from ovaries of normal bitches (> 6 months of age) were either vitrified in open pulled straw (OPS) using 20% ethylene glycol (EG) and 20% dimethyl sulfoxide (DMSO) as vitrification solution or exposed to vitrification solution without subjected to liquid nitrogen. After warming, oocytes were investigated for nuclear maturation following in vitro maturation (IVM), ultrastructural changes using transmission electron microscopy (TEM) and gene expression using RT-PCR. Fresh immature oocytes were used as the control group. Results The rate of resumption of meiosis in vitrified-warmed oocytes (53.4%) was significantly (P < 0.05) lower than those of control (93.8%) and exposure (91.4%) groups. However, there were no statistically significant differences among groups in the rates of GV oocytes reaching the maturation stage (metaphase II, MII). The ultrastructural alterations revealed by TEM showed that cortical granules, mitochondria, lipid droplets and smooth endoplasmic reticulum (SER) were affected by vitrification procedures. RT-PCR analysis for gene expression revealed no differences in HSP70, Dnmt1, SOD1 and BAX genes among groups, whereas Bcl2 was strongly expressed in vitrified-warmed group when compared to the control. Conclusion Immature canine oocytes were successfully cryopreserved, resumed meiosis and developed to the MII stage. The information obtained in this study is crucial for the development of an effective method to cryopreserve canine oocytes for establishment of genetic banks of endangered canid species. PMID:20565987

  2. Synthetic polymers improve vitrification outcomes of macaque ovarian tissue as assessed by histological integrity and the in vitro development of secondary follicles☆

    PubMed Central

    Ting, Alison Y.; Yeoman, Richard R.; Lawson, Maralee S.; Zelinski, Mary B.

    2013-01-01

    Ovarian tissue cryopreservation is the only proven option for fertility preservation in female cancer patients who are prepubertal or require immediate treatment. However it remains unclear which cryopreservation protocol is best in cases where the tissue may contain cancerous cells, as these should be matured in vitro rather than autografted. This study evaluated different cryoprotectant exposure times and whether the addition of synthetic polymers (Supercool X-1000, Z-1000 and polyvinylpyrrolidone [PVP K-12]) to the vitrification solution is beneficial to tissue morphology, cellular proliferation and subsequent in vitro function of secondary follicles. Pieces of macaque (n = 4) ovarian cortex were exposed to vitrification solution containing glycerol (25%, v/v) and ethylene glycol (25%, v/v) for 3 or 8 min, without (V3, V8) or with (VP3, VP8) polymers (0.2% [v/v] X-1000, 0.4% Z-1000 and 0.2% PVP). Fresh and vitrified tissues were fixed for histology and phosphohistone H3 (PPH3) analysis, or used for secondary follicle isolation followed by encapsulated 3D culture. Five-week follicle survival and growth, as well as steroid hormones (estradiol [E2], progesterone, androstenedione) were measured weekly. Morphology of the stroma and preantral follicles as well as PPH3 expression, was preserved in all vitrified tissues. Vitrification with polymers and shorter incubation time (VP3) increased in vitro follicle survival and E2 production compared to other vitrified groups. Thus, a short exposure of macaque ovarian tissue to a vitrification solution containing synthetic polymers preserves morphology and improves in vitro function of secondary follicles. PMID:22569078

  3. SECONDARY WASTE MANAGEMENT FOR HANFORD EARLY LOW ACTIVITY WASTE VITRIFICATION

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

    UNTERREINER BJ

    2008-07-18

    More than 200 million liters (53 million gallons) of highly radioactive and hazardous waste is stored at the U.S. Department of Energy's Hanford Site in southeastern Washington State. The DOE's Hanford Site River Protection Project (RPP) mission includes tank waste retrieval, waste treatment, waste disposal, and tank farms closure activities. This mission will largely be accomplished by the construction and operation of three large treatment facilities at the Waste Treatment and Immobilization Plant (WTP): (1) a Pretreatment (PT) facility intended to separate the tank waste into High Level Waste (HLW) and Low Activity Waste (LAW); (2) a HLW vitrification facilitymore » intended to immobilize the HLW for disposal at a geologic repository in Yucca Mountain; and (3) a LAW vitrification facility intended to immobilize the LAW for shallow land burial at Hanford's Integrated Disposal Facility (IDF). The LAW facility is on target to be completed in 2014, five years prior to the completion of the rest of the WTP. In order to gain experience in the operation of the LAW vitrification facility, accelerate retrieval from single-shell tank (SST) farms, and hasten the completion of the LAW immobilization, it has been proposed to begin treatment of the low-activity waste five years before the conclusion of the WTP's construction. A challenge with this strategy is that the stream containing the LAW vitrification facility off-gas treatment condensates will not have the option of recycling back to pretreatment, and will instead be treated by the Hanford Effluent Treatment Facility (ETF). Here the off-gas condensates will be immobilized into a secondary waste form; ETF solid waste.« less

  4. Comparison of ethylene glycol and propylene glycol for the vitrification of immature porcine oocytes.

    PubMed

    Somfai, Tamás; Nakai, Michiko; Tanihara, Fuminori; Noguchi, Junko; Kaneko, Hiroyuki; Kashiwazaki, Naomi; Egerszegi, István; Nagai, Takashi; Kikuchi, Kazuhiro

    2013-01-01

    Our aim was to optimize a cryoprotectant treatment for vitrification of immature porcine cumulus-oocyte complexes (COCs). Immature COCs were vitrified either in 35% ethylene glycol (EG), 35% propylene glycol (PG) or a combination of 17.5% EG and 17.5% PG. After warming, the COCs were in vitro matured (IVM), and surviving oocytes were in vitro fertilized (IVF) and cultured. The mean survival rate of vitrified oocytes in 35% PG (73.9%) was higher (P<0.05) than that in 35% EG (27.8%). Oocyte maturation rates did not differ among vitrified and non-vitrified control groups. Blastocyst formation in the vitrified EG group (10.8%) was higher (P<0.05) than that in the vitrified PG group (2.0%) but was lower than that in the control group (25.0%). Treatment of oocytes with 35% of each cryoprotectant without vitrification revealed a higher toxicity of PG on subsequent blastocyst development compared with EG. The combination of EG and PG resulted in 42.6% survival after vitrification. The maturation and fertilization rates of the surviving oocytes were similar in the vitrified, control and toxicity control (TC; treated with EG+PG combination without cooling) groups. Blastocyst development in the vitrified group was lower (P<0.05) than that in the control and TC groups, which in turn had similar development rates (10.7%, 18.1% and 23.3%, respectively). In conclusion, 35% PG enabled a higher oocyte survival rate after vitrification compared with 35% EG. However, PG was greatly toxic to oocytes. The combination of 17.5% EG and 17.5% PG yielded reasonable survival rates without toxic effects on embryo development.

  5. Plasma vitrification of asbestos fibers

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

    Camacho, S.L.

    Asbestos is a mineral in the form of long, thread-like fibers. Asbestos fibers have been among the best insulators of pipes, boilers, ducts, tanks, etc., in buildings, ships, and industrial furnaces. Over 150,000 metric tons of asbestos were consumed in the United States in 1984. The Environmental Protection Agency has declared asbestos fibers a known human carcinogen. And today, asbestos insulators are being replaced by manmade non-hazardous fibers. Millions of tons of replaced asbestos fiber insulators are in storage, awaiting the demonstration of effective alternative disposal technologies. Plasma vitrification has been demonstrated during May, June and July 1995 as amore » viable, cost-effective, safe technology for asbestos fiber disposal. A low-mass plasma arc heater is submerged under the waste asbestos insulating materials, and the intense heat of the plasma flame heats and melts the fibers. The by-product is dark, non-hazardous glass pellets. The vitrification process renders the asbestos waste safe for use as road construction aggregates or other fill materials. This paper will describe the results of start-up of a 1 ton-per-hour Plasma Mobile Asbestos Vitrification (MAV) Plant at a DOD Site in Port Clinton, Ohio. The Plasma MAV Plant is being demonstrated for the on-site disposal of 1.5 million pounds of Amosite asbestos fibers.« less

  6. Ovarian reserve and response to stimulation in women undergoing fertility preservation according to malignancy type.

    PubMed

    Lefebvre, Tiphaine; Mirallié, Sophie; Leperlier, Florence; Reignier, Arnaud; Barrière, Paul; Fréour, Thomas

    2018-05-02

    Does ovarian reserve and ovarian response to ovarian stimulation in women with cancer undergoing oocyte vitrification for fertility preservation vary according to the type of malignancy? Retrospective cohort study including 105 women aged between 18 and 40 years, who were referred for fertility preservation (oocyte vitrification) between 2013 and 2016. The women were divided into three groups: breast cancer, lymphoma or other cancer. All of them had been recently diagnosed with cancer, with gonadotoxic treatment scheduled, and had oocyte vitrification after ovarian stimulation with antagonist protocol. Baseline antral follicle count and anti-Müllerian hormone were no different between women with breast cancer, lymphoma or other cancer. The number of cancelled cycles for poor ovarian response was similar between the groups. The number of FSH units per mature oocyte, the number of mature oocytes (metaphase II) retrieved, and the oocyte maturity rate were not significantly different between the three groups. As the type of cancer does not seem to significantly affect ovarian reserve and ovarian response to ovarian stimulation, our results do not support the relevance of integrating this parameter when establishing ovarian stimulation protocol for oocyte vitrification cycle in women with cancer. Copyright © 2018 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  7. Vitrification of mouse embryos using the thin plastic strip method

    PubMed Central

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

    2012-01-01

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

  8. Experimental vitrification of human compacted morulae and early blastocysts using fine diameter plastic micropipettes.

    PubMed

    Cremades, N; Sousa, M; Silva, J; Viana, P; Sousa, S; Oliveira, C; Teixeira da Silva, J; Barros, A

    2004-02-01

    Vitrification of human blastocysts has been successfully applied using grids, straws and cryoloops. We assessed the survival rate of human compacted morulae and early blastocysts vitrified in pipette tips with a smaller inner diameter and solution volume than the previously described open pulled straw (OPS) method. Excess day 5 human embryos (n = 63) were experimentally vitrified in vessels. Embryos were incubated at 37 degrees C with sperm preparation medium (SPM) for 1 min, SPM + 7.5% ethylene glycol (EG)/dimethylsulphoxide (DMSO) for 3 min, and SPM + 16.5% EG + 16.5% DMSO + 0.67 mol/l sucrose for 25 s. They were then aspirated (0.5 microl) into a plastic micropipette tip (0.36 mm inner diameter), exposed to liquid nitrogen (LN(2)) vapour for 2 min before being placed into a pre-cooled cryotube, which was then closed and plunged into LN(2). Embryos were warmed and diluted using 0.33 mol/l and 0.2 mol/l sucrose. The survival rate for compacted morulae was 73% (22/30) and 82% (27/33) for early blastocysts. The survival rates of human compacted morulae and early blastocysts after vitrification with this simple technique are similar to those reported in the literature achieved by slow cooling and other vitrification protocols.

  9. Zinc supplementation of vitrification medium improves in vitro maturation and fertilization of oocytes derived from vitrified-warmed mouse ovaries.

    PubMed

    Geravandi, Shirin; Azadbakht, Mehri; Pourmoradi, Mahsa; Nowrouzi, Fatemeh

    2017-02-01

    Oocyte cryopreservation is an approach for fertility preservation for normal women and cancer patients facing chemo and radiotherapy. The present study evaluated the effect of adding zinc chloride to the vitrification medium used for whole mouse ovaries and then assessing the in vitro maturation and fertilization of oocytes when they were subsequently extracted from these vitrified ovarian tissues. Four vitrification solutions with 0, 100,150 and 200 μg/dl zinc (V0, V1, V2 and V3 respectively) were compared. The viability of oocytes isolated from ovaries vitrified-warmed in the highest concentration of zinc (V3) was significantly higher after 24 than in the control V0 group (72.99 vs 85.97). Progression to the MII stage, fertilization and cleavage by 48 h was also higher in the V3 than V0 control group (35.55 vs 44.73), (47.67 vs 63.74), (28.72 vs 43.03) (P < 0.05) respectively. These results indicate that supplementation of vitrification medium for intact ovaries with zinc can improve the oocyte viability and in vitro maturation-fertilization rate. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Local geology controlled the feasibility of vitrifying Iron Age buildings

    USGS Publications Warehouse

    Fabian B Wadsworth,; Michael J Heap,; Damby, David; Kai-Uwe Hess,; Jens Najorka,; Jérémie Vasseur,; Dominik Fahrner,; Donald B Dingwell,

    2017-01-01

    During European prehistory, hilltop enclosures made from polydisperse particle-and-block stone walling were exposed to temperatures sufficient to partially melt the constituent stonework, leading to the preservation of glassy walls called ‘vitrified forts’. During vitrification, the granular wall rocks partially melt, sinter viscously and densify, reducing inter-particle porosity. This process is strongly dependent on the solidus temperature, the particle sizes, the temperature-dependence of the viscosity of the evolving liquid phase, as well as the distribution and longevity of heat. Examination of the sintering behaviour of 45 European examples reveals that it is the raw building material that governs the vitrification efficiency. As Iron Age forts were commonly constructed from local stone, we conclude that local geology directly influenced the degree to which buildings were vitrified in the Iron Age. Additionally, we find that vitrification is accompanied by a bulk material strengthening of the aggregates of small sizes, and a partial weakening of larger blocks. We discuss these findings in the context of the debate surrounding the motive of the wall-builders. We conclude that if wall stability by bulk strengthening was the desired effect, then vitrification represents an Iron Age technology that failed to be effective in regions of refractory local geology.

  11. Preservation of human ovarian follicles within tissue frozen by vitrification in a xeno-free closed system using only ethylene glycol as a permeating cryoprotectant.

    PubMed

    Sheikhi, Mona; Hultenby, Kjell; Niklasson, Boel; Lundqvist, Monalill; Hovatta, Outi

    2013-07-01

    To study the preservation of follicles within ovarian tissue vitrified using only one or a combination of three permeating cryoprotectants. Experimental study. University hospital. Ovarian tissue was donated by consenting women undergoing elective cesarean section. Ovarian tissue was vitrified in closed sealed vials using either a combination of dimethyl sulfoxide, 1,2-propanediol, and ethylene glycol (EG), or only EG as permeating cryoprotectants. Ovarian tissue was vitrified with the use of two vitrification methods. Tissue from the same donor was used for comparison of two different solutions. The morphology of the follicles was evaluated after vitrification, warming, and culture by light microscopy and transmission electron microscopy. Apoptosis was assessed by immunohistochemistry for active caspase-3 in fresh and vitrified tissue. Light and electron microscopic analysis showed equally well preserved morphology of oocytes, granulosa cells, and ovarian stroma when either of the vitrification solutions was used. No apoptosis was observed in primordial and primary follicles. Using only EG as a permeating cryoprotectant in a closed tube gives as good ultrastructural preservation of ovarian follicles as a more complicated system using several cryoprotectants. Copyright © 2013 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  12. Multiphase, multi-electrode Joule heat computations for glass melter and in situ vitrification simulations

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

    Lowery, P.S.; Lessor, D.L.

    Waste glass melter and in situ vitrification (ISV) processes represent the combination of electrical thermal, and fluid flow phenomena to produce a stable waste-from product. Computational modeling of the thermal and fluid flow aspects of these processes provides a useful tool for assessing the potential performance of proposed system designs. These computations can be performed at a fraction of the cost of experiment. Consequently, computational modeling of vitrification systems can also provide and economical means for assessing the suitability of a proposed process application. The computational model described in this paper employs finite difference representations of the basic continuum conservationmore » laws governing the thermal, fluid flow, and electrical aspects of the vitrification process -- i.e., conservation of mass, momentum, energy, and electrical charge. The resulting code is a member of the TEMPEST family of codes developed at the Pacific Northwest Laboratory (operated by Battelle for the US Department of Energy). This paper provides an overview of the numerical approach employed in TEMPEST. In addition, results from several TEMPEST simulations of sample waste glass melter and ISV processes are provided to illustrate the insights to be gained from computational modeling of these processes. 3 refs., 13 figs.« less

  13. 40 CFR 71.25 - Permit content.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 16 2013-07-01 2013-07-01 false Permit content. 71.25 Section 71.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Permits for Early Reductions Sources § 71.25 Permit content. (a) Standard permit requirements. Each permit issued under...

  14. 40 CFR 71.6 - Permit content.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 16 2012-07-01 2012-07-01 false Permit content. 71.6 Section 71.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Operating Permits § 71.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include...

  15. 40 CFR 71.25 - Permit content.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 16 2014-07-01 2014-07-01 false Permit content. 71.25 Section 71.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Permits for Early Reductions Sources § 71.25 Permit content. (a) Standard permit requirements. Each permit issued under...

  16. 40 CFR 71.6 - Permit content.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 16 2013-07-01 2013-07-01 false Permit content. 71.6 Section 71.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Operating Permits § 71.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include...

  17. 40 CFR 71.25 - Permit content.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 16 2012-07-01 2012-07-01 false Permit content. 71.25 Section 71.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Permits for Early Reductions Sources § 71.25 Permit content. (a) Standard permit requirements. Each permit issued under...

  18. 40 CFR 71.6 - Permit content.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 16 2014-07-01 2014-07-01 false Permit content. 71.6 Section 71.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Operating Permits § 71.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include...

  19. 40 CFR 71.25 - Permit content.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 15 2010-07-01 2010-07-01 false Permit content. 71.25 Section 71.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Permits for Early Reductions Sources § 71.25 Permit content. (a) Standard permit requirements. Each permit issued under...

  20. 40 CFR 71.6 - Permit content.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 15 2010-07-01 2010-07-01 false Permit content. 71.6 Section 71.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL OPERATING PERMIT PROGRAMS Operating Permits § 71.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include...

  1. 40 CFR 97.23 - NOX Budget permit contents.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false NOX Budget permit contents. 97.23 Section 97.23 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.23 NOX Budget permit contents. (a) Each NOX Budget permit...

  2. 40 CFR 72.85 - Permit reopenings.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... REGULATION Permit Revisions § 72.85 Permit reopenings. (a) The permitting authority shall reopen an Acid Rain permit for cause whenever: (1) Any additional requirement under the Acid Rain Program becomes applicable... revoked to assure compliance with Acid Rain Program requirements. (b) In reopening an Acid Rain permit for...

  3. 40 CFR 72.85 - Permit reopenings.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... REGULATION Permit Revisions § 72.85 Permit reopenings. (a) The permitting authority shall reopen an Acid Rain permit for cause whenever: (1) Any additional requirement under the Acid Rain Program becomes applicable... revoked to assure compliance with Acid Rain Program requirements. (b) In reopening an Acid Rain permit for...

  4. 40 CFR 72.85 - Permit reopenings.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... REGULATION Permit Revisions § 72.85 Permit reopenings. (a) The permitting authority shall reopen an Acid Rain permit for cause whenever: (1) Any additional requirement under the Acid Rain Program becomes applicable... revoked to assure compliance with Acid Rain Program requirements. (b) In reopening an Acid Rain permit for...

  5. 40 CFR 72.85 - Permit reopenings.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... REGULATION Permit Revisions § 72.85 Permit reopenings. (a) The permitting authority shall reopen an Acid Rain permit for cause whenever: (1) Any additional requirement under the Acid Rain Program becomes applicable... revoked to assure compliance with Acid Rain Program requirements. (b) In reopening an Acid Rain permit for...

  6. 40 CFR 72.85 - Permit reopenings.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... REGULATION Permit Revisions § 72.85 Permit reopenings. (a) The permitting authority shall reopen an Acid Rain permit for cause whenever: (1) Any additional requirement under the Acid Rain Program becomes applicable... revoked to assure compliance with Acid Rain Program requirements. (b) In reopening an Acid Rain permit for...

  7. Bio-inspired cryo-ink preserves red blood cell phenotype and function during nanoliter vitrification.

    PubMed

    El Assal, Rami; Guven, Sinan; Gurkan, Umut Atakan; Gozen, Irep; Shafiee, Hadi; Dalbeyler, Sedef; Abdalla, Noor; Thomas, Gawain; Fuld, Wendy; Illigens, Ben M W; Estanislau, Jessica; Khoory, Joseph; Kaufman, Richard; Zylberberg, Claudia; Lindeman, Neal; Wen, Qi; Ghiran, Ionita; Demirci, Utkan

    2014-09-03

    Current red-blood-cell cryopreservation methods utilize bulk volumes, causing cryo-injury of cells, which results in irreversible disruption of cell morphology, mechanics, and function. An innovative approach to preserve human red-blood-cell morphology, mechanics, and function following vitrification in nanoliter volumes is developed using a novel cryo-ink integrated with a bioprinting approach. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Evaluation of alloy 690 process pot at the contact with borosilicate melt pool during vitrification of high-level nuclear waste

    NASA Astrophysics Data System (ADS)

    Sengupta, Pranesh; Kaushik, C. P.; Kale, G. B.; Das, D.; Raj, K.; Sharma, B. P.

    2009-08-01

    Understanding the material behaviour under service conditions is essential to enhance the life span of alloy 690 process pot used in vitrification of high-level nuclear waste. During vitrification process, interaction of alloy 690 with borosilicate melt takes place for substantial time period. Present experimental studies show that such interactions may result in Cr carbide precipitation along grain boundaries, Cr depletion in austenitic matrix and intergranular attack close to alloy 690/borosilicate melt pool interfaces. Widths of Cr depleted zone within alloy 690 is found to follow kinetics of the type x = 10.9 × 10 -6 + 1 × 10 -8t1/2 m. Based on the experimental results it is recommended that compositional modification of alloy 690 process pot adjacent to borosilicate melt pool need to be considered seriously for any efforts towards reduction and/or prevention of process pot failures.

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

    PubMed

    Gunnala, Vinay; Schattman, Glenn

    2017-02-01

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

  10. Emerging technologies in medical applications of minimum volume vitrification

    PubMed Central

    Zhang, Xiaohui; Catalano, Paolo N; Gurkan, Umut Atakan; Khimji, Imran; Demirci, Utkan

    2011-01-01

    Cell/tissue biopreservation has broad public health and socio-economic impact affecting millions of lives. Cryopreservation technologies provide an efficient way to preserve cells and tissues targeting the clinic for applications including reproductive medicine and organ transplantation. Among these technologies, vitrification has displayed significant improvement in post-thaw cell viability and function by eliminating harmful effects of ice crystal formation compared to the traditional slow freezing methods. However, high cryoprotectant agent concentrations are required, which induces toxicity and osmotic stress to cells and tissues. It has been shown that vitrification using small sample volumes (i.e., <1 μl) significantly increases cooling rates and hence reduces the required cryoprotectant agent levels. Recently, emerging nano- and micro-scale technologies have shown potential to manipulate picoliter to nanoliter sample sizes. Therefore, the synergistic integration of nanoscale technologies with cryogenics has the potential to improve biopreservation methods. PMID:21955080

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

    PubMed

    Boldt, Jeffrey

    2011-09-01

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

  12. 40 CFR 122.45 - Calculating NPDES permit conditions (applicable to State NPDES programs, see § 123.25).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Permit Conditions § 122.45 Calculating NPDES permit conditions..., and prohibitions, including those necessary to achieve water quality standards, shall unless...

  13. 40 CFR 70.6 - Permit content.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 16 2012-07-01 2012-07-01 false Permit content. 70.6 Section 70.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS § 70.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include the following...

  14. 40 CFR 70.6 - Permit content.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 16 2014-07-01 2014-07-01 false Permit content. 70.6 Section 70.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS § 70.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include the following...

  15. 40 CFR 70.6 - Permit content.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 16 2013-07-01 2013-07-01 false Permit content. 70.6 Section 70.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS § 70.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include the following...

  16. 40 CFR 70.6 - Permit content.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 15 2010-07-01 2010-07-01 false Permit content. 70.6 Section 70.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS § 70.6 Permit content. (a) Standard permit requirements. Each permit issued under this part shall include the following...

  17. 40 CFR 96.20 - General NOX Budget trading program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General NOX Budget trading program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.20 General NOX Budget trading program permit requirements. (a...

  18. 40 CFR 96.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.220 General CAIR SO2 Trading Program permit requirements. (a...

  19. 40 CFR 97.20 - General NOX Budget Trading Program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General NOX Budget Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.20 General NOX Budget Trading Program permit requirements. (a) For each NOX Budget source...

  20. 40 CFR 97.20 - General NOX Budget Trading Program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General NOX Budget Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.20 General NOX Budget Trading Program permit requirements. (a) For each NOX Budget source...

  1. 40 CFR 96.20 - General NOX Budget trading program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General NOX Budget trading program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.20 General NOX Budget trading program permit requirements. (a...

  2. 40 CFR 97.20 - General NOX Budget Trading Program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General NOX Budget Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.20 General NOX Budget Trading Program permit requirements. (a) For each NOX Budget source...

  3. 40 CFR 96.220 - General CAIR SO 2 Trading Program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General CAIR SO 2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.220 General CAIR SO 2 Trading Program permit requirements. (a...

  4. 40 CFR 97.20 - General NOX Budget Trading Program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General NOX Budget Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.20 General NOX Budget Trading Program permit requirements. (a) For each NOX Budget source...

  5. 40 CFR 96.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.220 General CAIR SO2 Trading Program permit requirements. (a...

  6. 40 CFR 96.220 - General CAIR SO 2 Trading Program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General CAIR SO 2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO 2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.220 General CAIR SO 2 Trading Program permit requirements. (a...

  7. 40 CFR 97.20 - General NOX Budget Trading Program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General NOX Budget Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.20 General NOX Budget Trading Program permit requirements. (a) For each NOX Budget source...

  8. 40 CFR 96.20 - General NOX Budget trading program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General NOX Budget trading program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.20 General NOX Budget trading program permit requirements. (a...

  9. 40 CFR 96.20 - General NOX Budget trading program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General NOX Budget trading program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.20 General NOX Budget trading program permit requirements. (a...

  10. 40 CFR 96.20 - General NOX Budget trading program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General NOX Budget trading program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO 2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.20 General NOX Budget trading program permit requirements. (a...

  11. 40 CFR 96.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.220 General CAIR SO2 Trading Program permit requirements. (a...

  12. Permit Fees

    EPA Pesticide Factsheets

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements.

  13. 50 CFR 15.23 - Permits for zoological breeding or display programs.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... OF WILDLIFE AND PLANTS WILD BIRD CONSERVATION ACT Permits and Approval of Cooperative Breeding Programs § 15.23 Permits for zoological breeding or display programs. (a) Application requirements for... 50 Wildlife and Fisheries 1 2011-10-01 2011-10-01 false Permits for zoological breeding or display...

  14. 50 CFR 15.23 - Permits for zoological breeding or display programs.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... OF WILDLIFE AND PLANTS WILD BIRD CONSERVATION ACT Permits and Approval of Cooperative Breeding Programs § 15.23 Permits for zoological breeding or display programs. (a) Application requirements for... 50 Wildlife and Fisheries 1 2010-10-01 2010-10-01 false Permits for zoological breeding or display...

  15. 40 CFR 96.22 - Information requirements for NOX Budget permit applications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Budget permit applications. 96.22 Section 96.22 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.22 Information requirements for NOX Budget permit applications...

  16. Title V Operating Permits

    EPA Pesticide Factsheets

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements.

  17. EPA Issued Operating Permits

    EPA Pesticide Factsheets

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements.

  18. 40 CFR 97.23 - NOX Budget permit contents.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false NOX Budget permit contents. 97.23... (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.23 NOX Budget permit contents. (a) Each NOX Budget permit will contain, in a format prescribed by the permitting...

  19. Air Permitting Delegations in EPA's New England Region

    EPA Pesticide Factsheets

    All Region 1 states have EPA-approved Title V operating permit programs. In addition, all Region 1 states have either SIP-approved NSR/PSD permitting programs or the state has been delegated as the permitting authority for NSR/PSD permits by EPA.

  20. 40 CFR 97.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.220 General CAIR SO2 Trading Program permit requirements. (a) For each CAIR SO2 source...

  1. 40 CFR 97.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.220 General CAIR SO2 Trading Program permit requirements. (a) For each CAIR SO2 source...

  2. 40 CFR 97.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.220 General CAIR SO2 Trading Program permit requirements. (a) For each CAIR SO2 source...

  3. 40 CFR 97.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.220 General CAIR SO2 Trading Program permit requirements. (a) For each CAIR SO2 source...

  4. 40 CFR 97.220 - General CAIR SO2 Trading Program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General CAIR SO2 Trading Program... (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.220 General CAIR SO2 Trading Program permit requirements. (a) For each CAIR SO2 source...

  5. Analysis of Hanford Cast Stone Supplemental LAW using Composition Adjusted SRS Tank 50 Salt Solution

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

    Crawford, C.; Cozzi, A.; Hill, K.

    Vitrification is the primary disposition path for Low Activity Waste (LAW) at the Department of Energy (DOE) Hanford Site. A cementitious waste form is one of the alternatives being considered for the supplemental immobilization of the LAW that will not be treated by the primary vitrification facility. Washington River Protection Solutions (WRPS) has been directed to generate and collect data on cementitious or pozzolanic waste forms such as Cast Stone.

  6. Review of vitreous islet cryopreservation

    PubMed Central

    Baicu, Simona

    2009-01-01

    Transplantation of pancreatic islets for the treatment of diabetes mellitus is widely anticipated to eventually provide a cure once a means for preventing rejection is found without reliance upon global immunosuppression. Long-term storage of islets is crucial for the organization of transplantation, islet banking, tissue matching, organ sharing, immuno-manipulation and multiple donor transplantation. Existing methods of cryopreservation involving freezing are known to be suboptimal providing only about 50% survival. The development of techniques for ice-free cryopreservation of mammalian tissues using both natural and synthetic ice blocking molecules, and the process of vitrification (formation of a glass as opposed to crystalline ice) has been a focus of research during recent years. These approaches have established in other tissues that vitrification can markedly improve survival by circumventing ice-induced injury. Here we review some of the underlying issues that impact the vitrification approach to islet cryopreservation and describe some initial studies to apply these new technologies to the long-term storage of pancreatic islets. These studies were designed to optimize both the pre-vitrification hypothermic exposure conditions using newly developed media and to compare new techniques for ice-free cryopreservation with conventional freezing protocols. Some practical constraints and feasible resolutions are discussed. Eventually the optimized techniques will be applied to clinical allografts and xenografts or genetically-modified islets designed to overcome immune responses in the diabetic host. PMID:20046679

  7. Successful slush nitrogen vitrification of human ovarian tissue.

    PubMed

    Talevi, Riccardo; Barbato, Vincenza; Fiorentino, Ilaria; Braun, Sabrina; De Stefano, Cristofaro; Ferraro, Raffaele; Sudhakaran, Sam; Gualtieri, Roberto

    2016-06-01

    To study whether slush nitrogen vitrification improves the preservation of human ovarian tissue. Control vs. treatment study. University research laboratory. Ovarian biopsies collected from nine women (aged 14-35 years) during laparoscopic surgery for benign gynecologic conditions. None. Ovarian cortical strips of 2 × 5 × 1 mm were vitrified with liquid or slush nitrogen. Fresh and vitrified cortical strips were analyzed for cryodamage and viability under light, confocal, and transmission electron microscopy. Compared with liquid nitrogen, vitrification with slush nitrogen preserves [1] follicle quality (grade 1 follicles: fresh control, 50%; liquid nitrogen, 27%; slush nitrogen, 48%); [2] granulosa cell ultrastructure (intact cells: fresh control, 92%; liquid nitrogen, 45%; slush nitrogen, 73%), stromal cell ultrastructure (intact cells: fresh control, 59.8%; liquid nitrogen, 24%; slush nitrogen, 48.7%), and DNA integrity (TUNEL-positive cells: fresh control, 0.5%; liquid nitrogen, 2.3%; slush nitrogen, 0.4%); and [3] oocyte, granulosa, and stromal cell viability (oocyte: fresh control, 90%; liquid nitrogen, 63%; slush nitrogen, 87%; granulosa cells: fresh control, 93%; liquid nitrogen, 53%; slush nitrogen, 81%; stromal cells: fresh control, 63%; liquid nitrogen, 30%; slush nitrogen, 52%). The histology, ultrastructure, and viability of follicles and stromal cells are better preserved after vitrification with slush nitrogen compared with liquid nitrogen. Copyright © 2016 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  8. The influence of cumulus cells during in vitro fertilization of buffalo (Bubalus bubalis) denuded oocytes that have undergone vitrification.

    PubMed

    Attanasio, Laura; De Rosa, Anna; De Blasi, Marina; Neglia, Gianluca; Zicarelli, Luigi; Campanile, Giuseppe; Gasparrini, Bianca

    2010-11-01

    The aim of this work was to evaluate whether providing a support of cumulus cells during IVF of buffalo denuded oocytes submitted to vitrification-warming enhances their fertilizing ability. In vitro matured denuded oocytes were vitrified by Cryotop in 20% EG + 20% of DMSO and 0.5 M sucrose and warmed into decreasing concentrations of sucrose (1.25 M-0.3M). Oocytes that survived vitrification were fertilized: 1) in the absence of a somatic support (DOs); 2) in the presence of bovine cumulus cells in suspension (DOs+susp); 3) on a bovine cumulus monolayer (DOs+monol); and 4) with intact bovine COCs in a 1:1 ratio (DOs+COCs). In vitro matured oocytes were fertilized and cultured to the blastocyst stage as a control. An increased cleavage rate was obtained from DOs+COCs (60.9%) compared to DOs, DOs+susp (43.6 and 38.4, respectively; P < 0.01) and DOs+monol (47.5%; P < 0.05). Interestingly, cleavage rate of DOs+COCs was similar to that of fresh control oocytes (67.8%). However, development to blastocysts significantly decreased in all vitrification groups compared to the control (P < 0.01). In conclusion the co-culture with intact COCs during IVF completely restores fertilizing capability of buffalo denuded vitrified oocytes, without improving blastocyst development. Copyright © 2010 Elsevier Inc. All rights reserved.

  9. Cold Test Operation of the German VEK Vitrification Plant

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

    Fleisch, J.; Schwaab, E.; Weishaupt, M.

    2008-07-01

    In 2007 the German High-Level Liquid Waste (HLLW) Vitrification plant VEK (Verglasungseinrichtung Karlsruhe) has passed a three months integral cold test operation as final step before entering the hot phase. The overall performance of the vitrification process equipment with a liquid-fed ceramic glass melter as main component proved to be completely in line with the requirements of the regulatory body. The retention efficiency of main radioactive-bearing elements across melter and wet off-gas treatment system exceeded the design values distinctly. The strategy to produce a specified waste glass could be successfully demonstrated. The results of the cold test operation allow enteringmore » the next step of hot commissioning, i.e. processing of approximately 2 m{sup 3} of diluted HLLW. In summary: An important step of the VEK vitrification plant towards hot operation has been the performance of the cold test operation from April to July 2007. This first integral operation was carried out under boundary conditions and rules established for radioactive operation. Operation and process control were carried out following the procedure as documented in the licensed operational manuals. The function of the process technology and the safe operation could be demonstrated. No severe problems were encountered. Based on the positive results of the cold test, application of the license for hot operation has been initiated and is expected in the near future. (authors)« less

  10. Effects of polyethylene glycol and a synthetic ice blocker during vitrification of immature porcine oocytes on survival and subsequent embryo development.

    PubMed

    Santos, Elisa Caroline da Silva; Somfai, Tamas; Appeltant, Ruth; Dang-Nguyen, Thanh Quang; Noguchi, Junko; Kaneko, Hiroyuki; Kikuchi, Kazuhiro

    2017-08-01

    We evaluated the effects of polyethylene glycol (PEG) and Supercool X-1000 (SC) as supplements during the vitrification of immature cumulus-enclosed porcine oocytes in a solution based on 17.5% ethylene glycol + 17.5% propylene glycol. After warming, the oocytes were subjected to in vitro maturation, fertilization and embryo culture. In Experiment 1, equilibration and vitrification solutions were supplemented with or without 2% (w/v) PEG (PEG+ and PEG-, respectively). The survival rate, cleavage and blastocyst development were similar between PEG+ and PEG- groups; however, all values were lower than those in the non-vitrified control. In Experiment 2, vitrification solution was supplemented with or without 1% (v/v) SC (SC+ and SC-, respectively). The percentages of survival and blastocyst development were similar between SC+ and SC- groups but lower than those in the non-vitrified control. The percentage of cleavage in the SC- group was significantly lower than the control and the SC+ groups, which were in turn similar to one another. In both experiments, the cell numbers in blastocysts were not significantly different among the non-vitrified and vitrified groups. In conclusion, PEG did not improve oocyte survival and embryo development, whereas SC improved the ability of surviving oocytes to cleave but not to develop into blastocysts. © 2016 Japanese Society of Animal Science.

  11. 40 CFR 122.64 - Termination of permits (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Transfer, Modification, Revocation and Reissuance, and Termination of Permits § 122.64...

  12. 40 CFR 122.61 - Transfer of permits (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Transfer, Modification, Revocation and Reissuance, and Termination of Permits § 122.61...

  13. 40 CFR 122.64 - Termination of permits (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Transfer, Modification, Revocation and Reissuance, and Termination of Permits § 122.64...

  14. 40 CFR 122.61 - Transfer of permits (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Transfer, Modification, Revocation and Reissuance, and Termination of Permits § 122.61...

  15. 40 CFR 97.21 - Submission of NOX Budget permit applications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Submission of NOX Budget permit... PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.21 Submission of NOX Budget permit applications. (a) Duty to apply. The NOX authorized account...

  16. 40 CFR 96.24 - Effective date of initial NOX Budget permit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Effective date of initial NOX Budget... PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.24 Effective date of initial NOX Budget permit. The initial NOX Budget permit...

  17. 40 CFR 96.21 - Submission of NOX Budget permit applications.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Submission of NOX Budget permit... PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.21 Submission of NOX Budget permit applications. (a) Duty to apply. The NOX...

  18. 40 CFR 270.60 - Permits by rule.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Special Forms of Permits § 270.60 Permits by... operator of a barge or other vessel which accepts hazardous waste for ocean disposal, if the owner or...

  19. 40 CFR 270.60 - Permits by rule.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Special Forms of Permits § 270.60 Permits by... operator of a barge or other vessel which accepts hazardous waste for ocean disposal, if the owner or...

  20. 40 CFR 270.43 - Termination of permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false Termination of permits. 270.43 Section 270.43 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Changes to Permit § 270.43...

  1. 40 CFR 270.43 - Termination of permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false Termination of permits. 270.43 Section 270.43 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Changes to Permit § 270.43...

  2. Historical Permit Fee Rates

    EPA Pesticide Factsheets

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements. Historical fee rates.

  3. 40 CFR 97.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.120 General CAIR NOX Annual Trading Program permit requirements. (a) For...

  4. 40 CFR 96.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.120 General CAIR NOX Annual Trading Program permit...

  5. 40 CFR 96.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.120 General CAIR NOX Annual Trading Program permit...

  6. 40 CFR 97.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.120 General CAIR NOX Annual Trading Program permit requirements. (a) For...

  7. 40 CFR 97.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.120 General CAIR NOX Annual Trading Program permit requirements. (a) For...

  8. 40 CFR 97.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.120 General CAIR NOX Annual Trading Program permit requirements. (a) For...

  9. 40 CFR 96.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO 2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.120 General CAIR NOX Annual Trading Program permit...

  10. 40 CFR 96.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.120 General CAIR NOX Annual Trading Program permit...

  11. 40 CFR 97.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.120 General CAIR NOX Annual Trading Program permit requirements. (a) For...

  12. 40 CFR 96.120 - General CAIR NOX Annual Trading Program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General CAIR NOX Annual Trading... AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.120 General CAIR NOX Annual Trading Program permit...

  13. 78 FR 20073 - Adequacy of Oregon's Municipal Solid Waste Landfill Permit Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-03

    ...] Adequacy of Oregon's Municipal Solid Waste Landfill Permit Program AGENCY: Environmental Protection Agency... Oregon's approved Municipal Solid Waste Landfill Program. On March 22, 2004, EPA issued final regulations... Oregon's Municipal Solid Waste Landfill permit program to allow for Research, Development, and...

  14. Impact of vitrification on human oocytes before and after in vitro maturation: A systematic review and meta-analysis.

    PubMed

    Mohsenzadeh, Mehdi; Salehi-Abargouei, Amin; Tabibnejad, Nasim; Karimi-Zarchi, Mojgan; Khalili, Mohammad Ali

    2018-05-21

    There are controversies regarding in vitro maturation (IVM) procedure, the time of storing frozen oocytes and maturation stage of vitrified oocytes and its impact on oocytes fertilization capability. The aim of this systematic review and meta-analysis was to evaluate the impact of vitrification on human oocytes during IVM procedure. A systematic review with meta-analysis was undertaken. Main search terms were those related key words. We searched Medline, Embase, Scopus and ISI web of science to detect English-language studies. The final search was performed on 27 January 2018. The original articles which studied laboratory outcomes after vitrification of MII or GV oocytes before or after IVM were included. Exclusion criteria were animal trials and the studies that performed cryopreservation using slow-freeze method. Oocyte maturation, survival, fertilization and cleavage rates were assessed. Bias and quality assessments were applied. 2476 articles were screened and after duplicates removing together with application of inclusion and exclusion criteria, 14 studies assessed for eligibility. Finally 5 studies included for analysis. All studies compared laboratory outcomes between oocytes that vitrified at the GV stage and those which firstly matured in vitro, and then vitrified. Meta-analysis showed that vitrification of oocytes at GV stage had a negative impact on maturation rate (RR = 1.28, 95% CI: 0.96-1.70); but not on cleavage rate (RR = 1.07, 95% CI: 0.70-1.64); fertilization rate (RR = 0.99, 95% CI: 0.85-1.14) and survival rate(RR = 1.01, 95% CI: 0.96-1.06). In general, Based on our results, oocyte vitrification decreases the maturation rate. In addition, survival, fertilization as well as cleavage rates did not significantly differ between the oocytes vitrified before IVM versus oocytes vitrified after IVM. Copyright © 2018. Published by Elsevier B.V.

  15. Survival of mouse oocytes after being cooled in a vitrification solution to -196°C at 95° to 70,000°C/min and warmed at 610° to 118,000°C/min: A new paradigm for cryopreservation by vitrification.

    PubMed

    Mazur, Peter; Seki, Shinsuke

    2011-02-01

    There is great interest in achieving reproducibly high survivals of mammalian oocytes (especially human) after cryopreservation, but the results to date have not matched the interest. A prime cause of cell death is the formation of more than trace amounts of intracellular ice, and one strategy to avoid it is vitrification. In vitrification procedures, cells are loaded with high concentrations of glass-inducing solutes and cooled to -196°C at rates high enough to presumably induce the glassy state. In the last decade, several devices have been developed to achieve very high cooling rates. Nearly all in the field have assumed that the cooling rate is the critical factor. The purpose of our study was to test that assumption by examining the consequences of cooling mouse oocytes in a vitrification solution at four rates ranging from 95 to 69,250°C/min to -196°C and for each cooling rate, subjecting them to five warming rates back above 0°C at rates ranging from 610 to 118,000°C/min. In samples warmed at the highest rate (118,000°C/min), survivals were 70% to 85% regardless of the prior cooling rate. In samples warmed at the lowest rate (610°C/min), survivals were low regardless of the prior cooling rate, but decreased from 25% to 0% as the cooling rate was increased from 95 to 69,000°C/min. Intermediate cooling and warming rates gave intermediate survivals. The especially high sensitivity of survival to warming rate suggests that either the crystallization of intracellular glass during warming or the growth by recrystallization of small intracellular ice crystals formed during cooling are responsible for the lethality of slow warming. Copyright © 2010 Elsevier Inc. All rights reserved.

  16. VITRIFICATION SYSTEM FOR THE TREATMENT OF PLUTONIUM-BEARING WASTE AT LOS ALAMOS NATIONAL LABORATORY

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

    R. NAKAOKA; G. VEAZEY; ET AL

    2001-05-01

    A glove box vitrification system is being fabricated to process aqueous evaporator bottom waste generated at the Plutonium Facility (TA-55) at Los Alamos National Laboratory (LANL). The system will be the first within the U.S. Department of Energy Complex to routinely convert Pu{sup 239}-bearing transuranic (TRU) waste to a glass matrix for eventual disposal at the Waste Isolation Pilot Plant (WIPP). Currently at LANL, this waste is solidified in Portland cement. Radionuclide loading in the cementation process is restricted by potential radiolytic degradation (expressed as a wattage limit), which has been imposed to prevent the accumulation of flammable concentrations ofmore » H{sub 2} within waste packages. Waste matrixes with a higher water content (e.g., cement) are assigned a lower permissible wattage limit to compensate for their potential higher generation of H{sub 2}. This significantly increases the number of waste packages that must be prepared and shipped, thus driving up the costs of waste handling and disposal. The glove box vitrification system that is under construction will address this limitation. Because the resultant glass matrix produced by the vitrification process is non-hydrogenous, no H{sub 2} can be radiolytically evolved, and drums could be loaded to the maximum allowable limit of 40 watts. In effect, the glass waste form shifts the limiting constraint for loading disposal drums from wattage to the criticality limit of 200 fissile gram equivalents, thus significantly reducing the number of drums generated from this waste stream. It is anticipated that the number of drums generated from treatment of evaporator bottoms will be reduced by a factor of 4 annually when the vitrification system is operational. The system is currently undergoing non-radioactive operability testing, and will be fully operational in the year 2003.« less

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

    PubMed

    Li, Bai-Quan; Feng, Chao-Hong; Wang, Min-Rui; Hu, Ling-Yun; Volk, Gayle; Wang, Qiao-Chun

    2015-11-20

    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 procedure that was previously reported by us. In both procedures, three types of shoot tip recovery were observed following cryopreservation: callus formation without shoot regrowth, leaf formation without shoot regrowth, and shoot regrowth. Three categories of histological observations were also identified in cross-sections of shoot tips recovered after cryopreservation using the two cryogenic procedures. In category 1, almost all of the cells (94-95%) in the apical dome (AD) were damaged or killed and only some cells (30-32%) in the leaf primordia (LPs) survived. In category 2, only a few cells (18-20%) in the AD and some cells (30-31%) in the LPs survived. In category 3, majority of the cells (60-62%) in the AD and some cells (30-33%) in the LPs survived. These data suggest that shoot regrowth is correlated to the presence of a majority of surviving cells in the AD after liquid nitrogen exposure. No polymorphic bands were detected by inter-simple sequence repeats or by random amplified polymorphic DNA assessments, and ploidy levels analyzed by flow cytometry were unchanged when plants recovered after cryoexposure were compared to controls. The droplet-vitrification procedure appears to be robust since seven genotypes representing four Malus species and one hybrid recovered shoots following cryopreservation. Mean shoot regrowth levels of these seven genotypes were 48% in the droplet-vitrification method, which were lower than those (61%) in the encapsulation-dehydration procedure reported in our previous study, suggesting the latter may be preferred for routine cryobanking applications for Malus shoot tips. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. 40 CFR 97.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General CAIR NOX Ozone Season Trading... TRADING PROGRAMS Permits § 97.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating permit or required, under...

  19. 40 CFR 97.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General CAIR NOX Ozone Season Trading... TRADING PROGRAMS Permits § 97.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating permit or required, under...

  20. 40 CFR 97.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General CAIR NOX Ozone Season Trading... TRADING PROGRAMS Permits § 97.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating permit or required, under...

  1. 40 CFR 97.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General CAIR NOX Ozone Season Trading... TRADING PROGRAMS Permits § 97.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating permit or required, under...

  2. 40 CFR 97.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General CAIR NOX Ozone Season Trading... TRADING PROGRAMS Permits § 97.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating permit or required, under...

  3. 40 CFR 96.23 - NOX Budget permit contents.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false NOX Budget permit contents. 96.23 Section 96.23 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.23 NOX Budget permit contents. (a)...

  4. 77 FR 16509 - Proposed Approval of Revision of Five California Clean Air Act Title V Operating Permits Programs

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-21

    ... Approval of Revision of Five California Clean Air Act Title V Operating Permits Programs AGENCY... the Operating Permits (Title V) programs of the Monterey Bay Unified Air Pollution Control District... been previously subject to Title V for other reasons to obtain a Title V permit. See ``Prevention of...

  5. Blood Banking in Living Droplets

    PubMed Central

    Shao, Lei; Zhang, Xiaohui; Xu, Feng; Song, YoungSeok; Keles, Hasan Onur; Matloff, Laura; Markel, Jordan; Demirci, Utkan

    2011-01-01

    Blood banking has a broad public health impact influencing millions of lives daily. It could potentially benefit from emerging biopreservation technologies. However, although vitrification has shown advantages over traditional cryopreservation techniques, it has not been incorporated into transfusion medicine mainly due to throughput challenges. Here, we present a scalable method that can vitrify red blood cells in microdroplets. This approach enables the vitrification of large volumes of blood in a short amount of time, and makes it a viable and scalable biotechnology tool for blood cryopreservation. PMID:21412411

  6. 40 CFR 123.25 - Requirements for permitting.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Requirements for permitting. 123.25 Section 123.25 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS STATE PROGRAM REQUIREMENTS State Program Submissions § 123.25 Requirements for permitting. (a) All State Programs under this part must have legal...

  7. Avoiding Title V permitting pitfalls

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

    Laswell, D.L.

    1993-04-01

    Title V of the 1990 Clean Air Act Amendments requires states to implement new air operating permit programs. States have a great deal of flexibility in developing their permit programs. Industry should work now to ensure that state programs contain the favorable aspects of the federal regulations and do not contain more stringent requirements that are not required under the Clean Air Act. This article outlines areas of the permit program that have the potential to handicap industry`s ability to expand.

  8. 40 CFR 96.23 - NOX Budget permit contents.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false NOX Budget permit contents. 96.23... (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.23 NOX Budget permit contents. (a) Each NOX Budget permit (including any draft or proposed...

  9. 40 CFR 96.25 - NOX Budget permit revisions.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false NOX Budget permit revisions. 96.25... (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.25 NOX Budget permit revisions. (a) For a NOX Budget source with a title V operating permit...

  10. 40 CFR 97.24 - NOX Budget permit revisions.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false NOX Budget permit revisions. 97.24... (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Permits § 97.24 NOX Budget permit revisions. (a) For a NOX Budget source with a title V operating permit, except as provided...

  11. 40 CFR 76.9 - Permit application and compliance plans.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... PROGRAMS (CONTINUED) ACID RAIN NITROGEN OXIDES EMISSION REDUCTION PROGRAM § 76.9 Permit application and... complete Acid Rain permit application (or, if the unit is covered by an Acid Rain permit, a complete permit... specify one or more other Acid Rain compliance options for NOX in accordance with the requirements of this...

  12. 40 CFR 72.73 - State issuance of Phase II permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.73 State issuance of Phase II permits... permit program under part 70 of this chapter and that has a State Acid Rain program accepted by the Administrator under § 72.71 shall be responsible for administering and enforcing Acid Rain permits effective in...

  13. 40 CFR 76.9 - Permit application and compliance plans.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... PROGRAMS (CONTINUED) ACID RAIN NITROGEN OXIDES EMISSION REDUCTION PROGRAM § 76.9 Permit application and... complete Acid Rain permit application (or, if the unit is covered by an Acid Rain permit, a complete permit... specify one or more other Acid Rain compliance options for NOX in accordance with the requirements of this...

  14. 40 CFR 76.9 - Permit application and compliance plans.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... PROGRAMS (CONTINUED) ACID RAIN NITROGEN OXIDES EMISSION REDUCTION PROGRAM § 76.9 Permit application and... complete Acid Rain permit application (or, if the unit is covered by an Acid Rain permit, a complete permit... or specify one or more other Acid Rain compliance options for NOX in accordance with the requirements...

  15. 40 CFR 72.73 - State issuance of Phase II permits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.73 State issuance of Phase II permits... permit program under part 70 of this chapter and that has a State Acid Rain program accepted by the Administrator under § 72.71 shall be responsible for administering and enforcing Acid Rain permits effective in...

  16. 40 CFR 76.9 - Permit application and compliance plans.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... PROGRAMS (CONTINUED) ACID RAIN NITROGEN OXIDES EMISSION REDUCTION PROGRAM § 76.9 Permit application and... complete Acid Rain permit application (or, if the unit is covered by an Acid Rain permit, a complete permit... specify one or more other Acid Rain compliance options for NOX in accordance with the requirements of this...

  17. 40 CFR 72.73 - State issuance of Phase II permits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.73 State issuance of Phase II permits... permit program under part 70 of this chapter and that has a State Acid Rain program accepted by the Administrator under § 72.71 shall be responsible for administering and enforcing Acid Rain permits effective in...

  18. 40 CFR 72.73 - State issuance of Phase II permits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.73 State issuance of Phase II permits... permit program under part 70 of this chapter and that has a State Acid Rain program accepted by the Administrator under § 72.71 shall be responsible for administering and enforcing Acid Rain permits effective in...

  19. 40 CFR 72.73 - State issuance of Phase II permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... (CONTINUED) PERMITS REGULATION Acid Rain Phase II Implementation § 72.73 State issuance of Phase II permits... permit program under part 70 of this chapter and that has a State Acid Rain program accepted by the Administrator under § 72.71 shall be responsible for administering and enforcing Acid Rain permits effective in...

  20. 40 CFR 76.9 - Permit application and compliance plans.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... PROGRAMS (CONTINUED) ACID RAIN NITROGEN OXIDES EMISSION REDUCTION PROGRAM § 76.9 Permit application and... complete Acid Rain permit application (or, if the unit is covered by an Acid Rain permit, a complete permit... or specify one or more other Acid Rain compliance options for NOX in accordance with the requirements...

  1. 40 CFR 270.275 - What information must I submit to the permitting agency to support my standardized permit...

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Federal laws required in § 270.3. (e) Solid waste management unit information required by § 270.14(d). (f... Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM RCRA Standardized Permits for Storage and Treatment Units...

  2. Survival and genetic stability of Dendranthema grandiflora Tzvelev shoot apices after cryopreservation by vitrification and encapsulation-dehydration.

    PubMed

    Martín, Carmen; González-Benito, M Elena

    2005-12-01

    The aim of this study was to compare the genetic stability of chrysanthemum (cv. Pasodoble) apices cryopreserved using two different methods: encapsulation-dehydration and vitrification. The assessment of the genetic stability was developed using RAPDs markers. Assessment of stability was evaluated in pot-cultivated mother plants (from which buds were excised for micropropagation), in shoots (leave tissue) from which apices were extracted for cryopreservation, and in shoots regenerated from cryopreserved apices 30 days after recovery and after further 3 months in culture. Throughout the process the origin of the apices (in vitro-shoot from which they were excised) was recorded. Twenty one regenerants cryopreserved by vitrification and 25 by encapsulation-dehydration were assessed. Only one cryopreserved regenerant from the encapsulation-dehydration method showed a different band pattern. These results support the necessity of monitoring the genetic stability of the regenerants obtained after cryopreservation, as this is a very useful technique for the conservation of plant genetic resources.

  3. Sodalite as a vehicle to increase Re retention in waste glass simulant during vitrification

    NASA Astrophysics Data System (ADS)

    Luksic, Steven A.; Riley, Brian J.; Parker, Kent E.; Hrma, Pavel

    2016-10-01

    Technetium (Tc) retention during Hanford waste vitrification can be increased if the volatility can be controlled. Incorporating Tc into a thermally stable mineral phase, such as sodalite, is one way to achieve increased retention. Here, rhenium (Re)-bearing sodalite was tested as a vehicle to transport perrhenate (ReO4-), a nonradioactive surrogate for pertechnetate (TcO4-), into high-level (HLW) and low-activity waste (LAW) glass simulants. After melting HLW and LAW simulant feeds, the retention of Re in the glass was measured and compared with the Re retention in glass prepared from a feed containing Re2O7. Phase analysis of sodalite in both these glasses across a profile of temperatures describes the durability of Re-sodalite during the feed-to-glass transition. The use of Re sodalite improved the Re retention by 21% for HLW glass and 85% for LAW glass, demonstrating the potential improvement in Tc-retention if TcO4- were to be encapsulated in a Tc-sodalite prior to vitrification.

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

    PubMed

    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.

  5. Dehydration Preparation of Mouse Sperm for Vitrification and Rapid Laser Warming.

    PubMed

    Paredes, E; Mazur, P

    Mice are fundamental models of study due to their ease of breeding, manipulation, and the well-studied genome. There has been extensive research focused on the cryopreservation of mouse germaplasm, as a way to help maintain the different transgenic mouse breeds. The first protocols for mouse sperm were developed in the 90's using slow cooling and a mixture of raffinose and glycerol. Since then, the rate of success reported remains highly variable. The Aim of this work is to study factors that are key for developing vitrification protocols for ultra-rapid laser warming of mouse sperm. Our results show that due to the exquisite sensitivity of sperm cells to osmotic excursions, our target levels of dehydration (~85% water content) cannot be achieved without causing a significant decrease in sperm motility and membrane fusion. It seems likely that mouse sperm vitrification is going to be difficult to develop due to the exquisite sensitivity of mouse sperm cells to handling and dehydration.

  6. HIGH INCIDENCE OF POLYSPERMIC FERTILIZATION IN BOVINE OOCYTES MATURED IN VITRO AFTER CRYOTOP VITRIFICATION.

    PubMed

    Hwang, In-Sul; Kwon, Dae-Jin; Im, Gi-Sun; Tashima, Kazuya; Hochi, Shinichi; Hwang, Seongsoo

    2016-01-01

    Vitrification with the Cryotop device is the most promising technique for oocyte cryopreservation, but the high post-warming morphological survival of bovine oocytes does not guarantee high developmental competence after in vitro fertilization (IVF). This study was designed to examine achievement of normal fertilization in bovine oocytes vitrified-warmed with the Cryotop device. Oocytes were matured in vitro and vitrified-warmed after complete removal of the cumulus layers. Distribution of cortical granules (CGs) was assessed by Lens culinaris agglutinin (LCA) lectin staining. Ten hours after IVF, presumptive zygotes were analyzed for pronuclear formation. Day-8 blastocysts were harvested and stained with Hoechst-33342 for total cell counting. Both yield and mean cell number of the blastocysts were impaired by Cryotop vitrification. Incidence of polyspermic fertilization was three-times higher in vitrified oocytes compared to fresh oocytes. No difference in CG distribution was found between vitrified and fresh oocytes. Polyspermic fertilization induced in vitrified-warmed bovine oocytes may be one of the possible causes responsible for their low developmental potential.

  7. Safeguardability of the vitrification option for disposal of plutonium

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

    Pillay, K.K.S.

    1996-05-01

    Safeguardability of the vitrification option for plutonium disposition is rather complex and there is no experience base in either domestic or international safeguards for this approach. In the present treaty regime between the US and the states of the former Soviet Union, bilaterial verifications are considered more likely with potential for a third-party verification of safeguards. There are serious technological limitations to applying conventional bulk handling facility safeguards techniques to achieve independent verification of plutonium in borosilicate glass. If vitrification is the final disposition option chosen, maintaining continuity of knowledge of plutonium in glass matrices, especially those containing boron andmore » those spike with high-level wastes or {sup 137}Cs, is beyond the capability of present-day safeguards technologies and nondestructive assay techniques. The alternative to quantitative measurement of fissile content is to maintain continuity of knowledge through a combination of containment and surveillance, which is not the international norm for bulk handling facilities.« less

  8. The role of troublesome components in plutonium vitrification

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

    Li, Hong; Vienna, J.D.; Peeler, D.K.

    1996-05-01

    One option for immobilizing surplus plutonium is vitrification in a borosilicate glass. Two advantages of the glass form are (1) high tolerance to feed variability and, (2) high solubility of some impurity components. The types of plutonium-containing materials in the United States inventory include: pits, metals, oxides, residues, scrap, compounds, and fuel. Many of them also contain high concentrations of carbon, chloride, fluoride, phosphate, sulfate, and chromium oxide. To vitrify plutonium-containing scrap and residues, it is critical to understand the impact of each component on glass processing and chemical durability of the final product. This paper addresses glass processing issuesmore » associated with these troublesome components. It covers solubility limits of chlorine, fluorine, phosphate, sulfate, and chromium oxide in several borosilicate based glasses, and the effect of each component on vitrification (volatility, phase segregation, crystallization, and melt viscosity). Techniques (formulation, pretreatment, removal, and/or dilution) to mitigate the effect of these troublesome components are suggested.« less

  9. Dewatering Treatment Scale-up Testing Results of Hanford Tank Wastes

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

    Tedeschi, A.R.; May, T.H.; Bryan, W.E.

    2008-07-01

    This report documents CH2M HILL Hanford Group Inc. (CH2M HILL) 2007 dryer testing results in Richland, WA at the AMEC Nuclear Ltd., GeoMelt Division (AMEC) Horn Rapids Test Site. It provides a discussion of scope and results to qualify the dryer system as a viable unit-operation in the continuing evaluation of the bulk vitrification process. A 10,000 liter (L) dryer/mixer was tested for supplemental treatment of Hanford tank low activity wastes, drying and mixing a simulated non-radioactive salt solution with glass forming minerals. Testing validated the full scale equipment for producing dried product similar to smaller scale tests, and qualifiedmore » the dryer system for a subsequent integrated dryer/vitrification test using the same simulant and glass formers. The dryer system is planned for installation at the Hanford tank farms to dry/mix radioactive waste for final treatment evaluation of the supplemental bulk vitrification process. (authors)« less

  10. An alternative approach to recovering valuable metals from zinc phosphating sludge.

    PubMed

    Kuo, Yi-Ming

    2012-01-30

    This study used a vitrification process (with good potential for commercialization) to recover valuable metals from Zn phosphating sludge. The involved vitrification process achieves two major goals: it transformed hazardous Zn phosphating sludge into inert slag and it concentrated Fe (83.5%) and Zn (92.8%) into ingot and fine particulate-phase material, respectively. The Fe content in the ingot was 278,000 mg/kg, making the ingot a potential raw material for iron making. The fine particulate-phase material (collected from flue gas) contained abundant Zn (544,000 mg/kg) in the form of ZnO. The content (67.7%) of ZnO was high, so it can be directly sold to refineries. The recovered coarse particulate-phase material, with insufficient amount of ZnO, can be recycled as a feeding material for Zn re-concentration. Therefore, the vitrification process can not only treat hazardous materials but also effectively recover valuable metals. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. Stabilization of biothreat diagnostic samples through vitrification matrices.

    PubMed

    Minogue, Timothy Devin; Kalina, Warren Vincent; Coyne, Susan Rajnik

    2014-06-01

    Diagnostics for biothreat agents require sample shipment to reference labs for diagnosis of disease; however high/fluctuating temperatures during sample transport negatively affect sample quality and results. Vitrification additives preserve sample integrity for molecular-based assay diagnostics in the absence of refrigeration by imparting whole molecule stability to a plethora of environmental insults. Therefore, we have evaluated commercially available vitrification matrices' (Biomatrica's CloneStable® and RNAStable®) ability to stabilize samples of Yersinia pestis and Venezuelan Equine Encephalitis Virus. When heated to 95°C in RNAStable®, Y. pestis had a 13-fold improvement in detection via real-time PCR compared to heated samples in buffer. VEEV, in RNAStable® at 55°C, had a ~10-fold improved detection versus heated samples in buffer. CloneStable® also preserved Y. pestis antigens for 7days after exposure to cycling temperatures. Overall, RNAStable® and CloneStable® respectively offered superior stabilization to nucleic acids and proteins in response to temperature fluctuations. Copyright © 2014. Published by Elsevier B.V.

  12. The role of frit in nuclear waste vitrification

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

    Vienna, J.D.; Smith, P.A.; Dorn, D.A.

    1994-04-01

    Vitrification of nuclear waste requires additives which are often vitrified independently to form a frit. Frit composition is formulated to meet the needs of glass composition and processing. The effects of frit on melter feed and melt processing, glass acceptance, and waste loading is of practical interest in understanding the trade-offs associated with the competing demands placed on frit composition. Melter feed yield stress, viscosity and durability of frits and corresponding waste glasses as well as the kinetics of elementary melting processes have been measured. The results illustrate the competing requirements on frit. Four frits (FY91, FY93, HW39-4, and SR202)more » and simulated neutralized current acid waste (NCAW) were used in this study. The experimental evidence shows that optimization of frit for one processing related property often results in poorer performance for the remaining properties. The difficulties associated with maximum waste loading and durability are elucidated for glasses which could be processed using technology available for the previously proposed Hanford Waste Vitrification Plant.« less

  13. 40 CFR 60.4120 - General Hg budget trading program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 6 2011-07-01 2011-07-01 false General Hg budget trading program... budget trading program permit requirements. (a) For each Hg Budget source required to have a title V... source covered by the Hg Budget permit, all applicable Hg Budget Trading Program requirements and shall...

  14. 40 CFR 144.34 - Emergency permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ....34 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) UNDERGROUND INJECTION CONTROL PROGRAM Authorization by Permit § 144.34 Emergency permits. (a) Coverage... unless a temporary emergency permit is granted; or (2) A substantial and irretrievable loss of oil or gas...

  15. 40 CFR 25.2 - Scope.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Hazardous Waste Program; the NPDES Permit Program; the Dredge and Fill Permit Program; and the Underground... programs. The requirements for public participation in State Hazardous Waste Programs, Dredge and Fill...

  16. 40 CFR 25.2 - Scope.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Hazardous Waste Program; the NPDES Permit Program; the Dredge and Fill Permit Program; and the Underground... programs. The requirements for public participation in State Hazardous Waste Programs, Dredge and Fill...

  17. Dynamics of intracellular phospholipid membrane organization during oocyte maturation and successful vitrification of immature oocytes retrieved by ovum pick-up in cattle.

    PubMed

    Aono, Akira; Nagatomo, Hiroaki; Takuma, Tetsuya; Nonaka, Rika; Ono, Yoshitaka; Wada, Yasuhiko; Abe, Yasuyuki; Takahashi, Masashi; Watanabe, Tomomasa; Kawahara, Manabu

    2013-05-01

    The objective was to determine if immature bovine oocytes with cumulus cells at the germinal vesicle (GV) stage could be vitrified by aluminum sheets (AS; pieces of sheet-like aluminum foil). Cleavage rates in fertilized oocytes previously vitrified by the AS procedure were higher than those vitrified by a nylon-mesh holder (NM) procedure (89.3 ± 2.1% vs. 65.0 ± 3.7%). Cleaved embryos derived from the AS but not from the NM procedures developed to blastocysts. Furthermore, to investigate the effects of vitrifying GV oocytes on cytoplasmic structure and on the ability to undergo cytoplasmic changes, the intracellular phospholipid membrane (IM) was stained with the lipophilic fluorescent dye, 3,3'-dioctadecyloxa-carbocyanine perchlorate. After vitrification by AS, the IM remained intact relative to that of oocytes vitrified by NM. During in vitro maturation, reorganization of the IM was also undamaged in oocytes vitrified by AS before oocyte maturation, and the IM within oocytes vitrified by the NM procedure was evidently impaired. Finally, vitrification (AS) was used for GV oocytes collected using the ovum pick-up method. A bull calf was born after in vitro production and subsequent embryo transfer. The vitrification techniques described herein should facilitate generation of viable in vitro production bovine blastocysts using oocytes recovered using the ovum pick-up method. Copyright © 2013 Elsevier Inc. All rights reserved.

  18. Fertility preservation treatment for young women with autoimmune diseases facing treatment with gonadotoxic agents.

    PubMed

    Elizur, S E; Chian, R C; Pineau, C A; Son, W Y; Holzer, H E G; Huang, J Y J; Gidoni, Y; Levin, D; Demirtas, E; Tan, S L

    2008-10-01

    To describe a case series of seven women with SLE and other systemic autoimmune rheumatic diseases (SARDs) who required cyclophosphamide therapy and underwent fertility preservation treatments. Of the seven patients reported here, five women had SLE with nephritis, the sixth had immune thrombocytopenia purpura (ITP) and the seventh had microscopic polyangiitis (MPA) with renal involvement. All women were nulliparous and younger than 35 yrs. Patients with SLE underwent in vitro maturation (IVM) of immature oocytes aspirated during a natural menstrual cycle followed by vitrification of the matured oocytes if a male partner was not available, or vitrification of embryos if one was available. The patient with ITP and the patient with MPA underwent gonadotropin ovarian stimulation followed by oocyte or embryo vitrification. All women completed fertility preservation treatment successfully and mature oocytes or embryos (36 and 13, respectively) were vitrified. No complications were associated with this treatment and cytotoxic therapy was initiated as scheduled in all cases. Oocyte or embryo cryopreservation should be considered for fertility preservation in young women with SARDs who face imminent gonadotoxic treatment. In patients, where gonadotropin ovarian stimulation is deemed unsafe, IVM of immature oocytes, aspirated during a natural menstrual cycle, followed by vitrification or fertilization of the mature oocytes, seems to be safe and feasible. For patients in whom hormonal ovarian stimulation is not contraindicated, this method may be considered depending on the urgency to start cytotoxic therapy.

  19. 40 CFR 270.61 - Emergency permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ....61 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Special Forms of Permits § 270.61 Emergency... an imminent and substantial endangerment to human health or the environment the Director may issue a...

  20. 40 CFR 270.61 - Emergency permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ....61 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Special Forms of Permits § 270.61 Emergency... an imminent and substantial endangerment to human health or the environment the Director may issue a...

  1. 40 CFR 270.61 - Emergency permits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ....61 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Special Forms of Permits § 270.61 Emergency... an imminent and substantial endangerment to human health or the environment the Director may issue a...

  2. 40 CFR 270.65 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM... may issue a research, development, and demonstration permit for any hazardous waste treatment facility which proposes to utilize an innovative and experimental hazardous waste treatment technology or process...

  3. 40 CFR 270.65 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM... may issue a research, development, and demonstration permit for any hazardous waste treatment facility which proposes to utilize an innovative and experimental hazardous waste treatment technology or process...

  4. 76 FR 12730 - Clean Air Act Operating Permit Program; Objection to State Operating Permit for U.S. Steel...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-08

    ... ENVIRONMENTAL PROTECTION AGENCY [Regional Docket Nos. V-2009-1, FRL-9276-7] Clean Air Act Operating Permit Program; Objection to State Operating Permit for U.S. Steel-Granite City Works AGENCY... operating permit issued by the Illinois Environmental Protection Agency to the U.S. Steel--Granite City...

  5. 40 CFR 270.40 - Transfer of permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 270.40 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Changes to Permit § 270.40... Act. (b) Changes in the ownership or operational control of a facility may be made as a Class 1...

  6. 40 CFR 270.40 - Transfer of permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 270.40 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Changes to Permit § 270.40... Act. (b) Changes in the ownership or operational control of a facility may be made as a Class 1...

  7. 40 CFR 52.1834 - Minor source permit to operate program.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 4 2014-07-01 2014-07-01 false Minor source permit to operate program. 52.1834 Section 52.1834 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... Minor source permit to operate program. Emission limitations and related provisions, which, in...

  8. 40 CFR 52.1834 - Minor source permit to operate program.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 4 2012-07-01 2012-07-01 false Minor source permit to operate program. 52.1834 Section 52.1834 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... Minor source permit to operate program. Emission limitations and related provisions, which, in...

  9. 40 CFR 52.1834 - Minor source permit to operate program.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 4 2013-07-01 2013-07-01 false Minor source permit to operate program. 52.1834 Section 52.1834 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... Minor source permit to operate program. Emission limitations and related provisions, which, in...

  10. 40 CFR 68.85 - Hot work permit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) CHEMICAL ACCIDENT PREVENTION PROVISIONS Program 3 Prevention Program § 68.85 Hot work permit. (a) The owner or operator shall issue a hot work permit for hot work operations conducted on or near a covered process. (b...

  11. 40 CFR 122.46 - Duration of permits (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Duration of permits (applicable to... ELIMINATION SYSTEM Permit Conditions § 122.46 Duration of permits (applicable to State programs, see § 123.25... in § 122.6, the term of a permit shall not be extended by modification beyond the maximum duration...

  12. Title V Petitions

    EPA Pesticide Factsheets

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements.

  13. Chapter 10 Human Oocyte Vitrification.

    PubMed

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

    2017-01-01

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

  14. 76 FR 55335 - Alternate Passenger Rail Service Pilot Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-09-07

    ... proceeding to develop a pilot program that permits a rail carrier or rail carriers that own infrastructure... develop a pilot program that permits a rail carrier or rail carriers that own infrastructure over which... that permits a rail carrier or rail carriers that own infrastructure over which Amtrak operates a...

  15. 77 FR 73316 - Approval of Air Quality Implementation Plans; California; Eastern Kern, Imperial, Placer, and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-10

    ... adopt and implement a SIP- approved Prevention of Significant Deterioration (PSD) permit program. We are...--Prevention of Significant Deterioration (PSD) Permit Program, PCAPCD Rule 518--Prevention of Significant Deterioration (PSD) Permit Program, and YSAQMD Rule 3.24-- Prevention of Significant Deterioration. The approval...

  16. 40 CFR 52.1834 - Minor source permit to operate program.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 4 2011-07-01 2011-07-01 false Minor source permit to operate program... Minor source permit to operate program. Emission limitations and related provisions, which, in accordance with Rule 33-15-14-03, are established as federally enforceable conditions in North Dakota minor...

  17. 40 CFR 52.1834 - Minor source permit to operate program.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 4 2010-07-01 2010-07-01 false Minor source permit to operate program... Minor source permit to operate program. Emission limitations and related provisions, which, in accordance with Rule 33-15-14-03, are established as federally enforceable conditions in North Dakota minor...

  18. 76 FR 270 - Alaska: Adequacy of Alaska Municipal Solid Waste Landfill Permit Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-04

    ...] Alaska: Adequacy of Alaska Municipal Solid Waste Landfill Permit Program AGENCY: Environmental Protection... approved Municipal Solid Waste Landfill (MSWLF) permit program. The approved modification allows the State..., EPA issued a final rule (69 FR 13242) amending the Municipal Solid Waste Landfill (MSWLF) criteria in...

  19. 75 FR 53220 - Adequacy of New Hampshire Municipal Solid Waste Landfill Permit Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-31

    ...] Adequacy of New Hampshire Municipal Solid Waste Landfill Permit Program AGENCY: Environmental Protection... approved municipal solid waste landfill (MSWLF) program. The approved modification allows the State to..., and demonstration (RD&D) permits to be issued to certain municipal solid waste landfills by approved...

  20. 77 FR 65875 - Adequacy of Arizona Municipal Solid Waste Landfill Permit Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-31

    ... Municipal Solid Waste Landfill Permit Program AGENCY: Environmental Protection Agency (EPA). ACTION: Notice... modification to Arizona's municipal solid waste landfill (MSWLF) permit program to allow the State to issue... amending the municipal solid waste landfill criteria at 40 CFR 258.4 to allow for Research, Development...

  1. 76 FR 303 - Alaska: Adequacy of Alaska's Municipal Solid Waste Landfill Permit Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-01-04

    ...] Alaska: Adequacy of Alaska's Municipal Solid Waste Landfill Permit Program AGENCY: Environmental... modification of its approved Municipal Solid Waste Landfill (MSWLF) permit program. On March 22, 2004, EPA..., Waste, and Toxics, U.S. EPA, Region 10, 1200 Sixth Avenue, Suite 900, Mailstop: AWT-122, Seattle, WA...

  2. 77 FR 54382 - Revisions of Five California Clean Air Act Title V Operating Permits Programs

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-05

    ... Five California Clean Air Act Title V Operating Permits Programs AGENCY: Environmental Protection... Permits (Title V) programs of the Monterey Bay Unified Air Pollution Control District (MBUAPCD), San Luis... thresholds in EPA's Tailoring Rule, which have not been previously subject [[Page 54383

  3. 36 CFR 13.934 - How will the superintendent manage the permit program?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... manage the permit program? 13.934 Section 13.934 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR NATIONAL PARK SYSTEM UNITS IN ALASKA Special Regulations-Denali National Park and Preserve Motor Vehicle Permits § 13.934 How will the superintendent manage the permit...

  4. 36 CFR 13.934 - How will the superintendent manage the permit program?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... manage the permit program? 13.934 Section 13.934 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR NATIONAL PARK SYSTEM UNITS IN ALASKA Special Regulations-Denali National Park and Preserve Motor Vehicle Permits § 13.934 How will the superintendent manage the permit...

  5. 36 CFR 13.934 - How will the superintendent manage the permit program?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... manage the permit program? 13.934 Section 13.934 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR NATIONAL PARK SYSTEM UNITS IN ALASKA Special Regulations-Denali National Park and Preserve Motor Vehicle Permits § 13.934 How will the superintendent manage the permit...

  6. 40 CFR 144.35 - Effect of a permit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 22 2010-07-01 2010-07-01 false Effect of a permit. 144.35 Section 144.35 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS (CONTINUED) UNDERGROUND INJECTION CONTROL PROGRAM Authorization by Permit § 144.35 Effect of a permit. (a) Except for...

  7. Current Regulations and Regulatory Actions

    EPA Pesticide Factsheets

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements.

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

    PubMed

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

    2016-04-01

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

  9. Other Agency Actions Related to Title V

    EPA Pesticide Factsheets

    This site will provide basic information on clean air permitting under the title V operating permits program, provide access to state and regional permitting programs, and maintain access to proposed and final regulatory requirements.

  10. Cryopreservation: Vitrification and Controlled Rate Cooling.

    PubMed

    Hunt, Charles J

    2017-01-01

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

  11. Comparison of different cryopreservation methods for horse and donkey embryos.

    PubMed

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

    2018-05-01

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

  12. Influence of Cryopreservation Solution on the In Vitro Culture of Skin Tissues Derived from Collared Peccary (Pecari tajacu Linnaeus, 1758).

    PubMed

    Borges, Alana A; Lira, Gabriela P O; Nascimento, Lucas E; Queiroz Neta, Luiza B; Santos, Maria V O; Oliveira, Moacir F; Silva, Alexandre R; Pereira, Alexsandra F

    2018-04-01

    Skin vitrification is a promising and alternative tool for the conservation of biodiversity, especially for wild mammals, such as collared peccaries. Several factors can affect the success of this procedure, such as the cryoprotectant solution used. Therefore, this study was carried out to compare the efficiency of various vitrification solutions for recovery of viable cells after in vitro culture of cryopreserved skin tissues derived from the collared peccary, aiming to study the application in biobanking, where cellular use is not immediately required. Then, Dulbecco's modified Eagle's medium (DMEM) composed of 2.2 g/L sodium bicarbonate and 10% fetal bovine serum (FBS) was supplemented with 3.0 M ethylene glycol (EG) or 3.0 M dimethyl sulfoxide (DMSO) or 1.5 M EG plus 1.5 M DMSO with or without sucrose (SUC; 0.25 M) to produce six solutions for solid-surface vitrification. After warming, skin tissues were cultured in vitro and recovered cells were analyzed for morphology, adhesion, subconfluence, and proliferative activity for developing the growth curve and determining the population doubling time (PDT), and viability by Trypan Blue. The vitrification did not alter the ability of the tissues to adhere to the culture dish, as well as the day of all explants with cell growth, subconfluence samples, subconfluence total time, and PDT (p > 0.05). Moreover, independent of the cryoprotectant solution used, the vitrification altered the day of all attached explants (p < 0.05). Nevertheless, for viability after the first passage, only the EG-SUC (86.9%) and DMSO-SUC (91.4%) groups maintained viable cell recovery similar to the nonvitrified group (96.3%, p > 0.05). Additionally, for viability after the third passage, only the EG-SUC group maintained the cell quality (88.3%), when compared with the nonvitrified (97.8%, p > 0.05). In conclusion, DMEM with 10% FBS, 3.0 M EG, and 0.25 M sucrose was the most efficient solution for vitrifying collared peccary skin tissues, leading to the in vitro culture of viable cells.

  13. 40 CFR Appendix A to Part 70 - Approval Status of State and Local Operating Permits Programs

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 16 2012-07-01 2012-07-01 false Approval Status of State and Local Operating Permits Programs A Appendix A to Part 70 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS Pt. 70, App. A Appendix A to Part 70—Approval Status of State and Local...

  14. 40 CFR Appendix A to Part 70 - Approval Status of State and Local Operating Permits Programs

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 16 2014-07-01 2014-07-01 false Approval Status of State and Local Operating Permits Programs A Appendix A to Part 70 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS Pt. 70, App. A Appendix A to Part 70—Approval Status of State and Local...

  15. 40 CFR Appendix A to Part 70 - Approval Status of State and Local Operating Permits Programs

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 16 2013-07-01 2013-07-01 false Approval Status of State and Local Operating Permits Programs A Appendix A to Part 70 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS Pt. 70, App. A Appendix A to Part 70—Approval Status of State and Local...

  16. 40 CFR Appendix A to Part 70 - Approval Status of State and Local Operating Permits Programs

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 15 2011-07-01 2011-07-01 false Approval Status of State and Local Operating Permits Programs A Appendix A to Part 70 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS Pt. 70, App. A Appendix A to Part 70—Approval Status of State and Local...

  17. 40 CFR Appendix A to Part 70 - Approval Status of State and Local Operating Permits Programs

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 15 2010-07-01 2010-07-01 false Approval Status of State and Local Operating Permits Programs A Appendix A to Part 70 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) STATE OPERATING PERMIT PROGRAMS Pt. 70, App. A Appendix A to Part 70—Approval Status of State and Local...

  18. 40 CFR 122.62 - Modification or revocation and reissuance of permits (applicable to State programs, see § 123.25).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE... limitation guideline, EPA approved or promulgated water quality standards, or the Secondary Treatment... regard to a water quality standard on which the permit condition was based; and (C) A permittee requests...

  19. 76 FR 26192 - Revisions to the California State Implementation Plan, Northern Sonoma County Air Pollution...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-06

    ...-approved Prevention of Significant Deterioration (PSD) permit programs. These revisions update the definitions used in the districts' PSD permit programs. DATES: This rule is effective on July 5, 2011 without... Deterioration (PSD) permit program. Both the NSCAPCD and MCAQMD are currently designated as attainment or...

  20. Spotiton: A prototype for an integrated inkjet dispense and vitrification system for cryo-TEM

    PubMed Central

    Jain, Tilak; Sheehan, Patrick; Crum, John; Carragher, Bridget; Potter, Clinton S.

    2012-01-01

    Over the last three decades, Cryo-TEM has developed into a powerful technique for high-resolution imaging of biological macromolecules in their native vitrified state. However, the technique for vitrifying specimens onto EM grids is essentially unchanged – application of ~ 3 µL sample to a grid, followed by blotting and rapid plunge freezing into liquid ethane. Several trials are often required to obtain suitable thin (few hundred nanometers or less) vitrified layers amenable for cryo-TEM imaging, which results in waste of precious sample and resources. While commercially available instruments provide some level of automation to control the vitrification process in an effort to increase quality and reproducibility, obtaining satisfactory vitrified specimens remains a bottleneck in the Cryo-TEM pipeline. We describe here a completely novel method for EM specimen preparation based on small volume (picoliter to nanoliter) dispensing using inkjet technology. A first prototype system (Spotiton v0.5) demonstrates feasibility of this new approach for specimen vitrification. A piezo-electric inkjet dispenser is integrated with optical real-time cameras (100 Hz frame rate) to analyze picoliter to nanoliter droplet profiles in-flight and spreading dynamics on the grid, and thus provides a method to optimize timing of the process. Using TEM imaging and biochemical assays we demonstrate that the piezo-electric inkjet mechanism does not disrupt the structural or functional integrity of macromolecules. These preliminary studies provide insight into the factors and components that will need further development to enable a robust and repeatable technique for specimen vitrification using this novel approach. PMID:22569522

  1. Recovery of valuable metals from electroplating sludge with reducing additives via vitrification.

    PubMed

    Huang, Ruth; Huang, Kuo-Lin; Lin, Zih-Yi; Wang, Jian-Wen; Lin, Chitsan; Kuo, Yi-Ming

    2013-11-15

    In this study, vitrification was applied to treat Ni-Cu electroplating sludge. The sludge was mixed with additives (limestone:cullet = 4:6) and then heated to 1450 °C. The cooled product could be separated into slag and ingot. An atomic absorption spectrometer was used to determine the metal levels of specimens and toxicity characteristic leaching procedure (TCLP) tests, whereas the crystalline and surface characteristics were examined using quantitative X-ray diffraction (XRD) analysis and scanning electron microscopy, respectively. With a glassy structure, the slag was mainly composed of Ca, Si, and Mg. The TCLP results of slags met the Taiwan regulated standards, suggesting that slag can be used for recycling purposes. With the aid of additives, the crystalline phase of slag was transformed form CaMgSiO4 into CsSiO3. The ingots were mainly composed of Ni (563,000-693,800 mg/kg), Cu (79,900-87,400 mg/kg), and Fe (35,000-43,600 mg/kg) (target metals) due the gravity separation during vitrification. At appropriate additives/sludge ratios (>0.2), >95% of target metals gathered in the ingot as a recoverable form (Ni-Fe alloy). The high Ni level of slag suggests that the ingot can be used as the raw materials for smelters or the additives for steel making. Therefore, the vitrification approach of this study is a promising technology to recover valuable metals from Ni-Cu electroplating sludge. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. Cryopreservation of Mexican fruit flies by vitrification: stage selection and avoidance of thermal stress.

    PubMed

    Rajamohan, A; Leopold, R A

    2007-02-01

    This report presents details of a vitrification methodology for the cryopreservation of embryos of the Mexican fruit fly, Anastrepha ludens. The overall summary of the data indicates that selecting the correct developmental stage for cryopreservation is the most important criterion. The key aspect in selection of the correct stage is to balance depletion of the gut yolk content against development of the embryonic cuticle. Embryogenesis was divided into four stages between 90 and 120 h after incubation at 21.7 degrees C. The classification was based on the intestinal yolk content and the initial development of mandibular-maxillary complex. Stages having low mid-gut yolk content and the appearance of mouth hooks were found to be the most suitable for cryopreservation. Embryos developing at 30 degrees C had premature cuticle formation relative to gut development and significantly lower hatching after cryopreservation. Vitrification of embryos by direct quenching in liquid nitrogen was less effective than quenching after annealing the samples in liquid nitrogen vapor. Quenched samples of vitrification solutions containing 1,2-ethanediol as the major component exhibited fractures. Fracturing occurred less frequently when the solutions were annealed and when containing polyethylene glycol. Hatching of vitrified embryos stored in liquid nitrogen for over 12 months was not statistically different from those held for only 15 min. Our protocol yielded normalized hatching rates that ranged as high as 61%. Selecting the exact stage for cryopreservation from a population of embryos obtained by collection from ovipositing females during a span of just 30 min resulted in nearly 80% of the embryos hatching into larvae.

  3. Predict the glass transition temperature of glycerol-water binary cryoprotectant by molecular dynamic simulation.

    PubMed

    Li, Dai-Xi; Liu, Bao-Lin; Liu, Yi-shu; Chen, Cheng-lung

    2008-04-01

    Vitrification is proposed to be the best way for the cryopreservation of organs. The glass transition temperature (T(g)) of vitrification solutions is a critical parameter of fundamental importance for cryopreservation by vitrification. The instruments that can detect the thermodynamic, mechanical and dielectric changes of a substance may be used to determine the glass transition temperature. T(g) is usually measured by using differential scanning calorimetry (DSC). In this study, the T(g) of the glycerol-aqueous solution (60%, wt/%) was determined by isothermal-isobaric molecular dynamic simulation (NPT-MD). The software package Discover in Material Studio with the Polymer Consortium Force Field (PCFF) was used for the simulation. The state parameters of heat capacity at constant pressure (C(p)), density (rho), amorphous cell volume (V(cell)) and specific volume (V(specific)) and radial distribution function (rdf) were obtained by NPT-MD in the temperature range of 90-270K. These parameters showed a discontinuity at a specific temperature in the plot of state parameter versus temperature. The temperature at the discontinuity is taken as the simulated T(g) value for glycerol-water binary solution. The T(g) values determined by simulation method were compared with the values in the literatures. The simulation values of T(g) (160.06-167.51K) agree well with the DSC results (163.60-167.10K) and the DMA results (159.00K). We drew the conclusion that molecular dynamic simulation (MDS) is a potential method for investigating the glass transition temperature (T(g)) of glycerol-water binary cryoprotectants and may be used for other vitrification solutions.

  4. Investigations on the heat transport capability of a cryogenic oscillating heat pipe and its application in achieving ultra-fast cooling rates for cell vitrification cryopreservation☆

    PubMed Central

    Han, Xu; Ma, Hongbin; Jiao, Anjun; Critser, John K.

    2010-01-01

    Theoretically, direct vitrification of cell suspensions with relatively low concentrations (~1 M) of permeating cryoprotective agents (CPA) is suitable for cryopreservation of almost all cell types and can be accomplished by ultra-fast cooling rates that are on the order of 106–7 K/min. However, the methods and devices currently available for cell cryopreservation cannot achieve such high cooling rates. In this study, we constructed a novel cryogenic oscillating heat pipe (COHP) using liquid nitrogen as its working fluid and investigated its heat transport capability to assess its application for achieving ultra-fast cooling rates for cell cryopreservation. The experimental results showed that the apparent heat transfer coefficient of the COHP can reach 2 × 105 W/m2·K, which is two orders of the magnitude higher than traditional heat pipes. Theoretical analyzes showed that the average local heat transfer coefficient in the thin film evaporation region of the COHP can reach 1.2 × 106 W/m2·K, which is approximately 103 times higher than that achievable with standard pool-boiling approaches. Based on these results, a novel device design applying the COHP and microfabrication techniques is proposed and its efficiency for cell vitrification is demonstrated through numerical simulation. The estimated average cooling rates achieved through this approach is 106–7 K/min, which is much faster than the currently available methods and sufficient for achieving vitrification with relatively low concentrations of CPA. PMID:18430413

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

    PubMed

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

    2014-03-01

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

  6. Successful vitrification of human amnion-derived mesenchymal stem cells.

    PubMed

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

    2008-08-01

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

  7. Investigations on the heat transport capability of a cryogenic oscillating heat pipe and its application in achieving ultra-fast cooling rates for cell vitrification cryopreservation.

    PubMed

    Han, Xu; Ma, Hongbin; Jiao, Anjun; Critser, John K

    2008-06-01

    Theoretically, direct vitrification of cell suspensions with relatively low concentrations ( approximately 1 M) of permeating cryoprotective agents (CPA) is suitable for cryopreservation of almost all cell types and can be accomplished by ultra-fast cooling rates that are on the order of 10(6-7) K/min. However, the methods and devices currently available for cell cryopreservation cannot achieve such high cooling rates. In this study, we constructed a novel cryogenic oscillating heat pipe (COHP) using liquid nitrogen as its working fluid and investigated its heat transport capability to assess its application for achieving ultra-fast cooling rates for cell cryopreservation. The experimental results showed that the apparent heat transfer coefficient of the COHP can reach 2 x 10(5) W/m(2).K, which is two orders of the magnitude higher than traditional heat pipes. Theoretical analyzes showed that the average local heat transfer coefficient in the thin film evaporation region of the COHP can reach 1.2 x 10(6) W/m(2).K, which is approximately 10(3) times higher than that achievable with standard pool-boiling approaches. Based on these results, a novel device design applying the COHP and microfabrication techniques is proposed and its efficiency for cell vitrification is demonstrated through numerical simulation. The estimated average cooling rates achieved through this approach is 10(6-7)K/min, which is much faster than the currently available methods and sufficient for achieving vitrification with relatively low concentrations of CPA.

  8. 40 CFR 68.85 - Hot work permit.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 16 2012-07-01 2012-07-01 false Hot work permit. 68.85 Section 68.85... ACCIDENT PREVENTION PROVISIONS Program 3 Prevention Program § 68.85 Hot work permit. (a) The owner or operator shall issue a hot work permit for hot work operations conducted on or near a covered process. (b...

  9. 40 CFR 68.85 - Hot work permit.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 16 2014-07-01 2014-07-01 false Hot work permit. 68.85 Section 68.85... ACCIDENT PREVENTION PROVISIONS Program 3 Prevention Program § 68.85 Hot work permit. (a) The owner or operator shall issue a hot work permit for hot work operations conducted on or near a covered process. (b...

  10. 40 CFR 68.85 - Hot work permit.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 16 2013-07-01 2013-07-01 false Hot work permit. 68.85 Section 68.85... ACCIDENT PREVENTION PROVISIONS Program 3 Prevention Program § 68.85 Hot work permit. (a) The owner or operator shall issue a hot work permit for hot work operations conducted on or near a covered process. (b...

  11. 40 CFR 68.85 - Hot work permit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 15 2011-07-01 2011-07-01 false Hot work permit. 68.85 Section 68.85... ACCIDENT PREVENTION PROVISIONS Program 3 Prevention Program § 68.85 Hot work permit. (a) The owner or operator shall issue a hot work permit for hot work operations conducted on or near a covered process. (b...

  12. 77 FR 59186 - Clean Air Act Operating Permit Program; Petition for Objection to State Operating Permit for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-26

    ... ENVIRONMENTAL PROTECTION AGENCY [Petition IV-2011-1; FRL-9734-2] Clean Air Act Operating Permit Program; Petition for Objection to State Operating Permit for Tennessee Valley Authority's Shawnee Fossil... Valley Authority for its Shawnee Fossil Plant (SFP) facility located in West Paducah, Kentucky. This...

  13. 76 FR 53452 - Clean Air Act Operating Permit Program; Response to Petition To Reopen the 2001 Title V Permit...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-26

    ... ENVIROMENTAL PROTECTION AGENCY [FRL-9457-3 ] Clean Air Act Operating Permit Program; Response to Petition To Reopen the 2001 Title V Permit for Reliant Portland Generating Station, Upper Mount Bethel Township, Northampton County, PA AGENCY: Environmental Protection Agency (EPA). ACTION: Notice of action...

  14. 40 CFR 270.13 - Contents of part A of the permit application.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Permit..., and disposing of hazardous waste, and the design capacity of these items. (j) A specification of the hazardous wastes listed or designated under 40 CFR part 261 to be treated, stored, or disposed of at the...

  15. 40 CFR 270.13 - Contents of part A of the permit application.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... WASTES (CONTINUED) EPA ADMINISTERED PERMIT PROGRAMS: THE HAZARDOUS WASTE PERMIT PROGRAM Permit..., and disposing of hazardous waste, and the design capacity of these items. (j) A specification of the hazardous wastes listed or designated under 40 CFR part 261 to be treated, stored, or disposed of at the...

  16. 40 CFR 49.158 - Synthetic minor source permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 1 2011-07-01 2011-07-01 false Synthetic minor source permits. 49.158... Federal Minor New Source Review Program in Indian Country § 49.158 Synthetic minor source permits. You may obtain a synthetic minor source permit under this program to establish a synthetic minor source for...

  17. 36 CFR 13.934 - How will the superintendent manage the permit program?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 36 Parks, Forests, and Public Property 1 2012-07-01 2012-07-01 false How will the superintendent manage the permit program? 13.934 Section 13.934 Parks, Forests, and Public Property NATIONAL PARK... National Park and Preserve Motor Vehicle Permits § 13.934 How will the superintendent manage the permit...

  18. 36 CFR 13.934 - How will the superintendent manage the permit program?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 36 Parks, Forests, and Public Property 1 2013-07-01 2013-07-01 false How will the superintendent manage the permit program? 13.934 Section 13.934 Parks, Forests, and Public Property NATIONAL PARK... National Park and Preserve Motor Vehicle Permits § 13.934 How will the superintendent manage the permit...

  19. 75 FR 36069 - Clean Air Act Operating Permit Program; Petition for Objection to a Federal Operating Permit for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-24

    ... ENVIRONMENTAL PROTECTION AGENCY [FRL-9167-5] Clean Air Act Operating Permit Program; Petition for Objection to a Federal Operating Permit for Waste Management of Louisiana L.L.C., Woodside Landfill and Recycling Center (WLRC), Walker, Livingston Parish, LA AGENCY: Environmental Protection Agency (EPA). ACTION...

  20. 76 FR 9772 - Adequacy of Arizona Municipal Solid Waste Landfill Permit Program

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-22

    ...) permit program to allow the State to issue research, development, and demonstration (RD&D) permits for... environment. The Director may provide a variance from existing requirements of MSWLF criteria for run-on... demonstrate that compliance with the RD&D permit will not increase risk to human heath and the environment...

  1. Implementing the NPDES program: An update on the WET ...

    EPA Pesticide Factsheets

    The U.S. EPA has utilized the Clean Water Act - National Pollutant Discharge Elimination System permitting program to protect waters of the U.S for over 40 years. NPDES permit effluent limitations serve as the primary mechanism for controlling discharges of pollutants to receiving waters. When developing effluent limitations for an NPDES permit, a permit writer must consider limits based on both the technology available to control the pollutants (i.e., technology-based effluent limits) and limits that are protective of the water quality standards of the receiving water (i.e., water quality-based effluent limits). WET testing is one of the water quality-based effluent limitation mechanisms available to permit writers that is useful in determining how the additive, synergistic and compounding effects of toxic effluents effect streams. This presentation will provide an overview of the current EPA NPDES permit program direction for increasing the efficacy of NPDES permits program administered by the U.S. EPA and States. The training implementation plan is expected to provide permit writers with a clearer understanding of WET requirements as established via the U.S. EPA WET test manuals, NPDES permitting regulatory authorities, and the WET science which has been long established. not applicable

  2. Description of waste pretreatment and interfacing systems dynamic simulation model

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

    Garbrick, D.J.; Zimmerman, B.D.

    1995-05-01

    The Waste Pretreatment and Interfacing Systems Dynamic Simulation Model was created to investigate the required pretreatment facility processing rates for both high level and low level waste so that the vitrification of tank waste can be completed according to the milestones defined in the Tri-Party Agreement (TPA). In order to achieve this objective, the processes upstream and downstream of the pretreatment facilities must also be included. The simulation model starts with retrieval of tank waste and ends with vitrification for both low level and high level wastes. This report describes the results of three simulation cases: one based on suggestedmore » average facility processing rates, one with facility rates determined so that approximately 6 new DSTs are required, and one with facility rates determined so that approximately no new DSTs are required. It appears, based on the simulation results, that reasonable facility processing rates can be selected so that no new DSTs are required by the TWRS program. However, this conclusion must be viewed with respect to the modeling assumptions, described in detail in the report. Also included in the report, in an appendix, are results of two sensitivity cases: one with glass plant water recycle steams recycled versus not recycled, and one employing the TPA SST retrieval schedule versus a more uniform SST retrieval schedule. Both recycling and retrieval schedule appear to have a significant impact on overall tank usage.« less

  3. Federal Register Notice: Final Rule Establishing Consolidated Permit Program Requirements Under Several Environmental Statutes

    EPA Pesticide Factsheets

    This final rule establishes consolidated permit program requirements governing the Hazardous Waste Management program under the Resource Conservation and Recovery Act (RCRA) and other related programs.

  4. In situ vitrification application to buried waste: Final report of intermediate field tests at Idaho National Engineering Laboratory

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

    Callow, R.A.; Weidner, J.R.; Loehr, C.A.

    This report describes two in situ vitrification field tests conducted on simulated buried waste pits during June and July 1990 at the Idaho National Engineering Laboratory. In situ vitrification, an emerging technology for in place conversion of contaminated soils into a durable glass and crystalline waste form, is being investigated as a potential remediation technology for buried waste. The overall objective of the two tests was to access the general suitability of the process to remediate waste structures representative of buried waste found at Idaho National Engineering Laboratory. In particular, these tests, as part of a treatability study, were designedmore » to provide essential information on the field performance of the process under conditions of significant combustible and metal wastes and to test a newly developed electrode feed technology. The tests were successfully completed, and the electrode feed technology successfully processed the high metal content waste. Test results indicate the process is a feasible technology for application to buried waste. 33 refs., 109 figs., 39 tabs.« less

  5. A sampling device with a capped body and detachable handle

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

    Jezek, Gerd-Rainer

    1997-12-01

    The present invention relates to a device for sampling radioactive waste and more particularly to a device for sampling radioactive waste which prevents contamination of a sampled material and the environment surrounding the sampled material. During vitrification of nuclear wastes, it is necessary to remove contamination from the surfaces of canisters filled with radioactive glass. After removal of contamination, a sampling device is used to test the surface of the canister. The one piece sampling device currently in use creates a potential for spreading contamination during vitrification operations. During operations, the one piece sampling device is transferred into and outmore » of the vitrification cell through a transfer drawer. Inside the cell, a remote control device handles the sampling device to wipe the surface of the canister. A one piece sampling device can be contaminated by the remote control device prior to use. Further, the sample device can also contaminate the transfer drawer producing false readings for radioactive material. The present invention overcomes this problem by enclosing the sampling pad in a cap. The removable handle is reused which reduces the amount of waste material.« less

  6. Compositional Models of Glass/Melt Properties and their Use for Glass Formulation

    DOE PAGES

    Vienna, John D.; USA, Richland Washington

    2014-12-18

    Nuclear waste glasses must simultaneously meet a number of criteria related to their processability, product quality, and cost factors. The properties that must be controlled in glass formulation and waste vitrification plant operation tend to vary smoothly with composition allowing for glass property-composition models to be developed and used. Models have been fit to the key glass properties. The properties are transformed so that simple functions of composition (e.g., linear, polynomial, or component ratios) can be used as model forms. The model forms are fit to experimental data designed statistically to efficiently cover the composition space of interest. Examples ofmore » these models are found in literature. The glass property-composition models, their uncertainty definitions, property constraints, and optimality criteria are combined to formulate optimal glass compositions, control composition in vitrification plants, and to qualify waste glasses for disposal. An overview of current glass property-composition modeling techniques is summarized in this paper along with an example of how those models are applied to glass formulation and product qualification at the planned Hanford high-level waste vitrification plant.« less

  7. 40 CFR 96.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false General CAIR NOX Ozone Season Trading... PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating...

  8. 40 CFR 96.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false General CAIR NOX Ozone Season Trading... PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating...

  9. 40 CFR 96.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false General CAIR NOX Ozone Season Trading... PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating...

  10. 40 CFR 96.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false General CAIR NOX Ozone Season Trading... PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating...

  11. 40 CFR 96.320 - General CAIR NOX Ozone Season Trading Program permit requirements.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false General CAIR NOX Ozone Season Trading... PROGRAMS FOR STATE IMPLEMENTATION PLANS Permits § 96.320 General CAIR NOX Ozone Season Trading Program permit requirements. (a) For each CAIR NOX Ozone Season source required to have a title V operating...

  12. 40 CFR 97.83 - Applying for NOX Budget opt-in permit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Applying for NOX Budget opt-in permit... PROGRAMS (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Individual Unit Opt-ins. § 97.83 Applying for NOX Budget opt-in permit. (a) Applying for initial NO X Budget opt...

  13. 40 CFR 96.83 - Applying for NOX Budget opt-in permit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Applying for NOX Budget opt-in permit... PROGRAMS (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Individual Unit Opt-ins § 96.83 Applying for NOX Budget opt-in permit. (a) Applying for...

  14. 77 FR 15267 - Clean Air Act Full Approval of Title V Operating Permits Program; Southern Ute Indian Tribe

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-15

    ... Full Approval of Title V Operating Permits Program; Southern Ute Indian Tribe AGENCY: Environmental Protection Agency (EPA). ACTION: Final rule. SUMMARY: The EPA is promulgating full approval of the Title V Operating Permits Program submitted by the Southern Ute Indian Tribe (Tribe). The Tribe's Title V Operating...

  15. 75 FR 22400 - Clean Air Act Operating Permit Program; Petition To Object to Title V Permit for Wheelabrator...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-28

    ... ENVIROMENTAL PROTECTION AGENCY [FRL-9142-6] Clean Air Act Operating Permit Program; Petition To Object to Title V Permit for Wheelabrator Baltimore, L.P., Baltimore City, MD AGENCY: Environmental Protection Agency (EPA). ACTION: Notice of final action. SUMMARY: Pursuant to section 505(b)(2) of the Clean...

  16. 40 CFR 96.85 - NOX Budget opt-in permit contents.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false NOX Budget opt-in permit contents. 96... (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS Individual Unit Opt-ins § 96.85 NOX Budget opt-in permit contents. (a) Each NOX Budget opt-in permit...

  17. 40 CFR 97.85 - NOX Budget opt-in permit contents.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 20 2010-07-01 2010-07-01 false NOX Budget opt-in permit contents. 97... (CONTINUED) FEDERAL NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS Individual Unit Opt-ins. § 97.85 NOX Budget opt-in permit contents. (a) Each NOX Budget opt-in permit will contain all elements...

  18. Vitrification of waste

    DOEpatents

    Wicks, G.G.

    1999-04-06

    A method is described for encapsulating and immobilizing waste for disposal. Waste, preferably, biologically, chemically and radioactively hazardous, and especially electronic wastes, such as circuit boards, are placed in a crucible and heated by microwaves to a temperature in the range of approximately 300 C to 800 C to incinerate organic materials, then heated further to a temperature in the range of approximately 1100 C to 1400 C at which temperature glass formers present in the waste will cause it to vitrify. Glass formers, such as borosilicate glass, quartz or fiberglass can be added at the start of the process to increase the silicate concentration sufficiently for vitrification.

  19. 75 FR 75463 - Clean Air Act Operating Permit Program; Petition To Object to Title V Permit for Luke Paper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-03

    ... ENVIROMENTAL PROTECTION AGENCY [FRL-9234-9] Clean Air Act Operating Permit Program; Petition To Object to Title V Permit for Luke Paper Company, Luke, MD AGENCY: Environmental Protection Agency (EPA). ACTION: Notice of final action. SUMMARY: Pursuant to section 505(b)(2) of the Clean Air Act (CAA), the...

  20. 43 CFR 3261.16 - Can my operations plan, drilling permit, and drilling program apply to more than one well?

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 43 Public Lands: Interior 2 2011-10-01 2011-10-01 false Can my operations plan, drilling permit... MINERALS MANAGEMENT (3000) GEOTHERMAL RESOURCE LEASING Drilling Operations: Getting a Permit § 3261.16 Can my operations plan, drilling permit, and drilling program apply to more than one well? (a) Your...

  1. 50 CFR 660.150 - Mothership (MS) Coop Program.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...) Accumulation limits—(i) MS permit usage limit. No person who owns an MS permit(s) may register the MS permit(s... permit owner complies with the accumulation limits. (ii) Catcher vessel usage limit. No vessel may catch...

  2. 40 CFR 122.6 - Continuation of expiring permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 22 2011-07-01 2011-07-01 false Continuation of expiring permits. 122.6 Section 122.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Definitions and...

  3. 40 CFR 122.6 - Continuation of expiring permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Continuation of expiring permits. 122.6 Section 122.6 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Definitions and...

  4. 40 CFR 122.5 - Effect of a permit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 22 2011-07-01 2011-07-01 false Effect of a permit. 122.5 Section 122.5 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Definitions and General...

  5. 40 CFR 122.5 - Effect of a permit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 21 2010-07-01 2010-07-01 false Effect of a permit. 122.5 Section 122.5 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Definitions and General...

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

    PubMed

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

    2013-11-01

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

  7. Vitamin K2 improves developmental competency and cryo-tolerance of in vitro derived ovine blastocyst.

    PubMed

    Sefid, Fatemeh; Ostadhosseini, S; Hosseini, S M; Ghazvini Zadegan, F; Pezhman, M; Nasr Esfahani, Mohammad Hossein

    2017-08-01

    Vitamin K2 (VK2), acts as an electron carrier in mitochondria and thereby effects reactive oxygen species (ROS) and ATP production. This study evaluates role of VK2 on in vitro developmental competency and cryo-survival of pre-implantation ovine embryos. Initially the optimal and beneficial concentration of VK2 on compaction and blastocyst formation rates was defined (0.1 μM). Subsequently, it was shown that 0.1 μM VK2, at blastocyst stage, reduces H2O2 production, increase the expression of mitochondrial related gene and improved embryos quality. We further assessed presence VK2 supplementation before and/or after vitrification of in vitro derived blastocysts. Our results reveal that presence of VK2 before and after vitrification improves rates of blastocysts re-expansion (88.19± 3.37% vs 73.68± 1.86%, P < 0.05) and hatching (49.55± 4.37% vs 32.7± 3.32%) compared to control group. These observation were consistent with reduction in H2O2 production and improved in expression of mitochondrial related genes. However, VK2 before or after vitrification, not only had no positive effect on these two parameters, but also significantly reduced these parameters. Therefore, in concordance with pervious report in bovine, we show that VK2 supplementation post genomic activation (Day 3-7) improved developmental competency of ovine in vitro derived embryos. We also showed that presence of VK2 after vitrification improves the cryo-survival of ovine embryos. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. The Dominance of Warming Rate Over Cooling Rate in the Survival of Mouse Oocytes Subjected to a Vitrification Procedure✰

    PubMed Central

    Seki, Shinsuke; Mazur, Peter

    2009-01-01

    The formation of more than trace amounts of ice in cells is lethal. The two contrasting routes to avoiding it are slow equilibrium freezing and vitrification. The cryopreservation of mammalian oocytes by either method continues to be difficult, but there seems a slowly emerging consensus that vitrification procedures are somewhat better for mouse and human oocytes. The approach in these latter procedures is to load cells with high concentrations of glass-inducing solutes and cool them at rates high enough to induce the glassy state. Several devices have been developed to achieve very high cooling rates. Our study has been concerned with the relative influences of warming rate and cooling rate on the survival of mouse oocytes subjected to a vitrification procedure. Oocytes suspended in an ethylene glycol-acetamide-Ficoll-sucrose solution were cooled to −196°C at rates ranging from 37°C/min to 1827°C/min between 20°C and −120°C, and for each cooling rate, warmed at rates ranging from 139°C/min to 2950°C/min between −70°C and −35°C. The results are unambiguous. If the samples were warmed at the highest rate, survivals were >80% over cooling rates of 187°C/min to 1827°C/min. If the samples were warmed at the lowest rate, survivals were near 0% regardless of the cooling rate. We interpret the lethality of slow warming to be a consequence of it allowing time for the growth of small intracellular ice crystals by recrystallization. PMID:19427303

  9. Forskolin improves the cryosurvival of in vivo-derived porcine embryos at very early stages using two vitrification methods.

    PubMed

    Gomis, J; Cuello, C; Sanchez-Osorio, J; Gil, M A; Parrilla, I; Angel, M A; Vazquez, J M; Roca, J; Martinez, E A

    2013-04-01

    This study was aimed to determine the effect of forskolin on the viability of in vivo-derived porcine embryos vitrified by the superfine open pulled straw (SOPS) or solid surface vitrification (SSV) methods at the 2-cell, 4-cell, and blastocyst stages. Zygotes, 2- to 4-cell embryos, and morulae were obtained from superovulated sows. After collection, embryos were cultured for 24h with 0 or 10 μM forskolin and then vitrified using the SOPS and SSV method, or not vitrified (fresh controls). Fresh and vitrified-warmed 2-cells, 4-cells, and blastocysts were cultured for additional 96 h, 72 h and 24 h, respectively. At the end of the culture, embryos were evaluated for progression to the blastocyst stage and total cell number. The vitrification method did not affect any of the parameters evaluated for any embryo stage. Forskolin increased (P<0.01) the blastocyst formation and the final developmental stage of vitrified 2- and 4-cell embryos. However, these embryos exhibited lower (P<0.003) blastocyst formation rates than their fresh counterparts. The total cell number and hatching rate were similar in both groups (vitrified and fresh) of 2- and 4-cell embryos. Vitrified blastocysts exhibited viabilities, final developmental stages, hatching rates, and total cell numbers that were similar to those of their fresh counterparts, regardless of the addition of forskolin. In conclusion, the SOPS and SSV methods are suitable for the cryopreservation of in vivo-derived 2- to 4-cell porcine embryos. Pre-treatment with forskolin for 24h before vitrification improves the cryotolerance of 2- and 4-cell porcine embryos. Copyright © 2013 Elsevier Inc. All rights reserved.

  10. Biophysical Characteristics of Successful Oilseed Embryo Cryoprotection and Cryopreservation Using Vacuum Infiltration Vitrification: An Innovation in Plant Cell Preservation

    PubMed Central

    Nadarajan, Jayanthi; Pritchard, Hugh W.

    2014-01-01

    Heterogeneity in morphology, physiology and cellular chemistry of plant tissues can compromise successful cryoprotection and cryopreservation. Cryoprotection is a function of exposure time × temperature × permeability for the chosen protectant and diffusion pathway length, as determined by specimen geometry, to provide sufficient dehydration whilst avoiding excessive chemical toxicity. We have developed an innovative method of vacuum infiltration vitrification (VIV) at 381 mm (15 in) Hg (50 kPa) that ensures the rapid (5 min), uniform permeation of Plant Vitrification Solution 2 (PVS2) cryoprotectant into plant embryos and their successful cryopreservation, as judged by regrowth in vitro. This method was validated on zygotic embryos/embryonic axes of three species (Carica papaya, Passiflora edulis and Laurus nobilis) up to 1.6 mg dry mass and 5.6 mm in length, with varying physiology (desiccation tolerances) and 80°C variation in lipid thermal profiles, i.e., visco-elasticity properties, as determined by differential scanning calorimetry. Comparisons between the melting features of cryoprotected embryos and embryo regrowth indicated an optimal internal PVS2 concentration of about 60% of full strength. The physiological vigour of surviving embryos was directly related to the proportion of survivors. Compared with conventional vitrification, VIV-cryopreservation offered a ∼ 10-fold reduction in PVS2 exposure times, higher embryo viability and regrowth and greater effectiveness at two pre-treatment temperatures (0°C and 25°C). VIV-cryopreservation may form the basis of a generic, high throughput technology for the ex situ conservation of plant genetic resources, aiding food security and protection of species from diverse habitats and at risk of extinction. PMID:24788797

  11. Oocyte cryopreservation: where are we now?

    PubMed

    Argyle, Catrin E; Harper, Joyce C; Davies, Melanie C

    2016-06-01

    Since the first live birth from oocyte cryopreservation three decades ago, oocyte cryopreservation has become an important component of ART. Cryopreservation techniques have evolved, leading to higher success rates and the introduction of oocyte cryopreservation into IVF clinics worldwide. Concurrently, there has been an increase in patient demand, especially for so-called 'social egg freezing' that allows women to preserve their fertility in anticipation of age-related fertility decline. This review addresses a need to evaluate the current status of oocyte cryopreservation. It explores current techniques and success rates, clinical applications, the rise of elective oocyte cryopreservation, and future implications. A search was performed using Web of Science and PubMed databases for publications between January 1980 and December 2015. Keywords used included 'egg freezing', 'oocyte freezing', 'oocyte cryopreservation', 'oocyte vitrification', and 'fertility preservation'. The success rate of oocyte cryopreservation has risen, and the increasing use of vitrification offers has improved outcomes, with IVF pregnancy rates now similar to those achieved with fresh oocytes. There are conflicting opinions about the comparative success rates of open and closed vitrification. Patients are accessing and receiving oocyte cryopreservation for a wide range of indications, and there has been a marked increase in patient numbers and oocyte cryopreservation cycles. Oocyte cryopreservation for circumventing age-related infertility is becoming more widely accepted. Oocyte cryopreservation is an established component of ART, with vitrification now being the cryopreservation technique of choice. Increasing numbers of women undergo oocyte cryopreservation for both medical and social reasons. It is important to continue auditing outcomes and reporting long-term follow-up of children born from frozen-thawed oocytes. © The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  12. CRYOPRESERVATION EFFECTS ON RECOMBINANT MYOBLASTS ENCAPSULATED IN ADHESIVE ALGINATE HYDROGELS

    PubMed Central

    Ahmad, Hajira F.; Sambanis, Athanassios

    2013-01-01

    Cell encapsulation in hydrogels is widely used in tissue engineering applications, including encapsulation of islets or other insulin-secreting cells in pancreatic substitutes. Use of adhesive, bio-functionalized hydrogels is receiving increasing attention, as cell-matrix interactions in 3-D can be important for various cell processes. With pancreatic substitutes, studies have indicated benefits of 3-D adhesion on the viability and/or function of insulin-secreting cells. As long-term storage of microencapsulated cells is critical for their clinical translation, cryopreservation of cells in hydrogels is actively being investigated. Previous studies have examined the cryopreservation response of cells encapsulated in non-adhesive hydrogels using conventional freezing and/or vitrification (ice-free cryopreservation), however, none have systematically compared the two cryopreservation methods with cells encapsulated within an adhesive 3-D environment. The latter would be significant, as evidence suggests adhesion influences cellular response to cryopreservation. Thus, the objective of this study was to determine the response to conventional freezing and vitrification of insulin-secreting cells encapsulated in an adhesive biomimetic hydrogel. Recombinant insulin-secreting C2C12 myoblasts were encapsulated in oxidized RGD-alginate and cultured 1 or 4 days post-encapsulation, cryopreserved, and assessed up to 3 days post-warming for metabolic activity and insulin secretion, and one day post-warming for cell morphology. Besides certain transient differences of the vitrified group relative to the Fresh control, both conventional freezing and vitrification maintained metabolism, secretion and morphology of the recombinant C2C12 cells. Thus, due to a simpler procedure and slightly superior results, conventional freezing is recommended over vitrification for the cryopreservation of C2C12 cells in oxidized RGD-modified alginate. PMID:23499987

  13. Recent advances in the cryopreservation of shoot-derived germplasm of economically important fruit trees of Actinidia, Diospyros, Malus, Olea, Prunus, Pyrus and Vitis.

    PubMed

    Benelli, Carla; De Carlo, Anna; Engelmann, Florent

    2013-01-01

    This paper presents the advances made over the last decade in cryopreservation of economically important vegetatively propagated fruit trees. Cryopreservation protocols have been established using both dormant buds sampled on field-grown plants and shoot tips sampled on in vitro plantlets. In the case of dormant buds, scions are partially dehydrated by storage at -5 °C, and then cooled slowly to -30 °C using low cooling rates (c.a. 1 °C/h) before immersion in liquid nitrogen. After slow rewarming and rehydration of samples, regrowth takes place either through grafting of buds on rootstocks or excision of apices and inoculation in vitro. In the case of shoot tips of in vitro plantlets, the cryopreservation techniques employed are the following: controlled rate cooling procedures involving slow prefreezing followed by immersion in liquid nitrogen or vitrification-based procedures including encapsulation-dehydration, vitrification, encapsulation-vitrification and droplet-vitrification. The current status of cryopreservation for a series of fruit tree species including Actinidia, Diospyros, Malus, Olea, Prunus, Pyrus and Vitis is presented. Routine application of cryopreservation for long-term germplasm storage in genebanks is currently limited to apple and pear, for which large cryopreserved collections have been established at NCGRP, Fort Collins (USA), using dormant buds and in vitro shoot tips, respectively. However, there are a growing number of examples of pilot scale testing experiments under way for different species in various countries. Progress in the further development and application of cryopreservation techniques will be made through a better understanding of the mechanisms involved in the induction of tolerance to dehydration and cryopreservation in frozen explants. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Effects of various freezing containers for vitrification freezing on mouse oogenesis.

    PubMed

    Kim, Ji Chul; Kim, Jae Myeoung; Seo, Byoung Boo

    2016-01-01

    In the present study, various freezing containers were tested for mouse embryos of respective developmental stages; embryos were vitrified and then their survival rate and developmental rate were monitored. Mouse two cell, 8 cell, and blastula stage embryos underwent vitrification freezing-thawing and then their recovery rate, survival rate, development rate, and hatching rate were investigated. EM-grid, OPS, and cryo-loop were utilized for vitrification freezing-thawing of mouse embryos. It was found that recovery rate and survival rate were higher in the group of cryo-loop compared to those of EM-grid (p < 0.05). Embryonic development rate, two cell embryos to blastocyst, as well as hatching rate were higher in the control group compared to the EM-grid group and OPS group (p < 0.05), yet no difference was noted between the control group and cryo-loop group. Development rate and hatching rate of eight cell morulae and blastocysts were all lower in the treatment groups than the control group whilst hatching rate of blastocysts was higher in the control group compared to the groups of EM-grid and OPS (p < 0.05); although the cryo-loop group was shown to be slightly higher than other groups, it was not statistically significant. In the study, we investigate effects of freezing containers on vitrified embryos of respective developmental stages; it was demonstrated that higher developmental rate was shown in more progressed (or developed) embryos with more blastomeres. There was however, no difference in embryonic development rate was shown amongst containers. Taken together, further additional studies are warranted with regards to 1) manipulation techniques of embryos for various vitrification freezing containers and 2) preventive measures against contamination via liquid nitrogen.

  15. The Effect of Vitrification and in vitro Culture on the Adenosine Triphosphate Content and Mitochondrial Distribution of Mouse Pre-Implantation Embryos

    PubMed Central

    Amoushahi, Mahboobeh; Salehnia, Mojdeh; HosseinKhani, Saman

    2013-01-01

    Background: The mitochondria are an important source of adenosine triphosphate (ATP) production in pre-implantation embryo. Therefore, the objective of this study was to investigate the effect of vitrification and in vitro culture of mouse embryos on their mitochondrial distribution and ATP content. Methods: The embryos at 2-PN, 4-cell and blastocyst stages were collected from the oviduct of stimulated pregnant mice and uterine horns. Then, the embryos were vitrified with the cryotop method using ethylene glycol and dimethylsulphoxide. After evaluating the survival rates of vitrified embryos, their development to hatching stages were assessed. The ATP content of collected in vivo and in vitro embryos at different stages was measured by luciferin-luciferase bioluminescence assay. The distribution of mitochondria was studied using Mito-tracker green staining under a fluorescent microscope. Results: The survival rates of vitrified embryos at 2-PN, 4-cell and early blastocyst stages were 84.3, 87.87 and 89.89%, respectively. The hatching rates in previous developmental stages in vitrified group were 57.44, 66.73 and 70.89% and in non-vitrified group were 66.32, 73.25 and 75.89%, respectively (P>0.05). The ATP content of in vivo or in vitro collected embryos was not significantly different in both vitrified and non-vitrified groups (P>0.05). Mitochondrial distribution of vitrified and non-vitrified 2-PN embryos was similar, but some clampings or large aggregation of mitochondria within the vitrified 4-cell embryos was prominent. Conclusions: Vitrification method did not affect the mouse embryo ATP content. Also, the cellular stress was not induced by this procedure and the safety of vitrification was shown. PMID:23748889

  16. Assessment of external heat transfer coefficient during oocyte vitrification in liquid and slush nitrogen using numerical simulations to determine cooling rates.

    PubMed

    Santos, M V; Sansinena, M; Zaritzky, N; Chirife, J

    2012-01-01

    In oocyte vitrification, plunging directly into liquid nitrogen favor film boiling and strong nitrogen vaporization. A survey of literature values of heat transfer coefficients (h) for film boiling of small metal objects with different geometries plunged in liquid nitrogen revealed values between 125 to 1000 W per per square m per K. These h values were used in a numerical simulation of cooling rates of two oocyte vitrification devices (open-pulled straw and Cryotop), plunged in liquid and slush nitrogen conditions. Heat conduction equation with convective boundary condition was considered a linear mathematical problem and was solved using the finite element method applying the variational formulation. COMSOL Multiphysics was used to simulate the cooling process of the systems. Predicted cooling rates for OPS and Cryotop when cooled at -196 degree C (liquid nitrogen) or -207 degree C (average for slush nitrogen) for heat transfer coefficients estimated to be representative of film boiling, indicated lowering the cooling temperature produces only a maximum 10 percent increase in cooling rates; confirming the main benefit of plunging in slush over liquid nitrogen does not arise from their temperature difference. Numerical simulations also demonstrated that a hypothetical four-fold increase in the cooling rate of vitrification devices when plunging in slush nitrogen would be explained by an increase in heat transfer coefficient. This improvement in heat transfer (i.e., high cooling rates) in slush nitrogen is attributed to less or null film boiling when a sample is placed in slush (mixture of liquid and solid nitrogen) because it first melts the solid nitrogen before causing the liquid to boil and form a film.

  17. Preliminary hazards analysis -- vitrification process

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

    Coordes, D.; Ruggieri, M.; Russell, J.

    1994-06-01

    This paper presents a Preliminary Hazards Analysis (PHA) for mixed waste vitrification by joule heating. The purpose of performing a PHA is to establish an initial hazard categorization for a DOE nuclear facility and to identify those processes and structures which may have an impact on or be important to safety. The PHA is typically performed during and provides input to project conceptual design. The PHA is then followed by a Preliminary Safety Analysis Report (PSAR) performed during Title 1 and 2 design. The PSAR then leads to performance of the Final Safety Analysis Report performed during the facility`s constructionmore » and testing. It should be completed before routine operation of the facility commences. This PHA addresses the first four chapters of the safety analysis process, in accordance with the requirements of DOE Safety Guidelines in SG 830.110. The hazards associated with vitrification processes are evaluated using standard safety analysis methods which include: identification of credible potential hazardous energy sources; identification of preventative features of the facility or system; identification of mitigative features; and analyses of credible hazards. Maximal facility inventories of radioactive and hazardous materials are postulated to evaluate worst case accident consequences. These inventories were based on DOE-STD-1027-92 guidance and the surrogate waste streams defined by Mayberry, et al. Radiological assessments indicate that a facility, depending on the radioactive material inventory, may be an exempt, Category 3, or Category 2 facility. The calculated impacts would result in no significant impact to offsite personnel or the environment. Hazardous materials assessment indicates that a Mixed Waste Vitrification facility will be a Low Hazard facility having minimal impacts to offsite personnel and the environment.« less

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

    PubMed

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

    2006-05-01

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

  19. Role of sodium ions in the vitrification process: glass matrix modification, slag structure depolymerization, and influence of metal immobilization.

    PubMed

    Kuo, Yi-Ming

    2014-07-01

    This study investigates the role of Na ions, a common flux, in the vitrification process. Artificial glass systems composed of Al2O3, CaO, and SiO2 with various Na concentrations were melted at 1450 degrees C. The specimens were cooled by air cooling and water quenching and the metal mobility was evaluated using a sequential extraction procedure. The X-ray diffraction analysis and scanning electron microscopy observations showed that Na ions governed the air-cooled slag's structure. Na ions initially depolymerized CaSiO3-linked chains into CaSiO3 chains, and further cut them into shorter and nonuniform ones, making the slag structure amorphous. With even more Na ions, CaSiO3 chains were divided into single SiO4 tetrahedrons and formed Na-related crystals (Na2Ca3Si2O8 and NaAlSiO4). The phase distributions of Al, Cr, Cu Mn, and Ni showed that Na has a positive effect on the immobilization of heavy metals at suitable concentrations, but a negative effect when in excess amounts. Implications: Vitrification has been widely used to treat hazardous materials. The Na-bearing additives were often used as a flux to improve the melting process. This study described the role of Na played in the vitrification process. The Na ions acted as glass modifier and depolymerize the chain structure of slag. With adequate addition amount of Na ions, the immobilization of heavy metals was improved. The results provided much information about the crystalline phase variation, metal mobility, and surface characteristics while Na serves as a flux.

  20. Ovarian tissue vitrification and heterotopic autologous transplantation in prepubertal Wistar rats.

    PubMed

    Wietcovsky, Leticia; Til, David; Salvador, Rafael Alonso; Amaral, Nicole Louise Lângaro; Senn, Alfred Paul; Amaral, Vera Lucia Lângaro

    2018-03-15

    To evaluate the efficiency of ovarian tissue heterotopic autografting after vitrification in prepubertal rats. Fragments of excised ovaries from prepubertal rats were used after assessing post-warming cellular viability, to determine the best vitrification protocol prior to retroauricular autografting. Pre-pubertal females (N=24) were castrated and divided into three group: Group 1 - fresh ovarian tissue transplantation; Group 2 - vitrified/warmed tissue transplantation; Group 3 - bilateral oophorectomy without transplantation. The ovarian fragments were exposed to solutions from the Ingamed® commercial kit, allocated in bacteriological loops and immersed in liquid nitrogen. Sixty days after transplantation, a vaginal mucus sample was collected for cytology tests, followed by sacrificing the animal, performing a cardiac puncture for collecting a blood sample to determine luteinizing hormone and estradiol levels, and excision of the transplanted fragment for histology tests. Vaginal cytology revealed that 87.5% of females from groups 1 and 2 had estrus while all females in Group 3 remained in diestrus. The mean LH value in groups 1 (0.08 mIU/mL) and 2 (0.34 mIU/mL) were statistically different from that of Group 3 (2.27 mIU/mL). E2 values did not differ between the groups. The histological analysis of Group 1 excised grafts versus those from Group 2 showed a higher percentage of primary follicles (62.5% vs. 12.5%), developing follicles (75% vs. 25%), corpus luteum (37.5% vs. 12.5%) and stromal region (100% vs. 87.5%). This study indicated that pre-pubertal ovarian tissue vitrification can be used to preserve fertility and to restore endocrine function in castrated rats.

  1. Efficient Term Development of Vitrified Ferret Embryos Using a Novel Pipette Chamber Technique1

    PubMed Central

    Sun, Xingshen; Li, Ziyi; Yi, Yaling; Chen, Juan; Leno, Gregory H.; Engelhardt, John F.

    2008-01-01

    Development of an efficient cryopreservation technique for the domestic ferret is key for the long-term maintenance of valuable genetic specimens of this species and for the conservation of related endangered species. Unfortunately, current cryopreservation procedures, such as slow-rate freezing and vitrification with open pulled straws, are inefficient. In this report, we describe a pipette tip-based vitrification method that significantly improves the development of thawed ferret embryos following embryo transfer (ET). Ferret embryos at the morula (MR), compact morula (CM), and early blastocyst (EB) stages were vitrified using an Eppendorf microloader pipette tip as the chamber vessel. The rate of in vitro development was significantly (P < 0.05) higher among embryos vitrified at the CM (93.6%) and EB (100%) stages relative to those vitrified at the MR stages (58.7%). No significant developmental differences were observed when comparing CM and EB vitrified embryos with nonvitrified control CM (100%) and EB (100%) embryos. In addition, few differences in the ultrastructure of intracellular lipid droplets or in microfilament structure were observed between control embryos and embryos vitrified at any developmental stage. Vitrified-thawed CM/EB embryos cultured for 2 or 16 h before ET resulted in live birth rates of 71.3% and 77.4%, respectively. These rates were not significantly different from the control live birth rate (79.2%). However, culture for 32 h (25%) or 48 h (7.8%) after vitrification significantly reduced the rate of live births. These data indicate that the pipette chamber vitrification technique significantly improves the live birth rate of transferred ferret embryos relative to current state-of-the-art methods.. PMID:18633142

  2. Secondary Waste Cast Stone Waste Form Qualification Testing Plan

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

    Westsik, Joseph H.; Serne, R. Jeffrey

    2012-09-26

    The Hanford Tank Waste Treatment and Immobilization Plant (WTP) is being constructed to treat the 56 million gallons of radioactive waste stored in 177 underground tanks at the Hanford Site. The WTP includes a pretreatment facility to separate the wastes into high-level waste (HLW) and low-activity waste (LAW) fractions for vitrification and disposal. The LAW will be converted to glass for final disposal at the Integrated Disposal Facility (IDF). Cast Stone – a cementitious waste form, has been selected for solidification of this secondary waste stream after treatment in the ETF. The secondary-waste Cast Stone waste form must be acceptablemore » for disposal in the IDF. This secondary waste Cast Stone waste form qualification testing plan outlines the testing of the waste form and immobilization process to demonstrate that the Cast Stone waste form can comply with the disposal requirements. Specifications for the secondary-waste Cast Stone waste form have not been established. For this testing plan, Cast Stone specifications are derived from specifications for the immobilized LAW glass in the WTP contract, the waste acceptance criteria for the IDF, and the waste acceptance criteria in the IDF Permit issued by the State of Washington. This testing plan outlines the testing needed to demonstrate that the waste form can comply with these waste form specifications and acceptance criteria. The testing program must also demonstrate that the immobilization process can be controlled to consistently provide an acceptable waste form product. This testing plan also outlines the testing needed to provide the technical basis for understanding the long-term performance of the waste form in the disposal environment. These waste form performance data are needed to support performance assessment analyses of the long-term environmental impact of the secondary-waste Cast Stone waste form in the IDF« less

  3. Site Safety and Health Plan (Phase 3) for the treatability study for in situ vitrification at Seepage Pit 1 in Waste Area Grouping 7, Oak Ridge National Laboratory, Oak Ridge, TN

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

    Spalding, B.P.; Naney, M.T.

    1995-06-01

    This plan is to be implemented for Phase III ISV operations and post operations sampling. Two previous project phases involving site characterization have been completed and required their own site specific health and safety plans. Project activities will take place at Seepage Pit 1 in Waste Area Grouping 7 at ORNL, Oak Ridge, Tennessee. Purpose of this document is to establish standard health and safety procedures for ORNL project personnel and contractor employees in performance of this work. Site activities shall be performed in accordance with Energy Systems safety and health policies and procedures, DOE orders, Occupational Safety and Healthmore » Administration Standards 29 CFR Part 1910 and 1926; applicable United States Environmental Protection Agency requirements; and consensus standards. Where the word ``shall`` is used, the provisions of this plan are mandatory. Specific requirements of regulations and orders have been incorporated into this plan in accordance with applicability. Included from 29 CFR are 1910.120 Hazardous Waste Operations and Emergency Response; 1910.146, Permit Required - Confined Space; 1910.1200, Hazard Communication; DOE Orders requirements of 5480.4, Environmental Protection, Safety and Health Protection Standards; 5480.11, Radiation Protection; and N5480.6, Radiological Control Manual. In addition, guidance and policy will be followed as described in the Environmental Restoration Program Health and Safety Plan. The levels of personal protection and the procedures specified in this plan are based on the best information available from reference documents and site characterization data. Therefore, these recommendations represent the minimum health and safety requirements to be observed by all personnel engaged in this project.« less

  4. Large Advanced Space Systems (LASS) computer-aided design program additions

    NASA Technical Reports Server (NTRS)

    Farrell, C. E.

    1982-01-01

    The LSS preliminary and conceptual design requires extensive iteractive analysis because of the effects of structural, thermal, and control intercoupling. A computer aided design program that will permit integrating and interfacing of required large space system (LSS) analyses is discussed. The primary objective of this program is the implementation of modeling techniques and analysis algorithms that permit interactive design and tradeoff studies of LSS concepts. Eight software modules were added to the program. The existing rigid body controls module was modified to include solar pressure effects. The new model generator modules and appendage synthesizer module are integrated (interfaced) to permit interactive definition and generation of LSS concepts. The mass properties module permits interactive specification of discrete masses and their locations. The other modules permit interactive analysis of orbital transfer requirements, antenna primary beam n, and attitude control requirements.

  5. Nome Offshore Mining Information

    Science.gov Websites

    Lands Coal Regulatory Program Large Mine Permits Mineral Property and Rights Mining Index Land potential safety concerns, prevent overcrowding, and provide for efficient processing of the permits and Regulatory Program Large Mine Permitting Mineral Property Management Mining Fact Sheets Mining Forms APMA

  6. 76 FR 29747 - State Program Requirements; Proposal To Approve Maine's Base National Pollutant Discharge...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-23

    ...'s Base National Pollutant Discharge Elimination System (NPDES) Permitting Program AGENCY... of Maine's Base NPDES Permitting Program in these territories as part of the administrative record to... Maine's base program as EPA approved it in 2001. Thus, the state's program would not include regulation...

  7. 50 CFR 15.21 - General application procedures.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... PLANTS WILD BIRD CONSERVATION ACT Permits and Approval of Cooperative Breeding Programs § 15.21 General... purposes only: Scientific research; zoological breeding or display programs; cooperative breeding programs...) A person wishing to obtain a permit under this subpart or approval of cooperative breeding programs...

  8. 50 CFR 15.21 - General application procedures.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... PLANTS WILD BIRD CONSERVATION ACT Permits and Approval of Cooperative Breeding Programs § 15.21 General... purposes only: Scientific research; zoological breeding or display programs; cooperative breeding programs...) A person wishing to obtain a permit under this subpart or approval of cooperative breeding programs...

  9. Office of River Protection Advanced Low-Activity Waste Glass Research and Development Plan

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

    Kruger, A. A.; Peeler, D. K.; Kim, D. S.

    2015-11-23

    The U.S. Department of Energy Office of River Protection (ORP) has initiated and leads an integrated Advanced Waste Glass (AWG) program to increase the loading of Hanford tank wastes in glass while meeting melter lifetime expectancies and process, regulatory, and product performance requirements. The integrated ORP program is focused on providing a technical, science-based foundation for making key decisions regarding the successful operation of the Hanford Tank Waste Treatment and Immobilization Plant (WTP) facilities in the context of an optimized River Protection Project (RPP) flowsheet. The fundamental data stemming from this program will support development of advanced glass formulations, keymore » product performance and process control models, and tactical processing strategies to ensure safe and successful operations for both the low-activity waste (LAW) and high-level waste vitrification facilities. These activities will be conducted with the objective of improving the overall RPP mission by enhancing flexibility and reducing cost and schedule.« less

  10. 40 CFR 144.31 - Application for a permit; authorization by permit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... (PSD) program under the Clean Air Act. (2) Name, mailing address, and location of the facility for... under SDWA. (iii) NPDES program under CWA. (iv) Prevention of Significant Deterioration (PSD) program...

  11. 30 CFR 778.17 - Permit term.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... MINING AND RECLAMATION OPERATIONS PERMITS AND COAL EXPLORATION SYSTEMS UNDER REGULATORY PROGRAMS PERMIT APPLICATIONS-MINIMUM REQUIREMENTS FOR LEGAL, FINANCIAL, COMPLIANCE, AND RELATED INFORMATION § 778.17 Permit...

  12. Vitrification of waste

    DOEpatents

    Wicks, George G.

    1999-01-01

    A method for encapsulating and immobilizing waste for disposal. Waste, preferably, biologically, chemically and radioactively hazardous, and especially electronic wastes, such as circuit boards, are placed in a crucible and heated by microwaves to a temperature in the range of approximately 300.degree. C. to 800.degree. C. to incinerate organic materials, then heated further to a temperature in the range of approximately 1100.degree. C. to 1400.degree. C. at which temperature glass formers present in the waste will cause it to vitrify. Glass formers, such as borosilicate glass, quartz or fiberglass can be added at the start of the process to increase the silicate concentration sufficiently for vitrification.

  13. Volatile species of technetium and rhenium during waste vitrification

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

    Kim, Dongsang; Kruger, Albert A.

    Volatile loss of technetium (Tc) during vitrification of low-activity wastes is a technical challenge for treating and immobilizing the large volumes of radioactive and hazardous wastes stored at the U.S. Department of Energy's Hanford Site. There are various research efforts being pursued to develop technologies that can be implemented for cost effective management of Tc, including studies to understand the behavior of Tc during vitrification, with the goal of eventually increasing Tc retention in glass. Furthermore, one of these studies has focused on identifying the form or species of Tc and Re (surrogate for Tc) that evolve during the waste-to-glassmore » conversion process. This information is important for understanding the mechanism of Tc volatilization. In this paper, available information collected from the literature is critically evaluated to clarify the volatile species of Tc and Re and, more specifically, whether they volatilize as alkali pertechnetate and perrhenate or as technetium and rhenium oxides after decomposition of alkali pertechnetate and perrhenate. The evaluated data ranged from mass spectrometric identification of species volatilized from pure and binary alkali pertechnetate and perrhenate salts to structural and chemical analyses of volatilized materials during crucible melting and scaled melter processing of simulated wastes.« less

  14. How thermal stress alters the confinement of polymers vitrificated in nanopores

    NASA Astrophysics Data System (ADS)

    Teng, Chao; Li, Linling; Wang, Yong; Wang, Rong; Chen, Wei; Wang, Xiaoliang; Xue, Gi

    2017-05-01

    Understanding and controlling the glass transition temperature (Tg) and dynamics of polymers in confined geometries are of significance in both academia and industry. Here, we investigate how the thermal stress induced by a mismatch in the coefficient of thermal expansion affects the Tg behavior of polystyrene (PS) nanorods located inside cylindrical alumina nanopores. The size effects and molecular weight dependence of the Tg are also studied. A multi-step relaxation process was employed to study the relationship between thermal stress and cooling rate. At fast cooling rates, the imparted thermal stress would overcome the yield stress of PS and peel chains off the pore walls, while at slow cooling rates, chains are kept in contact with the pore walls due to timely dissipation of the produced thermal stress during vitrification. In smaller nanopores, more PS chains closely contact with pore walls, then stronger internal thermal stress would be generated between core and shell of PS nanorod, which results in a larger deviation between two Tgs. The core part of PS shows lower Tg than bulk value, which can induce faster dynamics in the center region. A complex and important role stress plays is supposed in complex confinement condition, e.g., in nanopores, during vitrification.

  15. Adsorbing/dissolving Lyoprotectant Matrix Technology for Non-cryogenic Storage of Archival Human Sera

    NASA Astrophysics Data System (ADS)

    Solivio, Morwena J.; Less, Rebekah; Rynes, Mathew L.; Kramer, Marcus; Aksan, Alptekin

    2016-04-01

    Despite abundant research conducted on cancer biomarker discovery and validation, to date, less than two-dozen biomarkers have been approved by the FDA for clinical use. One main reason is attributed to inadvertent use of low quality biospecimens in biomarker research. Most proteinaceous biomarkers are extremely susceptible to pre-analytical factors such as collection, processing, and storage. For example, cryogenic storage imposes very harsh chemical, physical, and mechanical stresses on biospecimens, significantly compromising sample quality. In this communication, we report the development of an electrospun lyoprotectant matrix and isothermal vitrification methodology for non-cryogenic stabilization and storage of liquid biospecimens. The lyoprotectant matrix was mainly composed of trehalose and dextran (and various low concentration excipients targeting different mechanisms of damage), and it was engineered to minimize heterogeneity during vitrification. The technology was validated using five biomarkers; LDH, CRP, PSA, MMP-7, and C3a. Complete recovery of LDH, CRP, and PSA levels was achieved post-rehydration while more than 90% recovery was accomplished for MMP-7 and C3a, showing promise for isothermal vitrification as a safe, efficient, and low-cost alternative to cryogenic storage.

  16. Ovarian and oocyte cryopreservation.

    PubMed

    Lornage, Jacqueline; Salle, Bruno

    2007-08-01

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

  17. Metal behavior during vitrification of incinerator ash in a coke bed furnace.

    PubMed

    Kuo, Yi-Ming; Lin, Ta-Chang; Tsai, Perng-Jy

    2004-06-18

    In this study, municipal waste incinerator ash was vitrified in a coke bed furnace system and the behavior of metals was investigated. Coke and lime were added to provide heat which facilitated vitrification. Ash contributed more than 90% of metal (except for Ca) input-mass. Metal species with low boiling points accounted for the major fraction of their input-mass adsorbed by air pollution control devices (APCDs) fly ash. Among the remaining metals, those species with light specific weights in this furnace tended to be encapsulated in slag, while heavier species were mainly discharged by ingot. Meanwhile, the leachability of hazardous metals in slag was significantly reduced. The distribution index (DI) was defined and used as an index for distribution of heavy metals in the system. A high DI assures safe slag reuse and implies feasibility of recovering hazardous heavy metals such as Cr, Cu, Fe, Pb and Zn. The vitrification in a coke bed furnace proved to be a useful technology for the final disposal of MSW incinerator ash. The heavy metals are separated into the slag, ingot and fly ash, allowing safe reuse of the slag and possible recovery of the metals contained in the ingot and ash fractions.

  18. Corrosion assessment of refractory materials for high temperature waste vitrification

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

    Marra, J.C.; Congdon, J.W.; Kielpinski, A.L.

    1995-11-01

    A variety of vitrification technologies are being evaluated to immobilize radioactive and hazardous wastes following years of nuclear materials production throughout the Department of Energy (DOE) complex. The compositions and physical forms of these wastes are diverse ranging from inorganic sludges to organic liquids to heterogeneous debris. Melt and off-gas products can be very corrosive at the high temperatures required to melt many of these waste streams. Ensuring material durability is required to develop viable treatment processes. Corrosion testing of materials in some of the anticipated severe environments is an important aspect of the materials identification and selection process. Corrosionmore » coupon tests on typical materials used in Joule heated melters were completed using glass compositions with high salt contents. The presence of chloride in the melts caused the most severe attack. In the metal alloys, oxidation was the predominant corrosion mechanism, while in the tested refractory material enhanced dissolution of the refractory into the glass was observed. Corrosion testing of numerous different refractory materials was performed in a plasma vitrification system using a surrogate heterogeneous debris waste. Extensive corrosion was observed in all tested materials.« less

  19. Bench scale experiments for the remediation of Hanford Waste Treatment Plant low activity waste melter off-gas condensate

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

    Taylor-Pashow, Kathryn M.L.; Poirier, Michael; McCabe, Daniel J.

    The Low Activity Waste (LAW) vitrification facility at the Hanford Waste Treatment and Immobilization Plant (WTP) will generate an aqueous condensate recycle stream (LAW Off-Gas Condensate) from the off-gas system. The plan for disposition of this stream during baseline operations is to send it to the WTP Pretreatment Facility, where it will be blended with LAW, concentrated by evaporation and recycled to the LAW vitrification facility again. The primary reason to recycle this stream is so that the semi-volatile 99Tc isotope eventually becomes incorporated into the glass. This stream also contains non-radioactive salt components that are problematic in the melter,more » so diversion of this stream to another process would eliminate recycling of these salts and would enable simplified operation of the LAW melter and the Pretreatment Facilities. This diversion from recycling this stream within WTP would have the effect of decreasing the LAW vitrification mission duration and quantity of glass waste. The concept being tested here involves removing the 99Tc so that the decontaminated aqueous stream, with the problematic salts, can be disposed elsewhere.« less

  20. Investigation of variable compositions on the removal of technetium from Hanford Waste Treatment Plant low activity waste melter off-gas condensate simulant

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

    Taylor-Pashow, Kathryn M. L.; McCabe, Daniel J.; Pareizs, John M.

    The Low Activity Waste (LAW) vitrification facility at the Hanford Waste Treatment and Immobilization Plant (WTP) will generate an aqueous condensate recycle stream (LAW Off-Gas Condensate) from the offgas system. The plan for disposition of this stream during baseline operations is to send it to the WTP Pretreatment Facility, where it will be blended with LAW, concentrated by evaporation and recycled to the LAW vitrification facility again. The primary reason to recycle this stream is so that the semi-volatile 99Tc isotope eventually becomes incorporated into the glass. This stream also contains non-radioactive salt components that are problematic in the melter,more » so diversion of this stream to another process would eliminate recycling of these salts and would enable simplified operation of the LAW melter and the Pretreatment Facilities. This diversion from recycling this stream within WTP would have the effect of decreasing the LAW vitrification mission duration and quantity of glass waste. The concept being tested here involves removing the 99Tc so that the decontaminated aqueous stream, with the problematic salts, can be disposed elsewhere.« less

  1. Volatile species of technetium and rhenium during waste vitrification

    DOE PAGES

    Kim, Dongsang; Kruger, Albert A.

    2017-10-26

    Volatile loss of technetium (Tc) during vitrification of low-activity wastes is a technical challenge for treating and immobilizing the large volumes of radioactive and hazardous wastes stored at the U.S. Department of Energy's Hanford Site. There are various research efforts being pursued to develop technologies that can be implemented for cost effective management of Tc, including studies to understand the behavior of Tc during vitrification, with the goal of eventually increasing Tc retention in glass. Furthermore, one of these studies has focused on identifying the form or species of Tc and Re (surrogate for Tc) that evolve during the waste-to-glassmore » conversion process. This information is important for understanding the mechanism of Tc volatilization. In this paper, available information collected from the literature is critically evaluated to clarify the volatile species of Tc and Re and, more specifically, whether they volatilize as alkali pertechnetate and perrhenate or as technetium and rhenium oxides after decomposition of alkali pertechnetate and perrhenate. The evaluated data ranged from mass spectrometric identification of species volatilized from pure and binary alkali pertechnetate and perrhenate salts to structural and chemical analyses of volatilized materials during crucible melting and scaled melter processing of simulated wastes.« less

  2. Determining the dissolution rates of actinide glasses: A time and temperature Product Consistency Test study

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

    Daniel, W.E.; Best, D.R.

    1995-12-01

    Vitrification has been identified as one potential option for the e materials such as Americium (Am), Curium (Cm), Neptunium (Np), and Plutonium (Pu). A process is being developed at the Savannah River Site to safely vitrify all of the highly radioactive Am/Cm material and a portion of the fissile (Pu) actinide materials stored on site. Vitrification of the Am/Cm will allow the material to be transported and easily stored at the Oak Ridge National Laboratory. The Am/Cm glass has been specifically designed to be (1) highly durable in aqueous environments and (2) selectively attacked by nitric acid to allow recoverymore » of the valuable Am and Cm isotopes. A similar glass composition will allow for safe storage of surplus plutonium. This paper will address the composition, relative durability, and dissolution rate characteristics of the actinide glass, Loeffler Target, that will be used in the Americium/Curium Vitrification Project at Westinghouse Savannah River Company near Aiken, South Carolina. The first part discusses the tests performed on the Loeffler Target Glass concerning instantaneous dissolution rates. The second part presents information concerning pseudo-activation energy for the one week glass dissolution process.« less

  3. Ovine secondary follicles vitrified out the ovarian tissue grow and develop in vitro better than those vitrified into the ovarian fragments.

    PubMed

    Lunardi, Franciele Osmarini; de Aguiar, Francisco Leo Nascimento; Duarte, Ana Beatriz Graça; Araújo, Valdevane Rocha; de Lima, Laritza Ferreira; Ribeiro de Sá, Naiza Arcângela; Vieira Correia, Hudson Henrique; Domingues, Sheyla Farhayldes Souza; Campello, Cláudio Cabral; Smitz, Johan; de Figueiredo, José Ricardo; Ribeiro Rodrigues, Ana Paula

    2016-04-15

    Cryopreservation of preantral follicles is a promising technique to preserve female fertility. The aim of this study was to evaluate the effect of vitrification on the development of secondary follicles included in ovarian tissue or isolated after microdissection. An important end point included is the capacity of grown oocytes to resume meiosis. Sheep ovarian cortexes were cut into fragments and split into three different groups: (1) fresh (control): secondary follicles isolated without any previous vitrification; (2) follicle-vitrification (follicle-vit): secondary follicles vitrified in isolated form; and (3) tissue-vitrification (tissue-vit): secondary follicles vitrified within fragments of ovarian tissue (in situ former) and subsequently subjected to isolation. From the three groups, isolated secondary follicles were submitted to IVC for 18 days. After IVC, cumulus-oocyte complexes (COCs) were harvested from follicles. As an additional control group, in vivo grown, in vivo-grown COCs were collected from antral ovarian follicles. All, recovered COCs were matured and the chromatin configuration was evaluated. Data were analyzed by ANOVA, and the means were compared by Student-Newman-Keuls test, and by chi-square. Differences were considered to be significant when P < 0.05. Isolated preantral follicles from all treatments had normal morphology, antrum formation, and low follicle degeneration after IVC. The growth rate between control and follicle-vit did not differ (P > 0.05), and was higher (P < 0.05) than for tissue-vit. The percentage of follicles that decreased diameter during IVC was significantly higher in tissue-vit than the in follicle-vit. Recovery rate of oocytes from normal follicles was higher in follicle-vit than in tissue-vit. Furthermore, oocyte viability was lower in tissue-vit than other treatments, and follicle-vit did not differ from control and in vivo grown. The percentage of oocytes meiosis resuming was not different between treatments except for in vivo grown. After vitrification, only follicle-vit showed metaphase I oocyte. We conclude that secondary follicles vitrified after isolation displayed a better follicular growth rate, oocyte viability, percentage of oocytes reaching the metaphase I stage, and fewer follicles with decreased diameter after IVC. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Evaluation of Vitrification Processing Step for Rocky Flats Incinerator Ash

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

    Wigent, W.L.; Luey, J.K.; Scheele, R.D.

    In 1997, Pacific Northwest National Laboratory (PNNL) staff developed a processing option for incinerator ash at the Rocky Flats Environmental Technology Sites (RFETS). This work was performed with support from Los Alamos National Laboratory (LANL) and Safe Sites of Colorado (SSOC). A description of the remediation needs for the RFETS incinerator ash is provided in a report summarizing the recommended processing option for treatment of the ash (Lucy et al. 1998). The recommended process flowsheet involves a calcination pretreatment step to remove carbonaceous material followed by a vitrification processing step for a mixture of glass tit and calcined incinerator ash.more » Using the calcination pretreatment step to remove carbonaceous material reduced process upsets for the vitrification step, allowed for increased waste loading in the final product, and improved the quality of the final product. Figure 1.1 illustrates the flow sheet for the recommended processing option for treatment of RFETS incinerator ash. In 1998, work at PNNL further developed the recommended flow sheet through a series of studies to better define the vitrification operating parameters and to address secondary processing issues (such as characterizing the offgas species from the calcination process). Because a prototypical rotary calciner was not available for use, studies to evaluate the offgas from the calcination process were performed using a benchtop rotary calciner and laboratory-scale equipment (Lucy et al. 1998). This report focuses on the vitrification process step after ash has been calcined. Testing with full-scale containers was performed using ash surrogates and a muffle furnace similar to that planned for use at RFETS. Small-scale testing was performed using plutonium-bearing incinerator ash to verify performance of the waste form. Ash was not obtained from RFETS because of transportation requirements to calcine the incinerator ash prior to shipment of the material. Because part of PNNL's work was to characterize the ash prior to calcination and to investigate the effect of calcination on product quality, representative material was obtained from LANL. Ash obtained from LANL was selected based on its similarity to that currently stored at RFETS. The plutonium-bearing ashes obtained from LANL are likely from a RFETS incinerator, but the exact origin was not identified.« less

  5. Vitrification of ovarian tissue of Brazilian North-eastern donkeys (Equus asinus) using different cryoprotectants.

    PubMed

    Lopes, Kátia Regina F; Praxedes, Erica Camila G; Campos, Livia B; Bezerra, Marcelo B; Lima, Gabriela L; Saraiva, Márcia Viviane A; Silva, Alexandre R

    2018-05-29

    The aim of this study was to assess a vitrification protocol for asinine ovarian tissue, to preserve preantral follicles using different cryoprotectant solutions, composed of various concentrations (EG 3 M or 6 M) of dimethyl sulfoxide or ethylene glycol isolate, or as a combination (DMSO 3 M + EG 3 M). Ten pairs of ovaries from Brazilian north-eastern breed jennies were obtained through videolaparoscopy, and cortical fragments were submitted to a solid-surface vitrification (SSV) using each cryoprotectant solution. The ovarian tissue was evaluated for follicular morphology and viability, DNA integrity (TUNEL technique) and the presence of nucleolar organizing regions in granulosa cells (AgNOR technique). After thawing, the percentage of normal preantral follicles was significantly reduced in the vitrified ovarian tissue fragments compared to the fresh control (p < 0.05). When comparing treatments, the use of DMSO 3 M (81.7 ± 37.5%), EG 3 M (83.7 ± 27.4%) and the combination of both DMSO 3 M + EG 3 M (81.8 ± 46.8%) allowed a greater percentage of follicular survival in contrast to DMSO 6 M (69.8 ± 16.5%) and EG 6 M (72.3 ± 18.0%; p < 0.05). When vitrified using the DMSO + EG combination, a higher percentage (62.5 ± 29.1%) of viable follicles (trypan blue) was observed in relation to the other vitrification treatments (p < 0.05). The TUNEL technique identified that all treatments tested showed DNA fragmentation in the follicular cells, except in the case of the DMSO 3 M + EG 3 M treatment. When evaluating the presence of NORs, no significant differences were observed in the amount of NORs between the fresh and vitrified groups using DMSO 3 M + EG 3 M (p > 0.05). We concluded that the combination DMSO 3 M + EG was more efficient for the vitrification of ovarian tissue taken from Equus asinus, allowing adequate preservation of PAFs morphology, viability, DNA integrity and cell proliferative capacity. © 2018 Blackwell Verlag GmbH.

  6. 40 CFR 233.37 - Signatures on permit applications and reports.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Signatures on permit applications and... DUMPING 404 STATE PROGRAM REGULATIONS Program Operation § 233.37 Signatures on permit applications and... person designating the agent. In either case, the signature of the applicant or the agent will be...

  7. 40 CFR 233.34 - Making a decision on the permit application.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Making a decision on the permit... DUMPING 404 STATE PROGRAM REGULATIONS Program Operation § 233.34 Making a decision on the permit... each application outlining his decision and rationale for his decision. The determination shall be...

  8. 40 CFR 144.31 - Application for a permit; authorization by permit.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... Significant Deterioration (PSD) program under the Clean Air Act. (2) Name, mailing address, and location of... (PSD) program under the Clean Air Act. (v) Nonattainment program under the Clean Air Act. (vi) National...

  9. 40 CFR 144.31 - Application for a permit; authorization by permit.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... Significant Deterioration (PSD) program under the Clean Air Act. (2) Name, mailing address, and location of... (PSD) program under the Clean Air Act. (v) Nonattainment program under the Clean Air Act. (vi) National...

  10. 40 CFR 144.31 - Application for a permit; authorization by permit.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Significant Deterioration (PSD) program under the Clean Air Act. (2) Name, mailing address, and location of... (PSD) program under the Clean Air Act. (v) Nonattainment program under the Clean Air Act. (vi) National...

  11. 40 CFR 144.31 - Application for a permit; authorization by permit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Significant Deterioration (PSD) program under the Clean Air Act. (2) Name, mailing address, and location of... (PSD) program under the Clean Air Act. (v) Nonattainment program under the Clean Air Act. (vi) National...

  12. 75 FR 59711 - Audit Program for Texas Flexible Permit Holders

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-28

    ... ENVIRONMENTAL PROTECTION AGENCY [EPA-R06-OAR-2010-0510; FRL-9207-4] Audit Program for Texas... Act (CAA) voluntary audit compliance program for flexible permit holders in the State of Texas (hereinafter ``Audit Program''); response to public comments. SUMMARY: EPA is offering holders of Texas...

  13. Permitting program with best management practices for shale gas wells to safeguard public health.

    PubMed

    Centner, Terence J; Petetin, Ludivine

    2015-11-01

    The development of shale gas resources in the United States has been controversial as governments have been tardy in devising sufficient safeguards to protect both people and the environment. Alleged health and environmental damages suggest that other countries around the world that decide to develop their shale gas resources can learn from these problems and take further actions to prevent situations resulting in the release of harmful pollutants. Looking at U.S. federal regulations governing large animal operations under the permitting provisions of the Clean Water Act, the idea of a permitting program is proposed to respond to the risks of pollution by shale gas development activities. Governments can require permits before allowing the drilling of a new gas well. Each permit would include fluids and air emissions reduction plans containing best management practices to minimize risks and releases of pollutants. The public availability of permits and permit applications, as occurs for water pollution under various U.S. permitting programs, would assist governments in protecting public health. The permitting proposals provide governments a means for providing further assurances that shale gas development projects will not adversely affect people and the environment. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Final Amendments to Delegation of Authority Provisions in the Prevention of Significant Deterioration Permitting Program

    EPA Pesticide Factsheets

    finalized amendments to the New Source Review (NSR) Prevention of Significant Deterioration (PSD) permitting program that will allow the EPA to delegate administration of the program to interested and qualified tribal agencies.

  15. NPDES Permit Writers' Course

    EPA Pesticide Factsheets

    The objective of the NPDES permit writers' course is to provide the basic regulatory framework and technical considerations that support the development of wastewater discharge permits as required under the NPDES Permit Program.

  16. 40 CFR 130.12 - Coordination with other programs.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... PROGRAMS WATER QUALITY PLANNING AND MANAGEMENT § 130.12 Coordination with other programs. (a) Relationship... 208(e) of the Act, no NPDES permit may be issued which is in conflict with an approved Water Quality Management (WQM) plan. Where a State has assumed responsibility for the administration of the permit program...

  17. 76 FR 28223 - Clean Air Act Operating Permit Program; Petition for Objection to State Operating Permit for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-16

    ... Program; Petition for Objection to State Operating Permit for Tennessee Valley Authority--Paradise Fossil... for Air Quality to Tennessee Valley Authority (TVA) for its Paradise Fossil Fuel Plant located near... period. Petitioner submitted a petition regarding the Paradise Fossil Fuel Plant on January 9, 2010...

  18. 76 FR 10361 - Clean Air Act Operating Permit Program; Petition for Objection to State Operating Permit for...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-24

    ... July 14, 2010 response of the CDPHE, Air Pollution Control Division to the October 8, 2009 Order by EPA... ENVIRONMENTAL PROTECTION AGENCY [FRL-9270-5] Clean Air Act Operating Permit Program; Petition for... issued by the Colorado Department of Public Health and Environment (CDPHE). Specifically, the...

  19. 77 FR 59879 - Approval and Promulgation of Air Quality Implementation Plans; Ohio; PBR and PTIO

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-01

    ... its air pollution permit program more efficient. Approving these additions will make the Permits by... revisions will make Ohio's air permit program more efficient while continuing to protect human health and...-9714-6] Approval and Promulgation of Air Quality Implementation Plans; Ohio; PBR and PTIO AGENCY...

  20. 40 CFR 70.4 - State program submittals and transition.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... determine insignificant activities or emission levels for purposes of determining complete applications... any permit application, compliance plan, permit, and monitoring and compliance, certification report... take final action on an application for a permit, permit renewal, or permit revision within the time...

  1. 40 CFR 70.4 - State program submittals and transition.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... determine insignificant activities or emission levels for purposes of determining complete applications... any permit application, compliance plan, permit, and monitoring and compliance, certification report... take final action on an application for a permit, permit renewal, or permit revision within the time...

  2. 40 CFR 70.4 - State program submittals and transition.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... determine insignificant activities or emission levels for purposes of determining complete applications... any permit application, compliance plan, permit, and monitoring and compliance, certification report... take final action on an application for a permit, permit renewal, or permit revision within the time...

  3. Water Quality Trading Toolkit for Permit Writers

    EPA Pesticide Factsheets

    The Water Quality Trading Toolkit for Permit Writers is EPA’s first “how-to” manual on designing and implementing water quality trading programs. It helps NPDES permitting authorities incorporate trading provisions into permits.

  4. 43 CFR 4.1368 - Determination by the Administrative Law Judge.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Request for Review of Approval Or Disapproval of Applications for New Permits, Permit Revisions, Permit Renewals, the Transfer, Assignment Or Sale of Rights Granted Under Permit (federal Program; Federal Lands...

  5. 40 CFR 123.63 - Criteria for withdrawal of State programs.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... withdrawal of State programs. (a) In the case of a sewage sludge management program, references in this... under this part, including failure to issue permits; (ii) Repeated issuance of permits which do not... sludge management program, § 501.14 of this chapter). (5) Where the State fails to develop an adequate...

  6. 40 CFR 122.27 - Silvicultural activities (applicable to State NPDES programs, see § 123.25).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) WATER PROGRAMS EPA ADMINISTERED PERMIT PROGRAMS: THE NATIONAL POLLUTANT DISCHARGE ELIMINATION SYSTEM Permit Application and Special NPDES Program Requirements § 122.27... or after removal of bark held in self-contained bodies of water (mill ponds or log ponds) or stored...

  7. 40 CFR 72.25 - Objections.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., under the Acid Rain Program. In the event of such communication, the Administrator and the permitting... inaction under the Acid Rain Program. (c) Neither the Administrator nor any permitting authority will...

  8. 40 CFR 72.25 - Objections.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., under the Acid Rain Program. In the event of such communication, the Administrator and the permitting... inaction under the Acid Rain Program. (c) Neither the Administrator nor any permitting authority will...

  9. 40 CFR 72.25 - Objections.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., under the Acid Rain Program. In the event of such communication, the Administrator and the permitting... inaction under the Acid Rain Program. (c) Neither the Administrator nor any permitting authority will...

  10. 40 CFR 72.25 - Objections.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., under the Acid Rain Program. In the event of such communication, the Administrator and the permitting... inaction under the Acid Rain Program. (c) Neither the Administrator nor any permitting authority will...

  11. 40 CFR 72.25 - Objections.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., under the Acid Rain Program. In the event of such communication, the Administrator and the permitting... inaction under the Acid Rain Program. (c) Neither the Administrator nor any permitting authority will...

  12. Glass Waste Forms for Oak Ridge Tank Wastes: Fiscal Year 1998 Report for Task Plan SR-16WT-31, Task B

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

    Andrews, M.K.

    1999-05-10

    Using ORNL information on the characterization of the tank waste sludges, SRTC performed extensive bench-scale vitrification studies using simulants. Several glass systems were tested to ensure the optimum glass composition (based on the glass liquidus temperature, viscosity and durability) is determined. This optimum composition will balance waste loading, melt temperature, waste form performance and disposal requirements. By optimizing the glass composition, a cost savings can be realized during vitrification of the waste. The preferred glass formulation was selected from the bench-scale studies and recommended to ORNL for further testing with samples of actual OR waste tank sludges.

  13. 40 CFR 124.10 - Public notice of permit actions and public comment period.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... issued or is required to issue a RCRA, UIC, PSD (or other permit under the Clean Air Act), NPDES, 404... (CONTINUED) WATER PROGRAMS PROCEDURES FOR DECISIONMAKING General Program Requirements § 124.10 Public notice... Act for the same facility or activity (including EPA when the draft permit is prepared by the State...

  14. 77 FR 76479 - Clean Air Act Operating Permit Program: Order Responding to Petition for Objection to State...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-12-28

    ... ENVIRONMENTAL PROTECTION AGENCY [FRL-9765-4] Clean Air Act Operating Permit Program: Order Responding to Petition for Objection to State Operating Permit for U.S. Steel-- Granite City Works, Granite... Protection Agency (IEPA) for the U.S. Steel--Granite City Works (USGW) facility, located in Granite City...

  15. 77 FR 42493 - Clean Air Act Operating Permit Program; Petition for Objection to State Operating Permit for Cash...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-19

    ... Program; Petition for Objection to State Operating Permit for Cash Creek Generation, LLC--Cash Creek... issued by the Kentucky Division for Air Quality (KDAQ) to Cash Creek Generation, LLC for its Cash Creek Generation Station (Cash Creek) located near Owensboro in Henderson County, Kentucky. This Order constitutes...

  16. 40 CFR 74.12 - Opt-in permit contents.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... be included in the Acid Rain permit. (b) Scope. The opt-in permit provisions shall apply only to the... provision that participation of a combustion or process source in the Acid Rain Program may be terminated... deemed to be operating in compliance with the Acid Rain Program, except as provided in § 72.9(g)(6) of...

  17. 40 CFR 74.12 - Opt-in permit contents.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... be included in the Acid Rain permit. (b) Scope. The opt-in permit provisions shall apply only to the... provision that participation of a combustion or process source in the Acid Rain Program may be terminated... deemed to be operating in compliance with the Acid Rain Program, except as provided in § 72.9(g)(6) of...

  18. 40 CFR 74.12 - Opt-in permit contents.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... be included in the Acid Rain permit. (b) Scope. The opt-in permit provisions shall apply only to the... provision that participation of a combustion or process source in the Acid Rain Program may be terminated... deemed to be operating in compliance with the Acid Rain Program, except as provided in § 72.9(g)(6) of...

  19. 40 CFR 74.12 - Opt-in permit contents.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... be included in the Acid Rain permit. (b) Scope. The opt-in permit provisions shall apply only to the... provision that participation of a combustion or process source in the Acid Rain Program may be terminated... deemed to be operating in compliance with the Acid Rain Program, except as provided in § 72.9(g)(6) of...

  20. 75 FR 82254 - Action To Ensure Authority To Implement Title V Permitting Programs Under the Greenhouse Gas...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-12-30

    ...-AQ63 Action To Ensure Authority To Implement Title V Permitting Programs Under the Greenhouse Gas... this action, EPA is finalizing its proposed rulemaking to narrow EPA's previous approval of State title... raising the GHG thresholds that apply title V permitting to major sources in the affected States, this...

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